ANNUAL REPORT 2008 Outcome Line SBA-1 Improved Beans for the Developing World TABLE OF CONTENTS PAGE NO. Product Line LogFrame as in MTP 2008-2010 vi Research Highlights in 2008 xvi Progress Report Product 1: Beans with improved micronutrient concentration that have a positive impact on human health 1 Activity 1.1 Developing more nutritious bean varieties 1 1.1.1 1.1.2 1.1.3 1.1.4 1.1.5 Actvity 1.2 1.2.1 1.2.2 Development of new advanced lines from the program for the nutritional enhancement of Andean bush beans Selection of Mesoamerican lines for high minerals in cycle 2 of recurrent selection in Colombia Development of new sources for high iron: Selection among interspecific families Breeding of Andean beans to create lines with higher mineral content and superior agronomic traits in southern Africa Breeding micronutrient dense bean varieties in eastern Africa 11 15 Genotype x environment interaction 26 Multi-site evaluation of biofortified Andean lines, NUA35 and NUA56 Genotype x environment interactions for grain Fe and Zinc concentration 26 29 Activity 1.3 Associated traits: antinutrients 1.3.1 1.3.2 1 5 9 34 Inheritance of seed phosphorus and seed phytate content in a recombinant inbred line population of common bean Analysis of condensed tannins through HPLC in genotypes from an intergenepool bean population 34 36 Product 2: Beans that are more productive in smallholder systems of poor farmers 39 Activity 2.1 Developing germplasm tolerant to abiotic stresses 2.1.1 2.1.1.1 2.1.1.2 2.1.1.3 2.1.1.4 2.1.1.5 2.1.1.6 Drought resistance Evaluations of Mesoamerican lines segregating for drought resistance and tolerance to low soil P Evaluations of Mesoamerican lines segregating for drought resistance and bc-3 gene Evaluations of Andean lines for drought resistance and for yield potential under irrigation Development of drought tolerant Andean beans from inter and intra-genepool crosses Field evaluation of a common bean reference collection for drought tolerance Evaluation of Andean breeding lines for adaptation to drought stress i 39 39 40 43 46 51 55 63 2.1.1.7 Physiological evaluation of drought resistance in elite lines under field conditions 2.1.1.8 Physiological evaluation of drought resistance of 33 recombinant inbred lines (RILs) of DOR 364 x BAT 477 under terminal drought stress over two seasons 2.1.1.9 Physiological evaluation of drought resistance in recombinant inbred lines (RILs) of DOR 364 x BAT 477 under intermittent drought stress 2.1.1.10 Evaluation of drought resistance in recombinant inbred lines (RILs) of MD 23-24 x SEA 5 under intermittent drought stress 2.1.1.11 Evaluation of drought resistance and yield of 7 genotypes of Phaseolus vulgaris inoculated with and without Rhizobium etli strain CIAT 632 under greenhouse conditions 2.1.2 2.1.2.1 2.1.2.2 2.1.2.3 2.1.2.4 2.1.2.5 Aluminum resistance Evaluation in two environments, of interspecific lines selected under aluminum toxicity Improving phenotyping capacity to evaluate for aluminum resistance Qualitative indication of Al-induced citrate exudation in different Phaseoulus species using an Agarose-Aluminon method Phenotypic differences in aluminum resistance of landraces and bred lines Phenotyping for aluminum resistance in recombinant inbred lines (RILs) of DOR364 x BAT 477 Activity 2.2 Developing germplasm with resistance to insect pests: Bruchids and leafhopper 2.2.1 2.2.2 2.2.2.1 2.2.2.2 2.2.2.3 2.2.2.4 Screening for sources of resistance to major insect pests Developing germplasm resistant to insects Storage weevils (Zabrotes subfasciatus) Crosses to incorporate arcelin-based bruchid resistance into Andean beans Leafhopper (Empoasca kraemeri) Evaluation of Andean beans for resistance to leafhoppers Activity 2.3 Developing germplasm resistant to disease 2.3.1 2.3.2 2.3.3 2.3.4 2.3.5 2.3.6 2.3.7 2.3.8 Crosses to incorporate BCMV and CBB resistance into Andean beans Development and release of new red mottled bean varieties with multiple constraint resistance in eastern Africa Breeding red kidney bean varieties with multiple constraint resistance in eastern Africa Development and release of new Speckled Sugar bean varieties with multiple disease resistance in eastern Africa Breeding small and medium red bean varieties resistant to multiple stresses for smallholder producers in eastern Africa Development and release of brown and tan colored bean varieties with multiple stress resistance in eastern Africa Development and release of improved climbing bean varieties in eastern Africa Breeding for specific bean market classes within Southern Africa Bean Research Network (SABRN) ii 65 76 80 85 91 94 94 96 99 101 106 111 111 113 113 116 118 120 122 122 124 134 139 145 152 156 163 2.3.9 Developing bush and climbing bean lines with resistance to Pythium root rot, angular leaf spot, and bean common mosaic and necrotic viruses Activity 2.4 Yield potential: climbing beans 2.4.1 171 Detection of QTL for climbing ability and component traits in common bean Activity 2.5 Characterizing and monitoring pathogen and insect diversity 2.5.1 2.5.2 Monitoring of whitefly populations in the Andean zone Diversity, distribution and pathogenicity of Pythium species in Rwanda Activity 2.6 Developing integrated disease and pest management components 2.6.1 2.6.2 2.6.3 2.6.4 164 Reduction of pesticide use in common bean and snap bean crops through the development and implementation of IPM strategies in Colombia and Ecuador: on-farm surveys and baseline studies of pest resistance Continued monitoring of resistance to insecticides in Bemisia tabaci, on pepper hosts in Valle de Cauca Isolation and assessment of 2,4-diacetylphloroglucinol (DAPG/Phl)-producing fluorescent Pseudomonas strains for biological control of Pythium root rots Microsatellite analysis of common bean mixtures from SW Uganda 171 173 173 175 181 181 188 192 195 Product 3: Beans that respond to market opportunities 201 Activity 3.1 Development of large white beans for international markets 201 Activity 3.2 Breeding Navy and Large White bean varieties with multiple stress resistance in eastern Africa 202 Activity 3.3 Identification of a varietal candidate in Nicaragua with potential for international export 207 Activity 3.4 Progress in development of Snap and runner beans for smallholder production in East and Central Africa 208 Product 4: Strengthened institutions that enhance bean product development and delivery 213 Activity 4.1 Strengthened capacity of NARS: increasing the knowledge and skills of scientists and staff from NARIs, NGOs and Rural Service Providers 213 4.1.1 4.1.2 4.1.3 4.1.4 Degree and non-degree training in Latin America Degree and non-degree training in Africa Trips and attendance of Headquarters staff at meetings Trips and attendance of African staff at meetings iii 213 217 222 223 Activity 4.2 Strengthen international collaboration through networks (Intra- and internetwork collaboration), bi-lateral relations, and/or joint special projects 4.2.1 4.2.1.1 4.2.1.2 4.2.1.3 4.2.1.4 4.2.2 4.2.2.1 4.2.2.2 4.2.2.3 225 Projects developed in Africa List of ongoing special projects Regional research subprojects under SABRN Projects submitted, Proposals and Concept notes prepared New proposals approved Projects developed in Latin America List of ongoing special projects at Headquarters Projects submitted, Proposals, and Concept notes prepared New proposals approved 225 225 226 229 230 231 231 232 232 Activity 4.3 Supporting breeding programs in NARS, regional networks, farmers’ Associations, and CIALs with germplasm and technical knowledge 234 4.3.1 4.3.2 4.3.2.1 4.3.2.2 4.3.3 4.3.3.1 4.3.3.2 4.3.3.3 4.3.4 4.3.5 4.3.6 4.3.7 4.3.7.1 4.3.8 Nutritional analysis of Bolivian germplasm as support for the national genebank and breeding programs Selection by NARS of segregating populations for nutritional quality Progress in selection of cycle 2 populations in Central America Selection of populations in Brazil Evaluation of lines from the 1st cycle of selection in advanced yield trials and validation in Central America Nicaragua Honduras Brazil Evaluation of SU91 as a molecular marker for CBB resistance in common beans for Southern Africa Distribution of seed from CIAT Headquarters Distribution of germplasm within the ECABREN bean network Exchange of germplasm in Southern Africa Bean Research Network (SABRN) Southern Africa Regional Bean Yield Trial (SARBYT) Varietal releases in Latin America and Africa (2006-2008) Activity 4.4 Development of sustainable seed systems to support wide dissemination 4.4.1 4.4.2 4.4.3 4.4.4 4.4.5 4.4.6 4.4.6.1 4.4.6.2 4.4.6.3 4.4.7 4.4.7.1 4.4.7.2 4.4.7.3 Increasing Access to New and Existing Technologies Linking Participatory Variety Selection and Impact-oriented seed production and supply systems Marketing of small seed packs Skills and knowledge enhancement Backstopping to NARS and their partners Some lessons from keys approaches of PABRA to institutional strengthening with NARS and other partner organizations Partnership development Analysis of the effectiveness of capacity building in PABRA Seed systems Development of Seed Security Assessment Methodology Distinguishing between Seed security and food security Seed System Security Assessment Guide SSSA Uptake - International Public Good, Responding to Demand iv 234 236 236 239 239 239 240 241 242 244 248 249 250 253 255 255 255 256 257 259 260 260 261 266 268 268 269 272 Activity 4.5 Socio-economic activities 4.5.1 4.5.1.1 4.5.1.2 4.5.2 275 Targeting crop breeding and seed delivery efforts to enhance the impact on the livelihoods of the poor in drought-prone regions of sub-Saharan Africa Regional situational and outlook analysis Socio-economic baseline surveys Capacity building of enumerators for baseline study 275 275 275 277 Publications 278 Book Chapters Refereed Journals Non -Refereed Journals Workshops and Conferences Proceedings, Posters, and Others: Proceedings Posters Others Editorial contributions Donors Contracts Partners Collaborating with Headquarters Institutional Partners in Africa List of NARS and Collaborating Partners in Africa Project Staff List Acronyms and Abbreviations used 278 279 281 281 283 283 284 286 286 287 287 288 290 293 295 297 v PRODUCT LINE LOGFRAME IMPROVED BEANS FOR THE DEVELOPING WORLD: PRODUCT LINE SBA1 Rationale & Changes Rationale The common bean (Phaseolus vulgaris L.) is the world’s most important grain legume for direct human consumption. Its total production exceeds 12 million MT, of which 7 million MT are produced in tropical Latin America and Africa. Beans are the “poor man’s meat” and are particularly important in the diet of the underprivileged. Beans, like other legumes, supply proteins, carbohydrates, vitamins and minerals, and complement cereals, roots and tubers that compose the bulk of diets in most developing countries. Common bean is also one of the most diverse crops in terms of its cultivation methods and its uses. It serves as mature grain, as immature seed, and as a vegetable (both leaves and pods), and after harvest the stover is used as animal fodder. It is cultivated from sea level up to 3000 masl in monoculture, in association, or in rotations. The possibility of obtaining a harvest in as little as two months offers quick income, quick food supply, and also permits rotating with other crops or inter-planting among fruit trees or coffee before the primary crop produces income. At the other extreme are the aggressive climbing beans that subsistence farmers maintain in the garden for food security and continual harvest over a six month period. Apart from subsistence cultivation, beans have become increasingly commercial over the past thirty years in national, regional and international markets. In Central America beans are the #1 income generator among the traditional field crops. In Africa, farmers tap into regional bean markets in Nairobi, Kinshasa and Johannesburg. With the onset of globalization, the past decade has seen a growing international market that is now reported to reach 2.4 million MT. This heightens issues of equity for the small bean producers that have little other stable source of income, but some also see this as an opportunity. For example, bean represents 6% of external income for Ethiopia, and small farmers in Bolivia produce the large white and red mottled classes for export. Snap beans are a high value, labor intensive crop of small farmers in Kenya and the Andes. Besides the common bean, another four cultivated species are conserved in the CIAT gene bank, as well as wild relatives. This collection is the largest of the genus in the entire world, representing more than 35,000 accessions that have been declared as part of the designated collection before FAO. These other cultivated species fill niches that are unsuitable for the common bean, for example, P. acutifolius that thrives in desert environments. Our primary mission is to contribute to household and global food security by assuring an adequate supply of beans as a culturally acceptable and traditional staple; and to improve the income of small bean producers of Latin America and Africa, by making bean production more profitable. We also seek to improve human nutrition, both by augmenting the supply of beans, and by improvement of their nutritional value. Our products are designed to respond in particular to the needs of small, resource-poor bean farmers in Latin America and Africa. Thus, we seek to create solutions to biotic and abiotic production limitations that require minimal inputs, and in the case of improved germplasm, with good market potential. Our research strategy focuses on the exploitation of the vast genetic resources of bean that exist as a complex array of major and minor gene pools, races and sister species. CIAT’s gene bank with 41,000 accessions of common bean and related species is our most unique resource, and has been the source of vi genes for disease and insect resistance, abiotic stress tolerance, nutritional quality and yield potential. Most traits are still selected by conventional means in field sites (in some cases backed up by greenhouse evaluations) where most important diseases, edaphic constraints and drought can be manipulated for purposes of selection. However, Marker Assisted Selection (MAS) is employed selectively but strategically, in most cases for disease resistance genes. CIAT pioneered participatory selection with farmers and this practice is being extended and systematized. While most products are seed based, others involve agronomic practices or are knowledge based. Our research is strategic combined with both basic and applied elements, as called for by the particular challenge. Changes There have been no essential changes in relation to the MTP of 2007. However, in 2008 an agricultural economist, Dr. Enid Katungi came on board under the Tropical Legumes-II project, with base in Kampala, Uganda. CG System Priorities CIAT’s bean product line is housed principally under CG System Priority Area 2: Producing more and better food at lower cost through genetic improvements. Efforts are dedicated to improving yields through control of diseases and pests, tolerance to abiotic stresses (drought, aluminum toxicity and low soil fertility in particular), and expanding the adaptation range of climbing beans. The bean product line also places heavy emphasis on improvement of nutritional quality, especially through increase in iron and zinc content in the grain. There is potential to contribute to Priority Area 3A: Increasing income from fruits and vegetables, through the improvement of snap beans for both Africa and Latin America. The bean team collaborates with marketing specialists to create varieties with better market potential, including international export markets (Priority Area 5B). Finally, strengthening national institutions (Priority Area 5A) continues to be an important product, both in Africa where novel institutional arrangements and relations have been productive to achieve wide impact, and in Latin America where staff reductions have weakened national programs. On both continents national programs seek support to incorporate modern selection techniques. Impact Pathways Product 1 (Beans with improved micronutrient concentration that have a positive impact on human health) is targeted to small farmers and poor rural and urban consumers in Africa and Latin America. Targeting is developed in collaboration with nutritionists and with experts in GIS, to address human populations with nutritional deficiencies in iron and zinc. This product involves both small seeded germplasm that is often targeted to warmer climates or more difficult environments in Central America, Mexico, Venezuela, East Africa and Brazil. Large seeded germplasm is usually cultivated in more temperate climates in the Andean zone, the East African highlands and southern Africa, although in the African highlands small and large seeded types overlap, sometimes differentiated by soil fertility gradients within the farm, prevailing biotic constraints and household preferences. Improved germplasm is shared or developed jointly with NARS partners, who supply basic seed to a range of organizations interested in production of seed (local seed companies, NGO’s, CBO’s, women’s groups) who in turn distribute to farmers. NGOs and health workers play a special role in delivery. Benefits accrue to farmers/consumers through stable food supply of more nutritious beans for home consumption, and potentially to poor urban consumers. Assumptions for the successful delivery of these products include institutional and financial stability of partners, political stability, and institutional support. The role of CIAT is that of a primary research provider (of improved germplasm), at times a secondary research provider (backing up national bean improvement programs with technical expertise and training), and catalyser (to promote downstream alliances in the uptake chain). This product is complementary to those of CIMMYT and CIP. vii Beneficiaries of Product 2 (Beans that are more productive under low input agriculture of poor farmers) are in some cases researchers (both inside and outside of CIAT), and in some cases are bean producers. For example, molecular markers for resistance genes benefit researchers directly, and farmers indirectly as subsequent beneficiaries. Uptake pathway for such methodologies is direct communication through workshops and courses, and indirectly through publications, leading to benefits of more efficient and effective bean research. This assumes that partners are in a position to implement such technologies. On the other hand, crop management practices are of direct benefit to farmers as users, potentially across all bean ecosystems. Uptake chains for agronomic practices are similar to those for seed based technologies; results are communicated to NARS and other partners (NGO’s, CBO’s etc) who have successfully diffused practices to farmers, to the benefit of farmers who enjoy more stable productivity. Improved germplasm is diffused through many of the same channels as beans with improved nutritional value, with the exception that partners may have less specific interests, and may be more production oriented. The role of CIAT is that of primary source of research for development. Product 3 (Beans that respond to market opportunities) benefit small farmers in both Latin America and Africa. Farmers in Ethiopia have already benefited from tapping into export markets for canning beans, and other countries are positioning themselves to follow suite. In Central America exporters are seeking to fill a niche created by the Latin population in the USA. This is a demand-driven activity, and in large part has generated its own impact pathway. Exporters and international grain buyers have established market chains that give them access to export quality beans. CIAT’s role has been that of supplying germplasm in some cases, and in others to facilitate communication, and to give support in seed systems to avail quality seed to farmers of very specific varieties. Product 4, (Strengthened institutions that enhance product quality and delivery) seeks to benefit partners at multiple levels through facilitated interaction, including farmers who are at the end of the organizational chain. NGOs, government extension agencies, farmer organizations, local seed companies, and non-conventional seed actors such as women groups, people living with HIV/AIDS and tobacco companies all participate and benefit. The product will generate impact on target beneficiaries through their participation in development of innovations, knowledge and technologies in strategic alliances with multidisciplinary research teams and NGOs. Scaling out of innovations and best practices to areas with similar environments will be done through strategic alliances of research and development actors. The latter will use their network and other communications mechanism to adapt knowledge and results relevant to them. Scaling up regionally and internationally will be done through international NGOs, advocacy, and communication. The outcome is enhanced communication and complementarity of actors with resulting cost efficiencies, and in the case of technology diffusion, increased and diversified adoption. Another dimension of this product is support to NARS in development of projects, benefiting national program researchers and with the outcome of their integration into the product line research mode. This assumes a degree of consistency in partner personnel, while CIAT’s role is that of facilitator. International Public Goods The IPG of the bean product line include: • Improved germplasm with biotic and abiotic stress tolerance, and/or enhanced nutritional value, drawing upon the genetic resources of CIAT’s extensive gene bank, and 30 years of experience in bean improvement. CIAT’s geographical position and access to varied altitudes and research sites facilitates study and selection of germplasm. • Improved practices for the management of pests and diseases, including monitoring of pathogen populations with modern molecular tools developed at CIAT. • Knowledge and tools that contribute to the development and implementation of the above IPG’s. For example, molecular markers for useful traits, developed with CIAT’s in-house resources of genetic maps and markers. Knowledge of the structure of genetic resources housed viii • in the gene bank, and ways to exploit them. Screening methods to identify biotic and abiotic stress resistant genotypes. Participatory breeding methods with varying degrees of involvement of farmers, traders and other key actors. Methods for networking, both formal among official sector researchers, and less formal among a broader range of partners, with special emphasis on research partnerships and on effective and sustainable seed systems reaching a large number of households. Partners Most important partners and the respective person-years of professionals dedicated to bean research within the (several) products are: Product 1: NARS in Latin America, including those of Mexico (6), Guatemala (2.5), Honduras (2, including EAP-Zamorano), El Salvador (2), Cuba (2), Brazil (4) participate in the AgroSalud project to improve nutritional quality and productivity of bean. NARS in South America, including those of Colombia (5 between university staff, an NGO and the NARI), Bolivia (4 between university staff and a foundation) collaborate in the improvement of disease resistance of Andean bean with better nutritional quality, also under the AgroSalud project. NARS in East, Central and Southern Africa, including those of Kenya (5), Rwanda (6), Uganda (5), Malawi (1), Zimbabwe (1) are partners in the improvement of nutritional qualities in large seeded Andean beans. Linkage funds finance a project with one Canadian university, and with a partner in USDA. Product 2: Nicaragua (4.5) and Honduras (2) are partners in breeding for drought tolerance. NARS in East, Central and Southern Africa including those of Ethiopia (3), Kenya (2), Tanzania (3), Rwanda (4), Malawi (1), Zimbabwe (1) and DRCongo (4), participate in the improvement of productivity under low soil fertility and/or drought. The University of Hannover, Germany participates in a project to define physiological mechanisms of aluminum tolerance and drought resistance (2), which also includes Malawi (2) and Rwanda (4). Catholic University of Leuven (3) is a partner to improve nitrogen fixation technology. NARS in South America, including those of Colombia (5 between university staff, an NGO and the NARI), Bolivia (4 between university staff and a foundation) collaborate in the improvement of disease resistance of Andean bean. NARS in East, Central and Southern Africa, including those of Kenya (5), Rwanda (6), and Uganda (5) Tanzania (4) are partners in the development of disease resistance, medium altitude climbing beans (MAC), and productivity in large seeded Andean beans. NARS in Honduras (Zamorano) (1), Colombia (2), Uganda (3), Rwanda (4), and South Africa (2) share in the use of markers for MAS, especially for resistance. South Africa (3) participates in pathogen characterization, evaluation and validation of resistance sources. Agriculture and Agri-Food Canada (AAFC) is a partner in diagnosis and characterization of soil borne pathogens (especially Pythium species) using molecular techniques, and development of molecular based diagnostic assays for soil borne pathogens. Product 3: Partners in Latin America with specific attention to breeding market quality include NARS in Honduras and Nicaragua. NARS in Africa with active participation in canning beans include those of Ethiopia and Uganda. Partners in the development of snap beans include a university in Colombia, and one in Kenya. Product 4: NARS as above –plus a wide range of NGOS, CBOS, farmers’ groups, women’s groups, – totaling over 300 direct-link partnerships, to make users aware of technologies and to get these technologies widely disseminated. The ECABREN and SABRN bean networks coordinate nine NARS in East Africa and ten NARS in southern Africa, respectively. These networks participate in Products 1, 2, 3 and 4 with input from African NARS cited above, plus NARS in Burundi (3), Sudan (2), Zambia (1), Zimbabwe (1), Mozambique (3), Lesotho (3) and Swaziland (3). ix HarvestPlus Challenge Program: IFPRI, CIMMYT, and CIP are immediate collaborators in the CP and the AgroSalud (Latin American) nutritional improvement project, working in the same agro-ecological zones, while ICRISAT, IITA, IRRI, and ICARDA are indirect collaborators under HarvestPlus. ECABREN and SABRN networks in Africa also participate in HarvestPlus. Generation Challenge Program: Partners include EMBRAPA-Brazil (2), INTA-Cuba (1), Pairumani (an NGO) in Bolivia (2), National University in Colombia (2). Sub-Saharan Africa Challenge Program: ICIPE, AHI and NARS in Rwanda, Uganda and D.R. Congo are immediate partners. Product line Funding Budgeting 2007-2011 Year US Dollars (millions) 2007 (actual) 8.008 2008 (actual) 9.931 2009 (proposal) 7.597 x 2010 (plan) 7.702 2011 (plan) 7.812 IMPROVED BEANS FOR THE DEVELOPING WORLD: PRODUCT LINE SBA1 (2008-2010) Targets PRODUCT 1 Products Beans with improved micronutrient concentration that have a positive impact on human health NARS, farmers & consumers in Central America, the Caribbean, Brazil, East and Southern Africa Adoption of improved varieties by farmers Better nutritional status, especially of rural consumers • ~30 small seeded F3-derived F5 bush bean families developed with tropical adaptation, 60% more minerals, abiotic stress tolerance, and 2 biotic resistances for Central America (HarvestPlus) • NARS, NGO’s CBO’s, health workers, and farmers in target countries • Farmers incorporate high mineral and disease resistance lines into diverse production systems • Reduced levels of iron and zinc deficiency in bean consumers • 50 improved lines with varietal potential and 90 ppm iron (ie, 80% more iron) 15 new large seeded climbing beans with high mineral trait (HarvestPlus) Marker assisted selection for one nutritional trait (iron) tested Four fast track micronutrient dense bean varieties disseminated and promoted in two countries in eastern and southern Africa Two large seeded lines with 50% more iron enter formal varietal release process in eastern Africa • NARS, NGO’s CBO’s, health workers, and farmers in target countries • Adoption of micronutrient rich beans • Reduced levels of iron and zinc deficiency in bean consumers • NARS, NGO’s CBO’s, health workers and consumers • Adoption of micronutrient rich beans • Reduced levels of iron and zinc deficiency in bean consumers Product Targets 2008 Product Targets 2009 • • • Product Targets 2010 • Intended User xi Outcome Impact Products Intended User Outcome Impact Beans that are more productive in smallholder systems of poor farmers Breeders and pathologists in CIAT and NARS; farmers in E and S Africa, Andean zone, Caribbean Adoption of improved varieties by farmers; Best bet IDPM practices and genetic combinations for stable resistance deployed. More stable production, food availability and income • • • Targets PRODUCT 2 Product Targets 2008 • • • • Product Targets 2009 • • Product Targets 2010 • 5 molecular markers for detection, diagnosis and diversity studies of ALS and anthracnose pathogens made available At least 10 lines in major market classes combining resistance to Pythium root rots, BCMV and angular leaf spot An IPM system for whiteflies on snap beans refined and promoted in 2 major bean producing areas of the Andean zone • • NARS, NGO’s and farmers’ groups CIAT and NARS breeders NARIs researchers in LAC, Africa, IARCs • • An IDM system for bean root rots implemented and promoted in 2 major bean producing countries in Africa At least 40 lines combining drought resistance with resistance to BCMNV, root rots, and/or ALS available for testing in Africa 2 molecular markers linked to ALS and Pythium root rot implemented in MAS • Resistance genes for anthracnose or ALS introgressed into 5 BCMNV resistant climbing beans At least 10 genotypes combining drought resistance with aluminium resistance available for testing in Africa • • NARS breeders, NGO’s, CBOs, and farmer groups NARS pathologists, • • • • NARS breeders, NGO’s, CBOs, and farmer groups NARS soil scientists and agronomists xii • • Disease and pest characterization tools adopted by researchers Adoption of disease resistant lines in marginal environments Increased utilization of integrated management approaches. • Improved food security, & income. • More stable disease resistance in advanced lines leads to stable yield Resistant lines incorporated into improved systems Drought resistant lines with disease resistance used in drought prone areas in Africa Breeders improve efficiency of genetic improvement • Reduced yield losses from ALS, root rots and drought Farmers benefit from yield stability of high yield climbers Farmers benefit from stable yields in marginal areas • Improved food security, & income. Targets Products Beans that respond to market opportunities • • • Product Targets 2009 Product Targets 2010 • • Outcome Impact NARS in Africa and Latin America Adoption of commercial varieties by farmers, enhancing access to markets Higher income, especially for the poor and women farmers 10 lines of snap beans with confirmed resistance to Gemini virus in Colombia 1 variety released in Nicaragua for export market • • • At least 3 snap bean lines with resistance to rust and quality characteristics preferred in regional and export markets for Africa. 4 bean genotypes with very high commercial or export quality made available to farmers in 4 countries in Latin America and Africa • 5 canning bean lines with acceptable quality characteristics in yield trials in two countries in eastern Africa • PRODUCT 3 Product Targets 2008 Intended User NARS, NGOs, CBOs, farmer groups, seed producers Farmers reduce pesticide use, assuring production and profitability • NARS, NGOs, CBOs, farmer groups, seed producers NARS, NGOs, CBOs, farmer groups, seed producers xiii • Adoption of snap bean and reduced chemical use. • Farmers in marginal environments assure market access • Farmers improve yields and quality of product with improved varieties Less pesticide intoxication in rural communities and urban consumers Increased production and incomes. • Increased production and incomes. • Increased production and incomes. Targets PRODUCT 4 Products Strengthened institutions that enhance bean product development and delivery Intended User NARS in Africa and Latin America • • NARS, NGOs, CBOs, farmer groups, seed certification agencies, seed producers • UN, humanitarian and post-stress recovery organizations • PABRA • Product Targets 2008 • • • • Product Targets 2009 • One comprehensive methodology developed for assessing seed security and targeting responses in acute and chronic stress situations. Lessons from 3 case studies (approaches for partnership; capacity building; alternative seed delivery systems) of strategies for product development and delivery in PABRA analyzed. Protocols developed and adapted to facilitate application of MAS for disease resistance in 3 African countries Breeding programs for higher iron levels established in Honduras, Nicaragua, Bolivia, Venezuela, Kenya and Malawi A guide for mainstreaming and sustaining wider impact, developed and recommendations availed for 5 countries in East, Central and 4 countries in Southern Africa Three delivery channels strategies tested for reaching the poor and in marginal areas with new variety innovations and information At least 1 methodological frameworks/strategies for testing and evaluating multi-stakeholder networks and platforms (between private-public) for facilitating decentralized targeting for pro poor impact. • NARS, NGOs, Decentralized Local Governments, CBOs, farmer groups, seed certification agencies, seed producers ,agroprocessors, local financial institutions • UN, humanitarian and post-stress recovery organizations xiv Outcome Improved institutional performance by NARS, NGOs and other partners, reflected in more effective technology development and dissemination • Frameworks and methodologies for seed systems, PM&E, and MAS are in use by PABRA partners • Increased partner involvement in accessing technologies to a greater number of end users • Increased capacities of partner organizations / institutions to develop and promote integrated and decentralized strategies for reaching pro-poor farmers Impact More stable production, improved food availability, income and nutrition, especially for the poor and women farmers Products Targets Product Targets 2010 PRODUCT 5 Product Targets 2008 • Capacity to evaluate root systems in soil tubes established in Honduras and Nicaragua • Elements of Pro-poor seed delivery and production systems confirmed and such pro-poor seed enterprises established in 2 PABRA network countries. One strategy for wider utilization of non varietal bean technologies (IPM; soil management) developed and widely shared in 4 countries in Africa • • 1500 accessions conserved in long term storage and in back-up in CIMMYT 1000 samples of bean seed distributed • Bean scientists; other gene banks • Novel genes incorporated into breeding programs Another 1500 accessions conserved in long term storage and in back-up in CIMMYT Another 1000 samples of bean seed distributed A plan formulated to establish a database of evaluation data • Bean scientists; other gene banks • Novel genes incorporated into breeding programs Another 1500 accessions conserved in long term storage and in back-up in CIMMYT Another 1000 samples of bean seed distributed • Bean scientists; other gene banks • Novel genes incorporated into breeding programs • • • • Impact NARS, NGOs, CBOs, farmer groups, seed certification agencies, seed producers Breeders, geneticists, and other bean scientists; national gene banks • Product Targets 2010 • Outcome More than 35,000 accessions are conserved, documented and available for distribution • Product Targets 2009 Intended User xv Bean genetic resources are used directly or employed in breeding programs More stable production, improved food availability, income and nutrition RESEARCH HIGHLIGHTS IN 2008 Product 1: Beans with improved micronutrient concentration that have a positive impact on human health Activity 1.1 Developing more nutritious bean varieties Highlights: • • • • • • • • • More than 30 F3.5 small seeded Mesoamerican families were selected for high mineral concentration and some degree of drought resistance. Eighteen lines derived from interspecific crosses were coded as MIB (high mineral) lines, with levels of iron above those of the high iron check MIB 465. Some also have superior resistance to foliar pathogens. Over 200 pollinations were generated combining multiple sources of resistance to diseases and drought with high mineral (Fe and Zn) content in SABRN countries. Some good donor parent for drought resistance in common bean (SEA5, SEA15 and SER16) were also good parents for high Fe and Zn content. Several populations combining different market classes and disease or drought resistance, but also with high mineral (Fe and Zn) content have been generated, and some may have high Fe and or Zn content. Four bush and three climbing micronutrient dense bean varieties with high yield potential are pre-released for smallholder production in eastern Africa. This marks the first time biofortified mineral dense bean varieties are formally recommended for release following independent evaluations. Bioavailability of Fe in raw samples of fast track bean lines varies from 1.1% to 6.6%. Bioavailability of Zn in raw samples of fast track lines varies from 0.5 to 2.5%. Cooking enhances Fe and Zn bioavailability in beans by more than two fold. Freshly shelled beans have more bioavailable Fe and Zn compared with dry beans. Actvity 1.2 Genotype x environment interaction Highlights: • • • NUA35 and NUA56 were tested for yield potential and mineral accumulation across 15 sites in Latin America (72 replicates), showing that both lines have a 15 to 25 ppm differential iron advantage over CAL96. Significant genotype x environment interactions indicate that grain mineral concentration is influenced by soil type, soil nutrient status, moisture concentration and other environmental factors but the magnitude varies with genotypes. Some genotypes show high stability for mineral density. Fertilization regimes and other agronomic practices can be used to enhance expression of high mineral density traits. Activity 1.3 Associated traits: antinutrients Highlights: • The inheritance of seed phytate content was analyzed to determine if this anti-nutrient could be reduced and how it is related to seed phosphorus content. Quantitative inheritance was found xvi with several QTL explaining both traits independent of seed size. The results of this study show some genotypes with low levels of phytates which would seem to be sufficient for breeding attempts. The other anti-nutrient being analyzed is condensed tannins and an HPLC method was adapted to look at the tannin monomers that accumulate in genotypes from an inter-genepool population. Product 2: Beans that are more productive in smallholder systems of poor farmers Activity 2.1 Developing germplasm tolerant to abiotic stresses 2.1.1 Drought resistance Highlights: • • • • • • • Mesoamerican crosses among drought resistant parents that had expressed a degree of tolerance to low soil P availability, produced more than 20 lines that were excellent in drought resistance. Some lines subsequently showed adaptation to low P in Darién, producing grain of excellent quality under combined drought and low P stress. Another 15 Mesoamerican families combined drought tolerance and bc-3 gene for resistance to BCMNV. In an effort to incorporate drought tolerance in Andean bush beans we have created a series of 216 advanced drought Andean beans (DAB) lines from inter and intra-gene pool crosses involving 5 commercial genotypes from Southern Africa and 10 drought tolerance sources of which half were Andean and half were Mesoamerican to produce 46 populations. The lines represent large red, red mottled and cream mottled seed types. Selection has stressed bush bean architecture, adaptation to drought stress and yield potential under favorable conditions using alternate dry versus rainy season plantings. A reference collection of landraces from the CIAT core collection has been evaluated in the field for drought tolerance compared to a series of check genotypes. The reference collection was stratified into Andean and Mesoamerican gene pools and the association of drought tolerance with subgroups and common bean races was analyzed. Mid-elevation adaptation was tested in Darién for a series of SAB (drought resistant Andean) lines originally developed from crosses between the drought-resistant genotypes SAB 258, SAB 259, and ICA Quimbaya crossed with drought susceptible but commercial type genotypes ABA36, ABA58 and COS16. Results confirmed the genetic gain for drought tolerance and yield potential that has occurred in the breeding of the SAB lines compared to both their droughttolerant and susceptible parents. The same lines were tested in rainfed conditions in Palmira and several maintained their yield advantage over local checks. Certain SAB lines can be selected with greater stability across mid-elevation and lower-elevation sites based on this analysis. Field evaluation of elite lines at Palmira resulted in identification of five lines NCB 226, SEN 56, SER 113, SER 125 and SER 16 that were outstanding in their adaptation to drought stress conditions. The superior performance of these lines under drought stress was associated with higher values of harvest index, pod harvest index, leaf area index and canopy biomass. The SER lines that were developed in the last few years seem to combine the desirable traits for drought adaptation such as greater mobilization of photosynthates to seed with efficient use of water through stomatal control. Field evaluation of 33 RILs of the cross DOR 364 x BAT 477 at Palmira over two seasons under terminal drought stress conditions resulted in identification of two lines (BT 21138-17-1-1 and BT 21138-6-1-1) that were superior in their adaptation to drought stress conditions. The superior performance of these lines under drought stress was associated with higher values of harvest xvii • • • 2.1.2 index, pod harvest index and seed and pod number per area indicating the importance of greater mobilization of photosyntates to pods and to seeds under rainfed conditions. Field evaluation of 97 RILs of the cross DOR 364 x BAT 477 under intermittent drought stress resulted in identification of two RILs BT 21138-68-1-1 and BT 21138-74-1-1 that were outstanding in adaptation to intermittent drought stress conditions. The superior performance of these lines under intermittent drought stress was associated with higher values of harvest index, pod partitioning index, stem biomass reduction, seed number per area and pod number per area indicating the importance of greater mobilization of photosyntates to pods and seeds under rainfed conditions. Field evaluation of 121 RILs of the cross MD 23-24 x SEA 5 over 3 seasons resulted in identification of the lines MR 81 and MR 25 that were superior in adaptation to drought stress conditions. The superior performance of these lines was associated with higher vigor, higher values of pod harvest index, harvest index and seed number per area, highlighting the importance of the photosyntate mobilization to pods and seeds under intermittent drought stress. The response to inoculation with the strain Rhizobium etli CIAT 632 under drought stress was tested using 7 common bean genotypes. We found that Pinto Villa was better adapted to drought due to its ability to decrease stomatal conductance while Alubia cerrillos was more affected due to drought stress. Although there was no response to inoculation, the effect of terminal drought stress on nodulation was very marked on all 7 genotypes. Aluminum resistance Highlights: • • • • • • Lines derived from an interspecific cross of SER 16, a drought resistant line, by Phaseolus coccineus (G35346) yielded well under both rainfed and aluminum toxic conditions. Some actually yielded more in intermittent drought than SER 16. SER 16 proved to be an excellent common bean parent to cross with P. coccineus, perhaps due to its characteristic of excellent remobilization to grain. A filter paper-styrofoam sandwich germination method was developed to improve the phenotyping capacity for Al resistance in common bean and a primary root marking method was developed to evaluate short-term effects of Al on root elongation process. A method was adapted and validated for screening for aluminum resistance in common bean based on qualitative determination of aluminum-complexing compounds including citrate released from the roots. Phenotypic evaluation of 20 common bean genotypes for aluminium resistance confirmed the higher level of Al resistance of three Andean genotypes (ICA Quimbaya, BRB 198 and G5273) and identified one Mesoamerican genotype (G24601) also with higher level of Al resistance. Phenotypic evaluation of 97 RILs of DOR 364 x BAT 477 for aluminium resistance resulted in identification of a few RILs with low inhibition of root growth under high Al in solution. Two RILs (BT 21138-128-1-M-M-M and BT 21138- 2-1-1-M-M-M) were found to be outstanding in root growth both with and without aluminum in solution. Activity 2.2 Developing germplasm with resistance to insect pests: Bruchids and leafhopper Highlights: • • • New accessions from the gene bank were evaluated for insect resistance Resistance to Zabrotes subfasciatus was reconfirmed New breeding lines that have resistance to the leafhopper (Empoasca kraemeri) were identified xviii • Some Andean bean lines presented high level of tolerance to Empoasca and less yield loss Activity 2.3 Developing germplasm resistant to disease Highlights: • • • • • • • • • • • • A large number of crosses were made to pyramid insect (bruchid) and disease (BCMNV and CBB) resistance with drought tolerance or mid-elevation adaptation in Andean bush beans. The arcelin gene was used as the source of bruchid resistance and was effectively selected for in 115 different cross combinations. Meanwhile, 187 cross combinations were generated for disease resistance. These crosses are being advanced at our drought stress and mid-elevation sites in Colombia. Twelve new, large seeded red mottled bean varieties with multiple resistance to diseases and up to 30% better yield compared to commercial varieties released in six countries in eastern Africa. Thirteen new red kidney varieties combining multiple stress resistance with high yield potential and marketable grain characteristics are released in seven countries in east and central Africa. Eight new speckled sugar varieties with multiple disease resistance, marketable grain types and high yield potential (up to 24% over commercial checks) released for smallholder production in four countries in eastern Africa. Eight new small and medium red bean varieties combining multiple disease resistance, high yield potential and marketable grain characteristics released for smallholder production in three countries of eastern Africa. Eighteen new tan, brown and yellow seed varieties with multiple resistance to diseases and high yield potential released for production by smallholder farmers in four countries in eastern Africa Twenty-six new climbing bean varieties combining multiple resistances to diseases with high yield potential and marketable grain characteristics released for smallholder production in seven countries in east and central Africa. Several segregating populations and fixed lines in different market classes from South Africa, Malawi and Tanzania are available for distribution to interested NARS partners within SABRN and others in Africa From the regional breeding program 24 (brown/khaki), 73 (sugar) and 213 (red mottled) developed for resistance to angular leaf spot, or common bacterial blight or low soil fertility or a combination of these stresses were distributed to various NARS programs. Twenty-nine lines in various market classes which were developed for rust resistance or a combination of rust and angular leaf spot or rust and halo blight resistance by the NARS in South Africa were sent to the SABRN coordinator for seed increase and onward distribution to other interested NARS for the next planting season. Twenty four new lines with a resistance gene to Pythium root rot were identified and included in the Pythium root rot nursery. Forty lines combining resistance to Pythium root rot and angular leaf spot were identified and available for sharing with partners Activity 2.4 Yield potential: climbing beans Highlights: • QTL for growth habit and climbing ability were identified on six chromosomes, although many were located on B04. This illustrates the complexity of growth habit, and implicitly, of crop domestication as growth habit was reduced from climbing to bush type. xix Activity 2.5 Characterizing and monitoring pathogen and insect diversity Highlights: • Sixteen Pythium species were found to be associated with beans root rots in Rwanda and the cultivars CAL 96, RWR 617-97A, Urugezi and RWR 1668 were susceptible to all these species. G2331, AND 1062, MLB-40-89A, Vuninkingi, AND 1064 and RWR 719 were resistant. Activity 2.6 Developing integrated disease and pest management components Highlights: • • • • All of the bean-planted areas in Colombia and Ecuador included in the Fontagro-financed project “Reduction in the Use of Pesticides and Development of Resistance in Rice and Common Bean Crops in Colombia, Venezuela and Ecuador” were monitored for white fly populations. Whitefly species and biotypes, thrips and leafminers were identified, patterns of pesticide use for whitefly, thrips and leafminers registered, and levels of pesticide resistance in whitefly, thrips, and leafminer populations in Colombia quantified. Psuedomonas sp were isolated and shown to have antagonistic effects on Pythium spp Genetic diversity of varietal mixtures from southwest Uganda was characterized with indications of its potential value in the region. Product 3: Beans that respond to market opportunities Activity 3.1 Development of large white beans for international markets Highlights: • Crosses have been developed to combine alubia grain type (uniform, milky white, long cylindrical seed) with drought resistance. Activity 3.2 Breeding Navy and Large White bean varieties with multiple stress resistance in eastern Africa Highlights: • Fourteen new navy and large white bean varieties combining multiple resistance to diseases and abiotic stress factors, high yield potential and marketable grain characteristics released for smallholder production in four countries in eastern Africa Activity 3.3 Identification of a varietal candidate in Nicaragua with potential for international export Highlights: • The bean program of INTA-Nicaragua has selected a line for varietal release with the purpose of exporting grain to the USA. xx Activity 3.4 Progress in development of Snap and runner beans for smallholder production in East and Central Africa Highlights: • • Twenty bush and climbing snap bean lines with consumer preferred pod characteristics and resistance to diseases selected in four countries in eastern Africa. Nineteen new short-day runner bean lines with high pod yield potential selected making smallholder production under short-day conditions in eastern Africa feasible. Product 4: Strengthened institutions that enhance bean product development and delivery Activity 4.1 Strengthened capacity of NARS: increasing the knowledge and skills of scientists and staff from NARIs, NGOs and Rural Service Providers Highlights: • • • • • • • A total of forty-nine students conducted research activities related to their thesis work, of which twenty eight were at CIAT HQ, and twenty one in Africa. Of these, three PhD and one MSc students were as visiting researchers at HQ. In Latin America, two M.Sc. candidates, and two pre-graduate students completed their research theses. In Africa three PhD, four M.Sc. and one Bs. candidates completed their research theses. A total of thirty-three students continue their studies, as follows: six Ph.D. candidates in Africa and five in Latin America, eight M.Sc. candidates in Africa and four in Latin America, and ten pre-graduate in Latin America. Twelve visiting researchers coming from Colombia, Cuba, Denmark, Guatemala, Hungary, India, Panama and Zambia received training in different disciplines at headquarters. Several courses and workshops were held in Latin America and Africa During this reporting period there was a joint stakeholders meeting for PABRA partners, as well as a joint steering committee meeting for SABRN and ECABREN where they reflected on the progress over the past 4 years, and planned activities to achieve the milestones contributing towards achieving the goals in the final year of the project, as well as to plan for the next phase. A number of students have either registered or started their course work at various universities to sharpen their knowledge and skills in bean research for development. Two new students doing MSc in plant breeding enrolled at the University of Zambia and Penn State University, both from Malawi. Two other students had been accepted for Ph.D. programs at the University of Free State and Massey University – New Zealand. These scientists will add to the existing capacity for bean research in the region. Activity 4.2 Strengthen international collaboration through networks (Intra- and internetwork collaboration), bi-lateral relations, and/or joint special projects Highlights: • The Pan African Bean Research Alliance (PABRA) continued to provide funding support to research for development sub-projects within the SABRN xxi Activity 4.3 Supporting breeding programs in NARS, regional networks, farmers’ Associations, and CIALs with germplasm and technical knowledge Highlights: • More fixed lines with high yield potential and resistance to diseases and cultivars of commercial value for export market were distributed in a regional yield trial to various NARIs partners in different countries. • Early generation selections of interspecific crosses for high iron have been realized in ICTA, Guatemala and are pending shipment to CIAT for analysis. • Selections from EAP-Honduras have been analyzed and returned to the breeder there. • High iron lines are in validation trials in Nicaragua and a variety could be released in the course of 2009. Activity 4.4 Development of sustainable seed systems to support wide dissemination Highlights: • • • A strategy of marketing small seed packets as a profitable enterprise is being developed with a private seed company in Kenya and shows great promise for reaching thousands of bean growers. An analysis of the effectiveness of training in seed production suggests that participants have significantly improved both technical and communication skills. A seed security assessment methodology has found acceptance at the institutional level among important players such as FAO, USAID and important international NGO’s. Activity 4.5 Socio-economic activities Highlights: • A baseline study to determine the role of beans in drought prone areas of eastern Kenya has been completed. Beans are the second most important food after maize and are critical for food security. xxii EXECUTIVE SUMMARY ANNUAL REPORT 2008 Outcome Line SBA-1 Improved Beans for the Developing World TABLE OF CONTENTS PAGE NO. 1. Product Line LogFrame as in MTP 2008-2010 2 2. Improved beans for the Developing World - 2008 project output targets 12 3. Research Highlights in 2008 3.1. Drought resistance and yield potential in Andean beans 3.2 Baseline study on the role and importance of common bean in drought prone areas of East Africa 3.3 Application of MAS in support of the Ethiopian national bean improvement program 14 14 14 4. Project outcome 16 5. List of 2008 Publications (includes in press, in review and submitted) 5.1 Book chapters and books 5.2 Refereed and non-refereed journal articles 5.3 Workshop and conference papers 5.4 Proceedings, posters, abstracts and others 5.5 Editorial contribution 17 18 19 21 23 26 6. List of special projects 6.1 At Headquarters 6.1.1 New proposals approved in 2008 6.1.2 List of ongoing special projects in 2008 6.2 In Africa 6.2.1 New proposals approved in 2008 6.2.2 List of ongoing special projects in 2008 6.2.3 Regional research subprojects under SABRN 6.3 List of projects submitted, proposals, and concept notes prepared 6.3.1 At Headquarters 6.3.2 In Africa 7. Staff list 7.1 Staff at Headquarters 7.2 Staff in Africa 27 27 27 27 29 29 29 30 34 34 34 36 36 36 8. 36 Summary 2008 budget prepared by Finances 1 14 1. PRODUCT LINE LOGFRAME IMPROVED BEANS FOR THE DEVELOPING WORLD: PRODUCT LINE SBA1 Rationale & Changes Rationale The common bean (Phaseolus vulgaris L.) is the world’s most important grain legume for direct human consumption. Its total production exceeds 12 million MT, of which 7 million MT are produced in tropical Latin America and Africa. Beans are the “poor man’s meat” and are particularly important in the diet of the underprivileged. Beans, like other legumes, supply proteins, carbohydrates, vitamins and minerals, and complement cereals, roots and tubers that compose the bulk of diets in most developing countries. Common bean is also one of the most diverse crops in terms of its cultivation methods and its uses. It serves as mature grain, as immature seed, and as a vegetable (both leaves and pods), and after harvest the stover is used as animal fodder. It is cultivated from sea level up to 3000 masl in monoculture, in association, or in rotations. The possibility of obtaining a harvest in as little as two months offers quick income, quick food supply, and also permits rotating with other crops or inter-planting among fruit trees or coffee before the primary crop produces income. At the other extreme are the aggressive climbing beans that subsistence farmers maintain in the garden for food security and continual harvest over a six month period. Apart from subsistence cultivation, beans have become increasingly commercial over the past thirty years in national, regional and international markets. In Central America beans are the #1 income generator among the traditional field crops. In Africa, farmers tap into regional bean markets in Nairobi, Kinshasa and Johannesburg. With the onset of globalization, the past decade has seen a growing international market that is now reported to reach 2.4 million MT. This heightens issues of equity for the small bean producers that have little other stable source of income, but some also see this as an opportunity. For example, bean represents 6% of external income for Ethiopia, and small farmers in Bolivia produce the large white and red mottled classes for export. Snap beans are a high value, labor intensive crop of small farmers in Kenya and the Andes. Besides the common bean, another four cultivated species are conserved in the CIAT gene bank, as well as wild relatives. This collection is the largest of the genus in the entire world, representing more than 35,000 accessions that have been declared as part of the designated collection before FAO. These other cultivated species fill niches that are unsuitable for the common bean, for example, P. acutifolius that thrives in desert environments. Our primary mission is to contribute to household and global food security by assuring an adequate supply of beans as a culturally acceptable and traditional staple; and to improve the income of small bean producers of Latin America and Africa, by making bean production more profitable. We also seek to improve human nutrition, both by augmenting the supply of beans, and by improvement of their nutritional value. Our products are designed to respond in particular to the needs of small, resource-poor bean farmers in Latin America and Africa. Thus, we seek to create solutions to biotic and abiotic production limitations that require minimal inputs, and in the case of improved germplasm, with good market potential. Our research strategy focuses on the exploitation of the vast genetic resources of bean that exist as a complex array of major and minor gene pools, races and sister species. CIAT’s gene bank with 41,000 accessions of common bean and related species is our most unique resource, and has been the source of genes for 2 disease and insect resistance, abiotic stress tolerance, nutritional quality and yield potential. Most traits are still selected by conventional means in field sites (in some cases backed up by greenhouse evaluations) where most important diseases, edaphic constraints and drought can be manipulated for purposes of selection. However, Marker Assisted Selection (MAS) is employed selectively but strategically, in most cases for disease resistance genes. CIAT pioneered participatory selection with farmers and this practice is being extended and systematized. While most products are seed based, others involve agronomic practices or are knowledge based. Our research is strategic combined with both basic and applied elements, as called for by the particular challenge. Changes There have been no essential changes in relation to the MTP of 2007. However, in 2008 an agricultural economist, Dr. Enid Katungi came on board under the Tropical Legumes-II project, with base in Kampala, Uganda. CG System Priorities CIAT’s bean product line is housed principally under CG System Priority Area 2: Producing more and better food at lower cost through genetic improvements. Efforts are dedicated to improving yields through control of diseases and pests, tolerance to abiotic stresses (drought, aluminum toxicity and low soil fertility in particular), and expanding the adaptation range of climbing beans. The bean product line also places heavy emphasis on improvement of nutritional quality, especially through increase in iron and zinc content in the grain. There is potential to contribute to Priority Area 3A: Increasing income from fruits and vegetables, through the improvement of snap beans for both Africa and Latin America. The bean team collaborates with marketing specialists to create varieties with better market potential, including international export markets (Priority Area 5B). Finally, strengthening national institutions (Priority Area 5A) continues to be an important product, both in Africa where novel institutional arrangements and relations have been productive to achieve wide impact, and in Latin America where staff reductions have weakened national programs. On both continents national programs seek support to incorporate modern selection techniques. Impact Pathways Product 1 (Beans with improved micronutrient concentration that have a positive impact on human health) is targeted to small farmers and poor rural and urban consumers in Africa and Latin America. Targeting is developed in collaboration with nutritionists and with experts in GIS, to address human populations with nutritional deficiencies in iron and zinc. This product involves both small seeded germplasm that is often targeted to warmer climates or more difficult environments in Central America, Mexico, Venezuela, East Africa and Brazil. Large seeded germplasm is usually cultivated in more temperate climates in the Andean zone, the East African highlands and southern Africa, although in the African highlands small and large seeded types overlap, sometimes differentiated by soil fertility gradients within the farm, prevailing biotic constraints and household preferences. Improved germplasm is shared or developed jointly with NARS partners, who supply basic seed to a range of organizations interested in production of seed (local seed companies, NGO’s, CBO’s, women’s groups) who in turn distribute to farmers. NGOs and health workers play a special role in delivery. Benefits accrue to farmers/consumers through stable food supply of more nutritious beans for home consumption, and potentially to poor urban consumers. Assumptions for the successful delivery of these products include institutional and financial stability of partners, political stability, and institutional support. The role of CIAT is that of a primary research provider (of improved germplasm), at times a secondary research provider (backing up national bean improvement programs with technical expertise and training), and catalyser (to promote downstream alliances in the uptake chain). This product is complementary to those of CIMMYT and CIP. 3 Beneficiaries of Product 2 (Beans that are more productive under low input agriculture of poor farmers) are in some cases researchers (both inside and outside of CIAT), and in some cases are bean producers. For example, molecular markers for resistance genes benefit researchers directly, and farmers indirectly as subsequent beneficiaries. Uptake pathway for such methodologies is direct communication through workshops and courses, and indirectly through publications, leading to benefits of more efficient and effective bean research. This assumes that partners are in a position to implement such technologies. On the other hand, crop management practices are of direct benefit to farmers as users, potentially across all bean ecosystems. Uptake chains for agronomic practices are similar to those for seed based technologies; results are communicated to NARS and other partners (NGO’s, CBO’s etc) who have successfully diffused practices to farmers, to the benefit of farmers who enjoy more stable productivity. Improved germplasm is diffused through many of the same channels as beans with improved nutritional value, with the exception that partners may have less specific interests, and may be more production oriented. The role of CIAT is that of primary source of research for development. Product 3 (Beans that respond to market opportunities) benefit small farmers in both Latin America and Africa. Farmers in Ethiopia have already benefited from tapping into export markets for canning beans, and other countries are positioning themselves to follow suite. In Central America exporters are seeking to fill a niche created by the Latin population in the USA. This is a demand-driven activity, and in large part has generated its own impact pathway. Exporters and international grain buyers have established market chains that give them access to export quality beans. CIAT’s role has been that of supplying germplasm in some cases, and in others to facilitate communication, and to give support in seed systems to avail quality seed to farmers of very specific varieties. Product 4, (Strengthened institutions that enhance product quality and delivery) seeks to benefit partners at multiple levels through facilitated interaction, including farmers who are at the end of the organizational chain. NGOs, government extension agencies, farmer organizations, local seed companies, and non-conventional seed actors such as women groups, people living with HIV/AIDS and tobacco companies all participate and benefit. The product will generate impact on target beneficiaries through their participation in development of innovations, knowledge and technologies in strategic alliances with multidisciplinary research teams and NGOs. Scaling out of innovations and best practices to areas with similar environments will be done through strategic alliances of research and development actors. The latter will use their network and other communications mechanism to adapt knowledge and results relevant to them. Scaling up regionally and internationally will be done through international NGOs, advocacy, and communication. The outcome is enhanced communication and complementarity of actors with resulting cost efficiencies, and in the case of technology diffusion, increased and diversified adoption. Another dimension of this product is support to NARS in development of projects, benefiting national program researchers and with the outcome of their integration into the product line research mode. This assumes a degree of consistency in partner personnel, while CIAT’s role is that of facilitator. International Public Goods The IPG of the bean product line include: • Improved germplasm with biotic and abiotic stress tolerance, and/or enhanced nutritional value, drawing upon the genetic resources of CIAT’s extensive gene bank, and 30 years of experience in bean improvement. CIAT’s geographical position and access to varied altitudes and research sites facilitates study and selection of germplasm. • Improved practices for the management of pests and diseases, including monitoring of pathogen populations with modern molecular tools developed at CIAT. • Knowledge and tools that contribute to the development and implementation of the above IPG’s. For example, molecular markers for useful traits, developed with CIAT’s in-house resources of genetic maps and markers. Knowledge of the structure of genetic resources housed in the gene bank, and ways to exploit them. Screening methods to identify biotic and abiotic stress resistant 4 • genotypes. Participatory breeding methods with varying degrees of involvement of farmers, traders and other key actors. Methods for networking, both formal among official sector researchers, and less formal among a broader range of partners, with special emphasis on research partnerships and on effective and sustainable seed systems reaching a large number of households. Partners Most important partners and the respective person-years of professionals dedicated to bean research within the (several) products are: Product 1: NARS in Latin America, including those of Mexico (6), Guatemala (2.5), Honduras (2, including EAP-Zamorano), El Salvador (2), Cuba (2), Brazil (4) participate in the AgroSalud project to improve nutritional quality and productivity of bean. NARS in South America, including those of Colombia (5 between university staff, an NGO and the NARI), Bolivia (4 between university staff and a foundation) collaborate in the improvement of disease resistance of Andean bean with better nutritional quality, also under the AgroSalud project. NARS in East, Central and Southern Africa, including those of Kenya (5), Rwanda (6), Uganda (5), Malawi (1), Zimbabwe (1) are partners in the improvement of nutritional qualities in large seeded Andean beans. Linkage funds finance a project with one Canadian university, and with a partner in USDA. Product 2: Nicaragua (4.5) and Honduras (2) are partners in breeding for drought tolerance. NARS in East, Central and Southern Africa including those of Ethiopia (3), Kenya (2), Tanzania (3), Rwanda (4), Malawi (1), Zimbabwe (1) and DRCongo (4), participate in the improvement of productivity under low soil fertility and/or drought. The University of Hannover, Germany participates in a project to define physiological mechanisms of aluminum tolerance and drought resistance (2), which also includes Malawi (2) and Rwanda (4). Catholic University of Leuven (3) is a partner to improve nitrogen fixation technology. NARS in South America, including those of Colombia (5 between university staff, an NGO and the NARI), Bolivia (4 between university staff and a foundation) collaborate in the improvement of disease resistance of Andean bean. NARS in East, Central and Southern Africa, including those of Kenya (5), Rwanda (6), and Uganda (5) Tanzania (4) are partners in the development of disease resistance, medium altitude climbing beans (MAC), and productivity in large seeded Andean beans. NARS in Honduras (Zamorano) (1), Colombia (2), Uganda (3), Rwanda (4), and South Africa (2) share in the use of markers for MAS, especially for resistance. South Africa (3) participates in pathogen characterization, evaluation and validation of resistance sources. Agriculture and Agri-Food Canada (AAFC) is a partner in diagnosis and characterization of soil borne pathogens (especially Pythium species) using molecular techniques, and development of molecular based diagnostic assays for soil borne pathogens. Product 3: Partners in Latin America with specific attention to breeding market quality include NARS in Honduras and Nicaragua. NARS in Africa with active participation in canning beans include those of Ethiopia and Uganda. Partners in the development of snap beans include a university in Colombia, and one in Kenya. Product 4: NARS as above –plus a wide range of NGOS, CBOS, farmers’ groups, women’s groups, – totaling over 300 direct-link partnerships, to make users aware of technologies and to get these technologies widely disseminated. The ECABREN and SABRN bean networks coordinate nine NARS in East Africa and ten NARS in southern Africa, respectively. These networks participate in Products 1, 2, 3 and 4 with input from African NARS cited above, plus NARS in Burundi (3), Sudan (2), Zambia (1), Zimbabwe (1), Mozambique (3), Lesotho (3) and Swaziland (3). 5 HarvestPlus Challenge Program: IFPRI, CIMMYT, and CIP are immediate collaborators in the CP and the AgroSalud (Latin American) nutritional improvement project, working in the same agro-ecological zones, while ICRISAT, IITA, IRRI, and ICARDA are indirect collaborators under HarvestPlus. ECABREN and SABRN networks in Africa also participate in HarvestPlus. Generation Challenge Program: Partners include EMBRAPA-Brazil (2), INTA-Cuba (1), Pairumani (an NGO) in Bolivia (2), National University in Colombia (2). Sub-Saharan Africa Challenge Program: ICIPE, AHI and NARS in Rwanda, Uganda and D.R. Congo are immediate partners. Product line Funding Budgeting 2007-2011 Year US Dollars (millions) 2007 (actual) 8.008 2008 (actual) 9.931 2009 (proposal) 7.597 6 2010 (plan) 7.702 2011 (plan) 7.812 IMPROVED BEANS FOR THE DEVELOPING WORLD: PRODUCT LINE SBA1 (2008-2010) Targets PRODUCT 1 Products Beans with improved micronutrient concentration that have a positive impact on human health NARS, farmers & consumers in Central America, the Caribbean, Brazil, East and Southern Africa Adoption of improved varieties by farmers Better nutritional status, especially of rural consumers • ~30 small seeded F3-derived F5 bush bean families developed with tropical adaptation, 60% more minerals, abiotic stress tolerance, and 2 biotic resistances for Central America (HarvestPlus) • NARS, NGO’s CBO’s, health workers, and farmers in target countries • Farmers incorporate high mineral and disease resistance lines into diverse production systems • Reduced levels of iron and zinc deficiency in bean consumers • 50 improved lines with varietal potential and 90 ppm iron (ie, 80% more iron) 15 new large seeded climbing beans with high mineral trait (HarvestPlus) Marker assisted selection for one nutritional trait (iron) tested Four fast track micronutrient dense bean varieties disseminated and promoted in two countries in eastern and southern Africa Two large seeded lines with 50% more iron enter formal varietal release process in eastern Africa • NARS, NGO’s CBO’s, health workers, and farmers in target countries • Adoption of micronutrient rich beans • Reduced levels of iron and zinc deficiency in bean consumers • NARS, NGO’s CBO’s, health workers and consumers • Adoption of micronutrient rich beans • Reduced levels of iron and zinc deficiency in bean consumers Product Targets 2008 Product Targets 2009 • • • Product Targets 2010 • Intended User 7 Outcome Impact Products Intended User Outcome Impact Beans that are more productive in smallholder systems of poor farmers Breeders and pathologists in CIAT and NARS; farmers in E and S Africa, Andean zone, Caribbean Adoption of improved varieties by farmers; Best bet IDPM practices and genetic combinations for stable resistance deployed. More stable production, food availability and income • 5 molecular markers for detection, diagnosis and diversity studies of ALS and anthracnose pathogens made available At least 10 lines in major market classes combining resistance to Pythium root rots, BCMV and angular leaf spot An IPM system for whiteflies on snap beans refined and promoted in 2 major bean producing areas of the Andean zone • • An IDM system for bean root rots implemented and promoted in 2 major bean producing countries in Africa At least 40 lines combining drought resistance with resistance to BCMNV, root rots, and/or ALS available for testing in Africa 2 molecular markers linked to ALS and Pythium root rot implemented in MAS • Resistance genes for anthracnose or ALS introgressed into 5 BCMNV resistant climbing beans At least 10 genotypes combining drought resistance with aluminium resistance available for testing in Africa • Targets PRODUCT 2 Product Targets 2008 • • • • Product Targets 2009 • • Product Targets 2010 • • • NARS, NGO’s and farmers’ groups CIAT and NARS breeders NARIs researchers in LAC, Africa, IARCs • • • NARS breeders, NGO’s, CBOs, and farmer groups NARS pathologists, • • • • NARS breeders, NGO’s, CBOs, and farmer groups NARS soil scientists and agronomists 8 • • Disease and pest characterization tools adopted by researchers Adoption of disease resistant lines in marginal environments Increased utilization of integrated management approaches. • Improved food security, & income. • More stable disease resistance in advanced lines leads to stable yield Resistant lines incorporated into improved systems Drought resistant lines with disease resistance used in drought prone areas in Africa Breeders improve efficiency of genetic improvement • Reduced yield losses from ALS, root rots and drought Farmers benefit from yield stability of high yield climbers Farmers benefit from stable yields in marginal areas • Improved food security, & income. Targets Products Beans that respond to market opportunities • • • Product Targets 2009 Product Targets 2010 • • Outcome Impact NARS in Africa and Latin America Adoption of commercial varieties by farmers, enhancing access to markets Higher income, especially for the poor and women farmers 10 lines of snap beans with confirmed resistance to Gemini virus in Colombia 1 variety released in Nicaragua for export market • • • At least 3 snap bean lines with resistance to rust and quality characteristics preferred in regional and export markets for Africa. 4 bean genotypes with very high commercial or export quality made available to farmers in 4 countries in Latin America and Africa • 5 canning bean lines with acceptable quality characteristics in yield trials in two countries in eastern Africa • PRODUCT 3 Product Targets 2008 Intended User NARS, NGOs, CBOs, farmer groups, seed producers Farmers reduce pesticide use, assuring production and profitability • NARS, NGOs, CBOs, farmer groups, seed producers NARS, NGOs, CBOs, farmer groups, seed producers 9 • Adoption of snap bean and reduced chemical use. • Farmers in marginal environments assure market access • Farmers improve yields and quality of product with improved varieties Less pesticide intoxication in rural communities and urban consumers Increased production and incomes. • Increased production and incomes. • Increased production and incomes. Targets PRODUCT 4 Products Strengthened institutions that enhance bean product development and delivery Intended User NARS in Africa and Latin America • • NARS, NGOs, CBOs, farmer groups, seed certification agencies, seed producers • UN, humanitarian and post-stress recovery organizations • PABRA • Product Targets 2008 • • • • Product Targets 2009 • One comprehensive methodology developed for assessing seed security and targeting responses in acute and chronic stress situations. Lessons from 3 case studies (approaches for partnership; capacity building; alternative seed delivery systems) of strategies for product development and delivery in PABRA analyzed. Protocols developed and adapted to facilitate application of MAS for disease resistance in 3 African countries Breeding programs for higher iron levels established in Honduras, Nicaragua, Bolivia, Venezuela, Kenya and Malawi A guide for mainstreaming and sustaining wider impact, developed and recommendations availed for 5 countries in East, Central and 4 countries in Southern Africa Three delivery channels strategies tested for reaching the poor and in marginal areas with new variety innovations and information At least 1 methodological frameworks/strategies for testing and evaluating multi-stakeholder networks and platforms (between private-public) for facilitating decentralized targeting for pro poor impact. o NARS, NGOs, Decentralized Local Governments, CBOs, farmer groups, seed certification agencies, seed producers ,agroprocessors, local financial institutions • UN, humanitarian and post-stress recovery organizations 10 Outcome Improved institutional performance by NARS, NGOs and other partners, reflected in more effective technology development and dissemination o Frameworks and methodologies for seed systems, PM&E, and MAS are in use by PABRA partners • Increased partner involvement in accessing technologies to a greater number of end users • Increased capacities of partner organizations / institutions to develop and promote integrated and decentralized strategies for reaching pro-poor farmers Impact More stable production, improved food availability, income and nutrition, especially for the poor and women farmers Products Targets Product Targets 2010 PRODUCT 5 Product Targets 2008 • Capacity to evaluate root systems in soil tubes established in Honduras and Nicaragua • Elements of Pro-poor seed delivery and production systems confirmed and such pro-poor seed enterprises established in 2 PABRA network countries. One strategy for wider utilization of non varietal bean technologies (IPM; soil management) developed and widely shared in 4 countries in Africa • • 1500 accessions conserved in long term storage and in back-up in CIMMYT 1000 samples of bean seed distributed • Bean scientists; other gene banks o Novel genes incorporated into breeding programs Another 1500 accessions conserved in long term storage and in back-up in CIMMYT Another 1000 samples of bean seed distributed A plan formulated to establish a database of evaluation data • Bean scientists; other gene banks o Novel genes incorporated into breeding programs Another 1500 accessions conserved in long term storage and in back-up in CIMMYT Another 1000 samples of bean seed distributed • Bean scientists; other gene banks o Novel genes incorporated into breeding programs • • • • Impact NARS, NGOs, CBOs, farmer groups, seed certification agencies, seed producers Breeders, geneticists, and other bean scientists; national gene banks • Product Targets 2010 • Outcome More than 35,000 accessions are conserved, documented and available for distribution • Product Targets 2009 Intended User 11 Bean genetic resources are used directly or employed in breeding programs More stable production, improved food availability, income and nutrition 2. IMPROVED BEANS FOR THE DEVELOPING WORLD – 2008 OUTPUT TARGETS TARGETS 2008 Fully Achieved X 75% Achieved >50% Achieved <50% Achieved EXPLANATION Cancelled Deferred To be documented in 2008 Annual Report PRODUCT 1 • ~30 small seeded F3-derived F5 bush bean families developed with tropical adaptation, 60% more minerals, abiotic stress tolerance, and 2 biotic resistances for Central America (HarvestPlus) X Seven locus-specific microsatellite markers for ALS pathogen, which quickly distinguish between Andean and Mesoamerican pathogen groups were identified. Work on anthracnose was not pursued after the responsible pathologist left CIAT X Lines combining Pythium root rots and ALS and those with BCMVN in early generation. PRODUCT 2 • 5 molecular markers for detection, diagnosis and diversity studies of ALS and anthracnose pathogens made available • At least 10 lines in major market classes combining resistance to Pythium root rots, BCMV and angular leaf spot • An IPM system for whiteflies on snap beans refined and promoted in 2 major bean producing areas of the Andean zone X Partially in 2007 report with additional documentation in 2008 Annual Report 12 TARGETS 2008 Fully Achieved 75% Achieved >50% Achieved <50% Achieved Deferred X PRODUCT 3 • 10 lines of snap beans with confirmed resistance to Gemini virus in Colombia • EXPLANATION Cancelled X Weather conditions in Colombia did not permit the build up of the white fly vector to be able to evaluate lines in the field PRODUCT 4 • One comprehensive methodology developed for assessing seed security and targeting responses in acute and chronic stress situations. X A new line with commercial grain type is already in commercial production for export but is not officially released. To be documented in 2008 Annual Report • Lessons from 3 case studies (approaches for partnership; capacity building; alternative seed delivery systems) of strategies for product development and delivery in PABRA analyzed. X To be documented in 2008 Annual Report • Protocols developed and adapted to facilitate application of MAS for disease resistance in 3 African countries • Breeding programs for higher iron levels established in Honduras, Nicaragua, Bolivia, Venezuela, Kenya and Malawi 1 variety released in Nicaragua for export market X Protocols developed but adaptation in three countries delayed because of a delay in the start of Kirkhosue Trust supported projects (in 4 countries) which was to provide infrastructure and also support capacity development in collaboration with CIAT. This project start in 2009 To be documented in 2008 Annual Report X 13 3. RESEARCH HIGHLIGHTS IN 2008 We will highlight 3 areas of our current research portfolio: 3.1. Drought resistance and yield potential in Andean beans Contributors: S. Beebe, M. Blair, I. Rao, M. Grajales, C. Cajiao, F. Monserrate Breeding for drought resistance in the small seeded Mesoamerican beans has been successful, but the large seeded Andean beans have received less attention. In 2007 and 2008 advanced breeding lines with commercial Andean grain types were tested under drought, and in 2008 the same lines were evaluated in Palmira with irrigation, and at a mid-altitude site (1400 masl) in Darién under rainfed but favorable conditions. Several lines expressed an advantage of about 50% in the drought trials over check cultivars in three grain classes (large red; cream striped; and large white) while progress in the red mottled class was more modest. Furthermore, in the irrigated plots and in the mid-altitude site, where the Andean beans normally adapt especially well, some drought tolerant lines yielded as much as a ton more than the checks. This finding is comparable to that with Mesoamerican beans, whereby drought-selected lines expressed improved yield potential, a finding that has been attributed to better remobilization of biomass from vegetative parts to grain. The current results suggest a similar trend in Andean beans. Yield improvement has been especially difficult in Andean beans, and these results may indicate a means to overcome this long term bottleneck. 3.2 Baseline study on the role and importance of common bean in drought prone areas of East Africa Contributors: E. Katungi, L. Sperling, A. Farrow The bean program is undertaking massive diffusion of drought resistant varieties in drought prone areas of east Africa. A socio-economic baseline survey was conducted in semi-arid areas of Kenya (Eastern province) and Ethiopia (Oromia and Southern region) to contextualize this effort and to orient the breeding for drought resistance. A total 360 farming households in 18 villages, and 120 traders along the value chain were interviewed in the two countries. In Kenya farmers integrate a diversity of crops, cropping systems and farming management practices with local ecosystems and livelihoods to cope with drought. They dry-plant their crops, make terraces to harvest water, intercrop intensively, keep livestock, invest in social capital, work outside their farms for food or wage and undertake petty trade and handcraft but still experience an average of 5 months of inadequate food supply per year. Drought is ranked the most important constraint to livelihood improvement, causing about 70% yield loss in common beans when it occurs. Nevertheless, common bean is ranked the second most important food crop after maize, with about 70% of households growing from 3 to 10 varieties simultaneously, primarily for home consumption. Household characteristics, as well as consumption and production attributes are the driving factors that underlie variety choice and extent of planting. The breeding effort should target both categories of attributes. 3.3 Application of MAS in support of the Ethiopian national bean improvement program Contributors: M. Blair, H. Buendía, S. Beebe, T. Assefa, C. Cardona, J.M. Bueno The arcelin seed protein is the most effective resistance factor for the storage pest of common bean, Zabrotes subfasciatus (Boheman). Crosses were made between arcelin-containing RAZ lines and a series 14 of Andean and Mesoamerican beans with drought tolerance useful for Eastern and Southern Africa (Ethiopia, Kenya, Malawi, Tanzania and Zimbabwe). For Ethiopia, crosses were generated to incorporate arcelin into a drought tolerant background and then transfer that resistance/tolerance to the small white, Ethiopian variety ‘Awash Melka’. Double crosses were generated with Andean types including the Malawian release CIM9314-34, the Kenyan releases KAT B1 and KAT B9 and other African cultivars such as Canadian Wonder, CAL96 and CAL143. Marker assisted selection (MAS) is applied for the arcelin gene to facilitate the pyramiding of bruchid resistance with other biotic and abiotic stress resistances. MAS was carried out using microprep DNA. For Andeans, a total of 251 F 1 plants segregated for the arcelin locus, and of these, 236 amplified with the arcelin marker. For improvement of Awash Melka, a total of 498 F 1 plants segregated for the arcelin locus in seven different pedigrees. This latter work represents support to an Ethiopian Ph.D. candidate. This represents the first application of MAS for insect resistance in common bean. 15 PROJECT OUTCOME: 4. Managing Bean Root Rot - A constraint Associated with Intensification in Land Use Outcome statement: National program breeders and pathologists initiate breeding programs and select resistant lines based on information of pathogen distribution defined by CIAT pathologists. This outcome results from an output target in CIAT’s 2004-2007 MTP: “Pathogen distribution maps developed for ALS, anthracnose, Pythium and Fusarium.” Results meeting this target were reported in the 2005 Annual Report (pp. 182-185). It is also associated with the target, “Improved germplasm available to NARS, regional networks, and farmers, combining better yield with disease resistance”, by availing root rot resistant lines to partners in Africa (MTP 2004, 2005). Context: Intensified land use in the highlands of Eastern and Central Africa has been associated with the increased incidence of bean root rots, a devastating disease caused by a complex of soilborne pathogens, mainly Pythium species. In 2001, over 75% of farmers reported calamitous declines in bean production associated with root rots in a survey in western Kenya districts of Kakamega and Vihiga. These districts and those of southwestern Uganda and many parts of Rwanda are typical of regions affected by root rot: farm sizes are small (average 1-2.6 ha), population densities high (404 persons /km2 in Kakamega and 938 in Vihiga), and crop rotation near nil. CIAT identified major Pythium species prevalent in Kenya, Rwanda and Uganda on the basis of cultural and molecular techniques. Species distribution and prevalence were mapped, including at key root rot “hot spots”. This basic information was then used by breeders in East Africa to guide germplasm evaluations and varietal improvement programs. The regional breeder backstopping NARS breeding programs in East and Central Africa Bean Research Network (ECABREN) evaluated a range of germplasm representing different market classes using artificial inoculation of representative Pythium species and at a key “hot spot” in Western Kenya. A number of resistant germplasm such as AND 1055, NR 12793-8-1, NR 12631-7-1, RAB 475, DFA 52 and NM 12803-11 were identified (RF & CIAT Reports). Similarly NARS breeders from Kenya, Uganda, Rwanda, and southern Democratic Republic of Congo used the knowledge to evaluate nurseries and segregating populations for resistance to prevalent Pythium species (Musoni, et al. – in press; Otsyula, PhD thesis; Kimani et al, 2005; ECABREN Report; CIAT Annual reports) at respective “hot spots” (Vihiga, western Kenya; Kabale, southwest Uganda; Runyinya, Rwanda. Representative isolates (maintained at Kawanda, Uganda) were used to artificially screen germplasm from the three countries. A breeder from KARI, Kakamega, Kenya used the identified species to study the nature of resistance and mechanism of inheritance in selected sources of resistance (Otsyula, 2005 – Rockefeller meeting, PhD 2009 thesis). In addition he and his counterparts in Rwanda (ISAR) and Uganda (NARO) used the “hotspots” above and artificial inoculation of identified Pythium species to select resistant progenies from populations developed to improve root rot resistance in local bush (e.g. GLP-2, CAL 132, Urugezi) and climbing beans. Following extensive artificial inoculations with key Pythium species (P.ultimum var ultimum, P. salpingophorum, P. spinosum , P. torulosum, P. pachycaule) and evaluations under natural conditions at “hotspots” by CIAT and NARS partners in Uganda, Rwanda and Kenya, resistant germplasm was used to constitute a root rot nursery. About 80 entries were made available to several NARS partners in Africa (Kenya, Uganda, Rwanda, DRC, Ethiopia, Malawi, South Africa, and Cameroon) (CIAT Annual Reports). These partners in turn involved farmers to evaluate the materials. As a result in Uganda, two genotypes originally from Rwanda (RWR 2075 and RWR 1946) were highly appreciated by farmers and traders in evaluations over a 2 year period. The farmers gave them local names; RWR 1946 with a large dark red seed type was named “Murwanisa” meaning ‘resistant to harsh conditions’ and RWR 2075 ‘Muzahura’, meaning ‘restorer’ (Namayanja et al. Euphytica). These genotypes have been released in Uganda as NABE 13 (RWR 1946) and NABE 14 (RWR 2075), and have entered national performance trials in Kenya as well. In Kenya SCAM-CM80/15 has also been released. 16 5. LIST OF 2008 PUBLICATIONS (includes in press, in review and submitted) - see complete list 5.1 Book chapters and books (all in English) - 5.2 25 1 2 1 1 2 Papers in English: Papers in Spanish: 28 1 Proceedings, posters, abstracts, others - 5.5 Papers published in English: Papers in press in English: Papers in review in English: Papers accepted in English: Papers published in Spanish: Papers in review in Spanish: Workshop and conference papers - 5.4 6 4 Refereed and non-refereed journal articles - 5.3 Book chapters published: Book chapters in press: Proceedings: Posters: in English in English in Spanish Others: in English Media Campaign 13 10 4 4 Wires Online Broadcast Print Editorial Contributions - Scientific Committee of Agronomia Colombiana Journal Reviewed articles for: Crop Science Agroforestry Systems Acta Agronomica 17 5.1 BOOK CHAPTERS AND BOOKS Arora-Jonsson, Seema, Ballard, Heidi L., Buruchara, Robin, Casolo, Jennifer, Classen, Lauren, DeHose, Judy; Emretsson, Margareta; Fortmann, Louise; Halvarsson, Anne Lundgren; Halvarsson, Ewa; Humphries, Sally; Long, Jonathan; Murphree, Marshall W; Nemarundwe, Nontokozo; Olssen, Anne; Rhee, Steve; Ryen, Anna; Wilmsen, Carl; Wollenberg, Eva. 2008. Conclusions In Louise Fortmann (ed). Participatory Research in Conservation and Rural Livelihoods: Doing Science Together. Blackwell Publishing Ltd. Beebe, S.E., I.M. Rao, M.W. Blair and J.A. Acosta-Gallegos. 2008. Drought resistance phenotyping of common bean. Generation Challenge Program Special Issue on Phenotyping (in press). Buruchara., R.A. 2008. How Participatory Research Convinced a Skeptic. In Louise Fortmann (ed). Participatory Research in Conservation and Rural Livelihoods: Doing Science Together. Blackwell Publishing Ltd. Fortmann, L., H. Ballard, and L. Sperling. 2008. Change around the Edges: Gender Analysis, Feminist Methods and Sciences of Terrestrial Environments. In L. Schiebinger (ed). Gendered Innovations, Stanford University Press. Gepts, P., Aragao, F., Barros, E., Blair, M.W., Brondani, R, Broughton, W., Hernández, G., Kami, J., Lariguet, P., McClean, P., Melotto, M., Miklas, P., Pedrosa-Harand, A., Porch, T., Sánchez, F. 2008. Genomics of Phaseolus beans, a major source of dietary protein and micronutrients in the tropics. In P.H. Moore and R. Ming (eds) Genomics of Tropical Crops, Springer Publ., Chp 5. pp. 113-143. Jansa, J., A.Bationo, E. Frossard, I.M. Rao. 2008. Options for improving plant nutrition to increase common bean productivity in Africa. In: A. Bationo (ed) Fighting Poverty in Sub-Saharan Africa: The Multiple Roles of Legumes in Integrated Soil Fertility Management, Springer-Verlag, New York (in press). Kimani, P.M., Lunze Lubanga, Gideon Rachier and Vicky Ruganzu. 2009. Breeding common bean for tolerance to low fertility acid soils in East and Central Africa. In: Bationo, A. et al (eds). Innovations for the Green Revolution in Africa. Springer Verlag, Dordrecht, The Netherlands (accepted and in press). Mauyo, L. W. J. R. Okalebo, R. A. Kirkby, R. Buruchara, M. Ugen, and H. K. Maritim, 2007. Spatial pricing efficiency and regional market integration of cross-border beans (Phaseolus vulgaris) marketing in East Africa: The case of Western Kenya and Eastern Uganda. In p 1027-1033. Bationo et.al. (eds) Advances in Integrated Soil Fertility Management in Sub-Saharan Africa Nandwa, S.M., A. Bationo, S.N. Obanyi, I.M. Rao, N. Sanginga and B. Vanlauwe. 2008. Inter and intraspecific variation of legumes and mechanisms to access and adapt to less available soil phosphorus and rock phosphate. In: A. Bationo (ed) Fighting Poverty in Sub-Saharan Africa: The Multiple Roles of Legumes in Integrated Soil Fertility Management, Springer-Verlag, New York (in press). Teshale Assefa, H. Assefa and P.M. Kimani. 2007. Development of improved haricot bean germplasm for mid- and low altitude sub-humid ecologies of Ethiopia, pages 87-94. In: Food and Forage Legume of Ethiopia: Progress and Prospects. ICARDA, Allepo, Syria. 18 5.2 REFEREED AND NON-REFEREED JOURNAL ARTICLES REFEREED JOURNALS Akhter, A., M.S.H. Khan, E. Hiroaki, K. Tawaraya, I.M. Rao, P. Wenzl, S. Ishikawa and T. Wagatsuma. 2008. The greater contribution of low-nutrient tolerance to the combined tolerance under highaluminum and low-nutrient stresses for sorghum and maize in a solution culture simulating the nutrient status of tropical acid soils. Soil Science and Plant Nutrition (in press). Astudillo C., Blair, M.W. 2008. Evaluación del contenido de hierro y zinc en semilla y su respuesta al nivel de fósforo en variedades de fríjol colombianas. Agronomía Colombiana 26: 471-476. Beebe, S., I. M. Rao, C. Cajiao, and M. Grajales. 2008. Selection for drought resistance in common bean also improves yield in phosphorus limited and favorable environments. Crop Science 48: 582-592. Blair, M.W., Morales, F.J. 2008. Geminivirus resistance breeding in common bean. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources 3: 1-14. Blair, M.W., Buendía, H.F., Giraldo, M.C., Metais, .I, Peltier, D. 2008. Characterization of AT-rich microsatellites in common bean (Phaseolus vulgaris L.) Theor Appl Genet 118: 91-103. Blair, M.W., Porch, T., Cichy, K., Galeano, C.H., Lariguet, P., Pankurst, C., Broughton, W. 2008. Induced mutants in common bean (Phaseolus vulgaris), and their potential use in nutrition quality breeding and gene discovery. Israel Journal of Plant Sciences 55: 191 – 200. Checa, O.E., Blair, M.W. 2008. Mapping QTL for climbing ability and component traits in common bean (Phaseolus vulgaris L.) Molecular Breeding 22: 201-215. Dwivedi, S.L., Upadhyaya, H.D., Stalker, H.T., Blair, M.W., Bertioli, D., Nielen, S., Ortiz, R. 2008. Enhancing crop gene pools of cereals and legumes with beneficial traits using wild relatives. Plant Breeding Reviews 30: 179-230. Garzón, L.N., Ligaretto, G., Blair, M.W. 2008. Molecular marker assisted backcrossing of anthracnose resistance into Andean climbing beans (Phaseolus vulgaris L.) Crop Science 48:562-570. López-Marín, H.D., I.M. Rao and M.W. Blair. 2008. Quantitative trait loci for aluminum toxicity resistance in common bean (Phaseolus vulgais L.). Theoretical and Applied Genetics (in review). Mauyo, L. W., J. R. Okalebo, R. A. Kirkby, R. Buruchara, M. Ugen and R.O. Musebe. 2007. Legal and institutional constraints to Kenya-Uganda cross-border bean marketing. African Journal of Agricultural Research Vol. 2 (11), pp. 578-582 Mauyo, L.W., J. R. Okalebo, R. A. Kirkby, R. Buruchara, M. Ugen, C.T. Mengist, V.E. Anjichi and R.O. Musebe. 2007. Technical efficiency and regional market integration of cross-border bean marketing in western Kenya and eastern Uganda. African Journal of Business Management pp. 077-084 McGuire, S. and Sperling, L. 2008. Leveraging farmers’ strategies for coping with stress: seed aid in Ethiopia. Global Environmental Change, Vol 18 (4): 679-688. 19 Montoya, C.A., Leterme, P., Beebe, S., Souffrant, W.B., Mollé, D., and Lalle`s, J.P. 2008. Phaseolin type and heat treatment influence the biochemistry of protein digestion in the rat intestine. British Journal of Nutrition, 99, 531–539. Montoya, C.A., Leterme, P., Victoria, N.F., Toro, O., Souffrant, W.B., Beebe, S., and Lallès, J.P. 2008. Susceptibility of Phaseolin to in Vitro Proteolysis Is Highly Variable across Common Bean Varieties (Phaseolus vulgaris). J. Agric. Food Chem., 56, 2183–2191. Mwang'ombe, A.W., Wagara, N. Kimenju, J.W., Buruchara, R.A. 2007. Occurrence and Severity of Angular Leaf Spot of Common Bean in Kenya as Influenced by Geographical Location, Altitude and Agroecological Zones. Plant Pathology Journal. 6: 235-241 Odeny, D.A., S. M. Githiri and P.M. Kimani. 2009. Inheritance of resistance to fusarium wilt in pigeonpea, cajanus cajan (L.) Millsp. J. Animal and Plant Sciences 2: 89-95. Polanía, J., I.M. Rao, S. Beebe, and R. García. 2008. Desarrollo y distribución de raices bajo estrés por sequía en frijol común usando tubos con suelo en condiciones de invernadero. Agronomía Colombiana (in review). Rangel, A.F., I M. Rao and W.J. Horst. 2008. Cellular distribution and binding state of aluminum in root apices of common bean (Phaseolus vulgaris L.) genotypes differing in aluminum resistance. Physiologia Plantarum (published online on 5 November 2008). Rao, I.M., P. Wenzl, A. Arango, J. Miles, T. Watanabe, T. Shinano, M. Osaki, T. Wagatsuma, G. Manrique, S. Beebe, J. Tohme, M. Ishitani, A. Rangel and W. Horst. 2008. Advances in developing screening methods and improving aluminum resistance in common bean and Brachiaria. Braz. J. Agric. Res. (in review). Remans, R., S. Beebe, M.W. Blair, G. Manrique, I.M. Rao, A. Croonenborghs, R.T. Gutierrez, M. ElHoweity, J. Michiels and J. Vanderleyden. 2008. Detection of quantitative trait loci for root responsiveness to auxin producing plant growth promoting bacteria in common bean (Phaseolus vulgaris L.). Plant and Soil 302:149-161. Rivera, M., E. Amézquita, I. Rao and J. C. Menjivar. 2008. Análisis de la variabilidad especial y temporal del contenido de humedad en el suelo de diferentes sistemas de uso de suelo. Acta Agronómica (in review). Rubyogo, J.C., L. Sperling , R. Muthoni and R. Buruchara. Bean seed delivery in sub-Saharan Africa: the power of partnerships. peer reviewed journal: Society and Natural Resources (accepted July 2008). Forthcoming 2009. Schlueter, J.A., Goicoechea, J.L., Collura, K., Gill, N., Lin, J-Y., Yu, Y., Vallejos, E., Muñoz, M., Blair, M.W., Tohme, J, Tomkins, J., McClean, P., Wing, R., Jackson, S.A. 2008. BAC-end sequence analysis and a draft physical map of the common bean (Phaseolus vulgaris L.) genome. Tropical Plant Biology 1: 40-48. Sperling, L., H.D. Cooper, and T. Remington. 2008. Moving toward more effective seed aid Journal of Development Studies, Vol 44(4):586-612. 20 Wagara, I.N. A. W. Mwangombe, J. W. Kimenju, and R. A. Buruchara, 2007. Variation in aggressiveness of Phaeoisariopsis griseola and angular leaf spot development in common bean J. Trop. Microbiol. Biotechnol. 3:3-13 Zhang, X., Blair M.W., Wang, S. 2008. Genetic diversity of Chinese Common bean (Phaseolus vulgaris L.) landraces assessed with simple sequence repeat (SSR) markers. Theor Appl Genet 117:629–640. NON-REFEREED JOURNALS Blair, M.W., Buendía, H.F., Díaz, L.M., Díaz, J.M., Giraldo, M.C., Tovar, E., Duque, M.C., Beebe, S.E., Debouck, D.G. 2008. Utilization of microsatellite markers in diversity assessments for common bean. Annual Report of the Bean Improvement Cooperative 51: 12-13. Blair, M.W., Caldas, G.V., Muñoz, C., Bett, K.E. 2008. Evaluation of condensed tannins in tepary bean genotypes. Annual Report of the Bean Improvement Cooperative 51: 130-131. Blair, M.W., Iriarte, G., Beebe, S.E. 2008. Utilization of wild accessions to improve common bean (Phaseolus vulgaris) varieties for yield and other agronomic characteristics. Grain Legumes 50: 8-9. Blair, M.W., Namayanja, A., Kimani, P., Checa, O., Cajiao, C., Kornegay, K. 2008. Development and testing of mid-elevation, commercial-type, Andean climbing beans. Annual Report of the Bean Improvement Cooperative 51: 124-125. Porch, T.G., Blair, M.W., Lariguet, P., Broughton, W. 2008. Mutagenesis of common bean genotype BAT 93 for the generation of a mutant population for TILLING. Annual Report of the Bean Improvement Cooperative 51: 16-17. 5.3 WORKSHOP AND CONFERENCE PAPERS Blair, M.W. 2008. Advances in Common Bean Genomics. Presented at IV International Congress on Legume Genetics and Genomics, in Vallarta, Mexico, 7-12 Dec. Blair, M.W., A. Asfaw, G. Makunde. 2008. Advances for the common bean TL1 project. Presented at Tropical Legumes I meeting, 2 July. Blair, M.W. Bean Genomics/ Genetics at CIAT. 2008. Presented at INIA- Quilamapu, Chile, 23 Jan. Blair, M.W. 2008. Breeding medium – large seeded Andean beans for high minerals. Presented at Harvest Plus bean meetings in Bukavu, DR Congo, 8 Oct., and Butare, Rwanda, 12 Oct. Blair, M.W. 2008. Genômica do Feijoeiro no CIAT. IX Congresso Nacional de Pesquisa de Feijão, in Campinas, Brazil, 21 Oct. Blair, M.W. 2008. Improving common bean productivity for drought prone environments in sub-Saharan Africa. GCP Annual Research Meeting in Bangkok, Thailand, 15-20 Sept. Blair, M.W., and Beebe, S. 2008. Marcadores Moleculares para el Mejoramiento de Frijol Común. Primer Congreso Internacional y Feria de Frijol in Celaya, Guanajuato, México, 22 May. 21 Blair, M.W. 2008. Microsatellite diversity of cultivated common bean (Phaseolus vulgaris L.). - CIAT internal seminar, 23 April. Blair, M.W. 2008. Population structure in cultivated common bean (Phaseolus vulgaris L.). IV International Conference on Legume Genomics and Genetics in Vallarta, Mex., 6 Dec. Blair, M.W. 2008. Potential of the Common Bean reference collection (diversity structure and drought tolerance performance assessment). ADOC meeting – ICRISAT, Hyedrabad, AP, India, 10-12 Sept. Blair, M.W. 2008. Race structure and relationships among “ecotypes” in cultivated common bean (Phaseolus vulgaris L.). Plant and Animal Genome, San Diego, California, 11-16 Jan. Buruchara, R. A. 2008. Contributing towards reducing hunger and poverty in Africa: CIAT’s approach, experience and opportunities. Presentation at JIRCAs, Tokyo, Japan, May 2008 Buruchara, R. A. 2008. ISFM-based crop production systems for major impact zones in sub-Saharan Africa. Presentation at the Round Table |Meeting on Agricultural Research for African Development May, 2008, University of Tokyo. Kimani, P.M., S. Beebe, M. Blair, R. Chirwa and I. Rao. 2008. Improving productivity of common bean and incomes for the poor in marginal environments of sub-Saharan Africa: Overview of TL I and II projects. Drought phenotyping workshop, 4-17 May 2008 Lilongwe, Malawi. Kimani, P. M., G. Mbugua and P. Okwiri. 2008. Breeding beans for drought resistance in East and Central Africa region. Drought phenotyping workshop, 4-17 May 2008 Lilongwe, Malawi. Kimani, P.M. 2008. Characterisation in drought testing sites in East and Central Africa. Drought phenotyping workshop, 4-17 May 2008, Lilongwe, Malawi. Kimani, P.M. 2008. Future breeding for drought resistance in eastern Africa. Drought phenotyping workshop, 4-17 May 2008, Lilongwe, Malawi. Kimani, P.M., R. Chirwa, A. Namayanja, C. Ruradama, S. Gebeyehu, N. Mbikayi and Lodi Lama. 2008. Breeding better bean varieties for African farmers: Achievements and Future directions. PABRA Stakeholders Workshop, 21-25 January 2008, Kampala, Uganda Kimani, P.M., S. Beebe and M. Blair. 2008. Breeding Micronutrient Dense Bean Varieties in East and Central Africa. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M., S. Beebe, Nkonko Mbikayi and M. Blair. 2008. Screening bean germplasm for micronutrients. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. 2008. Genotype x environment interactions for micronutrient density and variety release. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. 2008. Breeding micronutrient dense beans in ECABREN: Objectives, activities and Milestones. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. 22 Kimani, P.M., Ben Okonda, S. Beebe and J.P. Keter. 2008. Influence of fertilization with inorganic macroelements on micronutrient density and agronomic traits in common bean genotypes. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. and R. Chirwa. 2008. Future Breeding for Drought Resistance, Better Nutrition and Health in PABRA. Pan African Bean Research Alliance Annual workshop, 20-24 October 2008, Lilongwe, Malawi Kimani, P.M. 2008. New Research Directions in PABRA: Implications for WECABREN. IRADWECABREN Collaborative Bean Research Program Workshop, 16-21 November 2008, Bafoussam, Cameroon. Kimani, P.M. 2008. Agronomic management for maximising micronutrient density in beans. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. 2008. Improving food security and quality for low input farmers in the East African Highlands: Lessons Learnt. Nutribean Review and Planning Workshop, 24-27 August 2008, Nyeri, Kenya. Rubyogo, J.C., and L. Sperling, 2008. Developing seed systems in Africa . In Robert Chambers, Ian Scoones and John Thompson eds. Farmer First Revisited : Farmer Participatory Research and Development Twenty Years on. Workshop held Institute of Development Studies, University of Sussex, Brighton, UK. 12-14 December, Sussex: IDS Sperling , L , S. Nagoda and A. Tveteraas. 2008. Moving from emergency seed aid to seed security linking relief with development. Workshop organized by the Drylands Coordination Group Norway and Caritas Norway, in collaboration with Norad and The Norwegian Ministry of Foreign Affairs. Oslo, Norway, DCG Proceedings No. 24, 14 May. 5.4 PROCEEDINGS, POSTERS, ABSTRACTS AND OTHERS PROCEEDINGS Chirwa, R. M., M. Pyndji and R. Buruchara. 2008. CIAT-PABRA Management and Organization – An assessment of strengths, weaknesses, opportunities and threats. A paper presented at a PABRA Stakeholders Workshop, Kampala, Uganda , 15-20 January Chirwa, R. M, R. Buruchara. 2008. CIAT’s Pan Africa Bean Research Alliance (PABRA) – An Overview. A paper presented at a Grain Legumes CRSP inception Workshop, Barcelona, Spain, 29 Feb. - 4 March Chirwa, R. M., J. M. Bokosi and E. Mazuma. 2008. Use of Marker Assisted Selection in Developing Bean Varieties for multiple disease resistance in Malawi. A paper presented at a Meeting organized by Kirkhouse Trust in Kampala, Uganda 6-7 March Chirwa, R. M. 2008. The Status of Southern Africa Bean Research Network – Progress Towards Achieving Targets in the Current Phase. A paper presented at the PABRA Steering Committee Meeting, Lusaka, Zambia, 17-19 March . Chirwa, R. M., D. Fourie and G. Makunde. 2008. Bean breeding for drought resistance in SABRN. A paper presented at the TL-II training workshop held at MIM, Lilongwe, Malawi, 5-16 May 23 Chirwa, R. M. 2008. Future bean breeding for drought resistance in SABRN. A paper presented at the TL-II training workshop held at MIM, Lilongwe, Malawi, 5-16 May Chirwa, R. M., E. Mazuma and J. C. Rubyogo. 2008. Getting back to basics: creating impact -oriented bean seed delivery systems for the poor (and others) in Malawi. A paper presented at the PVS training Workshop for NARS partners, Mponela, Malawi, 26-27 May Chirwa, R. M., H. Tefera and M. Siambi. 2008. Current Status of the Legume Industry: Bean, Soybean, Groundnut & Goal of the Legume Platform. Presented at the 1st RIU-Legume Platform Meeting Held at NASFAM Conference Room, Lilongwe, Malawi, 5th June Gomonda, R.W.J, I.M.G Phiri, R. Chirwa and C. Mwale. 2008. Improving Soil Fertility: Key Programmes, Strategies and Challenges in Malawi. Presented at the Soil Health Program launch workshop, held at Windsor Golf Hotel, Nairobi Kenya 16-18 June Chirwa, R. M., J. C. Rubyogo, L. Sperling, E. Mazuma, M. Amane and C. Madata. 2008. Getting back to basics: creating impact -oriented bean seed delivery systems for the poor (and others) in Malawi, Mozambique and Tanzania - A progress report. A paper presented at the McKnight’s Legumes CCRP community of practice workshop held at Hotel VIP, Maputo, Mozambique, 6-9 Oct. Chirwa, R. M. 2008. The status of bean research activities in the SABRN. A paper presented at the SABRN/ECABREN joint SC meeting held at Lilongwe Hotel, Lilongwe, 22-24 Oct. Chirwa, R.M, C. Mwale, A. R. Saka, and Ian Kumwenda. 2008. Alliance for a Green Revolution in Africa - Soil Health Program Business Planning Process. A Country Report for Malawi. October Horst, W.J., A.F. Rangel, D. Eticha, M. Ishitani and I.M. Rao. 2008. Aluminum toxicity and resistance in Phaseolus vulgaris – physiology drives molecular biology. Proceedings of the 7th International Symposium on Plant-Soil Interactions at Low pH, Guangzhou, China, 17-21 May. POSTERS Asfaw, A., M.W. Blair. 2008. Population Genetic Structure of Common Bean (Phaseolus vulgaris L.) Landraces from Ethiopia and Kenya. Plant Animal Genome, San Diego, California, 11-17 Jan. Becerra, V., M. Paredes, C. Rojo, M.W. Blair, J. Tay. 2008. Morphological, agronomical and genetic characterization of a core collection of common bean (Phaseolus vulgaris L.): Race Chile. IV International Conference on Legume Genomics and Genetics, Chillán, Chile, 21-26 Jan. Blair, M.W., H.F. Buendía, L. Díaz, J.M. Díaz, M.C. Giraldo, E. Tovar, M.C. Duque, S.E. Beebe, D. Debouck. 2008. Microsatellite marker diversity in common bean (Phaseolus vulgaris L.). Plant Animal Genome, San Diego, California, 11-17 Jan. Checa, O.E., M.W. Blair. 2008. Mapping QTL for climbing ability and component traits in common bean (phaseolus vulgaris L.) – CIAT posters. Diaz, A., G.V. Caldas, M.W. Blair, 2008, Cuantificación de taninos condensados e identificación de QTLs asociados a su contenido en una población de frijol comun (P. vulgaris). Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. 24 Kimani, P.M., John Nderitu and Levi Akundabweni. 2008. Towards Vision 2030: New Bean Varieties for improved productivity, food and nutrition security and wealth creation. 9-12 November 2008, Strategy for Revitalising Agriculture, Second National Workshop, Safari Park Hotel, Nairobi (Award winning poster presentation). Presented to H.E the President, H.E. Vice-President and Hon Minister for Agriculture. Kimani, P.M., A. Mwang’ombe and J. W. Kimenju. 2008. New Varieties from University of Nairobi Bean Program. 9-12 November 2008, Strategy for Revitalising Agriculture, Second National Workshop, Safari Park Hotel, Nairobi (Award Winning poster presentation). Presented to H.E the President, H.E. Vice-President and Hon Minister for Agriculture. Lozano, M.A., G.V. Caldas, M.W. Blair. 2008. Cuantificación de fitatos por espectroscopía visible en 16 genotipos de una población de fríjol común (P. vulgaris l.) sembrada en suelos con alto y bajo fósforo. Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Makumba, W., R. Chirwa, J.C. Rubyogo, R. S. Weldesemayat and M. Jonasse. 2008. Improving smallholders food security, nutrition and income through increased production and marketing of climbing beans in Malawi and Mozambique – presented in Mozambique Ortiz, D., H. Pachón, M.W. Blair, D. Gutiérrez, C. Araujo, J. Restrepo. 2008. Evaluación del valor nutricional de micronutrientes en una receta típica (fríjol sancochado) preparada con fríjoles nutricionalmente mejorados. Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Ortiz, D., H. Pachón, M.W. Blair, D. Gutiérrez , C. Araujo, J. Restrepo. 2008. Evaluación de la calidad proteica de recetas preparadas con cultivos de maíz mejorado nutricionalmente. Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Papp, P., T. Gollenar, L. Holly, M.L. Warburton, M.W. Blair, G.B. Kiss. 2008. Evaluation of allelic diversity in maize and common bean germplasm, GCP annual meeting, Thailand, 15-20 Sept. Rubyogo J.C., F. Tembo., R. Chirwa. E. Mazuma, M. Amane. and C. Madata. 2008. Collaborative research program for creating impact oriented bean seed delivery systems for the poor in Malawi, Mozambique and Tanzania – presented in Mozambique Yang, Z.B., D. Eticha, I.M. Rao and W. Horst. 2008. The interaction between aluminum toxicity and drought stress in common bean (Phaseolus vulgaris L.). Poster paper presented at the Annual Meeting of the German Society of Plant Nutrition in Limbergerhof/Speyer, Germany, 23-24 Sept. OTHERS International Newsletters Sperling, L. and S. McGuire, 2008 Seed aid in Ethiopia. Anthropology News 49(7):52 Guides and Handbooks Buruchara, R. A., C. Mukankusi and K. Ampofo. Pests and Diseases of Common Bean and their Management in Africa. Handbook for Small Scale Seed Producers (in Press) Sperling, Louise, 2008. When Disaster Strikes: A Guide to Assessing Seed System Security. Cali, Colombia: International Center for Tropical Agriculture 25 Brochures PABRA Outlook: Issue 3. Media Campaign May/June 2008: Seed Aid, with, CIAT Communications unit, CG Communication Unit and Burness Communications. Based on Seed AID work of L. Sperling, Tom Remington and other partners Wires Asian News International (India) Reuters (Nature….) (which linked to Science) Broadcast BBC Network Africa South African Broadcasting Corporation (SABC) Channel Africa Print Hindustan Times (India) New Vision (Uganda) Bistandaktuelt (Norway) Online Africa Science News Service Agricultural Biodiversity Blog Andhranews.net (India) DailyIndia.com KTIC Rural Radio Online Malaysia Sun Online Nature News NewKerala.com (India) Star Online (Malaysia) Thaindian.com (India) TopNews.in (India) Webindia123.com 5.5 EDITORIAL CONTRIBUTION I.M. Rao served on the scientific committee of the editorial board of the journal, Agronomia Colombiana, and a reviewer to the journals: Crop Science, Agroforestry Systems and Acta Agronomica. 26 6. LIST OF SPECIAL PROJECTS 6.1 AT HEADQUARTERS 6.1.1 New proposals approved in 2008 Title Donor Biofortificación del Frijol Común (Phaseolus Vulgaris L.) en Panamá con Micronutrientes” SENACYT – Panama Improved beans for Africa and Latin America DFID, UK Characterization of bean diversity in Central Europe 12,000 Amount to Partners (US $) - Available in 2008 (US$) 7,000 2008 120,690 - 120,690 GCP 2008-2009 9,000 - 9,000 Dry bean improvement and marker assisted selection for diseases and abiotic stresses in Central America and the Caribbean” GCP 2008-2009 40,120 - 40,120 Capacity Building Needs regarding the Tropical Legume I (TLI) Project BMGF grant to GCP 2008-2009 5,904 5,904 Obtención y evaluación de Phaseolus vulgaris y Zea mays tolerantes a la sequía CYTED, Spain 2008-2009 $1,000,000 - 29,906 Development of a handling system of Bemisia tabaci in paprika and pepper in the Cauca Valley Gracias MADR 2008-2011 58,288 Improvement of Chitti bean in Iran. SPII, Iran Iranian government 2008 18,423 - 18,423 Funding period Total amount Amount to Partners (US $) Available in 2008 (US$) 64.276 125.152 US153,907 US303,233 6.1.2 Funding period Total amount 2008-2011 16.560 List of ongoing special projects in 2008 Title Donor Reducing pesticide use and pesticide resistance in rice and beans in the Andean zone FONTAGRO 2006-2009 224.000 Fighting Drought and Aluminium Toxicity: Integrating Genomics, Phenotypic Screening and BMZ 2006-2009 € 1,100,000 27 Title Donor Funding period Total amount Amount to Partners (US $) Available in 2008 (US$) Participatory Research with Women and Small-Scale Farmers to Development Stress-Resistant Common Bean and Brachiaria for the Tropics Biofortified Crops for Improved Human Nutrition – Harvest Plus Challenge Program (Yearly contracts) Gates Foundation World Bank DANIDA, Denmark 2003-2008 305,000 50,000 255,000 Combating hidden hunger in Latin America: Biofortified crops with improved vitamin A, essential minerals and quality protein (AgroSalud) CIDA 2004-2010 20,000,000 123,855 254,894 Integrated management of whiteflies in the tropics DFID 2005 - 2008 259.788 7.849 22.864 Increasing Food Security and Rural Incomes in Eastern, Central and Southern Africa through Genetic Improvement of Bush and Climbing Beans (Headquarters component) RF 2005-2008 US 254,000 - 10,750 Nutritional Improvement of the important pulse legume, the common bean, through the reduction of seed tannin content, for the benefits of people' diet in Africa and Latin America CIDA/Univ. of Saskatchewan 2007-2010 CAD 225,000 US 32,102 US 34,503 TL1: Improving tropical legume productivity for marginal environments in sub-Saharan Africa (Headquarters component) BMGF grant to GCP 2007-2010 115,000 473,944 TL2: Enhancing grain legumes productivity, production and income of poor farmers in droughtprone areas of sub-Saharan Africa and South Asia (HQ component) BMGF grant to CGIAR 2007-2010 1,104.056 197,701 Variedades de fríjol tolerantes al estrés abiótico de la baja fertilidad y la sequía, y a la sostenibilidad productiva y alimentaria de Centroamérica Red-SICTA, SDC 2007- 2008 - 45,450 28 1,867,328 3,454.802 246,100 6.2 IN AFRICA 6.2.1 New proposals approved in 2008 Title Donor Supporting Nutrition and health, Food security, Environmental Stresses and Market Challenges that contribute to improve livelihood and create income resource poor small holder families in Sub –Saharan Africa SDC Funding period 2009-2011 Total Amount US 3.2 million Amount to partners US$ 2,221,384 Available in 2008 US$ 978,616 Amount to Partners (US $) Available in 2008 (US$) 115,000 601,250 502,866 6.2.2 List of ongoing special projects in 2008 Title Donor Funding period Total amount TL1: Improving tropical legume productivity for marginal environments in sub-Saharan Africa (African component) BGMF 2007-2010 115,000 TL2: Enhancing grain legumes’ productivity, production and the incomes of poor farmers in drought-prone areas of subSaharan Africa and South Asia: Seed Systems (African component) BGMF 2007-2010 2,866.084 Getting back to basics: creating impact-oriented bean seed delivery systems for the poor in Malawi, Mozambique and Tanzania McKnight Foundation 2007-2010 US$ 400,000 300,000 100,000 Improved Smallholder food Security, Nutrition and Income through Increased Production and Marketing of Climbing Beans. McKnight Foundation 2007-2010 US$ 400,000 300,000 100,000 Fighting Drought and Aluminium Toxicity: Integrating Genomics, Phenotypic Screening and Participatory Research with Women and Small-Scale Farmers to Development Stress-Resistant Common Bean and Brachiaria for the Tropics BMZ 2006-2009 Increasing Food Security and Rural Incomes in Eastern, Central and Southern Africa through Genetic Improvement of Bush and Climbing Beans (African component) RF 2005-2008 1, 368,000 million seed systems 29 US 63,185 US 254,000 - 76,739 Total amount Available in 2008 (US$) Donor Supporting improved nutrition, food security and community empowerment for poverty alleviation – PABRA SDC 2007-2008 US 944,616 944,616 Supporting improved nutrition, food security and community empowerment for poverty alleviation – PABRA III CIDA 2003-2008 US5,298.787 2,231,057 6.2.3 Funding period Amount to Partners (US $) Title Regional research subprojects under SABRN Activity Value Country 1.1.1 Complete germplasm collection, characterization and mineral analysis for all accessions 1000 3000 DRC Zambia 1.1.2 Conduct multi-location evaluations and national performance trials 650 500 1500 600 800 1500 1000 Angola DRC Malawi Mozambique Swaziland Zambia Zimbabwe 1.1.3 Analyze candidate varieties for minerals and protein in some countries in SABRN 1000 1000 400 Angola Malawi Mozambique 500 300 500 500 Malawi Mozambique Zambia Zimbabwe 1.1.5 Conduct DUS in applicable and present for release: 3 countries in SABRN 500 1000 800 500 DRC Malawi Mozambique Zimbabwe 1.1.6 produce breeder seed in countries that have released varieties 900 1000 400 1000 800 800 Angola Malawi Mozambique Swaziland Zambia Zimbabwe 1.1.4 Develop descriptors for candidate varieties 30 Activity 1.2.1 On-farm evaluations using PVS Value 2250 2800 1500 2600 3500 1500 3000 1000 1.2.2 Develop descriptors for the new bean varieties Country Angola DRC Malawi Mozambique South Africa Swaziland Zambia Zimbabwe 700 700 Zambia Zimbabwe 1.2.3 Produce breeders' seed for the new and old bean varieties 3000 500 DRC Zimbabwe 1.2.4 Rejuvinate BILFA, Drought and disease nurseries 1700 3000 1000 1700 1500 Angola DRC Malawi Mozambique Zimbabwe 1.2.8 Combine resistance and select for pyramid (ALS, CBB) in ZA and BSM (MW and ZW) 1500 7500 Malawi Zambia 1.2.10 Selection and testing of climbing beans adapted to mid-altitude (1200 -500 masl) 1500 850 2000 2000 Angola Mozambique Zambia Zimbabwe 1.3.1 Identify export market potential including enhancing competitiveness of beans in SABRN 500 500 Malawi Zimbabwe 1.3.2 Conduct a bean cross-border trade study across South TZ, South DRC and Zambia 3000 1500 DRC Zambia 1.4.2 Continue with backcrossing programme to improve commercial cultivars - Southern Africa 1000 4000 Malawi South Africa 1.4.3 Strengthen capacity for application of MAS - Bunda 4000 Malawi 1.4.6 Produce adequate seed for all breeding materials 1500 1500 1000 Malawi Zambia Zimbabwe 1.4.7 Production of foundation seed with partners 2000 3700 2000 1000 DRC Mozambique Zambia Zimbabwe 31 Activity Value Country 2.1.1 Validate effectiveness and farmers' acceptance and gender perceptions of promising ISFM and IPDM options with farmers 1850 1000 500 1000 1000 Angola Malawi Mozambique Zambia Zimbabwe 2.1.2 Disseminate and promote accepted options with partners for technologies in 10 all countries 1500 500 1500 2000 Malawi Mozambique Swaziland Zimbabwe 2.1.3 Perform cost-benefit tradeoffs analyses and adoption potential of these technologies 1000 1000 Malawi Zimbabwe 3.1.1 Organize, train and technically backstop community seed producers to bulk seeds 2000 2500 1000 DRC Mozambique Zimbabwe 3.1.2 Update number, type location and activities of service providers 250 1000 1000 1000 Angola DRC Mozambique Zimbabwe 3.2.3 Facilitate production of promotional and information publications (including publications for SABRN website), transilations in each network 950 1000 1000 1000 1000 1000 1000 Angola DRC Malawi Mozambique Swaziland Zambia Zimbabwe 5.5.2 Conduct participatory formulation and evaluation of a basket of diets for improved nutrition - using biofort products 2000 2000 1500 1000 1000 DRC Malawi Swaziland Zambia Zimbabwe 32 Activity Value Country 6.1.4. Conduct training workshops on nutrition assessment and linking nutrition support with agricultural extension 800 1000 Swaziland Zimbabwe 8.1.1 Inventory by year products (varietal and non-varietal), promotional materials 1750 3500 Angola Mozambique TOTAL 140050 HarvestPlus funded activities Activity Value Country 1. Germplasm collection 4000 4000 3000 Malawi Tanzania Zimbabwe 2. Evaluation of fast trucks lines in various countries 2000 2000 2000 2000 2000 2000 2000 Angola Lesotho Malawi DRC Tanzania Zambia Zimbabwe 3. Breeding for high Fe combining with other biotic and abiotic stresses 2000 2000 2000 2000 Malawi South Africa Tanzania Zimbabwe 4. Supplies and small equipment: reagents, computer, printer 5000 SABRN TOTAL 40000 33 6.3 LIST OF PROJECTS SUBMITTED, PROPOSALS, AND CONCEPT NOTES PREPARED 6.3.1 AT HEADQUARTERS Funding period Total amount US 2008-2011 $889,350 Title Donor Comments Extracting the best from a desert species: Mining tepary bean for drought tolerance GCP Concept note not selected for full proposal development Basal root architecture and drought tolerance in common bean GCP Concept note and full proposal approved An integrated experimental and modeling approach to optimize soil water use under limited water GCP Concept note not selected for full proposal development A cross-legume phenotyping effort to identify common traits for superior adaptation to drought GCP Concept note under review 2009-2011 $459,020 Improving tolerance to drought stress in crops WUN Seed grant under review 2009 $48,000 2008-2011 2008-2011 $ 345,000 $905,060 6.3.2 IN AFRICA Title Impact and development of Conservation Agriculture techniques in developing countries Donor European commission Comments Collaborators are: University of Applied Sciences Eberswalde, Germany; International Food Policy Research Institute (IFPRI), USA International, University of Ghana and Makerere University Participating CIAT technical team include: Enid Katungi and Roger Kirby 34 Funding period 3 years Total amount US 220,000 (CIAT’s budget only) Title Donor Supporting Nutrition and health, Food security, Environmental Stresses and Market Challenges that contribute to improve livelihood and create income resource poor small holder families in Sub –Saharan Africa.. CIDA Enhancing productivity, nutrition and incomes through improved marketable climbing bean and biofortified bean varieties Government of Kenya Improving Food and Nutrition Security, and Incomes of Smallholder Farmers in East and Central Africa through increased access to Markets and Technology Innovation Climbing out from poverty: Realizing the benefits from high yield potential of Climbing beans for smallholder farmers in Africa Use of marker Assisted Selection in Developing Multiple Disease Resistant Bean Varieties in Malawi - Comments Funding period Total amount US 2009-2013 7.8 million In review 2009-2011 $110,000 Belgium Development Cooperation (BADC) Unsuccessful 2008-2011 $3,148,632 JIRCA Presented to donor in Jan 2008 Kirk House Trust 35 Under review by donor (second round) 2009-12 150,000 7. STAFF LIST (INCLUDING % TIME ASSIGNMENT) 7.1 STAFF AT HEADQUARTERS Stephen Beebe, PhD, Breeder, Geneticist, Project Manager (70% SBA-1, 30% SBA-6) Matthew Blair, PhD, Germplasm Characterization Specialist, Bean Breeder (70% SBA-6, 30% SBA-1) Francisco Morales, PhD, Virologist (30% SBA-1, 50% PE-1) Idupulapati Rao, PhD, Plant Nutritionist, Physiologist (50% SBA-1, 50% SBA-3) 7.2 STAFF IN AFRICA Robin Buruchara, Ph.D., Plant Pathologist/CIAT Africa Coordinator (stationed in Kampala, Uganda - 90% SBA-1, 10% PA-2) Rowland Chirwa, PhD, Plant Breeder/SABRN Coordinator (stationed in Lilongwe, Malawi 100% SBA-1) Enid Katungi, PhD, Agricultural economist (stationed in Kampala, Uganda - 100% SBA-1) Paul Kimani, PhD, Plant Breeder for ECABREN (University of Nairobi/CIAT, stationed in Nairobi, Kenya - 75% SBA-1) Rachel Muthoni, BSc, MPA, Monitoring and Evaluation Specialist, (stationed in Kampala, Uganda - 100% SBA-1) Jemimah Njuki, PhD, ERI Specialist, (stationed in Zimbabwe – 44% SBA-1, 56% TSBF-1) Martha Nyag’aya,, MSc, Nutrition (stationed in Kampala, Uganda – 90% SBA-1, 10% TSBF-1) Mukishi Pyndji, PhD, Plant Pathologist, ECABREN Coordinator (stationed in Arusha, Tanzania - 100% SBA-1) Jean Claude Rubyogo, MSc, Seed System Specialist (stationed in Malawi – 100% SBA-1) Louise Sperling, PhD, Social Scientist, (stationed in Rome, Italy - 80% SBA-1, 20% SBA-6) 8. SUMMARY 2008 BUDGET PREPARED BY FINANCES: ACTUAL EXPENDITURES 2008 Outcome Line SBA-1: Beans SOURCE Unrestricted Core Bean Program Total US$ (%) 120,901 743,185 7% 35,500 35,500 0% 156,401 778,685 8% 2,874,851 6,988,201 70% 35,450 254,592 290,042 3% 312,089 429,099 741,188 7% HQ + LAC Africa 622,284 Restricted Core Japan Sub-total Core 622,284 - Biotech Restricted Special Projects Generation Challenge Program Harvest Plus 1,045,811 3,067,539 Sub Total Restricted 1,393,349 3,067,539 3,558,543 8,019,431 81% Direct Expenditures 2,015,634 3,067,539 3,714,943 8,798,116 89% Non Research Cost 259,492 394,914 478,261 1,132,667 11% Total Expenditures 2,275,126 3,462,453 4,193,204 9,930,783 100% 36 Product 1: Beans with improved micronutrient concentration that have a positive impact on human health Activity 1.1 Developing more nutritious bean varieties Highlights: • • • • • • • • • 1.1.1 More than 30 F3.5 small seeded Mesoamerican families were selected for high mineral concentration and some degree of drought resistance. Eighteen lines derived from interspecific crosses were coded as MIB (high mineral) lines, with levels of iron above those of the high iron check MIB 465. Some also have superior resistance to foliar pathogens. Over 200 pollinations were generated combining multiple sources of resistance to diseases and drought with high mineral (Fe and Zn) content in SABRN countries. Some good donor parent for drought resistance in common bean (SEA5, SEA15 and SER16) were also good parents for high Fe and Zn content. Several populations combining different market classes and disease or drought resistance, but also with high mineral (Fe and Zn) content have been generated, and some may have high Fe and or Zn content. Four bush and three climbing micronutrient dense bean varieties with high yield potential are pre-released for smallholder production in eastern Africa. This marks the first time biofortified mineral dense bean varieties are formally recommended for release following independent evaluations. Bioavailability of Fe in raw samples of fast track bean lines varies from 1.1% to 6.6%. Bioavailability of Zn in raw samples of fast track lines varies from 0.5 to 2.5%. Cooking enhances Fe and Zn bioavailability in beans by more than two fold. Freshly shelled beans have more bioavailable Fe and Zn compared with dry beans. Development of new advanced lines from the program for the nutritional enhancement of Andean bush beans Rationale: Iron deficiency anemia and other micronutrient deficiencies affect large numbers of people worldwide and biofortification is an approach to address this problem by breeding for higher micronutrient content in staple food crops. Legumes are a good source of iron and other essential micronutrients that are found only in low amounts in the cereals or root crops. Also beans and other grain legumes are usually consumed whole, thus conserving their nutritional content. An ongoing project has shown that bean seeds are variable in the amount of minerals (iron, zinc and other elements) that they contain and that these traits are likely to be inherited quantitatively. Over the past 4 years we have developed an initial set of Andean bush beans that have higher mineral content and red mottled seed type and named this set the NUA (Nutritional Andean) lines. These genotypes have now been widely distributed and a few of the advanced lines are near varietal release; their one drawback being a limited number of genetic parents used to develop the lines (predominantly CAL96 as recurrent parent and G14519 as high mineral parent). The objective of this research and varietal development program has been to create a new generation of NUA lines based on triple, double and multiple crosses using the original NUA lines as well as additional sources in various breeding combinations. This report summarizes the nutritional analysis of the F6 derived lines which are currently in yield testing before shipment to partner countries in Eastern and Central Africa and further testing in Colombia. Most of the new genotypes aim to combine the high iron trait into red mottled background but we also derive some large red and cream mottled genotypes. 1 Materials and Methods: Advanced lines were developed from the following donor parents for the high mineral trait: G14519, G23823E, G21242, NUA35, NUA56, BID29, BID115, through a series of backcrosses, simple crosses, triple crosses, double crosses and multiple crosses with the commercial parents: CAL96, CAL143, CAL144, PVA773, AND277 and AFR612 (all red mottled); as well as AFR298, RADICAL SAN GIL, RADICAL CERINZA, RED CANADIAN WONDER (G6592) (all large red seeded); SUG131 (cream mottled), KABLANKETI (G22454) (purple stippled), and DORE DE KIRUNDO (G21715) (large yellow). Some combinations contained the angular leaf spot resistance sources G5686 and MEX54 to complement those crosses with AND277 and CAL143 which also provide some resistance to the disease. After initial crosses and generation advance, single plant selections were made in the F3 and F6 generations. Seed multiplication for one generation was used to obtain seed for iron and zinc analysis which was carried out with atomic absorption spectroscopy (AAS) in CIAT analytical lab. Advanced lines were also selected for highly acceptable architecture, growth habit, yield potential and seed types. Results and Discussion: The selected genotypes were numbered consecutively beginning with the numbering of the previous NUA lines developed from the initial crosses for the biofortification program. A total of 490 advanced lines were developed with these listed as NUA101 through NUA591. The new NUA lines were organized based on pedigree and information on seed type was used to create separate nurseries for red mottled, large red and cream mottled genotypes for planting in Darién 2008B (Andic Dystrudept, pH 5.5, 1500 masl, average temperature 19°C, 500 mm during the crop cycle) season. Due to heavy rains throughout the season the planting was delayed until December which was a better period for planting into moist soils. In addition, a new plot was selected based on its soil type, which contained higher concentrations of iron and zinc compared with previous plots used. A total of 76 pedigrees were represented by the 490 selected NUA lines and these were divided into backcrosses, triple, double and simple cross derived genotypes. Table 1 shows the summary of genotypes per pedigree, cross number, NUA codes and range of iron and zinc content (measured as parts per million or ppm) within each pedigree as well as the seed colors of the F6:7 advanced lines derived from each cross. For example, lines NUA101 to NUA265 (165 in total) were derived from backcrosses with AFR298, AFR612, RED CANADIAN WONDER (G6592), CAL144, PVA773, SUG131, DORE DE KIRUNDO (G21715) using the high mineral parents G14519 (Mesoamerican) and G21242 (Andean). Lines NUA266 to NUA411 (145 in total) were derived from triple crosses using a wider range of high iron parents (NUA35, NUA56, G23823E, BID29 and BID115) in the genetic background of commercial type parents AFR612, SUG131, PVA773, CAL96, CAL143 and CAL144 (these last two genotypes being sister lines with different yield and adaptation potential). Lines NUA412 to NUA442 (31 lines in total) were derived from double crosses involving some of the same high iron parents (G23823E, BID29, BID115) and the large red seeded genotypes RADICAL SAN GIL, RADICAL CERINZA, AFR298, RED CANADIAN WONDER (G6592) as well as KABLANKETI (G22454). Finally, the lines NUA443 to NUA591 (143 in total), were derived from simple crosses between high iron genotypes NUA35, NUA56, G23823E, G21242, BID29 and BID115 with commercial red mottled and large red genotypes AFR298, AFR612, AND277, PVA773, KABLANKETI (G22454), G5686, MEX54, CAL96, CAL144, SUG131 and RADICAL CERINZA. 2 Table 1. Development of NUA lines from the program for the nutritional enhancement of Andean bush beans. NUA number 101 - 140 141 - 161 162 - 181 182 - 198 199 - 209 210 - 220 221 - 231 232 - 241 242 - 248 249- 250 251 - 255 256 - 258 258 - 260 261 - 262 263 - 264 265 266 - 310 311 - 328 329 - 340 341 - 351 352 - 363 364 - 371 372 - 379 380 - 385 386 - 391 392 - 396 397 - 400 401 - 403 404 - 405 406 - 407 408 - 409 410 411 Genotype Backcross AFR612 X (AFR612 X G14519) CAL144 X (CAL144 X G14519) AFR612 X (AFR612 X G21242) SUG131 X (SUG131 X G21242) G21715 X (G21715 X G21242) AFR298 X (AFR298 X G14519) CAL144 X (CAL144 X G21242) PVA773 X (PVA773 X G14519) SUG131 X (G14519 X SUG131) SUG131 X (SUG131 X G14519) AFR298 X (AFR298 X G21242) G6592 X (G14519 X G6592) G21715 X (G14519 X G21715) G22454 X (G14519 X G22454) G22454 X (G22454 X G21242) G22454 X (G22454 X G14519) Triple Cross NUA35 X (AFR612 X BID115) NUA56 X (BID29 X SUG131) NUA35 X (PVA773 X BID29) NUA56 X (CAL143 X G23823E) NUA35 X (AFR612 X G23823E) NUA35 X (CAL96 X G23823E) NUA56 X (AFR612 X BID115) NUA35 X (BID29 X CAL144) NUA56 X (PVA773 X BID29) NUA56 X (AFR612 X G23823E) NUA35 X (SUG131 X G23823E) NUA35 X (CAL143 X G23823E) NUA35 X (CAL144 X G23823E) NUA56 X (CAL144 X G23823E) NUA56 X (SUG131 X G23823E) NUA56 X (PVA773 X G23823E) NUA56 X (BID115 X PVA773) Total of lines Iron range (ppm) Zinc range (ppm) Color type of beans 40 21 20 17 11 11 11 10 7 2 5 3 2 2 2 1 73 - 41 84 - 53 66 - 44 88 - 64 69 - 53 66 - 45 90 - 59 75 - 63 81 - 67 65 64 - 52 60 - 48 61 65 - 64 56 - 53 67 31 - 18 49 - 23 33 - 20 33 - 25 31 - 25 37 - 27 45 - 32 37 - 25 36 - 26 25 28 - 25 35 - 27 25 39 - 27 25 29 Red mottled and red Purple mottled, red mottled, red and purple Red, red mottled and purple mottled Cream mottled and red mottled Yellow Red Red mottled, purple mottled and cream mottled Red, red mottled Red mottled and pink mottled Cream mottled and red mottled Red Red Yellow Purple mottled Purple mottled Purple mottled 45 18 12 11 12 8 7 6 6 5 4 3 2 2 2 1 1 92 - 54 75 - 48 77 - 63 84 - 51 81 - 67 86 - 65 71 - 49 83 - 61 83 - 54 86 - 62 105 - 79 98 - 62 84 - 83 80 - 60 66 71 72 32 - 22 30 - 24 30 - 25 34 - 22 30 - 25 31 - 25 29 - 23 31 - 23 26 - 22 31 - 22 33 - 30 27 - 24 29 - 27 24 - 21 27 - 21 28 28 Purple mottled, red mottled and purple Cream mottled, purple mottled, red, red mottled Red, red mottled and purple mottled Purple mottled and red mottled Purple mottled and red mottled Red mottled and purple mottled Red mottled and purple mottled Purple mottled and red mottled Red mottled Red and red mottled Purple mottled and red mottled Purple and red mottled Purple mottled Red mottled and purple mottled Purple mottled Purple mottled Purple mottled 3 Table 1. cont’d. NUA number 412 - 417 418 - 422 423 - 427 428 - 431 432 - 435 436 - 437 438 - 439 440 - 441 442 443 - 463 464 - 484 485 - 494 495 - 504 505 - 513 514 - 521 522 - 528 529 - 535 536 - 542 543 - 547 548 - 555 556 - 559 560 - 563 564 - 566 567 - 568 569 - 570 571 - 572 573 - 574 575 - 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 Genotype Double Cross (RAD.SANGIL X G23823E) X (BID115 X RAD.SANGIL) (RAD.CERINZA X G23823E)X(RAD.CERINZA X BID115) (AFR298 X G23823E) X (BID115 X AFR298) (RAD.SANGIL X G23823E) X (RAD.SANGIL X BID115) (AFR298 X G23823E) X (AFR298 X BID29) (G6592 X G23823E) X (G6592 X BID115) (G22454 X G23823E) X (BID29 X G22454) (AFR298 X G23823E) X (AFR298 X BID115) (G6592 X G23823E) X (BID115 X G6592) Single Cross AFR612 X BID115 AND277 X NUA56 BID115 X G22454 MEX54 X NUA56 BID29 X CAL144 SUG131 X G23823E RADICALCERINZA X BID115 AFR298 X G21242 AFR298 X BID29 BID29 X SUG131 BID115 X SUG131 or SUG131 X BID115 CAL143 X G23823E PVA773 X BID29 G5686 X NUA35 RADICALCERINZA X G23823E G6592 X G23823E AFR298 X G23823E AFR298 X BID115 BID29 X G21242 BID29 X G22454 BID29 X G23823E G21715 X G23823E G6592 X BID115 CAL96 X G23823E PVA773 X BID29 CAL144 X G23823E AFR612 X G23823E BID115 X AFR298 AFR298 X G21242 RADICALSANGIL X G21242 AFR298 X G21242 RADICALCERINZA X G21242 AND277 X G23823E AND277 X NUA35 Total of lines Iron range (ppm) Zinc range (ppm) Color type of beans 6 5 5 4 4 2 2 2 1 74 - 48 73 - 61 81 - 64 78 - 69 63 - 54 79 - 75 76 - 60 71 - 62 75 35 - 24 31 - 27 39 - 31 32 - 26 33 - 25 30 - 28 31 - 28 37 - 33 32 Red Red Red Red Red Red Red Red Red 21 21 10 10 9 8 7 7 6 4 8 4 4 3 2 2 2 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 63 - 43 71 - 37 68 - 48 70 - 47 75 - 50 90 - 65 60 - 44 74 - 51 85 - 56 67 - 50 58 - 51 75 - 52 62 - 48 78 - 62 70 - 68 70 - 58 72 - 54 57 - 50 75 - 72 61 66 40 54 66 49 56 51 75 0 46 67 62 79 43 28 - 18 40 - 18 37 - 27 41 - 23 56 - 27 44 - 37 34 - 28 39 - 30 32 - 27 34 - 29 34 - 24 27 - 22 31 - 29 38 - 29 34 - 30 27 - 25 37 - 34 32 - 29 37 26 33 20 24 26 28 35 33 35 0 14 35 35 28 21 Red and red mottled Purple mottled and purple Red Purple mottled Purple mottled , red and red mottled Rosado mottled and cream mottled Red Red and cream mottled Red Red and cream mottled Red, cream mottled and red mottled White and red mottled Red and red mottled Purple mottled White Red Purple Red Red and purple mottled Purple mottled Red Cream mottled Red Purple mottled Red mottled Purple mottled Cream Pink mottled Purple Cream mottled Purple Cream mottled Red Red mottled 4 In terms of iron and zinc content the genotypes varied between 90 and 40 ppm iron and 35 to <10 ppm zinc in backcrosses, 98 to 43 ppm iron and 28 to <10 ppm zinc in triple crosses, 105 to 49 ppm iron and 34 to <10 ppm zinc in double crosses and 90 to 37 ppm iron and 36 to 10 ppm zinc in simple crosses. In each case some lower zinc advanced lines were kept because the corresponding iron levels were high or vice versa and for all the lines there are plans to confirm iron and zinc content either through NIRS or through a repetition of AAS analyses. These results show that there is large variability for iron and zinc content in the germplasm pool for Andean bush beans after improvement for mineral levels through the nutritional breeding. We were also pleased to find that high iron and zinc levels seem to be combining well with very commercial seed type and with better plant architecture and yield potential which was somewhat deficient in the first set of NUA lines. In terms of seed colors, the commercial types could be classed into 145 large red, 127 dark red mottled, 125 light red mottled, 44 cream mottled, 19 purple speckled, 13 yellow, 7 pink mottled , 4 large white and 4 beige types. This set of NUA lines will provide a basis for additional breeding in each of these seed classes as the previous NUA lines were all of the red mottled seed class. Within the red mottled seed class, variability for seed brightness and tone will also be useful for ensuring that future combinations of breeding crosses have variability for this important commercial class which is preferred as a predominant type in Eastern and Southern Africa as well as parts of the Andean region. Future Plans: The present genotypes will be useful for studying mineral accumulation in large-seeded Andean beans and will be essential for further biofortification breeding efforts. They also may be useful for bioefficacy and bioavailability studies since they all show advantageous production characteristics which will allow them to be mass produced for nutritional promotion program. However, further testing to evaluate GxE interaction and to select the best NUA lines from this set are needed. Currently the yield potential of the genotypes is being tested at one mid-elevation site and seed multiplication is underway for shipment to the ISAR, INERA, ECABREN and SABRN breeding programs. It is expected that regional trials will be conducted with the best genotypes and that farmer preferences will be considered in the selection of the best candidate biofortified varieties prior to consumer testing for cooking time and culinary quality. Contributors: 1.1.2 M.W. Blair, F. Monserrate, C. Astudillo, A. Hoyos, Y. Viera, A. Hincapié (SBA-1) Selection of Mesoamerican lines for high minerals in cycle 2 of recurrent selection in Colombia Rationale: Improvement of the nutritional value of common beans has become one of the central themes of the CIAT program, both in Latin America and Africa. The CIAT Bean program continues to be fully committed to developing agronomically acceptable bean lines with varietal potential and with higher levels of iron and zinc. In the case of Mesoamerican beans, our goal is to produce lines that are also drought resistant to make them attractive to farmers. Materials and Methods: F2 populations of double (4 parent) crosses combining iron, drought tolerance and disease resistance were evaluated for drought tolerance in July 2007. 531 selected F1.2 populations were subsequently planted in Popayán in October 2007 for inoculation with the anthracnose pathogen. 1610 individual plants were harvested from 129 populations with superior performance, and planted in Quilichao for evaluation of ALS resistance. Bulk remnant seed of the individual selections was evaluated with NIRS, permitting the elimination of 66 of the 129 populations based on low iron. Of the remaining F4 families in Quilichao, 295 were selected based on field performance and evaluated for iron and zinc by 5 atomic absorption, permitting the selection of 71 F3.5 families for evaluation in the 2008 drought nursery. This selection scheme is outlined in Figure 1. 531 drought-selected F1.2 families 129 disease-selected F1.3 families 1610 F4 families from ind. plant selections 66 eliminated 129 F1.4 bulks, NIRS 295 selected F4 families, field Atom. Abs. 71 F5 families Correlation (NIRS F4; A.A. F5) = 0.49 Figure 1. Combining field selection, NIRS and Atomic Absorption The 71 selected families were planted in 3 different lattice designs. 867 individual plant selections were taken within the plots of the most promising families based on visual evaluation of productivity for planting plant-to-row, and the grains of these were weighed and added to the plot yield. Minerals were analyzed by atomic absorption of bulked seed of selections, and by NIRS of individual plants, and both data were used in the selection of F6 families. Results and Discussion: The correlation between iron estimated by NIRS in F3.4 and atomic absorption in F3.5 was 0.49. While only moderate, this in fact is quite good considering that, 1) the population was severely truncated between the F4 and F5 generations, reducing the genetic variability on which the correlation was calculated; 2) the F4 generation represented a bulk of selected plants, and thus the value of NIRS reflects an average of the variability within the family as compared to the F5 families; 3) the environments of the F4 and F5 generations were very different. Drought was intermittent in 2008 and the crop suffered moderate drought stress. Nonetheless several of the selected lines out-produced the commercial check, Tio Canela which averaged 1781 kg ha-1 over the three trials (Table 2). Tio Canela is well adapted to intermittent drought, and thus a yield which is superior to that of Tio Canela is quite promising. 6 Table 2. Families selected for high iron, drought tolerance and resistance to foliar pathogens # fam. Sel. COL Fe, F3.5 bulk Fe, F 3. 6 bulk, Drt Range in Fe, F6 sel. (NIRS) Zn, F3.5 bulk Zn, F3.6 bulk Drt kg ha-1 Drt ANT ALS (NCB 226xRCB 591)F1 X (SXB 597xMIB 497)F1/-MC4P-MQ-1C 9 Lt. rd 71 66 57-69 32 29 2300 1/4 4 (NCB 226xRCB 591)F1 X (SXB 597xMIB 497)F1/-MC5P-MQ-1C 3 rd 66 70 58-65 33 29 2089 1/4 4 2 cr 71 67 67-68 30 28 2268 2 3 cr str 68 63 67-77 31 28 1997 1 cr 78 72 71 31 27 4 cr str 93 72 64-79 34 3 cr 87 71 69-74 8 cr 86 73 7 cr str 75 4 cr 7 Cross code Pedigree 16168-017 16168-017 16181-017 16181-019 16181-020 16181-020 16181-020 16181-020 16181-020 16181-020 16181-020 16181-020 16181-025 16181-025 16187-021 16188-006 16188-008 16188-009 (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC23P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC20P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC2P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC7P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC11P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC12P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC13P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC17P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC19P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC21P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC2P-MQ-1C (SXB 405xSEN 37)F1 X (SMB 3xMIB 602)F1/-MC20P-MQ-1C (SXB 403xSEN 53)F1 X (SMB 6xMIB 601)F1/-MC2P-MQ-1C (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-12PMQ-1C (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-14PMQ-1C (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-4P-MQ1C bgm-1 W12 3 + + 1/4 6 + + 2080 1 5 + + 28 1681 1 5 + + 34 29 2351 1 5 + + 68-88 31 29 2329 1 4 + + 62 68-92 32 29 1678 1 5 + + 76 65 66-80 33 29 2281 1 3 + + cr str 86 76 70-84 31 30 1743 1 5 + + 6 cr 76 62 67-78 31 26 2166 1 5 + + 1 cr str 79 69 68 40 27 1940 2/4 6 - + 2 cr 85 67 72-82 35 28 1977 2/4 6 - + 8 bl 97 63 63-84 40 35 2092 _1/5 6 1 bl 78 53 75 35 28 2456 1 5 8 bl 75 58 68-80 33 27 2389 1/3 3 2 bl 73 66 68-81 32 26 2099 2 1 16188-009 (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-11PMQ-1C 2 bl 79 56 73-81 32 26 2040 2 1 16188-025 (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-1P-MQ1C 1 bl 69 79 67 35 28 2176 5 5 16188-025 (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-4P-MQ1C 11 bl 73 66 71-100 31 27 2268 5 5 16188-025 (SEN 52xSEN 67)F1 X (MIB 487xMIB 601)F1/-MC-9P-MQ1C 14 bl 100 78 69-88 36 33 1965 5 6 16198-044 (SER 119xSEN 46)F1 X (MIB 499xMIB 602)F1/-MC-1P-MQ1C 9 bl 90 83 68-90 36 28 1453 1 3 119)F1 X 602)F1/-MC- 5 cr str 84 64 71-79 38 27 1660 1/5 4 119)F1 X 602)F1/-MC- 4 cr str 86 54 73-79 41 26 2056 1/5 2 16204-010 16204-010 (SXB 407xSER (MIB 499xMIB 4P-MQ-1C (SXB 407xSER (MIB 499xMIB 5P-MQ-1C 7 Table 2. cont’d. Cross code 16204-010 16204-010 # fam. Sel. COL Fe, F3.5 bulk Fe, F 3. 6 bulk, Drt Range in Fe, F6 sel. (NIRS) Zn, F3.5 bulk Zn, F3.6 bulk Drt kg ha-1 Drt ANT ALS 119)F1 X 602)F1/-MC- 14 pk sp 72 61 71-101 35 28 2117 1/5 3 119)F1 X 602)F1/-MC- 6 cr str 73 73 68-90 34 30 1519 1/5 6 Pedigree (SXB 407xSER (MIB 499xMIB 7P-MQ-1C (SXB 407xSER (MIB 499xMIB 12P-MQ-1C 16204-010 (SXB 407xSER 119)F1 X (MIB 499xMIB 602)F1/-MC15P-MQ-1C 6 cr str 77 68 76-89 33 32 1565 1/5 5 16204-010 (SXB 407xSER 119)F1 X (MIB 499xMIB 602)F1/-MC16P-MQ-1C 1 cr str 74 69 61 32 26 2351 1/5 3 16204-016 (SXB 407xSER 119)F1 X (MIB 499xMIB 602)F1/-MC2P-MQ-1C 15 rd 69 69 68-87 24 28 2102 1/5 6 5 rd 71 60 75-92 28 27 1962 1/5 6 9 rd 94 70 69-80 29 27 2052 1/5 6 7 cr str 71 67 60-73 36 27 1867 1/3 5 1 cr str 85 71 80 38 29 1715 1/3 3 1 bl 74 71 69 36 27 1960 1 6 6 bl 74 61 70-82 40 27 2363 2 5 16204-016 16204-016 16218-009 16218-009 16246-020 16291-002 (SXB 407xSER 119)F1 X (MIB 499xMIB 602)F1/-MC14P-MQ-1C (SXB 407xSER 119)F1 X (MIB 499xMIB 602)F1/-MC16P-MQ-1C (SXB 415xAQB 608)F1 X (SMB 3xMIB 499)F1/-MC8P-MQ-1C (SXB 415xAQB 608)F1 X (SMB 3xMIB 499)F1/-MC13P-MQ-1C (SER 76xRCB 589)F1 X (MIB 499xMIB 602)F1/-MC-8P-MQ1C (SER 42xRCB 593)F1 X (MIB 487xMIB 601)F1/-MC-4P-MQ1C 16291-002 (SER 42xRCB 593)F1 X (MIB 487xMIB 601)F1/-MC-7P-MQ1C 2 rd 74 52 71 32 23 2409 2 1 16291-036 (SER 42xRCB 593)F1 X (MIB 487xMIB 601)F1/-MC-11PMQ-1C 3 bl 76 60 70-82 39 31 1792 1 3 DOR 500 55 59 53 36 28 CAL 96 42 60 51 25 22 MIB 465 70 86 90 50 39 bgm-1 W12 + - COL=Color; Lt rd=light red; rd=red; bl=black; cr=cream; cr str=cream striped; Drt= drought; ANT=anthracnose (1-9 scale); ALS=angular leaf spot (1-9 scale); bgm-1 and W12=QTL for resistance to BGMV. Among the 867 F5 individual selections made within the 71 F3.5 families under evaluation in drought, a total of 201 F6 lines were maintained, derived from 38 of the F3.5 families (Table 2). Both NIRS and atomic absorption evaluation suggest that some of these may be drawing close to the level of iron in the high iron check, MIB 465. Most have acceptable levels of resistance to anthracnose, several are resistant or intermediate to ALS, and a number of them are segregating genes for resistance to BGYMV (pending evaluation). Very few have acceptable red grain color, but a number are black seeded, and others are cream or cream-striped (carioca) type. It must be noted that the values of iron represented here tend to be lower than the real values. Accuracy of the atomic absorption and NIRS values is monitored using uniform, universal checks of bean flour (DOR 500 and MIB 465) in each evaluation. These samples were evaluated by ICP in Adelaide and in Cornell, giving respective values of 64 and 102 ppm iron. Our values of both NIRS and atomic absorption tend to be about 16% below these values. Crosses from cycle 2 of the recurrent selection scheme were also sent to Central America in 2007. These have been selected on site. We have analyzed grain of 96 selections from Zamorano, Honduras; we are 8 awaiting the release of some 97 samples from Nicaragua from Colombian quarantine; and we are expecting the shipment of 54 samples from El Salvador and 99 from Guatemala. A second set of crosses within cycle 2 of recurrent selection were planted as F1.2 populations in drought in July, 2008, and in Popayán as F1.3 populations for anthracnose evaluation. These are undergoing the same selection process described above, with the possible modification of NIRS evaluation of individual plants, if this procedure proves to be effective. Collaborators: S. Beebe, C. Cajiao, M. Grajales, M.L. Cortés, A.F. Guerrero 1.1.3 Development of new sources for high iron: Selection among interspecific families Rationale: In previous reports we have informed about the utilization of Phaseolus polyanthus and Phaseolus coccineus as sources of high iron to improve common bean. In some trials the former species has presented as much as 127 ppm in iron, or perhaps 20 ppm higher than that found in common bean. Since these two species cross readily with P. vulgaris, they were used to improve the levels of minerals in common bean. These species are also resistant to several fungal pathogens of bean, and although often used as sources of these traits to improve common bean, this is the first attempt to use them as a source of nutritional traits. Materials and Methods: High mineral accessions G35575 (P. polyanthus) and G35999 (P. coccineus) were crossed to three common bean genotypes: FEB 226 (carioca), CAL 96 (calima type), and G2333 (Mesoamerican climber). Only crosses of FEB 226 with G35575 have proved to be promising and are reported here. Populations were selected from the F2 to the F4 generation using pedigree selection. In each generation individual plant selections were made and seed was sampled from each individual plant within a family for evaluation as a bulk for mineral concentration. All mineral concentrations were estimated by atomic absorption in CIAT’s Analytical Services laboratory, using both uniform checks in the field, and laboratory checks of known mineral concentration. In the course of 2008 these families reached the F5 generation. Results and Discussion: Over several seasons some families and lines have maintained an advantage of 40 ppm over the low iron checks (Table 3). For example, MIB 755 presented levels of iron of 85, 72 and 80 ppm versus DOR 500 with 45, 39, and 43 ppm. Furthermore in the Popayán environment the iron values tend to be compressed such that the range in values is narrower, and the high iron check, MIB 465 (which normally presents 85-90 ppm iron) expresses only 65-75 ppm. The interspecific lines have presented 10-15 ppm more than the high iron check! It is fully possible that if these genes can be introduced into other genetic backgrounds that give adaptation to other environments, this range of iron expression could be even wider. These lines could well represent levels of iron that meet the goal of 95100 ppm. In two preliminary tests in Palmira, some of the lines continued to express high iron, as high as 100 ppm (data not shown). However, data from Palmira must be taken with caution in this case, as the lines tend not to adapt well in Palmira (as expected). Under higher temperatures most tend to be late maturing and with poor pod load, which may augment their iron concentration in this environment. Several lines also express levels of zinc of 45-56 ppm in Popayán, which are above that of the high mineral check, MIB 465 with 38-36 ppm (Table 3). This is also very exciting, considering that the levels of zinc in the newest lines from intraspecific crosses have improved modestly at best (Table 2 Section 1.1.2). 9 Table 3. Selected interspecific families with high iron and zinc, derived from crosses of common bean line FEB 226 and P. polyanthus. Fe, ppm, F 4. 6 Fe, ppm F4.5 Fe, ppm F 3. 4 Fe, ppm F 2. 3 Zn, ppm F 4. 6 Zn, ppm F 4. 5 Zn, ppm F 3. 4 Zn, ppm F2.3 62 60 81 63 40 36 43 31 74 72 85 63 50 43 43 31 74 72 85 63 55 43 43 31 82 72 85 63 49 43 43 31 81 72 85 63 48 43 43 31 81 72 85 63 41 39 43 31 79 72 85 63 41 39 43 31 80 72 85 63 44 39 43 31 68 55 86 63 43 34 50 31 63 55 86 63 40 34 50 31 65 55 86 63 41 34 50 31 FEB 226 X (FEB 226xG 35575-2P)F1/-12P4P-12P-MP 70 55 86 63 43 34 50 31 FEIN 15545-19 FEB 226 X (FEB 226xG 35575-2P)F1/-12P4P-13P-MP 75 55 86 63 47 34 50 31 FEIN 15545-19 FEIN 15545-18 FEB 226 X (FEB 226xG 35575-2P)F1/-13P2P-11P-MP FEB 226 X (FEB 226xG 35575-2P)F1/-10P2P-1P-MP 71 58 80 63 45 33 45 31 68 70 97 81 56 44 56 40 763 FEIN 15545-18 FEB 226 X (FEB 226xG 35575-2P)F1/-10P2P-5P-MP 80 70 97 81 52 44 56 40 764 FEIN 15546- 5 FEB 226 X (FEB 226xG 35575-5P)F1/-3P8P-6P-MP 63 68 71 71 52 42 41 43 765 FEIN 15546- 5 FEB 226 X (FEB 226xG 35575-5P)F1/-3P8P-7P-MP 71 68 71 71 49 42 41 43 Field checks CAL 96 DOR 500 FEB 226 MIB 465 39 43 48 63 39 39 45 75 . 45 . 63 42 37 44 63 31 33 33 46 25 29 29 38 . 31 . 43 28 31 31 42 MIB 466 64 63 . 55 37 31 . 37 Lab checks DOR 500, universal ch.* MIB 465, universal ch.* 59 59 59 25 22 25 93 98 71 37 35 40 MIB Cross code Identification FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEIN 15545-19 FEB 226 X (FEB 3P-8P-MP FEB 226 X (FEB 1P-2P-MP FEB 226 X (FEB 1P-3P-MP FEB 226 X (FEB 1P-4P-MP FEB 226 X (FEB 1P-7P-MP FEB 226 X (FEB 18P-2P-MP FEB 226 X (FEB 18P-3P-MP FEB 226 X (FEB 18P-4P-MP FEB 226 X (FEB 4P-1P-MP FEB 226 X (FEB 4P-8P-MP FEB 226 X (FEB 4P-10P-MP 759 FEIN 15545-19 760 748 749 750 751 752 753 754 755 756 757 758 761 762 226xG 35575-2P)F1/-1P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-10P226xG 35575-2P)F1/-12P226xG 35575-2P)F1/-12P226xG 35575-2P)F1/-12P- * Universal checks DOR 500 and MIB 465 have true iron values of 64 and 102 ppm respectively, as determined by ICP in Waite Laboratory, Adelaide, AU, and in Cornell University. Most lines have a carioca grain type, and although not yet ready for deployment in the warm tropics where carioca is used in Brazil, they could have utility in areas of highland Africa where BCMNV is less prevalent. They have shown excellent yield potential in Popayán, as well as improved disease resistance compared to the common bean parent, FEB 226 (Figure 2). Collaborators: S. Beebe, C. Cajiao, M. Grajales, M.L. Cortés 10 Figure 2. 1.1.4 Common bean line FEB 226 (left) compared with MIB 755 (right), a high iron interspecific progeny of FEB 226 and P. polyanthus. Breeding of Andean beans to create lines with higher mineral content and superior agronomic traits in southern Africa Rationale: Following on the breeders’ workshop which was held in Kampala, Uganda 2005, several countries within SABRN started breeding activities to improve Andean beans for higher mineral content and superior agronomic traits. During this reporting period, various countries continued to generate crosses combining multiple sources of resistance to diseases, high mineral content and drought. Materials and Methods: Crosses were generated by various NARS programs (South Africa, Tanzania, Zambia and Zimbabwe) and the regional breeding program in Malawi. The female parental sources representing the major market classes, as well as the donor parents for resistance to targeted diseases and for high Fe and Zn content were identified during the breeders training workshop in Kampala. Results and Discussion: In Tanzania, they noted that the first set of high micronutrient density NUA and other lines were not well adapted due to susceptibility to diseases, particularly ALS. The NUA lines were also low yielding compared to the standard bean varieties in southern highlands of Tanzania. To improve the Fe and Zn content in bean varieties, a crossing program was initiated. Over 50 pollinations were made to build up disease resistance and to get better grain type. The progenies were grown as bulk population at F3 generation and F4 seed has been harvested. Selected lines under disease pressure at F4 11 generation were crossed to NUA 45 and NUA 56 during the mid-August 2008 planting. In addition 40 pollinations were made between varieties Kirundo and Uyole 04 and also between Kirundo and OR-BR – a local breeding line which is resistant to several diseases at Uyole Agricultural Research Institute. The progenies were at F3 generation, and sufficient seed had been harvested to plant F4 population, which will be evaluated under disease pressure. The selected best progenies will be crossed to NUA45 and NUA56 for high micronutrient content. In Zimbabwe, over 25 new crosses were initiated where they used three NUA lines as male donor parents for high mineral content - NUA45, NUA59 and NUA35 to improve the following female parents: Red Canadian Wonder, DRK68, CIM9314-17, CAL143, VTTT923/10-3, MG38, AND897, DRK134, RAA34, purple, brown and G17722 – representing the red kidney, red mottled, sugar, purple and brown market classes. Purple and brown were local landraces with good attributes (resistance to drought, good pod load, good market class). The crosses also included parents with multiple traits for disease resistance. The progenies from the earlier crosses were at F4 generation. In Zambia, the breeding program had generated over 40 crosses combining attributes from various landrace varieties (Kablanketi, Solowezi Rose, Pembela, Lusaka Yellow) which had good market classes and some released varieties which had good disease resistance (Lyambayi and Lukupa) with high mineral content from AND620, NUA45 and NUA59. In Malawi the regional breeding program made 80 new 2-way crosses, resulting in 320 seeds at F1 generation, combining preferred market classes and disease resistance, using MEX54 and AND277 as sources of angular leaf spot (ALS) resistance and XAN159 or VAX6 or RMX27 as sources of resistance to common bacterial blight (CBB). These will be grown in a screen house, and will be crossed either as F1xF1 to combine more than one resistance genes for diseases or to AND620 and NUA56 as sources of high Fe and Zn. In addition there were 88 families in F1:2 generation and 606 F2:3 generation originating from 3-way crosses combining good parental lines for ALS (MEX54, CAL143) or CBB (XAN159, VAX6 or RMX27) and high Fe & Zn (NUA56 and AND620) were planted in single row plots 4 m long (41 plants) at Chitedze and Bembeke to evaluate them for CBB and ALS respectively. These populations were in various market classes: Calima (CAL113), Khaki (Nasaka) and purple (Kablanketi). Both sites were hit by drought, because the rains cut off early this year, and disease pressure, especially for CBB at Chitedze was not significant, but more importantly, grain harvest was very limited, such that selection was not justified. Remnant seed of these families has been grown at Bwanje, this June 2008 under irrigation to generate F3 and F4 families, which will be further evaluated during the coming rainy season. Grain samples from select plants at F4 generation will be evaluated for high Fe and Zn. In South Africa, 57 new 3-way crosses were made, realizing 225 seeds at F1 generation, combining preferred bean market class, disease resistance and high micronutrient content. The focus was on improving the nutritional value of red speckled sugar (RSS) with resistance to rust, angular leaf spot (ALS) and common bacterial blight (CBB) and halo blight (HB) and using AND620 and NUA56 as donor parents for high Fe and Zn content. Apart from the new crosses, some countries had advanced generations of segregating populations from the earlier crosses, which combined parental sources for various diseases and high mineral content. The F1:2 segregating populations, combining CBB resistance and high Fe and Zn content, as well as those combining ALS, rust x Fe and Zn progenies were evaluated for disease resistance in the field during summer 2008. For CBB resistance, 178 single plant progeny rows (F2:3 generation) were planted in June 2008 at Makhatini Research Station, KwaZulu-Natal and similarly under inoculation inoculated. CBB resistant single plants were selected and 315 single row plots (F3:4), were planted at the ARC-GCI in December 2008. For ALS and rust resistance 330 single plant progeny rows (F2:3 generation) with AND620 and NUA56 as donor parental lines and progeny rows from bulked F2 populations were planted during June 2008 at Makhatini Research Station, KwaZulu12 Natal. After further single plant selection, 548 (F3:4), were selected and harvested and planted in single progeny rows at Cedara Research Station during in December 2008. In addition, there were some plants which were selected from progeny rows with SEA5, SEA15 and SER16 as donor parents for drought as well as high Fe and Zn content. These together with the redundant plants from progeny rows of AND620 and NUA56 as donor parents were harvested in bulk at Cedara in 2008 and retained for Fe and Zn analyses. Forty one (41) samples of grain each consisting of 40 seeds (± 10-15g each), preferably in RSS and cranberry types were selected for analyses at the ARC-soil, climate and Water Institute (ISCW), Pretoria, South Africa. In addition 19 samples of parental lines and cultivars were included as checks. Results from the analyses showed that some of the progenies like P743, P748, P750, P751 and P780 had higher Fe and Zn content than one or both of their parents (Table 4). It was also worth noting that some bred varieties like OPS-RS4 from South Africa showed good levels of Fe content. In addition, lines used in crosses as good sources of resistance to drought, SEA15, SER16 and SEA5 also showed to have good levels of Fe and Zn, indicating that we might have more sources of parents for high Fe and Zn, and that some parents are both good sources for drought as well as Fe and Zn Table 4. Identity P743 P748 P780 P750 P751 P744 P747 P764 P749 P759 P768 P763 P772 P760 P767 P757 P771 P746 P761 P745 P758 P756 P769 P753 P755 P762 P766 P773 P765 P752 P782 Fe and Zn content data from selected progeny and parental lines used in generating the breeding for high Fe and Zn content. Line/Cross PC3834 PC3840 PC3870 PC3841 PC3847 PC3834 PC3840 PC3853 PC3841 PC3852 PC3853 PC3853 PC 3864 PC3852 PC3853 PC3848 PC 3864 PC3835 PC3853 PC3835 PC3848 PC3848 PC3860 PC3847 PC3848 PC3853 PC3853 PC3865 PC3853 PC3847 PC3870 Pedigree PC 3674/AND620 PC 3677/AND620 PC3725/NUA56 PC 3677/NUA56 PC 3681/AND620 PC 3674/AND620 PC 3677/AND620 PC3684/NUA56 PC 3677/NUA56 PC3684/AND620 PC3684/NUA56 PC3684/NUA56 PC3714/AND620 PC3684/AND620 PC3684/NUA56 PC 3681/NUA56 PC3714/AND620 PC 3674/NUA56 PC3684/NUA56 PC 3674/NUA56 PC 3681/NUA56 PC 3681/NUA56 PC3711/SEA56 PC 3681/AND620 PC 3681/NUA56 PC3684/NUA56 PC3684/NUA56 PC3714/NUA56 PC3684/NUA56 PC 3681/AND620 PC3725/NUA56 13 Nutrient content (ppm) Fe Zn 154 44 149 38 117 34 114 43 113 39 112 45 108 34 108 36 104 33 102 37 102 36 100 29 100 41 99 40 99 34 96 33 96 37 95 32 95 31 94 41 94 36 93 37 93 34 92 36 92 31 92 35 92 38 92 37 91 32 90 35 90 32 Table 4. cont’d. Identity P754 P770 P778 P781 P774 P775 P777 P776 P779 P783 P793 P792 P789 P791 P799 P784 P787 P790 P794 P796 P797 P802 P788 P800 P795 P785 P786 P798 P801 Line/Cross PC3847 PC3861 PC3870 PC3870 PC3865 PC3869 PC3870 PC3869 PC3870 PC3870 SEA 5 SER 16 OPS-RS4 SEA 15 PC3834 NUA 56 SEDERBERG PAN 116 PC 3725 PC 3681 PC 2526-BC2 (14) RS6 KRANSKOP-HR1 KRANSKOP PC3714 AND 620 JENNY PC3677 RS5 Pedigree PC 3681/AND620 PC3711/SEA5 PC3725/NUA56 PC3725/NUA56 PC3714/NUA56 PC3725/AND620 PC3725/NUA56 PC3725/AND620 PC3725/NUA56 PC3725/NUA56 Nutrient content (ppm) Fe Zn 88 88 88 88 87 87 82 81 79 76 190 127 125 116 116 106 101 100 99 96 96 93 91 91 89 88 84 82 82 Contributor: R. Chirwa Collaborators: A. Liebenberg, M. Liebenberg, D. Fourie, J. Bokosi, C. Madata, G. Makunde, K. Muimui, S. Beebe, M. Blair, R. Buruchara, P. Kimani 14 38 36 30 31 36 30 34 36 33 28 42 38 35 40 37 31 35 31 30 31 37 38 35 36 27 33 35 37 32 1.1.5 Breeding micronutrient dense bean varieties in eastern Africa Introduction. Development and utilization of biofortified varieties is regarded as probably the most effective and sustainable and potentially long-lasting strategy for reducing micronutrient deficiencies in Africa. Micronutrient malnutrition is now recognized as one of the most serious health challenges facing vast sectors of Africa’s population particularly resource-poor women and children (Kimani et al., 2001). Major deficiencies include iron, zinc, vitamins and protein. Micronutrient deficiency is often referred as ‘hidden hunger’ because the problem does not show any easily recognizable symptoms in the early stages, until it is almost too late, when considerable, and often irreversible damage has occurred. Prevalence of iron deficiency anaemia (IDA) varies from 8% in Ethiopia, 67% in Tanzania, to 69% in Burundi. The main cause of these deficiencies is a diet rich in energy but poor in proteins, minerals and vitamins. The problem is further aggravated by widespread poverty, which makes it difficulty for the vast majority to access the more expensive animal based products, which are rich in vitamins and minerals. Limited knowledge on the nutritional value of locally available foodstuffs and changing eating habits that regard traditional vegetables and other non-staples as ‘old-fashioned’ have further worsened the problem. The preferred foods include cereal-based products (sifted maize meal, milled rice), white potatoes and cassava that are generally low in micronutrients. A three-pronged approach has been followed in alleviating the micronutrient deficiency problem in Africa. These are: supplementation of vulnerable groups with micronutrients, fortification of common processed foods, and dietary improvement. Mineral supplementation is effective for easy-to-reach vulnerable groups with access to medical facilities. In East and Central Africa, this constitutes a very small group. It requires a large capital input, an elaborate and costly distribution network and patience compliance. It has to be carried out on a regular basis. This approach often leaves out those hard-to-reach or practically unreachable at-risk groups as well as other household and community members not targeted to receive any kind of supplementation. In Africa, the latter group constitutes the majority who are located in rural communities with limited access to medical facilities and too ill from the effects of the deficiency. Fortification of common foods has had a limited degree of success in Africa because of the underdeveloped food industry and lack of effective legislation. For example, at present food fortification programs are operational only in two of the ten ASARECA member countries in East and Central Africa: Kenya (vitamin A and iron in wheat flour, maize flour, millet porridge, margarine and cooking oil) and Uganda (iron in wheat flour). The number is equally small in southern Africa. Food fortification is effective for small affluent communities mostly in urban areas and households with a capacity to purchase fortified foods on a regular basis. This again leaves out the majority of urban poor and rural communities. Dietary improvement is probably the most effective and sustainable strategy for reducing micronutrient deficiencies in Africa. This approach aims to increase dietary availability, regular access and consumption of mineral-rich foods in at - risk and micronutrient-deficient groups of populations. It involves development and promoting enhanced consumption of culturally acceptable, mineral rich grains, vegetables and root crops. Program Objectives: A regional program led by PABRA, and based at the University of Nairobi and the Malawi National Bean program in partnership with CIAT and HarvestPlus was initiated in 2001. The objectives of this program were to: (i) Characterize the variation of grain iron and zinc concentration in east, central and southern Africa, (ii) identify potential parents for further breeding work, and iii) determine whether there are regions with high diversity for this trait and complement existing collection at Genetic Resources Unit at CIAT, Colombia, iv) identify lines which could be fast-tracked as mineral dense lines for cultivation by farmers in regions with severe Fe and Zn malnutrition, v) assess the cooking, bioavailability of Fe and Zn and other organoleptic characteristics of promising mineral dense lines. 15 Variety development. Major steps in development of common bean varieties rich in micronutrients has involved collection and screening of varieties, landraces, germplasm accessions and advanced lines for micronutrients, participatory evaluation of promising lines for adaptability and agronomic traits in observation and preliminary on-farm and on-station trials, advancement and further characterization of selected lines in advanced and multi-location yield trials to generate data required for formal release, and production of breeder seed to facilitate production and dissemination of commercial seed to end users. Genotypes with high concentration of micronutrient but lacking in agronomic traits and susceptible to major diseases entered hybridization programs to generate segregating populations and select for lines combining mineral density with resistance to biotic and abiotic stress factors, marketable grain types and high yield potential. We previously highlighted the regional strategy adopted in the development of micronutrient rich bean varieties (CIAT, 2007). This report highlights key milestones reached by the regional program towards the development and release of micronutrient dense varieties for smallholder farmers in eastern, central and west Africa between 2003 and 2008. Materials and Methods: Bean germplasm was collected in nine countries in east and central Africa. Collections included landraces, varieties, introductions, germplasm accessions and breeding lines held by national bean programs and gene banks. A germplasm collection, characterization and conservation protocol was developed and distributed to bean programs in east, central and southern Africa. Samples were analyzed at the University of Nairobi (Kenya), and subsequently at CIAT (Colombia), Cornell University (USA), University of Copenhagen (Denmark) and Sokoine University (Tanzania) to facilitate cross-lab comparison (CIAT, 2001, 2002, 2003 and 2004). Thirty-eight fast track lines were identified and distributed for regional evaluation across agro-ecological zones in more than 15 countries in east, central and southern Africa, and later in West Africa (CIAT, 2004, 2007). Fast track lines were also evaluated for anti-nutritional factors (tannins and phytates), cooking time, mineral retention and other organoleptic characteristics (CIAT, 2005, 2006, 2007). Genotypes with high micronutrient concentration but lacking in preferred agronomic traits and susceptible to major diseases were entered in hybridization program at Kabete to generate segregating populations and select for lines combining mineral density with resistance to biotic and abiotic stress factors, marketable grain types and high yield potential. Mineral analyses were performed used ashing and wet digestion techniques (Zarcinas et al., 1983). Normal agronomic practices were followed in field trials. Data was analyzed using Genstat and/or SAS statistical software. Results and Discussion: Germplasm screening. Although PABRA partners had a target of 2000 accessions, more than 2853 germplasm accessions were collected in nine countries in east and central Africa in the last five years (Table 5). They included landraces, old varieties and some breeding lines. Mineral analyses showed that there was considerable genetic variation for grain iron and zinc concentration among the local germplasm, which could facilitate development of mineral rich bean varieties. For example, analysis of 300 lines from D. R. Congo, Kenya, Uganda, Rwanda and Tanzania showed that iron concentration varied from 40 to 105 ppm. Sixty-six lines had more than 90 ppm indicating that potential of increasing iron concentration in the grain by more than 90%. Analyses of four landraces collected in central and southern Tanzania showed that iron concentration was lowest in white beans (73 ppm) and highest in grey (114 ppm) and yellow beans (120 ppm). Kidney beans had intermediate levels of iron (87 ppm). Bean showed higher iron concentration compared to whole maize (19 ppm), dehulled maize (7 ppm), cassava (5 ppm), cassava flour (5 ppm), cocoyam (4 ppm), potato (3ppm) sweet potato (5 ppm) and cooking bananas (5 ppm), which are widely consumed in the region. Mineral concentration in grey and yellow beans was higher than beef (98 ppm) and comparable to fish (124 ppm). This implied that beans are among the most important sources of iron in local diets, and is critical to the vast majority who have limited access to animal sources. 16 Table 5. Germplasm accessions collected, characterized and analyzed for mineral density in East and Central Africa, 2003-2008. Country Target Number collected 27 Number characterized 0 Number analyzed for Fe and Zn 0 Burundi 0 DRC-East DRC-West 700 100 500 150 500 150 300 10 Ethiopia Kenya Madagascar Tanzania Sudan Rwanda Uganda Total 0 400 0 400 0 1100 600 2000 5 362 52 177 8 1100 500 2853 5 362 52 141 8 1037 125 2380 5 282 0 12 5 900 180 1684 Although mineral density was affected by the growing conditions (soil type, soil nutrients) and methods of analyses, results showed that most of the lines consistently showing high levels of micronutrients originated from the Great Lakes region especially D. R. Congo and Rwanda. Genotypes with high levels of iron included Maharagi Soja, Gofta, AND 620, MLB 49 89A, HRS 545, Nakaja, VCB 87013, Nain de Kyondo, TY 3396-13, PVA 8, Nguaku Nguaku, Urugezi, Lib 1, Roba-1 and Mwamfutala. All except Gofta, Roba-1 (Ethiopia) and HRS 545 (Sudan) originated from the Great Lakes Region. Blair et al. (2007) also reported Nakaja and Urugezi as a high Fe lines. Lines with high levels of zinc included VNB 81010, MLB 49 89A, AND 620, LIB 1, Ranjonoby, Naindeky, Nakaja, Jesca, K131, K132 and Kiang’ara. When these lines were grown under uniform low N conditions in the greenhouse at Kabete, they showed lower levels of Fe (21-81 ppm) and Zn (8-38 ppm) compared with results with original samples. This suggested possible genotype x environment interaction for these traits, or possibly contamination, which is common in work with minerals. Large seeded Andean varieties from SABRN and ECABREN regions had higher Fe concentration than the small seeded Mesoamerican varieties from the same region. Finally, a total of 38 lines were selected for next stage of variety development. These were dubbed ‘fast track’ lines because they combined high mineral density with other important traits preferred by farmers and consumers, and would therefore require less time for testing before being released as varieties. Selection of Mineral dense Climbing Varieties. Although the current initiative to develop mineral dense bean lines initially focused on bush bean with marketable grain characteristics, it became necessary to develop a parallel program for the more productive biofortified climbing bean types. Climbing beans typically give a three to four fold yield increase per unit area and can be ideal in urban and rural areas with high population pressure and declining land size. To realize this objective, CIAT introduced advanced climbing bean lines selected for high mineral density (referred to as NUVs) for preliminary observations in primary evaluation sites in east, central and southern Africa. The objective of these trials was to conduct preliminary adaptability trials, and to increase seed for distribution to national programs and communities for further participatory evaluation and eventual release as commercial varieties. In East and Central Africa region, two sets of climbing beans have been introduced and evaluated. The first set of climbing beans originated from fast track nursery which comprised of local landraces, released varieties, and advanced breeding from the national bean programs. This nursery was created in 2001 but new accessions have been added in the last eight years. Genotypes in this nursery were screened for grain iron 17 and zinc concentration. Genotypes with iron levels above 70 ppm and zinc levels above 30 ppm were evaluated in participatory on-farm and on-station trials for agronomic traits and consumer acceptance. The second set of climbing bean lines were NUV lines. They were introduced in eastern Africa in 2007 from CIAT, Colombia. Field evaluation of these lines started in 2007. First track climbing bean nursery was distributed to nine countries in eastern Africa (ECABREN) from 2003, and to central and west Africa (WECABREN) in 2005, 2006 and 2007. The nursery had type III (semi-climbers) and type IV (climbers). The status of evaluation is shown in Table 6. Table 6. Status of evaluation of fast track mineral dense climbing beans lines in eastern Africa, December 2008. Country Burundi D. R. Congo (east) D. R. Congo (west) Ethiopia Kenya Madagascar Rwanda Sudan Tanzania Uganda Cameroon Central African Republic Guinea Conakry Congo (Brazzaville) Observation trials Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Preliminary yield trials Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Advanced yield trials Yes Yes Yes Yes In progress National multilocation trials In progress In progress In progress Yes Candidates for release 4 4 3 2* In progress Yes Yes * Madagascar does not have a formal variety release program yet. The two lines (VNB 81010 and M211) were selected from fast track nursery. FOFIFA bean program considers them as pre-releases and seed production with partners started in 2008. Results show that considerable progress has been made in Burundi, D. R. Congo, Kenya, Madagascar and Cameroon. In Rwanda, one line (G59/1-2) has been selected for advanced yield trials because it combined high yield potential (2.5 to 3.5 t ha-1) with mineral density (110 ppm Fe). More than 2400 kg of seed was produced with partners in Ruhengeri region. In Cameroon, TY 3396-12 with type III growth habit is among the most promising lines from the fast track nursery. In Burundi selection was done in multilocation trials in on-farm and on-station trials in high, medium and low altitude locations. Five climbing bean lines (Nakaja, VNB 81010, VCB 81012, Kiangara and G59/1-2) were selected in 2008 as candidates for final evaluation and seed production. In D. R. Congo, ten climbing bean lines from fast track nursery were selected in participatory variety selection conducted at three high altitude locations in eastern part of the country. They included (in decreasing order of preference) VCB 81013, G59/1-2, AND 10, LIB 1, MLV 224/97B, VCB 81012, VNB 81010, Kiang’ara, MLV 59/97A and MLV 06-90B. Low mineral check M211 was selected in Madagascar, Burundi and D. R. Congo because of its good productivity. In western DRC, four climbers which combined high mineral density and agronomic potential were selected (Table 7). 18 Table 7. Grain iron, zinc and protein concentration of four climbing bean genotypes adapted to low and medium altitudes selected in western D. R. Congo. Line G59/1-2 VCB 81013 LIB 1 Kiangara Source: Mbikayi and Lodi Lama, 2008 Fe (ppm) 90 95 94 80 Zn (ppm) 45 20 52 20 Protein (%) 22.9 20.8 20.2 In Kenya, seven climbing bean candidates from the fast track nursery were evaluated in national performance trials (NPT) conducted at eight locations for two seasons between 2007 and 2008. Regional important cultivars, Vunikingi, Umbano and recent releases (MAC 13, MAC 34 and MAC 64) were used as checks. Four candidate varieties were pre-released (MV 14, MV17, MV18 and MV19). MV 19 was the best yielding. The pre-released lines were evaluated for a second round of NPT in 2008. Table 8 shows a summary of the combined results over the two years. Three candidate varieties (MV 19, MV17 and MV 14) were recommended for full release in December 2008. This marks the first time bean genotypes selected for micronutrient density reached full release status after independent, multilocation evaluation for agronomic traits. These results also confirmed that it is possible to combine high mineral density with good agronomic potential. Table 8. Yield of four mineral dense climbing bean lines evaluated in national performance trials at seven locations in Kenya over two years. Line Status MV 19 MV17 MV14 MV18 MAC 34 MAC 13 MAC 64 Vunikingi Candidate Candidate Candidate Candidate Check Check Check Check Mean yield across environments (kg ha-1) 2230 2200 2150 1560 1530 1190 1180 1110 Yield over best check (%) 45.8 44.1 40.1 2.2 Source: KEPHIS, 2008 Evaluation of NUVs. 264 NUV lines were planted in observation trial at Mwea during the 2007 long rain season. Grain yield varied from 420 to more than 3000 kg ha-1. Yield was adversely affected by moisture stress during the flowering and pod filling growth stages. Twenty-five lines had yields of more than 3 t ha-1 despite the stress. Some of the best performing lines included NUV 16, NUV 35, NUV 41, NUV 71, NUV 72, NUV 73, NUV 90 and NUV99. This trial also facilitated seed increases. The lines are currently being evaluated in replicated trials at Kabete, Laikipia, Thika and Mwea. Mineral analyses of grain from the observation trial is in progress. 19 Status of biofortified fast track bush lines: Considerable progress was made during the year in evaluation of fast track bush lines in eastern Africa. Table 9 shows the status of these lines in the region. Table 9. Status of evaluation of fast track mineral dense bush bean lines, December 2008. Country Burundi D. R. Congo (east) D. R. Congo (west) Ethiopia Kenya Madagascar Rwanda Sudan Tanzania Uganda Cameroon Central African Republic Guinea Conakry Congo (Brazzaville) Observation trials Yes Yes Yes Preliminary yield trials Yes Yes Yes Advanced yield trials Yes Yes Yes National multilocation trials In progress In progress Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes In progress Yes Candidates for release 3 4 - Yes Yes In Burundi, fast track lines were evaluated in advanced yield trials at three locations during 2008 A and B seasons. The trial sites were Moso (1250m), Mparambo (800 m) and Murongwe (1500m), representing the major agroecological zones for bean production. Eight lines were selected based on yield, reaction to diseases and adaptation and better performance compared with the local check variety Mukungungu. The selected bush lines were GLP 2, PVA 8, MLB49-89 A, MLB40-89A, Maharagi Soja , Gofta and Nguaku – Nguaku. However, Nguaku Nguaku showed poor performance at the low altitude site due to weak stems and prostrate growth habit, excessive vegetative growth and poor pod load. These lines will be evaluated in on-farm trials in 2009. In eastern D. R. Congo, eleven lines were selected based on their performance at five locations and farmer evaluations. The selected lines were BRB 194, CODMLB 005, Selian 97, COD MLB 007, Nguaku Nguaku, CODMLB 001, AFR 708, CODMLB 003, Maharagi soja, M’Sole and CODMLB078. BRB 194 was selected by 61% of the participating farmers compared with 52 % for CODMLB 005, 48% for Selian 97, 46% for CODMLB 007 and 14.8% for the check variety. Table 10 shows fast track lines selected in western D. R. Congo. These lines showed wide adaptation from 500 to 1500 m above sea level. HM 21-7 originated from Bilfa nursery and is adapted to low fertility soils and drought. Seed of these lines is being increased. In Kenya, eight bush lines were submitted for national performance trials in 2007 and 2008. Candidate lines were evaluated by the Kenya Plant Health Inspectorate (KEPHIS) and the national variety release committee in ‘highland’ and ‘lowland’ test sites. ‘Lowland’ sites included Kabete, Embu, Kaguru, Katumani and Thika. ‘Highland’ sites included Chepkoilel, Kakamega, Kisii, Mabanga and Njoro. Four lines (AND 620 (MN1), Gofta (MN3), TY 3396-12(MN5) and NUA 1(MN9) were pre-released in 2008 based on better performance compared with checks. The performance of these lines in the national performance trials over two years (2007 and 2008) is presented in Table 11. 20 Table 10. Grain iron, zinc and protein concentration of seven fast track bush bean genotypes adapted to low and medium altitudes selected in western D. R. Congo. Line HM 21-7 Nguaku Nguaku BRB 194 CODMLB 078 Maharagi Soja AND 620 CODMLB 007 Fe (ppm) 90 85 Zn (ppm) 25 20 Protein (%) 20.2 20.6 75 55 97 122 40 30 20 23 26 20 22.7 19.4 19.7 Source: Mbikayi and Lodi Lama, 2008 Table 11. Grain yield of eight mineral dense lines selected from the Fast track nursery at 10 sites over two years in Kenya. Genotype MN 1 (AND 620) MN 3 (Gofta) MN 6 MN 9 (NUA 1) MN 5 (TY 3396-12) MN 10 MN 2 MN 11 GLP 92 GLP 1127 GLP 2 Lowland sites Highland sites Mean yield (kg ha-1) Yield over best check (%) Yield over mean of checks Mean yield (kg ha-1) Yield over best check Yield over mean of checks 1150 1130 1130 1010 880 950 880 510 950 800 830 20.9 18.8 18.0 6.1 -7.3 -0.1 -7.4 -46.6 33.8 31.5 30.6 17.5 2.6 10.6 2.5 -40.3 2030 1960 2800 1890 1150 1210 1260 1420 1380 1530 1280 32.6 28.7 80.6 23.3 -24.8 -20.5 -7.0 -46.6 45.1 45.1 97.7 35.0 2.6 10.6 2.5 1.8 Source: KEPHIS, 2008. GLP 92 (Mwitemania), GLP 1127 (Mwezi Moja) and GLP 2 (Rosecoco) were checks. In Rwanda, four bush varieties from the fast track nursery were selected in advanced yield trials. These were AND 620, Maharagi Soja, MLB 49 – 89 A and MLB 40– 89A. These lines had mean grain yield of 1 to 1.6 t ha-1 in multilocation testing. Seed of the four lines and Gofta was produced in Gitarama, Ruhengeri, Kigali-Ngali, Bugesera and Kibungo regions with partners (NGOs, Ministry of Health and communities) and on-station. In Tanzania, 11 lines selected from the fast track nursery were evaluated in uniformity cultivar multilocational trials (UCT).They included GLP 2, Nain de Kyondo, PVA 8, OBA 1, Kirundo, K132, K131, Ranjonomby, Lingot Blanc, RWR 10 and Zebra. Selection from Segregating Populations. Mineral dense lines derived from fast track nursery, which are susceptible to diseases (anthracnose, angular leaf spot, BCMV, BCMNV and rust) were entered in crossing block to correct the deficiencies. Selection for new combinations with mineral density, resistance to diseases and preferred grain types started at INERA-Mulungu (D. R. Congo), Selian 21 Agricultural Research Institute (Tanzania) and Kabete (Kenya). At Kabete 8 NUA red mottled lines generated from crosses between CAL96 and mineral dense parents were analyzed for minerals and protein concentration (Table 12). Results showed iron concentration varied from 45 to 75 ppm. Zinc concentration varied from 25 to 50 ppm. At Kabete, seven new populations combining resistance to diseases and marketable grain types were advanced from F2.5 to F2.6 and F2.7 . Selected lines showed considerable variation for iron, zinc and protein concentration (Table 13). Several lines with high iron, zinc and protein concentration were identified. Grain iron concentration varied 30 to 105 ppm in lines selected from KAB 2 population, 55 to 125 in KAB 5, 30 to 130 ppm in KAB 6, 30 to 115 in KAB 10, 40 to 115 in KAB 11, 35 to 100 ppm in KAB 12, and 50 to 115 ppm in KAB 13 derived lines. Zinc concentration varied from 10 to 55 ppm among the 300 lines. Protein concentration varied from 17.4% to 28.5%. These results suggested that varieties combining high micronutrient density, resistance to diseases and marketable grain types can be developed from these populations. Table 12. Iron, zinc and protein concentration of advanced NUA lines, runner bean parental lines and their F1, F2 populations and F3 families in Kenya. Genotype NUA 1 NUA 2 NUA 3 NUA 4 NUA 5 NUA 6 NUA 7 NUA 8 Fe (ppm) 65 75 70 50 45 60 65 55 Zn (ppm) 50 45 45 40 25 35 50 35 Protein (%) 24.1 24.8 26.5 22.9 24.1 24.3 25.2 22.9 Runner beans Nyeri 1 Kinangop 2 Kinangop 4 (KIN 4) White Emergo Kinangop 1 (KIN 1) White Emergo x KIN 4 F1 105 60 45 70 60 60 45 30 25 20 35 30 19.3 19.7 19.8 18.9 21.1 21.5 White Emergo x KIN 4 F2 White Emergo x Kenya Local (F2) Kenya Local x White Emergo (F3) White Emergo F2 White Emergo x KIN 1 (F2) 110 80 60 90 65 25 35 20 20 25 17.2 15.3 15.5 17.9 17.4 Table 13. Population KAB 02 KAB 05 KAB 06 KAB 10 KAB 11 KAB 12 KAB 13 Total Iron, zinc and protein concentration in F2.6 lines selected from seven bean populations in Kenya. No. of lines 67 16 70 47 30 25 26 281 Fe (ppm) 30-120 55-125 30-130 30-115 40-115 35-100 50-115 22 Zn (ppm) 10-40 10-35 10-55 10-45 10-40 10-40 10-60 Protein (%) 18-26 19-24 17-24 19-28 20-28 19-24 21-29 Nutritional evaluation. Seed of the 38 fast track lines was sent to Sokoine University for planting to generate leaves, pods, green shelled beans for mineral analyses. At the University of Nairobi two studies were conducted to assess micronutrient concentration in bean leaves. In the first study, seed and leaves of 72 bean lines of diverse origins grown in 40 farmers fields in Kisii district, Kenya was analyzed for iron , zinc and protein concentration. Results showed that iron concentration in leaves was much higher than in the seeds. Leaf iron concentration varied from 397 ppm in Awash 1 to 2498 ppm in Ngwinurare, with a mean of 1118 ppm. Seed iron concentration varied from 50 ppm (MCM 2001) to 108 ppm (M’Sole). However, the levels of zinc were comparable to that of seeds. Leaf zinc concentration varied from 20 ppm (Sugar 73) to 67 ppm (Ngwinurare). It is significant to note that Ngwinurare is a popular variety in Rwanda. Its leaves and grain are being used by communities in five districts participating in the ‘Agriculture, Nutrition and Health collaborative project’. The results suggested that bean leaves which are widely consumed in the region can make a significant contribution to micronutrient nutrition. Preliminary results from the Rwandese project indicate improved micronutrient health status in participating communities. In the second study, leaves of the 38 lines fast track lines were analyzed for iron, zinc and protein concentration. Results showed that leaf iron concentration varied from 236 ppm in cv. ‘Zebra’ to 1961 ppm in Kirundo. Leaf zinc concentration varied from 17 ppm in cv. Nguaku Nguaku to 94 ppm in cv. Kiangara. Crude protein varied from 23.7% in Red Wolaita to 35.6% in TY 3396. These results seemed to confirm the first study that bean leaves have much higher iron levels compared to the grain. Results on cooking time, nutrient retention, taste, water absorption have been reported (CIAT, 2007). Bioavailability of Fe and Zn in bean varieties: Trials were conducted to determine bioavailability of iron and zinc in dry beans and green shelled beans of the 38 fast track lines. The in vitro studies were conducted in partnership with Sokoine University of Agriculture, Tanzania. Results showed that bioavailability of iron varied with genotypes (Figures 3 and 6). Iron bioavailability was lowest in Roba (1.1%) and highest in Maharagi Soja (6.6%). Cooking enhanced bioavailability. Bioavailability of Fe in cooked samples varied from 3.9% for VCB 81012 to 6.8% for cooked samples of Maharagi Soja. Bioavailability of zinc also varied with genotypes (Figures 4 and 5). It was lowest in OBA (0.5%) and highest in Ituri Matata (2.5%). Cooking enhanced bioavailability of zinc. Bioavailability of zinc in cooked samples of dry bean varied from 0.4% (M’Mafutala) to 3.9% for G59/1-2. Higher bioavailability was observed in green shelled beans compared to dry mature grains (Figures 3, 4, 5 and 6). Seed production and Dissemination. Several countries increased seed of fast track lines to facilitate local trials and germplasm exchange with other countries and dissemination to farmers. The regional program in Kenya increased seed of biofortified varieties at Kabete, Thika, Ol Jorok and Laikipia. Seeds were shipped to western and eastern D. R. Congo, Rwanda, KARI-Katumani, Tanzania (Selian Agric Research Institute), Denmark, Sokoine University, Madagascar and Cameroon. More than 9000 kg of five fast track lines (Maharagi Soja, Ngwinurare, AND 620, MLB 40-89A and MLB 49-89A) were produced and distributed to communities in six districts (Ruhuha, Ruhengeri, Gihara, Kigali, Kigoma and Rwamagana) in Rwanda in partnership with the ATDT Health and Agriculture project. Lagrotech Seed Company increased seed of biofortified varieties to facilitate on-farm trials and multilocation on-station trials in western Kenya. D. R. Congo increased seed to meet local needs and shipment to Burundi. Contributors: Paul Kimani, S. Beebe, M. Blair (CIAT) and Peter Mamiro (Sokoine University of Agriculture) Collaborators: Nkonko Mbikayi (D. R. Congo), A. Namayanja (Uganda), Rael Karimi( KARIKatumani), and Bean teams at Kabete, CIAT-Colombia, FOFIFA (Madagascar), ISAR (Rwanda), Martha Nyagaya (CIAT), Lagrotech Seed Company (Kenya). 23 Bioavailable Iron from raw and cooked green shelled beans 18.0 16.0 14.0 Iron (%) 12.0 10.0 8.0 6.0 4.0 2.0 0.0 0 5 10 15 20 25 30 35 40 Variety % Bioavailable Iron (Fe) Cooked Green Shelled Beans Figure 3. % Bioavailable Iron (Fe) Raw Green Shelled Beans Bioavailable iron from raw and cooked green shelled bean varieties. Bioavailable Zinc from raw and cooked green shelled beans 7.0 6.0 Zinc (%) 5.0 4.0 3.0 2.0 1.0 0.0 0 5 10 15 20 25 30 35 Variety % Bioavailable Zinc (Zn) Raw Green Shelled Beans Figure 4. % Bioavailable Zinc (Zn) Cooked Green Shelled Beans Bioavailable zinc from raw and cooked green shelled bean varieties. 24 40 Zinc (%) Bioavailability of zinc from raw and cooked beans 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 0 5 10 15 20 25 30 35 40 Variety % Bioavailable Zinc (Zn) from Raw Beans % Bioavailable Zinc (Zn) from Cooked Beans Figure 5. Bioavailable zinc from raw and cooked dry beans of 38 fast track lines. Bioavailability of iron from raw beans 8 Iron (%) 6 4 2 0 0 5 10 15 20 25 30 Figure 6. Bioavailable iron from raw and cooked beans of 38 fast track lines. References CIAT. 2001. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2002. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2003. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2004. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2005. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2006. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2007. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia Zarcinas, B.A., B. Cartwright, and L.R. Spoucer. 1987. Nitric acid digestion and multi-elemental analysis of plant material by inductively coupled plasma spectrometry. Communications in Soil Science and Plant analysis. 18 : 131-146. Kimani, P.M. and E. Karuri. 2001. Potential of micronutrient dense bean cultivars in sustainable alleviation of Fe-Zn malnutrition in Africa. In: Novel Plant breeding Approaches to fight micronutrient deficiencies. International Center for Tropical Agriculture (CIAT), Bill and Melinda Gates Foundation and the Micronutient Initiative. CIAT, Cali, Colombia. (CD) 25 Activity 1.2 Genotype x environment interaction Highlights: • NUA35 and NUA56 were tested for yield potential and mineral accumulation across 15 sites in Latin America (72 replicates), showing that both lines have a 15 to 25 ppm differential iron advantage over CAL96. • Significant genotype x environment interactions indicate that grain mineral concentration is influenced by soil type, soil nutrient status, moisture concentration and other environmental factors but the magnitude varies with genotypes. Some genotypes show high stability for mineral density. • Fertilization regimes and other agronomic practices can be used to enhance expression of high mineral density traits. 1.2.1 Multi-site evaluation of biofortified Andean lines, NUA35 and NUA56 Rationale: Biofortified genotypes of common bean hold promise for improving nutritional status in many countries but require agronomic testing to determine their yield and adaptation potential. Andean biofortified beans from the NUA series have generated interest in various countries of Africa and Latin America and are potential releases for Bolivia, Colombia, Malawi and Zimbabwe. Two sister lines (NUA35 and NUA56) were selected for their high iron and zinc content. Both of these were developed from the high iron source genotype G14519 in backcrosses with the recurrent parent CAL96 and have good red mottled seed type. These genotypes have now been tested over a large number of sites and seasons and this report outlines the iron and zinc levels found in them. Materials and Methods: Two genotypes, NUA35 and NUA56 were grown across 15 sites and analyzed uniformly for seed mineral content with atomic absorption spectrophotometry (AAS). Before analysis and to reduce variability, seed for each experiment was hand harvested and processed to avoid soil contamination and then shipped to CIAT for analysis where grain was cleaned with a damp cloth. Seed mineral content was evaluated for 4 g of grain dried overnight in an oven at 37ºC. Each sample was then ground into a fine powder using a modified Retsch mill with a teflon capsule chamber and zirconium grinding balls. Mineral content is reported in parts per million (ppm), equivalent to mg of iron or zinc per kg of seed. Results and Discussion: Mean seed yield for NUA35 at a moderate-elevation/rainfall site in Colombia (Darién at 1459 masl with 20°C seasonal average temperature, 1328 mm average yearly rainfall, with soil pH 5.5-6.0 and 5 to 8 % organic matter) was 1099 kg ha-1 with a range from 632 to 1787 kg ha-1 over 6 seasons (given bimodal yearly rainfall at this sites, the line was tested over three and a half years over both March - June and September - January growing seasons). The mean seed yield of NUA56 at this same site was 1076 kg ha-1 with a range of 704 to 2095 kg ha-1 over 5 seasons. In comparison, control genotypes AFR612 and CAL96 yield between 700 and 1800 kg ha-1 at this site. At a high rainfall site in Colombia (Popayán at 1725 masl with 17°C seasonal average temperature, 1991 mm average yearly rainfall with soil pH 5.5-6.0 and 10 to 20 % organic matter), mean seed yield for NUA35 was 1521 kg ha-1 with a range from 719 to 2810 kg ha-1 while mean seed yield for NUA56 was 1324 kg ha-1 with a range from 526 to 2839 kg ha-1. At a lower elevation site in Colombia (Palmira at 967 masl with 24°C seasonal average temperature, 1043 mm average yearly rainfall, soil pH 7.0-7.5 and 2 to 4 % organic matter), mean yields for NUA35 and NUA56 were 946 and 810 kg ha-1, respectively. Meanwhile, in Santa Cruz, Bolivia (Andres Ibañez at 398 masl with 24°C seasonal average temperature and 1406 mm average yearly rainfall with soil pH 6.0-6.5 and <2 % organic matter), average yields for 26 NUA35 and NUA56 were 1265 and 1452 kg ha-1, respectively, both surpassing the average yield of the control genotype, CAL96, with a yield of 997 kg ha-1 but beneath the average yield of a local check, ‘Rojo Oriental’ a released variety based on CIAT line PVA773, with 2000 kg ha-1. Nutrition quality evaluations for seed iron and zinc content (Table 14) showed that the NUA lines always produced more than the average amount of iron for common bean which is 55 ppm; and close to the average amount of zinc for the crop which is 35 ppm. While both lines were high in iron, NUA56 had somewhat higher average seed iron content (81 ppm) than NUA35 (76 ppm) across the 15 sites where the genotypes were tested; while the opposite was true for average seed zinc content where NUA35 had 34 ppm and NUA56 had 33 ppm. Considering that Andean beans usually have lower than average seed zinc content, this amount of zinc can be considered moderate within the genepool. Table 14 also shows that iron levels in the NUA lines responded to environmental effects with variability for iron from 53 to 101 ppm for NUA35 and 61 to 112 for NUA56, with highest concentrations reached in Santa Cruz, Bolivia and Yacuanquer, Colombia. In comparison, zinc content was fairly stable varying from 28 to 43 ppm for NUA35 and from 25 to 43 ppm for NUA56 with the same sites mentioned for iron producing high seed concentrations of zinc. The iron differential of the two germplasm lines compared to the recurrent parent CAL96 was usually 15 to 20 ppm with up to 25 ppm difference for NUA35 in Santa Cruz, Bolivia and 37 ppm for NUA56 in Yacuanquer, Colombia; while the zinc differential was less substantial. Conclusions and Future Plans: NUA35 is being considered for release in Colombia and Bolivia so the results presented here are timely. Promotion will continue by FIDAR, IPRA/CIAT and partners. The seed color of the NUA lines is very acceptable as they have the same deep red color that is noteworthy of CAL96 the recurrent parent used for development of the NUA lines evaluated so far. The environmental factors that affect iron accumulation at the different sites used for multiplication or testing of NUA lines will be further studied. Contributors: M.W. Blair, F. Monserrate, C. Astudillo, S. Beebe (SBA-1, CIAT) G. Hyman (GIS, CIAT), J. Restrepo, P. Ojeda (FIDAR), J. Ortubé, V. Choque, J. Padilla (UAGRM, Bolivia) 27 Table 14. Seed iron and zinc concentration1 (in ppm) for CAL96 (released variety) versus NUA35 and NUA56 (high iron advance lines) grown over 15 sites in Latin America. CAL 96 Location (Department, Country) Seasons Replications A. Ibañez (Santa Cruz,Bolivia) San Juan (Santa Cruz, Bolivia) Darién (Valle, Colombia) Palmira (Valle, Colombia) Yotoco (Valle, Colombia) Vijes (Valle, Colombia) Sandoná (Nariño, Colombia) Yacuanquer (Nariño, Colombia) Consacá (Nariño, Colombia) Popayán (Cauca, Colombia) Quilichao (Cauca, Colombia) Caldono (Cauca, Colombia) Puriscal (San Jose, Costa Rica) 1 1 6 4 1 1 1 2 3 5 3 2 1 1 1 18 12 1 1 3 4 4 15 3 2 1 Quesada (Jutiapa, Guatemala) 1 3 Chinantenango (Guatemala) Average across sites 1 33 3 72 NUA 35 NUA 56 Latitude (dd°mm') Altitude (masl) 17°42.51' S 17°57.00' S 3°55.73' N 3°30.26' N 3°59.88' N 3°43.99' N 1°15.71' N 1°8.66' N 1°13.51' N 2°31.36' N 3°44.44' N 2°48.10' N 9°51.00" N 14°17.85' N 14°49.33' N NA 398 1557 1459 967 1459 1564 1674 2032 2277 1725 993 1508 1083 75 92 56 53 63 65 62 65 50 50 57 47 0.18 0.15 0.12 0.03 0.11 0.01 0.16 0.22 0.08 - 32 33 23 23 30 27 28 28 23 24 32 24 0.14 0.18 0.07 0.04 0.08 0.01 0.18 0.12 0.03 - 99 101 69 70 71 53 83 83 67 73 64 79 70 0.16 0.16 0.02 0.17 0.23 0.11 0.23 0.14 - 41 34 28 29 43 31 33 37 30 30 29 35 34 0.12 0.11 0 0.12 0.23 0.14 0.18 0.04 - 95 100 61 75 82 112 81 67 78 84 0.18 0.22 0.09 0.03 0.11 0.13 0.12 - 34 30 25 27 36 43 36 25 29 34 0.15 0.12 0.16 0.03 0.03 0.19 0.19 - 963 69 0.15 30 0.06 95 0.08 38 0.13 89 0.09 39 0.14 1801 NA 65 58 0.21 0.18 29 26 0.19 0.17 92 76 0.16 0.14 39 34 0.19 0.12 75 81 0.11 0.13 32 33 0.18 0.15 Iron (ppm) Avg. CV 1 Zinc (ppm) Avg. CV Iron (ppm) Avg. CV Zinc (ppm) Avg. CV Iron (ppm) Avg. CV Zinc (ppm) Avg. CV Iron and zinc content were determined by atomic absorption spectrophotometry by the CIAT analytical services lab. To avoid variability seed for each experiment was hand harvested and processed to avoid contamination and then shipped to CIAT for analysis where seed mineral content was evaluated by grinding 4 g of grain from each sample into a fine powder using a modified Retsch mill with a teflon capsule chamber and zirconium grinding balls. Powder was transferred to 25 ml plastic tubes and analyzed for both iron and zinc concentration measured in parts per million (ppm) with a wet digestion method. NA = not applicable. 28 1.2.2 Genotype x environment interactions for grain Fe and Zinc concentration Rationale: Biofortification seeks to alleviate micronutrient deficiencies through the development, production and consumption of mineral rich varieties on-farm and across agricultural regions. Bean programs in east, central and southern Africa recently embarked on developing nutrient rich and stable bean varieties which can contribute to alleviation of micronutrient malnutrition in the region (CIAT, 2002). Initial activities focused on screening available germplasm for genetic variation in iron and zinc (CIAT, 2003). These studies showed that considerable variation exists to increase seed iron concentration by more than 80% and zinc by 50%. Thirty-eight promising lines were selected following screening of landraces, advanced lines and varieties grown in eastern Africa (CIAT, 2005). However, stability of micronutrient density in bean cultivars across environments is not well known. Productivity and stability in the diverse bean growing environments is influenced by many environmental factors. Soil fertility factors are among the most important influences. Plants require considerable amounts of macronutrients such as phosphorus, nitrogen and potassium. Comprehensive agronomic approaches including specific fertilization strategies to enhance seed nutrient concentration have yet to be pursued (Rengel at al, 1999). However, fertilization aimed at increasing grain nutrient density to allow good establishment when the seed is sown in nutrient-deficient soil has been reported occasionally. Limited literature indicates that fertilization with inorganic and organic forms of micronutrients has potential to increase their concentration in grain. For example, Marschner (1995) showed that concentrations of Zn and Fe in cereal grain increased with an increase in Zn or Fe fertilizer additions. Addition of macronutrients (N, P and K) which promote root and shoot development, can increase the uptake of all nutrients required by the plant (Constant and Seldrick, 1991; Rengel et al, 1999). Soils in many bean growing environments in East, Central and Southern Africa are deficient in soil phosphorus, nitrogen and potassium and are acidic (pH below 6.5). Recent information indicates that concentration of iron and zinc in seeds is influenced by several quantitative loci (CIAT, 2005). It would therefore be expected that mineral density traits are determined not only by the genotype, but also may be influenced by environmental factors and the differential response of genotypes to agronomic management, soil and climatic factors. To test this hypothesis, studies were conducted to determine the influence of soil type, season, inorganic soil amendments (phosphorus, nitrogen, potassium, iron and zinc) on seed iron and zinc concentration and the associated genotype x environment interactions. Effects of N, P and K fertilization were reported in 2006. In 2008 trials were planted to confirm earlier results. Materials and Methods: Ten bean lines were grown at four levels of N, P, K fertilization at three locations in Kenya during the short rain season (November to February) and long rain season (AprilAugust). The bean lines included nine with high levels of iron and/or zinc, and a check (M211). P treatments were 0, 25, 50, 75 kg P ha-1. Source of P was triple super phosphate (46% P2O5) fertilizer. The four N levels were 0, 50, 100 and 150 kg N ha-1. Potassium was applied at 0, 50, 100 and 150 kg ha-1. Potassium fertilizer in the form of potassium sulphate (K2SO4-50%K2O) was preferred to potassium chloride (KCL) because the SO4-2 ion enhances phosphorus. Fertilizer was applied in the furrows and thoroughly mixed with the soil before planting. The factorial experiments were laid out a split-plot with three replicates. Varieties were the main plots and N, P, K and lime levels, the subplots. A plot consisted of 3 m rows. Spacing was 45 cm between rows and 10 cm within rows. The trial was conducted at Thika (1548 masl) and Kabete (Field 16, 1849 masl) during the short rain season, and at Kakamega (1583 masl) and Kabete (Field 10, 1794masl) in the long rain season. Soils at Kabete are humic nitisols (FAO, 1990; Jaetzold and Schmidt, 1983), slightly acidic (pH 5.5) and deficient in available phosphorus and nitrogen (Mwaura, 1995). Soils at Thika are eutric nitosols and acrisols, and low in nitrogen and phosphorus. At Kakamega soils are Dystro-Mollic nitisols, low in phosphorus (MOA, 1987; Siderius and Muchena, 1977). Soil samples were collected at each site before planting. Leaf samples were collected before flowering. Seed samples for mineral analyses were taken at harvest. Ground samples were digested with hydrogen peroxide, sulphuric acid and salicylic acid following methods of Novozamsky et al. (1983) and 29 Okalebo et al. (2002), and read on atomic absorption spectrophotometer (Perkin-Elmer Corporation, USA). Data was collected on phenology, disease incidence, 100-seed mass and grain yield following the CIAT standard scale (Schoohoven and Pastor-Corrales, 1987). Genstat (2005) software was used for analyses of variance. Results and Discussion: Results showed that there significant location, season, treatment and genotypic effects on the grain iron and zinc concentration. Significant genotype x environment interactions were detected. N effects. Fertilization with N up to 100 kg N ha-1 increased grain Fe and Zn concentration in all four environments. This result is similar to that obtained in 2006. However, grain mineral density varied with locations and seasons. Figures 7 and 8 illustrate genotypic differences averaged over N treatments. P Effects. Phosphorus fertilization significantly (P<0.05) increased seed Fe concentration. Results showed that grain iron concentration increased with P fertilizer application up to 50 kg P ha-1 rate for bean lines grown in Kabete Field 16, Thika and Kabete Field 10. However, at Kakamega, grain iron concentration increased with P fertilization up to 25 kg P ha-1. Application of P at 75 kg P ha-1 rate led to a decline in seed Fe concentration in all the sites for the two seasons. This is similar to the pattern observed in 2006, although in that year iron continued to increase at 75 kg ha-1 except at Kakamega. Genotype x environment interaction effects on grain Fe concentration are shown in Figure 9. K effects. Mean seed Fe increased but not significantly with increasing levels of K up to 150 kg K ha-1. In 2006 also, response of grain iron to K application was minimal. AND 620 had the highest grain iron concentration when grown at Kabete 16 (105 ppm), while the lowest grain concentration was observed for TY 3396-12 grown at Thika (Figure 10). Zn concentration increased significantly (P<0.05) with increasing levels of K up to 150 kg K ha-1. In 2006 an increase was also noted, but not at the highest level of K application. VNB 81010 showed the highest Zn concentration (34.3 ppm) when grown in Kabete Field 10, while lowest Zn levels were recorded from Roba-1 (19.6) grown in Kabete Field 16 (Figure 11). The results obtained indicate that adequate levels of K have the potential to increase bean yields, seed iron and zinc concentrations. However, the significant genotype x environment interactions suggest that grain mineral density varies with locations and genotypes. In general, results in 2008 were consistent with previous results. N and P gave the most dramatic increases in grain iron, while K application gave modest increases in seed zinc. Contributors: Paul Kimani, Ben Okonda, J. K. Keter and S. Beebe Collaborators: Bean Team at Kabete, Kenya. 30 160 Iron, ppm 140 120 Kab F16 100 Thika 80 Kab F10 60 Kak 40 20 0 AND 620 GLP 2 Gofta M211 Maharagi MLB 49Soja 89A Nakaja Roba-1 TY 339612 VNB 81010 Genotype Kab F10= Kabete Field 10, Kab F16= Kabete Field 16, Kak= Kakamega. Average of four levels of N fertilization. Figure 7. Genotypic x environment interactions for seed iron concentration of ten bean lines grown in four environments at four N levels for two seasons. 40 35 Zinc, ppm 30 Kab F16 Thika 25 20 Kab F10 15 Kak 10 5 0 AND 620 GLP 2 Gofta M211 Maharagi MLB 49- Nakaja Soja 89A Roba-1 TY 3396- VNB 12 81010 Genotype Kab F10= Kabete Field 10, Kab fertilization. Figure 8. F16= Kabete Field 16, Kak= Kakamega. Average of four levels of N Genotypic x environment interactions for grain zinc concentration of ten bean genotypes grown at four N levels in four locations for two seasons. 31 Iron, ppm 140 120 100 Kab F16 80 Thika 60 Kab F10 Kak 40 20 0 AND 620 GLP 2 Gofta M211 Maharagi MLB 49Soja 89A Nakaja Roba-1 TY 339612 VNB 81010 Genotype Kab F16= Kabete Field 16, short rain season; Kab F10= Kabete Field 10, long rain season; Thika= KARI-Thika, short rain, and Kak= KARI-Kakamega, long rain. Figure 9. Genotypic x environment interactions for seed iron concentration of 10 bean lines grown at four P levels in four locations over two seasons. 120 Iron, ppm 100 80 Kab F16 Thika 60 Kab F10 Kak 40 20 0 AND 620 GLP 2 Gofta M211 Maharagi MLB 49Soja 89A Nakaja Roba-1 TY 339612 VNB 81010 Genotype Kab F16= Kabete Field 16, short rain season; Kab F10= Kabete Field 10, long rain season; Thika= KARI-Thika, short rain season and Kak= Kakamega, long rain. Figure 10. Genotypic x environment interactions for seed iron concentration of ten bean genotypes grown at four potassium levels in four environments for two seasons. 32 40 Zinc, ppm 35 30 Kab F16 25 Thika Kab F10 20 15 Kak 10 5 0 AND 620 GLP 2 Gofta M211 Maharagi MLB 49- Nakaja Soja 89A Roba-1 TY 3396- VNB 12 81010 Genotype Kab F16= Kabete Field 16, short rain season; Kab F10= Kabete Field 10, long rain season; Thika= Thika, short rain season, and Kak=Kakamega, long rain season. Figure 11. Genotypic x environment interactions for seed zinc concentration of ten bean lines grown at four potassium levels in four environments over two seasons References: CIAT. 2003. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2004. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2005. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia Constant, K.M. and Seldrick, W.F. 1991. An outlook for fertilizer demand, supply, and trade. World Bank Techn. Paper No. 137, Asia Techn. Dept. series. Washington, D.C. Marschner, H. 1995. Mineral nutrition of higher plants. 2ed. Acad. Press, London, p 889. Rengel, Z., Batten, G.D. and Crowley, D.E. 1999. Agronomic approaches for improving the micronutrient density in edible portions of field crops. Field Crops Research 60: 27 – 40. Schoonhoven A van and Pastor Corrales, M.A. 1987. Standard system for the evaluation of bean germplasm. CIAT, Cali, Colombia. 33 Activity 1.3 Associated traits: antinutrients Highlights: • The inheritance of seed phytate content was analyzed to determine if this anti-nutrient could be reduced and how it is related to seed phosphorus content. Quantitative inheritance was found with several QTL explaining both traits independent of seed size. The results of this study show some genotypes with low levels of phytates which would seem to be sufficient for breeding attempts. The other anti-nutrient being analyzed is condensed tannins and an HPLC method was adapted to look at the tannin monomers that accumulate in genotypes from an inter-genepool population. 1.3.1 Inheritance of seed phosphorus and seed phytate content in a recombinant inbred line population of common bean Rationale: Phytates are an important anti-nutritional component of legume seeds because they chelate mineral uptake in human digestion. Phytates can also bind certain charged proteins making them less digestible as well and the lack of phytase production in monogastric digestive systems prevents phytates from being hydrolyzed and utilized by humans. On the other hand phytates are important as a seed supply of phosphorus and as a health-promoting factor in some human populations susceptible to diseases such as heart disease and certain cancers. It is notable that phytate levels are often correlated with total seed phosphorus (P) and are the main storage form of P in plant seeds with phytates representing 65% or more of the P present in cereal or legume grain; and therefore both seed P and phytates are characteristics that should be considered jointly. From this perspective our goal has been to understand the inheritance of phytate content and its relationship with seed phosphorus in common bean seeds. The objective of this research was to evaluate quantitative trait loci (QTL) for seed phosphorus and phytate content in an intergenepool (G2333 x G19839) recombinant inbred line population of common bean Materials and Methods: Plant material: An inter-genepool recombinant inbred line population derived from the cross of G2333 (Mesoamerican, type IV climbing bean from Mexico) by G19839 (Andean, type III bush bean from Peru) and consisting of 84 F5:8 lines was grown in two experiments in Popayán, Colombia in the 2004 growing season on soils that are inceptisols with a native P content of 2 ppm which is considered deficient. The two experiments differed in P fertilization: a total of 200 kg ha–1 of 10-30-10 N-P-K fertilizer was applied for a medium phosphorus treatment and 400 kg ha–1 was applied for a high phosphorus treatment. The two levels of phosphorus fertilization were used since P supply is thought to influence seed phytate content. All other agronomic management except for P supply was the same for the two trials with plants grown on trellises and plot size consisting of double rows that were 3 m in length and 2 m wide. Both medium and high P experiments were randomized complete block designs with two repetitions each and included the parents as control genotypes. Seed P and phytate analysis: Seed was hand harvested from each plot at full maturity, dried to 12% humidity prior to storage at 4ºC and used in seed phytate and total phosphorus analysis. Seed P and phytate content were quantified with spectro-photometric methods based on acid digestion with molybdenum blue and Wade reagents, respectively, and net seed P and net phytate content were calculated on a per seed basis using seed weights for each experiment. Data analysis: Analyses of variance were conducted for the seed phosphorus and seed phytate concentration traits in the RIL genotypes and parents across the two environments (medium P and high P fertilization) using SAS with all effects considered random and each term assumed to be independent. 34 The means for each genotype in each environment were used for quantitative trait locus (QTL) analysis with the probability of a QTL being present expressed in terms of LR (likelihood ratio) values. Results and Discussion: The molybdenum blue / Wade reagent method was found to be rapid as a quantification technique for total phytates, compared to more expensive, time consuming and multi-step analyses implemented for common beans with high pressure liquid chromatography (HPLC). In addition, the solid phase extraction column was found to be highly reproducible and coefficients of variation for the genotypes with this method were less than 5%. The analyses of variance showed significant differences between RIL genotypes for seed weight, total seed phosphorus, percentage seed phytate, net seed phytate and net seed phosphorous (Table 15). Total seed P in the RILs varied from 2.8 to 6.1 g kg-1 and phytates varied from 0.29 to 1.78 % across fertilization levels. Calculations of net phytate and net P content were used to evaluate the amount of phytate or phosphorus per seed rather than on the percentage bases as described above. This was justified by the fact that we analyzed a Mesoamerican x Andean inter genepool population that segregated widely for seed size and by the hypothesis that larger seeds serve to store greater amounts of nutrients than smaller seeds but that nutrient requirements are similar for seedling establishment. Given the larger average seed size of G19839 (0.62 to 0.73 g seed-1), this parent had up to 100% higher net phytate and net P content than G2333 (0.29 to 0.31 g seed-1) under both soil P levels. Table 15. Range for seed phytate content, total seed phosphorus (P), seed weight, net seed phytate and net seed P content in recombinant inbred line population G2333 x G19839 grown in two experiments in Popayán under high (HP) and medium phosphorus (MP) soil fertilization. RILs Trait P level Seed Phytate (%) Mean HP 0.93 ± 0.31 MP 0.94 ± 0.48 -1 Total Seed P (g kg ) HP 4.22 ± 0.52 MP 4.23 ± 0.45 Seed Weight (g) HP 0.40 ± 0.08 MP 0.39 ± 0.08 -1 Net Phytate Content (mg seed ) HP 0.37 ± 0.15 MP 0.38 ± 0.22 -1 Net P Content (mg seed ) HP 1.69 ± 0.41 MP 1.69 ± 0.39 * and *** , significance at probability levels of 95% and 99.9%. Range PRILs 0.29 - 1.78 0.29 - 1.76 2.75 - 6.06 3.11 - 5.95 0.24 - 0.67 0.25 - 0.65 0.88 - 9.26 0.87 - 8.26 0.78 - 2.97 0.94 - 2.90 * * *** *** *** *** *** *** *** *** Population histograms for percentage total seed phosphorus, percentage phosphorus, net phytate content, net seed P content and seed size in the G2333 x G19839 RILs were normally distributed in both environments and there was no evidence of kurtosis or skewing in any of the histograms. These results suggest that all of the traits measured were inherited in a quantitative manner. In each case, parental means tended to be less distinct than the lowest and highest seed P or phytate containing RILs suggesting that transgressive segregation was important in the inheritance of the traits and that both parents contributed positive and negative alleles for the traits. A total of six QTL were found for total or net seed P while three were found for percentage or net seed phytates. In addition six QTL were found for seed weight. QTL for seed P and percent phytates were located independently. Meanwhile the QTL for net seed P or phytate content were related to seed weight QTL. The QTL were of moderate effect and the phytate and seed P QTL were independent of each other and of QTL for seed size. 35 Conclusions and Future Plans: The ability to select QTL for seed phosphorus or phytate separately is important since seed phosphorus is important for plant growth while phytate is a major factor influencing the bioavailability of iron, zinc and calcium. The results of this study show that some genotypes have levels of phytates which would seem to be sufficiently low for breeding attempts. Furthermore, the results suggest that bean plants can adapt to different initial supplies of phosphorus and that seed P and phytate levels in common bean can be modified through plant breeding. We plan to use the methodology developed here to evaluate additional populations and to consider marker assisted selection strategies for reducing phytates while maintaining seed P levels. We are also exploring candidate genes for phytate content as potentially suitable markers for this work in the future. Contributors: 1.3.2 Blair MW, Sandoval TA, Caldas GV (SBA-2-CIAT), Beebe SE (SBA-1, CIAT) , Páez MI (Univalle) Analysis of condensed tannins through HPLC in genotypes from an inter-genepool bean population Introduction: Seed coat color in P. vulgaris is determined by the amount and presence of flavonol glycosides, tannins and anthocyanins. These compounds are synthesized by the flavonoid pathway and although the pathway is well characterized in some species, in common bean the genes are not yet well known. On the other hand, extensive genetic analyses have identified specific Mendelian genes that control seed coat pattern and color. However, it has not been possible to identify the genes responsible for producing specific flavonoid compounds. This means that until now the relationship between genes which control the enzymes in the pathway and the Mendelian genes for seed coat color is not clear. With this in mind and because of our previous studies on QTL mapping for tannin content, we decided to begin analyses in tannin composition on some genotypes from the DOR364 x G19833 bean population. Materials and Methods: Plant Materials: We chose 20 genotypes from the DOR364 x G19833 population with contrasting seed colors ranging from red to yellow to brown. The population was harvested in Darién 2006 with three repetitions by genotype. The genotypes were contrasting in tannin content. Samples of 10 seeds each were analyzed for each genotype. The seeds were peeled and the seed coats were ground for analysis. HPLC analysis: Condensed tannins were extracted using 70% acetone and converted to anthocyanidins by the butanol-HCl method in triplicates. For the HPLC analysis we dried 1mL of each sample after the reaction with butanol-HCl in a sample concentrator. The dried samples were re-dissolved in a methanolHCl 1% solution, filtered (Millipore PTFE, 0.45uM) and placed in a vial for the injection. Separation of anthocyanidins (resulting from the de-polymerizing of the tannins via butanol-HCl method) was achieved using a 8 x 100 mm Nova-Pack C18 column (4um, Waters). The solvents were A, 100% Methanol and B, 5% acetic acid. The gradient consisted of: 40% B for 1 min, 30% B for 1.3 min, 35% B for 20 sec, 40% B for 1.3 min, 60% B for 2 min. Detection was carried out in a UV detector (Shimadzu CL-10A) using 530nm as wavelength. Data Analysis: The standards that we used for the identification were the three anthocyanidins, cyanidin chloride, delphinidin chloride and pelargonidin chloride (Supplied by Apin Chem Ltd, Abington, UK). The control sample was the standard cyanidin chloride and the data were analyzed by Statistix v.8.0. Overall, we collected 567 datapoints and these were expressed as area percentage of each anthocyanidin and also as seed coat percentage. 36 Results and Discussion: The chromatographic analysis of the samples treated with butanol-HCl, showed the existence of 3 principal anthocyanidins (Figure 12). The retention times for delphinidin, cyanidin and pelargonidin were 2.18, 2.9 and 3.5 minutes respectively. Analysis of variance for each trait indicated the existence of significant variability among the chosen genotypes (Table 16). This analysis was based in the data expressed as percentage of anthocyanidin in seedcoat. The highest anthocyanidin in content was cyanidin, followed by delphinidin and then perlargonidin, indicating that in the analyzed genotypes the most representative monomer was catequin. Pelargonidin was significant only in a few cases. Cyanidin Delphinidin Pelargonidin Figure 12. Chromatogram of a bean sample through HPLC analysis for the quantification of anthocyanidins. Table 16. Analysis of variance for three types of anthocyanidins in common bean seed coats of 20 genotypes and one control. Trait Delphinidin Cyanidin Pelargonidin Source DF SS MS F P Genotype 20 0.43896 0.02195 2.58** 0.0048 Error 42 0.35693 0.00850 Total 62 0.79590 Genotype 20 0.05079 0.00254 3.04** 0.0012 Error 42 0.03506 0.00083 Total 62 0.08585 Genotype 20 0.04274 0.00214 1.83** 0.0487 Error 42 0.04892 0.00116 Total 62 0.09166 ** Asterisks indicate significance at the 0.05 probability level 37 Overall, the red genotypes had less amount of cyanidin and were the highest in pelargonidin content, whereas the other colors had similar content in the three anthocyanidins. However, according to our results, the proportion of anthocyanidins related to the type of monomer in the tannin polymer changed notably among field repetitions. Thus, in some genotypes it was possible to observe traces of pelargonidin in just one of the field repetitions. In terms of anthocyanidin content in the seed coat, the average for cyanidin was 0.19%, for delphinidin was 0.13% and for pelargonidin was 0.04%. A sample of cyanidin chloride was run during each analysis as a control, the standard error and coefficient of variation was 0.07 and 3.7% respectively, indicating the suitability and reproducibility of the method. Conclusions and Future Plans: The differences between genotypes evidence the existence of segregation in the population for tannin composition; therefore the next step is the evaluation of the entire population in order to carry out QTL analysis. The information about the regions which control condensed tannins, will be more specific and along with the anthocyanin analysis we could establish regions in the genome, specific for steps in the pathway and at the same time a type of association between them and the Mendelian genes for seed coat color. Future activities are QTL analysis for tannin monomer composition and anthocyanin content in the DOR364 x G19833 population. Collaborators: A.M. Díaz, G.V. Caldas, M.W. Blair 38 Product 2: Beans that are more productive in smallholder systems of poor farmers Activity 2.1 Developing germplasm tolerant to abiotic stresses 2.1.1 Drought resistance Highlights: • Mesoamerican crosses among drought resistant parents that had expressed a degree of tolerance to low soil P availability, produced more than 20 lines that were excellent in drought resistance. Some lines subsequently showed adaptation to low P in Darién, producing grain of excellent quality under combined drought and low P stress. • Another 15 Mesoamerican families combined drought tolerance and bc-3 gene for resistance to BCMNV. • In an effort to incorporate drought tolerance in Andean bush beans we have created a series of 216 advanced drought Andean beans (DAB) lines from inter and intra-gene pool crosses involving 5 commercial genotypes from Southern Africa and 10 drought tolerance sources of which half were Andean and half were Mesoamerican to produce 46 populations. The lines represent large red, red mottled and cream mottled seed types. Selection has stressed bush bean architecture, adaptation to drought stress and yield potential under favorable conditions using alternate dry versus rainy season plantings. • A reference collection of landraces from the CIAT core collection has been evaluated in the field for drought tolerance compared to a series of check genotypes. The reference collection was stratified into Andean and Mesoamerican gene pools and the association of drought tolerance with subgroups and common bean races was analyzed. • Mid-elevation adaptation was tested in Darién for a series of SAB (drought resistant Andean) lines originally developed from crosses between the drought-resistant genotypes SAB 258, SAB 259, and ICA Quimbaya crossed with drought susceptible but commercial type genotypes ABA36, ABA58 and COS16. Results confirmed the genetic gain for drought tolerance and yield potential that has occurred in the breeding of the SAB lines compared to both their droughttolerant and susceptible parents. The same lines were tested in rainfed conditions in Palmira and several maintained their yield advantage over local checks. Certain SAB lines can be selected with greater stability across mid-elevation and lower-elevation sites based on this analysis. • Field evaluation of elite lines at Palmira resulted in identification of five lines NCB 226, SEN 56, SER 113, SER 125 and SER 16 that were outstanding in their adaptation to drought stress conditions. The superior performance of these lines under drought stress was associated with higher values of harvest index, pod harvest index, leaf area index and canopy biomass. The SER lines that were developed in the last few years seem to combine the desirable traits for drought adaptation such as greater mobilization of photosynthates to seed with efficient use of water through stomatal control. • Field evaluation of 33 RILs of the cross DOR 364 x BAT 477 at Palmira over two seasons under terminal drought stress conditions resulted in identification of two lines (BT 21138-17-1-1 and BT 21138-6-1-1) that were superior in their adaptation to drought stress conditions. The superior performance of these lines under drought stress was associated with higher values of harvest index, pod harvest index and seed and pod number per area indicating the importance of greater mobilization of photosyntates to pods and to seeds under rainfed conditions. • Field evaluation of 97 RILs of the cross DOR 364 x BAT 477 under intermittent drought stress resulted in identification of two RILs BT 21138-68-1-1 and BT 21138-74-1-1 that were outstanding in adaptation to intermittent drought stress conditions. The superior performance of these lines under intermittent drought stress was associated with higher values of harvest index, pod partitioning index, stem biomass reduction, seed number per area and pod number per area 39 • • 2.1.1.1 indicating the importance of greater mobilization of photosyntates to pods and seeds under rainfed conditions. Field evaluation of 121 RILs of the cross MD 23-24 x SEA 5 over 3 seasons resulted in identification of the lines MR 81 and MR 25 that were superior in adaptation to drought stress conditions. The superior performance of these lines was associated with higher vigor, higher values of pod harvest index, harvest index and seed number per area, highlighting the importance of the photosyntate mobilization to pods and seeds under intermittent drought stress. The response to inoculation with the strain Rhizobium etli CIAT 632 under drought stress was tested using 7 common bean genotypes. We found that Pinto Villa was better adapted to drought due to its ability to decrease stomatal conductance while Alubia cerrillos was more affected due to drought stress. Although there was no response to inoculation, the effect of terminal drought stress on nodulation was very marked on all 7 genotypes. Evaluations of Mesoamerican lines segregating for drought resistance and tolerance to low soil P Rationale: Abiotic stresses tend to be associated with marginal environments and seldom occur individually. Rather, co-occurrence may result in interactions and greater yield losses than when stresses occur singly. This is especially the case when drought and low soil fertility co-occur. It appears that nutrient deficiencies may limit root development and result in greater susceptibility to drought. It is therefore desirable to combine resistance or tolerance to drought and to low soil fertility, especially low soil P, which is the most common edaphic constraint of beans. Some progress had been made in this regard already, both in the small red class and in carioca type. In the genotypes reported here, we sought to obtain greater expression of multiple stress tolerance and in a greater number of genotypes. Materials and methods: Double crosses of four parents were created involving drought resistant parents, especially those with small red grain, with lines that had been observed to perform well under fertility stress. Sources of low P tolerance included: SXB’s 407, 409, 415, 418 (all derived from crosses for Brazil including carioca parents); small reds SER’s 118 and 119; NCB 226 (black seeded); and BFB’s 611 and 613 (from low P crosses). Other parents carried the bc-3 gene for BCMNV and had also been selected under drought stress (RCB’s 588, 589, 591, 592). F2 populations were selected under drought in 2007 as F1.2 families, and as F1.3 families in Popayán. F3.4 families were selected in Santander de Quilichao under inoculation with angular leaf spot and moderate fertility stress. The F3.5 bulks were returned to the drought nursery in August, 2008 in two lattice designs: a 5 x 5 and a 6 x 6. Yield data under drought stress are reported here. Data on response to the NL-3 (necrotic) strain of BCMNV were obtained in the greenhouse. Results and Discussion: Although most trials in the drought nursery in 2008 suffered only intermittent, moderate drought, these two trials were planted late and matured in September when days were sunny and drought stress was more reliable. These trials received an estimated 134 mm of water, from one furrow irrigation and 99 mm of rainfall. Thus, although stress was still moderate, it was more severe on these trials than in others in the main planting. Yields of checks were similar in the two trials: average yields of Tio Canela were 1463 kg ha-1, and those of SER 16 were 1959 kg ha-1 (Tables 17 and 18). About 40% of the lines yielded significantly more than the Tio Canela check. Among lines that out yielded Tio Canela, two lines in Table 17 presented evidence of the bc-3 gene combined with drought resistance, and four lines in Table 18 appeared to be segregating this gene. This gene is especially important in Africa, where multiple stress tolerance is a high priority. These lines were planted under drought and low P stress in Darién, and individual plants were selected. These will be evaluated under drought stress in the upcoming season. Collaborators: S. Beebe, M. Grajales, C. Cajiao, M. Castaño, A. Guerrero 40 Table 17. Yield under drought of 23 lines developed to combine drought resistance with low fertility tolerance. Cross code SBCZ 16238-030 SBCZ 16238-017 SBCZ 16238-019 SBCZ 16238-024 SBCZ 16261-004 SBCZ 16238-003 SBCZ 16238-017 SBCZ 16284-006 SBCZ 16238-019 SBCZ 16236-066 SBCZ 16274-081 SBCZ 16238-003 SBCZ 16238-009 SBCZ 16279-034 SBCZ 16261-025 SBCZ 16261-004 SBCZ 16238-019 SBCZ 16238-031 SBCZ 16234-031 SBCZ 16279-034 SBCZ 16284-006 SBCZ 16236-004 SCTZ 16268-25 Pedigree (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-1P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-3P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-9P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-7P-MQ SER 16 (SER 48xNCB 226)F1 X (SER 119xRCB 234)F1/-MC-5P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-2P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-5P-MQ (SER 97xNCB 226)F1 X BFB 611/MC-7P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-11P-MQ (SER 89xRCB 234)F1 X (SXB 407xSER 118)F1/-MC-1P-MQ (SER 119xNCB 226)F1 X (SER 113xRCB 590)F1/-MC-2P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-1P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-3P-MQ (SXB 418xNCB 226)F1 X BFB 613/MC-4P-MQ (SER 48xNCB 226)F1 X (SER 119xRCB 234)F1/-MC-3P-MQ (SER 48xNCB 226)F1 X (SER 119xRCB 234)F1/-MC-3P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-7P-MQ (SXB 415xSER 125)F1 X (SER 89xRCB 588)F1/-MC-7P-MQ (SER 176xRCB 591)F1 X (SXB 407xSER 118)F1/-MC-11P-MQ (SXB 418xNCB 226)F1 X BFB 613/MC-5P-MQ (SER 97xNCB 226)F1 X BFB 611/MC-10P-MQ (SER 89xRCB 234)F1 X (SXB 407xSER 118)F1/-MC-2P-MQ (SER 118xBCB 587)F1 X (SXB 409xAQB 609)F1/-MC-1P-MQ TIO CANELA 75 COL ALS BCMNV DTF DTM g/100s kg ha-1 kg ha-1 d-1 rd 1 9N 32 63 28 2313 36.5 bl 3 5N 33 65 28 2118 32.6 rd 4 8N 33 64 26 2056 31.9 Cr 6 10_O 32 32 64 60 26 21 2038 2029 31.8 33.7 rd 4 9N 32 64 25 2018 31.6 rd 5 10N 34 64 23 2001 31.3 bl 4 8N 33 65 26 1960 30.3 rd 4 8N 33 63 24 1947 30.8 rd 4 8N 32 64 24 1942 30.3 rd 4 10N 33 63 26 1860 29.7 rd 2 3N_ 6_O 35 64 24 1858 29.0 rd 5 7N 33 63 25 1835 29.0 rd 5 9N 33 65 27 1825 28.0 pk 2 4N 35 62 21 1819 29.2 rd 4 8N 33 63 23 1805 28.5 rd 4 10N 33 66 25 1796 27.3 rd 4 8N 33 65 26 1790 27.7 rd 3 9N 33 64 26 1698 26.6 pk 4 5N 33 64 20 1649 25.8 rd 5 7N 32 63 25 1627 25.9 rd 4 6N 33 62 22 1603 25.8 rd Cr Str 5 9N 33 62 25 1564 25.1 4 5N 38 38 67 65 23 19 1522 1414 22.8 21.7 1.2 1.8 1.5 394 12.1 LSD (0.05) COL=Color; rd=red; bl=black; pk=pink; cr=cream; ALS= angular leaf spot (1-9 scale); BCMNV=bean common mosaic necrotic virus; N=necrotic reaction; 0=no symptoms; g/100s=grams per 100 seed. 41 Table 18. Code SBCZ 16257-33 SBCZ 16245-01 SBCZ 16257-33 SBCF 16231-006 SBCZ 16257-21 SBCZ 16253-006 SBCZ 16234-031 SBCZ 16253-008 SCFZ 16265-002 SBCZ 16245-01 SBCF 16231-005 SBCF 16231-006 SBCF 16231-002 SBCZ 16234-004 SBCZ 16253-008 SBCF 16231-002 SBCZ 16234-031 SBCZ 16253-040 SBCZ 16253-014 SBCF 16231-006 SBCZ 16257-21 SBCZ 16245-01 SBCF 16231-005 SBCZ 16253-008 SBCZ 16253-040 SBCZ 16253-040 SBCZ 16253-006 SBCF 16231-015 SCFZ 16265-002 SBCF 16231-002 SBCF 16231-005 SBCZ 16257-21 SBCZ 16257-21 SBCF 16231-005 Yield under drought of 34 lines developed to combine drought resistance with low fertility tolerance. Pedigree (SER 48xRCB 234)F1 X (SER 118xNCB 226)F1/-MC-2P-MQ (SER 76xRCB 589)F1 X (SXB 407xSER 119)F1/-MC-4P-MQ (SER 48xRCB 234)F1 X (SER 118xNCB 226)F1/-MC-1P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-14P-MQ (SER 48xRCB 234)F1 X (SER 118xNCB 226)F1/-MC-9P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-5P-MQ SER 16 (SER 176xRCB 591)F1 X (SXB 407xSER 118)F1/-MC-2P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-6P-MQ (SER 118xRCB 590)F1 X (SMR 3xMIB 499)F1/-MC-15P-MQ (SER 76xRCB 589)F1 X (SXB 407xSER 119)F1/-MC-3P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-1P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-4P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-11P-MQ (SER 176xRCB 591)F1 X (SXB 407xSER 118)F1/-MC-1P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-2P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-8P-MQ (SER 176xRCB 591)F1 X (SXB 407xSER 118)F1/-MC-12P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-2P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-1P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-9P-MQ (SER 48xRCB 234)F1 X (SER 118xNCB 226)F1/-MC-5P-MQ (SER 76xRCB 589)F1 X (SXB 407xSER 119)F1/-MC-5P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-2P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-4P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-23P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-12P-MQ (SER 102xRCB 592)F1 X (SXB 415xSER 119)F1/-MC-3P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-5P-MQ (SER 118xRCB 590)F1 X (SMR 3xMIB 499)F1/-MC-13P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-21P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-6P-MQ (SER 48xRCB 234)F1 X (SER 118xNCB 226)F1/-MC-12P-MQ TIO CANELA 75 (SER 48xRCB 234)F1 X (SER 118xNCB 226)F1/-MC-4P-MQ (SER 155xRCB 591)F1 X (SER 118xSXB 409)F1/-MC-4P-MQ LSD (0.05) COL ALS BCMNV DTF DTM g/100s kg ha-1 kg ha-1 d-1 rd 4 7N_ 2_O 33 65 27 2024 31.1 rd 3 9N 33 64 27 1992 31.0 rd 5 6N 33 65 28 1984 30.5 rd 5 8N 35 64 30 1967 30.6 rd 3 7N 34 63 29 1928 30.7 rd 5 4N_ 3_O 34 33 65 63 24 22 1908 1889 29.3 30.0 rd 5 6N_ 2_O 33 66 28 1887 28.6 rd 3 6N 33 65 31 1859 28.6 rd 4 5N_ 3_O 33 64 31 1856 28.9 rd 3 7N 34 64 28 1836 28.5 rd 5 6N 35 65 29 1823 27.9 rd 3 9N 33 65 26 1818 27.9 rd 6 9N 34 66 25 1802 27.3 Sd 5 9_O 35 65 28 1785 27.3 rd 3 8N 34 64 27 1785 27.9 rd 4 9N 34 64 26 1764 27.5 rd 4 7N 33 64 24 1749 27.3 rd 5 9N 34 65 27 1742 26.6 rd 4 6N 34 66 26 1715 26.1 rd 6 8N 35 64 27 1714 26.7 rd 4 6_O 34 64 32 1690 26.4 rd 3 8N 34 65 28 1677 25.9 Sd 4 8N 36 67 24 1651 24.8 rd 3 7N 34 65 30 1650 25.4 rd 4 7N 35 65 24 1641 25.1 rd 4 10N 34 66 27 1636 24.7 rd 4 1N_ 5_O 32 64 27 1614 25.1 rd 3 8N 35 65 26 1593 24.6 rd 4 2N_ 6_O 35 65 31 1581 24.2 rd 5 9N 34 65 24 1532 23.6 rd 5 3N_ 2_O 35 66 27 1523 23.2 rd 4 9_O 34 36 64 68 32 20 1515 1512 23.5 22.3 rd 5 1N_ 6_O 32 64 30 1505 23.5 rd 5 8N 35 1.0 67 1.4 27 2.0 1500 332 22.4 5.0 COL=Color; rd=red; Sd=rojo de seda light red; ALS= angular leaf spot (1-9 scale); BCMNV=bean common mosaic necrotic virus; N=necrotic reaction; 0=no symptoms; g/100s=grams per 100 seed. 42 2.1.1.2 Evaluations of Mesoamerican lines segregating for drought resistance and bc-3 gene Rationale: At the outset of the Bean Program in the 1970’s, BCMV was highlighted as a priority, and in Latin America the dominant I gene was amply deployed for this problem. Over time it has become necessary to broaden the genetic base of the BCMV resistance with the bc-3 gene, on the one hand to avoid certain problems of genetic linkage to dark grain color, and on the other hand to address the threat of necrotic strains of BCMNV in Africa and elsewhere. As drought resistance has become one of the central priorities of the bean breeding programs, we have sought to develop a gene pool with drought resistance and key genes including the bc-3. The present report informs on the performance of families derived from populations segregating for drought resistance and bc-3 gene. Materials and Methods: Simple crosses were created between elite drought resistant lines with the I gene and sources of bc-3 gene, and well as crosses among the elite lines. Individual plants were selected from populations of simple crosses in F3 generation, seed was bulked in the F4 generation, and trials were planted in the drought season as F5 families in two 6 x 6 lattices. Reaction to the NL-3 strain of BCMNV was obtained in greenhouse inoculations. Results and Discussion: These trials received an estimated 170 mm of water (see section 1 above) but soil structure was not favorable. Root development may have been limited, contributing to greater stress. About half of the lines yielded significantly more than the Tio Canela check, which averaged 1279 kg ha-1 in these two trials (Tables 19 and 20). Only two of the families with superior drought yields were homozygous for the bc-3 gene (as evidenced by the reaction of “0” in Tables 19 and 20), but at least 12 families showed evidence of segregation of bc-3, requiring additional selection to purify lines for bc-3. Such lines will augment a growing number of such genotypes that are creating a gene pool based on drought and in which the bc-3 gene must become the basis for BCMNV resistance. Collaborators: S. Beebe, M. Grajales, C. Cajiao, M. Castaño, A. Guerrero Table 19. Yield under drought of 33 families derived from populations segregating for drought resistance and bc-3 resistance to BCMNV. Code SC 16040 SC 16045 SC 16050 SC 16040 SC 16050 SC 16041 SC 16040 SC 16041 SC 16039 SC 16040 SC 16040 SC 16045 SC 16045 SC 16040 Pedigree SER 48xRCB 593/-MC22C-MC SER 118xNCB 226/-MC14C-MC SER 113xRCB 590/-MC1C-MC SER 48xRCB 593/-MC7C-MC SER 113xRCB 590/-MC11C-MC SER 48xNCB 226/-MC16C-MC SER 48xRCB 593/-MC8C-MC SER 48xNCB 226/-MC1C-MC SER 48xRCB 234/-MC4C-MC SER 48xRCB 593/-MC20C-MC SER 48xRCB 593/-MC23C-MC SER 118xNCB 226/-MC15C-MC SER 118xNCB 226/-MC20C-MC SER 48xRCB 593/-MC4C-MC COL BCMNV DTF DTM g/100s kg ha-1 kg ha-1 d-1 rd 9N 35 64 28 2167 33.9 rd 11N 33 63 24 2062 32.7 rd 6N_ 2O 34 65 25 2022 31.4 rd 8N_ 1O 33 63 27 2002 31.8 rd 8N 34 65 27 1957 29.9 rd 7N_ 2O 37 67 25 1947 28.9 rd 8N 34 65 25 1911 29.3 rd 6N_ 3O 33 62 23 1908 30.7 rd 1N_ 6O 32 61 26 1883 30.8 rd 7N_ 3O 35 64 26 1877 29.6 rd 7N_ 1O 34 66 28 1862 28.4 rd 8N 35 68 27 1860 27.5 rd 10N 41 70 26 1859 26.7 rd 7N_ 3O 33 63 25 1856 29.6 43 Table 19. cont’d. Code Pedigree COL BCMNV DTF DTM g/100s kg ha-1 kg ha-1 d-1 rd 5N_ 3O 33 64 26 1848 28.9 rd 10N 33 63 21 1836 29.0 rd 6_O 33 64 27 1832 28.5 32 61 21 1812 29.6 SC 16040 SC 16047 SC 16039 SER 48xRCB 593/-MC33C-MC SER 119xNCB 226/-MC9C-MC SER 48xRCB 234/-MC14C-MC Test 1 SER 16 rd SC 16045 SER 118xNCB 226/-MC4C-MC rd 10_O 33 66 24 1772 27.1 SC 16039 SER 48xRCB 234/-MC16C-MC rd 2N_ 1M_ 7_O 33 62 24 1730 27.7 SC 16045 SER 118xNCB 226/-MC8C-MC rd 9N 35 63 23 1730 27.2 SC 16040 SER 48xRCB 593/-MC28C-MC rd 8N_ 1_O 33 62 28 1693 27.1 SC 16041 SER 48xNCB 226/-MC13C-MC rd 7N 34 67 26 1674 25.0 SC 16047 SER 119xNCB 226/-MC7C-MC rd 8N 34 65 24 1665 25.8 SC 16040 SER 48xRCB 593/-MC19C-MC rd 6N_ 2_O 33 63 27 1660 26.5 SC 16040 SER 48xRCB 593/-MC25C-MC rd 7N_ 1_O 34 63 28 1659 26.3 SC 16040 SER 48xRCB 593/-MC3C-MC rd 8_O 34 64 25 1659 25.9 SC 16041 SER 48xNCB 226/-MC12C-MC rd 6N_ 2_O 33 64 27 1657 25.8 SC 16050 SER 113xRCB 590/-MC4C-MC rd 6N 33 64 25 1656 25.9 SC 16045 SER 118xNCB 226/-MC5C-MC rd 10_O 38 70 22 1549 22.0 SC 16050 SER 113xRCB 590/-MC6C-MC rd 11N 41 70 25 1541 22.1 SC 16040 SER 48xRCB 593/-MC12C-MC rd 1N_ 9_O 33 63 27 1531 24.2 SC 16040 SER 48xRCB 593/-MC5C-MC rd 10_O 33 62 26 1511 24.3 Test 3 DOR 390 bl 40 70 18 1480 21.1 Test 2 TIO CANELA 75 rd 39 70 18 1471 21.1 SC 16045 SER 118xNCB 226/-MC17C-MC LSD (0.05) rd 39 1.0 65 1.4 24 2.0 1417 332 21.7 5.0 10_O COL=Color; rd=red; bl=black; BCMNV=bean common mosaic necrotic virus; N=necrotic reaction; 0=no symptoms; g/100s=grams per 100 seed. 44 Table 20. Yield under drought of 34 families derived from populations segregating for drought resistance and bc-3 resistance to BCMNV. SD Code Pedigree 16012 SER 101xSEN 53/-MC-4C-MC COL BCMNV DTF DTM g/100s kg ha-1 kg ha-1 d-1 bl N 34 65 22 1922 29.4 SD 16006 SXB 403xSEN 53/-MC-4C-MC bl N 33 61 23 1815 29.6 SCFZ 16073 RCB 584xMIB 487/-MC-7C-MC bl 10_O 35 62 24 1760 28.6 SC 16047 SER 119xNCB 226/-MC-2C-MC bl 11N 34 64 23 1683 26.1 SC 16047 SER 119xNCB 226/-MC-6C-MC bl 11N 33 63 23 1656 26.3 SC 16054 SER 42xRCB 593/-MC-32C-MC rd 10_O 33 61 25 1642 26.8 SD 15597 SER 119xSEN 46/-MC-12C-MC bl N 33 64 23 1627 25.6 SC 16041 SER 48xNCB 226/-MC-3C-MC bl 2N_ 8_O 33 63 28 1619 25.7 SC 16054 SER 42xRCB 593/-MC-16C-MC rd 10N 33 64 25 1603 24.9 SC 16054 SER 42xRCB 593/-MC-22C-MC rd 8N 36 67 25 1572 23.4 SD 15597 SER 119xSEN 46/-MC-8C-MC bl N 33 62 22 1562 25.3 SCFZ 16073 RCB 584xMIB 487/-MC-5C-MC bl 8N_ 2_O 34 64 24 1557 24.3 SD 16011 SER 119xSEN 46/-MC-4C-MC bl N 34 64 23 1543 24.1 SC 16054 SER 42xRCB 593/-MC-29C-MC rd 4N_ 4_O 33 63 24 1528 24.1 SBCZ 15999 NCB 226xRCB 588/-MC-2C-MC bl 5_O 34 68 27 1517 22.1 SD 16011 SER 119xSEN 46/-MC-1C-MC bl N 35 68 23 1516 22.6 SC 16054 SER 42xRCB 593/-MC-2C-MC rd 1N_ 9_O 34 63 26 1500 24.1 SC 16050 SER 113xRCB 590/-MC-18C-MC rd 9N 34 65 26 1499 23.1 SC 16054 SER 42xRCB 593/-MC-17C-MC rd 10_O 34 65 24 1488 22.9 SC 16054 SER 42xRCB 593/-MC-7C-MC rd 1N_ 9_O 33 63 25 1487 23.4 SC 16051 SER 97xNCB 226/-MC-19C-MC bl 10N 33 62 26 1482 23.8 SBCZ 15999 NCB 226xRCB 588/-MC-8C-MC bl 5_O 34 65 25 1468 22.6 SC 16054 SER 42xRCB 593/-MC-27C-MC rd 5N_ 5_O 33 65 26 1460 22.4 SD 15597 SER 119xSEN 46/-MC-14C-MC rd N 34 66 23 1411 21.4 SD 15597 SER 119xSEN 46/-MC-10C-MC bl N 33 63 22 1400 22.3 SD 15597 SER 119xSEN 46/-MC-1C-MC rd N 36 66 23 1400 21.3 SD 15597 SER 119xSEN 46/-MC-6C-MC bl N 34 63 24 1364 21.6 SD 15597 SER 119xSEN 46/-MC-13C-MC rd N 33 63 23 1345 21.3 SC 16054 SER 42xRCB 593/-MC-28C-MC rd 9_O 37 68 23 1327 19.7 SD 15597 SER 119xSEN 46/-MC-11C-MC bl N 33 62 24 1318 21.2 SC 16054 SER 42xRCB 593/-MC-9C-MC rd 9N 33 63 25 1310 20.7 SC Test 16054 1 SER 42xRCB 593/-MC-1C-MC SER 16 rd rd 4N_ 3_O 33 32 65 63 28 21 1306 1287 20.2 20.4 SC 16045 SER 118xNCB 226/-MC-34C-MC bl 10_O 37 68 22 1237 18.1 SC Test 16054 2 SER 42xRCB 593/-MC-11C-MC TIO CANELA 75 LSD (0.05) rd rd 8N 37 39 1.0 65 68 1.4 23 19 2.0 1191 1187 332 18.3 17.6 5.0 COL=Color; rd=red; bl=black; BCMNV=bean common mosaic necrotic virus; N=necrotic reaction; 0=no symptoms; g/100s=grams per 100 seed. 45 2.1.1.3 Evaluations of Andean lines for drought resistance and for yield potential under irrigation Rationale: In 2007 we reported on the yield of Andean lines under rainfed conditions. In that year different trials performed quite differently according to their position in the field, although several new lines were promising. To have more confidence in the drought reaction of these lines, the trials were repeated in 2008 in the drought season. In 2008 the same lines were also planted under irrigated conditions to establish their yield potential in a favorable environment in Palmira. Materials and Methods: Yield trials were established in Palmira (1000 masl; 26oC average temperature) as described in previous years, in lattice designs of 4 x 4, 5 x 5, or 6 x 6 in three replications. Two row plots 3.75 m long were planted in July under rainfed conditions to induce drought stress. Irrigation was applied twice up to 20 days after planting, after which all additional moisture resulted from rainfall. An irrigated treatment was established with the same design in which plots were furrow irrigated as needed. Pests were controlled as needed. Results and Discussion: Excessive rain prior to planting made field preparation extremely difficult in the 2008 drought season, and the soil structure in the drought treatement was especially poor. As a result plant development was sub-optimal, and root penetration might have been limited. As with other trials in this field, the crop received an estimated 170 mm of water during the crop cycle, but in this field the crop appeared to suffer severe drought stress. Compared to the irrigated treatment, yield reduction averaged 48%. Furthermore, weeds were plowed down only shortly before planting, and the fresh organic matter generated a severe attack of Sclerotium rolfsii. The local check ‘Calima’ fared badly in these conditions and was the poorest yielding in every trial where it appeared. In spite of agronomic problems, yields were quite acceptable in several lines, approaching 1.5 MT or more in some materials (Tables 21 to 25). In every trial except for that of the red-mottled types, at least one line significantly out yielded the drought resistant check, ICA Quimbaya. Some lines performed well in both years, for example: red seeded SAB 680; white seeded SAB 711; cream striped SAB’s 627, 684, 686, and 702; and red mottled SAB 641. Several drought resistant lines also yielded well in irrigated conditions. For example, red-seeded SAB’s 623 and 671 yielded very well in drought, and comparably to ICA Quimbaya with irrigation, but were four days earlier to mature (Table 21). Cream-striped SAB’s 685 and 686 yielded well in both drought and irrigation, and were 3-5 days earlier to mature than the checks (Table 24). However, in general the yield advantage in relation to checks was narrower in the red mottled class than in other grain types in both 2007 and 2008. These results serve to confirm the advantage under rainfed conditions of the selected Andean genotypes. Furthermore, when compared to the results under rainfed but favorable conditions in Darién (section 2.1.1.6) and irrigated conditions in Palmira (this section), in which yields of these lines were superior to checks, this suggests that selection for drought resistance has increased yield potential. This is the same pattern that has been observed in Mesoamerican beans. However, in determinate Andean beans it is especially significant, since yield improvement in these types has always been a particular challenge. Physiological analysis will serve to determine if the drought resistant Andean types also present improved remobilization to grain, as in the case of the Mesoamerican drought selected lines. It is important to note that the sources of drought resistance included in these Andean lines, SAB’s 258 and 259, have Mesoamerican genes derived from race Durango which has served as drought resistance source in much breeding work. Collaborators: S. Beebe, I. Rao, M. Grajales, C. Cajiao, A. Guerrero 46 Table 21. Yield of selected red-seeded Andean lines under rainfed and irrigated conditions in Palmira, July-September 2008, and comparison to rainfed yield in 2007. Drought 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 15455-5 15453-6 15455-14 15455-14 15455-14 15455-14 15455-14 15455-14 (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-34C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-27C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-5C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-20C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-8C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-31C-MC (ABA 58 x ICA QUIMBAYA)F1xSAB 258-MC-28CMC-MC-32C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-4C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-17C-MC-MC-4C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-12C-MC ICA QUIMBAYA (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-7C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-17C-MC-MC-17C-MC (ABA 36 x ICA QUIMBAYA) x SAB 258-MC-10C-MC-MC-15C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-15C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-17C-MC-MC-3C-MC COS 16 (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-9C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-19C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-17C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-28C-MC-MC-3C-MC (ABA 36 x ICA QUIMBAYA) x SAB 258-MC-10C-MC-MC-6C-MC -1 -1 Irrigated -1 -1 kg ha-1 (2007) kg ha-1 DTM g 100s kg ha kg ha d SAB 680 60 36 1651 27,6 1571 2138 63 SAB 623 60 33 1501 25,0 1476 2631 63 SAB 671 60 38 1450 24,2 1576 2568 63 SAB 677 60 36 1322 22,1 1465 2533 62 SAB 672 61 34 1295 21,5 1339 2605 65 SAB 678 59 35 1265 21,6 1536 2592 62 SAB 679 61 38 1243 20,4 1790 2696 63 SAB 670 61 30 1234 20,4 1473 2756 63 SAB 669 59 37 1213 20,4 1176 2298 64 SAB 674 60 33 1200 20,0 1403 2275 63 70 39 1183 16,7 1178 2606 67 SAB 621 59 36 1135 19,3 1528 2684 62 SAB 732 59 31 1121 18,9 1290 2383 63 SAB 667 61 42 1109 18,3 1340 1972 63 SAB 622 61 31 1084 17,6 1548 2824 65 SAB 668 61 31 1081 17,8 1149 2294 63 64 30 1079 16,9 973 2280 68 SAB 673 58 31 1075 18,5 1371 2268 62 SAB 676 59 36 1067 18,2 1315 2412 62 SAB 675 61 35 1027 17,0 1635 2376 62 SAB 620 61 32 1022 16,9 1199 2430 63 DTM SAB 666 60 34 1016 16,9 1171 2170 63 15455-5 (ABA 58 x ICA QUIMBAYA) x SAB 258-MC-17C-MC-MC-14C-MC SAB 731 59 29 923 15,5 1031 2411 63 15453-6 (ABA 36 x ICA QUIMBAYA) x SAB 258-MC-10C-MC-MC-4C-MC SAB 665 60 33 854 14,3 1242 1816 64 15455-14 CALIMA 67 43 667 9,9 1325 1394 63 LSD (0.05) 3.1 3.4 411 6.7 561 1.8 15453-6 47 Table 22. Yield of selected white-seeded Andean lines under rainfed conditions in Palmira, JulySeptember 2008, and comparison to rainfed yield in 2007. Irrigated Drought 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-13C-MC-MC-13C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-13C-MC-MC-16C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-13C-MC-MC-4C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-13C-MC-MC-17C-MC 15453-24 (ABA 36 x ICA QUIMBAYA) x SAB 258MC-21C-MC-MC-15C-MC 15453-24 (ABA 36 x ICA QUIMBAYA) x SAB 258MC-21C-MC-MC-18C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-32C-MC-MC-24C-MC 15455-2 (ABA 58xICA QUIMBAYA) x SAB 258MC-13C-MC-MC-9C-MC 15453-24 (ABA 36xICA QUIMBAYA) x SAB 258MC-21C-MC-MC-24C-MC 15455-2 15455-2 (ABA 58xICA QUIMBAYA) x SAB 258MC-13C-MC-MC-14C-MC ABA 36 (ABA 58 x ICA QUIMBAYA) F1 x SAB 258MC-32C-MC-MC-3C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-32C-MC-MC-26C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-12C-MC-MC-6C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-12C-MC-MC-14C-MC 15453-24 (ABA 36 x ICA QUIMBAYA) x SAB 258MC-21C-MC-MC-19C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-32C-MC-MC-21C-MC ICA QUIMBAYA (ABA 58 x ICA QUIMBAYA) x SAB 258MC-17C-MC-MC-5C-MC SAB 737 SAB 711 SAB 708 SAB 712 SAB 703 SAB 704 SAB 718 SAB 709 SAB 705 SAB 738 SAB 714 SAB 719 SAB 736 SAB 707 SAB 734 SAB 716 15453-24 (ABA 36 x ICA QUIMBAYA) x SAB 258MC-21C-MC-MC-26C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-13C-MC-MC-12C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-32C-MC-MC-15C-MC 15453-24 (ABA 36 x ICA QUIMBAYA) x SAB 258MC-21C-MC-MC-30C-MC 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-32C-MC-MC-22C-MC SAB 634 SAB 706 SAB 710 SAB 715 SAB 735 SAB 717 15455-2 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-17C-MC-MC-1C-MC SAB 713 15455-2 LSD (0.05) -1 -1 -1 -1 kg ha-1 (2007) kg ha-1 DTM DTM g 100s kg ha kg ha d 63 32 1446 23,0 1368 2496 69 61 27 1439 23,8 1577 3029 68 61 32 1370 22,6 1546 2268 68 60 33 1363 23,0 1546 2611 69 60 34 1307 21,8 1129 2101 63 60 31 1279 21,1 1292 2146 64 59 31 1274 21,3 1267 2516 64 61 30 1273 20,9 1593 2805 67 60 37 1243 20,8 1303 1815 62 63 25 1215 19,4 1384 2296 65 65 33 1210 18,6 1240 2491 71 59 30 1209 20,5 1138 21231 63 59 31 1127 19,1 1300 2767 63 60 27 1126 18,8 1415 2754 62 59 27 1115 19,1 1526 2507 61 60 35 1095 18,5 1121 2188 63 60 31 1051 17,6 1214 2311 62 64 38 1037 16,2 1298 2489 66 61 27 1017 16,5 1199 2504 64 59 36 986 16,7 1239 2270 63 62 28 973 15,7 1654 2758 67 59 29 950 16,3 1115 2339 63 60 34 934 15,5 1416 1958 63 60 30 921 15,2 1177 2324 63 63 29 854 13,4 1519 2689 70 1.8 3.0 388 6.3 305 2.8 48 Table 23. Yield of selected red-mottled Andean lines under rainfed conditions in Palmira, JulySeptember 2008, and comparison to rainfed yield in 2007. 15455-5 15455-2 15455-5 15455-5 15459-4 15455-5 15459-11 15455-14 15455-5 15459-10 15459-4 15459-4 15459-4 15459-4 15455-14 15455-2 15455-5 15459-4 15453-8 15459-4 15455-5 15453-6 15459-4 15453-6 15459-4 15455-5 15453-8 15455-5 15455-14 15455-14 15453-8 15459-4 15455-5 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-12C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-1C-MC-MC-4C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-17C-MC-MC-7C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-19C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-15C-MC-MC-3C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-28C-MC-MC-6C-MC ICA QUIMBAYA (COS 16 x ICA QUIMBAYA) x SAB 258/MC-2C-MC-MC-1C-MC COS 16 (ABA 58 x ICA QUIMBAYA) x SAB 258MC-4C-MC-MC-4C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-2C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-17C-MC-MC-10C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-15C-MC-MC-16C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-8C-MC-MC-14C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-15C-MC-MC-17C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-8C-MC-MC-4C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-33C-MC-MC-4C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-11C-MC-MC-1C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-10C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-8C-MC-MC-11C-MC (ABA 36 x ICA QUIMBAYA) x SAB 258MC-1C-MC-MC-4C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-8C-MC-MC-6C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-3C-MC (ABA 36xICA QUIMBAYA) x SAB 258MC-7C-MC-MC-6C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-15C-MC-MC-6C-MC (ABA 36 x ICA QUIMBAYA) x SAB 258MC-7C-MC-MC-14C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-8C-MC-MC-19C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-4C-MC (ABA 36 x ICA QUIMBAYA) x SAB 258MC-7C-MC-MC-15C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-28C-MC-MC-7C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-17C-MC-MC-5C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-17C-MC-MC-7C-MC (ABA 36 x ICA QUIMBAYA) x SAB 258MC-7C-MC-MC-4C-MC (COS 16 x ICA QUIMBAYA) x SAB 258MC-8C-MC-MC-3C-MC (ABA 58 x ICA QUIMBAYA) x SAB 258MC-36C-MC-MC-11C-MC CALIMA DTF DTM g 100s-1 kg ha-1 kg ha-1 d-1 kg ha-1 (2007) SAB 651 31 59 34 1157 19,9 2012 SAB 641 32 61 33 1009 16,3 2135 SAB 643 31 59 28 952 16,0 1670 SAB 652 31 59 34 948 16,0 1973 SAB 618 30 60 31 926 15,5 2180 SAB 644 31 59 29 920 15,7 1716 33 64 37 898 14,1 2318 30 59 34 881 15,1 1723 33 64 29 878 13,7 1777 SAB 653 30 59 31 857 14,7 1748 SAB 646 30 59 31 855 14,3 2203 SAB 619 32 61 35 830 13,7 1595 SAB 662 30 60 37 805 13,6 1709 SAB 659 31 61 39 789 13,0 2358 SAB 663 31 60 32 748 12,5 1802 SAB 617 31 63 37 724 11,6 1838 SAB 616 30 58 28 718 12,4 2104 SAB 642 31 59 26 706 12,1 1935 SAB 649 31 58 34 697 12,0 2040 SAB 658 31 62 37 682 10,9 2054 SAB 638 32 61 35 682 11,1 1993 SAB 657 32 61 37 680 11,3 2131 SAB 647 31 59 29 651 11,0 1929 SAB 636 30 60 37 651 11,0 1965 SAB 661 30 59 37 645 10,9 1656 SAB 637 32 61 37 633 10,3 1859 SAB 660 31 63 36 615 9,9 1961 SAB 648 31 60 34 604 10,0 1834 SAB 640 31 61 37 602 9,8 2127 SAB 645 31 59 30 573 9,5 2152 SAB 654 30 58 31 566 9,7 1442 SAB 655 32 59 29 555 9,4 1399 SAB 639 32 62 35 554 8,9 1909 SAB 656 32 63 39 540 8,6 1902 SAB 650 32 61 34 528 8,7 2272 33 1.3 63 1.9 39 2.6 436 341 7,0 5.7 1897 SAB 664 LSD (0.05) 49 Table 24. Yield of selected cream-striped Andean lines under rainfed amd irrigated conditions in Palmira, July-September 2008, and comparison to rainfed yield in 2007. Drought 15459-2 15459-2 15459-2 15459-2 15459-2 15460-7 15459-2 15459-19 15459-2 15460-7 15454-2 15459-2 15459-2 15454-2 15459-2 15459-2 15459-2 15459-2 15459-2 15459-2 15459-2 15459-19 15459-2 15459-2 CALIMA 15459-2 15459-2 15459-2 15459-2 15459-19 15459-19 15459-19 15459-19 15459-2 (COS 16 x ICA QUIMBAYA) x SAB 258MC-13C-MC-MC-19C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-36C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-26C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-22C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-21C-MC (COS 16 x ICA QUIMBAYA) x SAB 259MC-16C-MC-MC-30C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-4C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -11C-MC-MC-4C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-11C-MC (COS 16 x ICA QUIMBAYA) x SAB 259-MC -16C-MC-MC-28C-MC (ABA 36 x ICA QUIMBAYA) x SAB 259MC-8C-MC-MC-8C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-31C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-4C-MC (ABA 36xICA QUIMBAYA)F1xSAB 259/MC-8C-MC-MC-2C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-7C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-25C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-26C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-32C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-35C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -13C-MC-MC-8C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-3C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -11C-MC-MC-13C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-7C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-29C-MC CALIMA (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-6C-MC ICA QUIMBAYA (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-23C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-9C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-13C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -11C-MC-MC-7C-MC COS 16 (COS 16 x ICA QUIMBAYA) x SAB 258-MC -11C-MC-MC-3C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -11C-MC-MC-12C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -11C-MC-MC-8C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC -19C-MC-MC-21C-MC LSD (0.05) Irrigated kg ha kg ha d kg ha-1 (2007) 31 1326 21.9 2356 2427 63 61 33 1283 21.2 1708 2657 64 SAB 686 60 33 1260 21.2 2155 2900 65 SAB 627 60 35 1218 20.3 2123 2848 64 SAB 685 60 34 1212 20.2 2093 3303 66 SAB 702 61 32 1206 19.6 2124 2349 66 SAB 628 60 34 1197 20.1 2037 2705 65 SAB 697 61 34 1175 19.5 1467 2432 63 SAB 626 61 31 1159 19.1 2387 2760 65 SAB 701 61 33 1152 18.8 1719 3136 67 SAB 624 62 36 1146 18.4 1858 2957 67 SAB 733 60 32 1142 19.1 1448 2892 63 SAB 682 60 33 1126 18.7 2032 2547 64 SAB 681 61 38 1125 18.2 2009 2782 68 SAB 625 61 31 1120 18.5 1757 2637 63 SAB 630 61 33 1105 18.3 2061 2422 65 SAB 696 61 35 1081 17.8 2013 2568 65 SAB 687 60 30 1061 17.7 2005 3283 64 SAB 688 61 30 1059 17.6 1663 3177 64 SAB 683 60 32 1051 17.5 2111 2934 65 SAB 690 60 34 1014 16.8 2073 2979 66 SAB 633 59 33 1012 17.4 1546 2436 63 SAB 692 61 35 1007 16.7 2382 2860 64 SAB 631 61 33 1002 16.3 2327 2626 66 63 37 974 15.4 1718 1549 68 59 33 962 16.0 2460 3114 64 DTM g 100s SAB 684 61 SAB 689 SAB 691 -1 -1 -1 -1 kg ha-1 DTM 64 40 962 15.1 2131 2217 70 SAB 695 62 30 953 15.5 1776 2507 65 SAB 693 61 34 915 15.0 2297 2395 66 SAB 629 60 33 869 14.4 2537 2963 65 SAB 698 59 32 858 14.3 1672 2114 62 63 29 851 13.5 1742 2418 69 SAB 632 59 33 838 14.2 1404 2105 62 SAB 700 58 34 832 14.3 1620 1525 62 SAB 699 58 31 804 14.0 1400 1755 62 SAB 694 60 35 791 13.1 2337 2480 63 1.8 2.9 349 5.9 775 1.6 50 Table 25. Yield of selected Andean lines of several colors under rainfed conditions in Palmira, JulySeptember 2008, and comparison to rainfed yield in 2007. 15460-7 15460-7 (COS 16 x ICA QUIMBAYA) x SAB 259-MC-16C-MC-MC-34C-MC (COS 16 x ICA QUIMBAYA) x SAB 259-MC-16C-MC-MC-33C-MC DTF DTM g 100s-1 kg ha-1 kg ha-1 d-1 kg ha-1 (2007) 31 61 36 1060 17,4 1017 32 32 62 62 35 29 974 973 15,9 15,7 1110 SAB 721 32 60 34 944 15,8 1125 62 62 33 37 937 909 15,2 14,8 1610 SAB 728 32 32 SAB 727 33 62 29 885 14,3 1183 SAB 726 33 62 31 827 13,2 1293 SAB 720 31 59 35 800 13,6 1133 SAB 724 30 59 36 762 12,9 1065 69 60 37 36 751 713 10,8 11,9 1227 SAB 725 33 31 SAB 739 31 60 37 695 11,6 1137 SAB 723 31 59 35 661 11,3 977 SAB 722 31 60 33 595 9,9 947 1004 SAB 730 SAB 729 COS 16 15459-11 (COS 16 x ICA QUIMBAYA) x SAB 258-MC-2C-MC-MC-12C-MC 15459-11 SAB 560 (COS 16 x ICA QUIMBAYA) x SAB 259-MC-16C-MC-MC-19C-MC (ABA 58 x ICA QUIMBAYA) x SAB 259-MC-7C-MC-MC-9C-MC (ABA 58 x ICA QUIMBAYA) x SAB 259-MC-7C-MC-MC-4C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC-2C-MC-MC-2C-MC 15459-11 (COS 16 x ICA QUIMBAYA) x SAB 258-MC-5C-MC-MC-6C-MC 15460-7 15456-4 15456-4 15459-11 15459-11 15459-11 15459-11 ICA QUIMBAYA (COS 16 x ICA QUIMBAYA) x SAB 258-MC-5C-MC-MC-7C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC-5C-MC-MC-4C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC-5C-MC-MC-5C-MC (COS 16 x ICA QUIMBAYA) x SAB 258-MC-5C-MC-MC-1C-MC CALIMA LSD (0.05) 2.1.1.4 33 64 40 559 8,8 1.4 2.7 2.6 249 4.0 942 973 1007 Development of drought tolerant Andean beans from inter and intra-genepool crosses Rationale: Common bean suffers from drought stress in a large part of Eastern and Southern Africa where Andean large-seeded beans are the preferred types among the majority of the countries and populations represented (Wortman et al., 1998). Furthermore, although common bean is often grown on marginal soils in drought-prone areas where yields are low, they are still the second source of protein and third source of calories in the African highlands (Asfaw et al., 2007). Therefore breeding for drought tolerance in Andean beans is a priority from the perspective of both agricultural productivity and food security. With this in mind we have developed a series of advanced lines from inter- and intra-genepool crosses using 10 sources of drought tolerance and 5 commercial cultivars from the region. These lines have been called Drought Andean Bean (DAB) lines and are now being tested in a yield trial under drought stress at CIAT. Materials and Methods: A North Carolina design II was used to generate 49 experimental F2 populations from 5 large seeded varieties popular in Southern Africa and parts of Eastern Africa (Red Canadian Wonder, CAL143, SUG131, PAN147, Natal Sugar) and drought tolerant sources including 5 males of Andean origin (RAA21, SEQ1003, SAB259, ICA Quimbaya [=AFR 298], ICA Palmar) and 5 males of Mesoamerican origin (SER8, SER16, SER22, SEC16, SEQ11) as described in further detail in Makunde et al. (2007). One cross between Red Canadian Wonder and SER22 failed due to dwarf lethality but all other F1s produced F2 seed in Cali, Colombia in September 2005. Generations were advanced at 51 two sites in Colombia, alternating between dry season plantings in Palmira and Darién, and pedigree selection was used to make selections. Final single plant selections were made in the F5 generation and two sets of lines were made: one derived from inter-genepool crosses (AxM) and one from intragenepool crosses (AxA). The F5:6 lines were then planted for agronomic evaluation in Darién (Andic Dystrudept, pH = 5.5) in the case of the AxA lines and Palmira (Aquic Haplustoll, pH 7.0) in the case of the AxM lines in the June – September 2008 dry season under rainfed conditions, where the experiments consisted in 1354 and 492 lines respectively and these were interspersed with the parents as drought control genotypes. Rainfall during the season in Palmira was 163 mm with minimum temperatures of 17 to 25 °C and maximum temperatures of 25 to 33 °C; while in Darién, rainfall during the season was 280 mm, with average temperatures that fluctuated between 13 and 26°C. Only those advanced lines with favorable field production characteristics (earliness, compact growth habit, yielding ability and agronomic suitability) were harvested, each as F5:7 bulks, and these were in turn evaluated for yield (g plant-1), seed weight (g 100seed-1) and seed color. The resulting best selections were coded as DAB lines and prepared for further analysis. Results and Discussion: During the 2008 dry season a total of 1846 F5:6 selections were planted over the two sites selected for testing of the AxA and AxM progeny and of these 1390 were harvested (75%). In the case of the AxA progeny (Table 26), 898 lines out of the 1354 single plant selections were harvested representing 60% of the total planted, while in the case of the AxM progeny (Table 27), almost all 492 genotypes were harvested, representing 97% of the genotypes. Comparison of each cross combination showed that the pedigree resulting in the best average yields among the AxA cross combinations were G4523(ICA PALMAR) x NATAL SUGAR, G4523(ICA PALMAR) x RED CANADIAN WONDER and RAA21 x PAN127 with yields greater than 15 g / plant equivalent to 3000 kg ha-1. In contrast the cross producing progeny with the lowest yields was AFR298 x NATAL SUGAR with average yield of 9.7 g/plant or 1950 kg ha-1. Combining ability was best therefore for G4523 and worst for NATAL SUGAR and PAN127 and this was reflected in the number of selections advanced per cross combination. Average seed weight was highest with selections from ICA QUIMBAYA and RAA21, the two large-red seeded parents and was lower for G4523, SAB259 and SEQ1003 which were the mottled parents. Among the AxM cross combinations, SER8 and SER22 were the best female combiners, SER16 and SEQ11 were intermediate and SEC16 was the worst, but mostly in combination with SUG131 and RED CANADIAN WONDER as male parents. NATAL SUGAR and PAN127 were intermediate combiners and CAL 143 was the worst combiner for the plantings in Palmira, mostly due to an undesirable green stem characteristic reflecting less mobilization of photosynthates to grain leading to lower yields. Despite this many well-adapted progeny were found from the cross of SER22 x CAL143 with average yields among the harvested genotypes of 2821 kg ha-1. Lowest average yields were found for SEC 16 x NATAL SUGAR with 1168 kg ha-1. Future Plans: A total of 216 lines were selected for seed multiplication and coding as DAB lines consisting in 162 from the AxA experiment and 54 from the AxM experiment. The AxA derived lines included 41 from the crosses with ICA QUIMBAYA, 46 from crosses with G4523, 32 from crosses with SAB259, 6 from crosses with SEQ1003 and 28 from crosses with RAA21. Meanwhile the AxM derived lines included 26 from crosses with SER22, 13 from crosses with SER16, 8 from crosses with SEQ11, 7 from crosses with SER8 and none from crosses with SEC16. Grain types in the AxM lines tended to be smaller in seed weight (around 30 g 100seed-1) compared to those from the AxA crosses (48 to 57 g 100seed-1), as would be expected given the genepool combinations made. It was surprising that SER22 x SUG131, produced some larger seeded progeny which is why this combination was overrepresented in those that produced DAB lines from the AxM crosses. The new DAB lines will be tested in additional environments and in replicated trials. As a first set of drought trials, the DAB lines have been planted as a 52 replicated yield trial in the January – March 2009 dry season under rainfed conditions in Palmira. These trials consist in 4 lattice design experiment each with three replications. Three of the lattices contain the 162 AxA derived lines while a fourth lattice contains the 54 AxM lines. The three Andean lattices are distinguished by seed color with large-red, red mottled and cream mottled experiments planted separately and are planted only in drought treatment. The AxM lattice has been planted under drought and irrigated treatments. Control genotypes for the lattices include the Andean DIACOL Calima and the Mesoamerican Tio Canela. Table 26. Selections made from the Andean x Andean cross combinations. 2769 2433 1950 2162 2257 2314 Number of Harvested Lines (June - Sept 08) 111 56 28 33 105 228 2478 3125 2655 3188 2861 107 33 11 37 188 30 27 30 37 - 50 50 48 50 49 13 12 5 16 46 259 x CAL 143 259 x SUG 131 259 x NATAL SUGAR 259 x PAN 127 259 x RC WONDER Total 2940 2561 2057 2066 2047 2334 38 31 41 36 31 177 28 23 23 26 20 - 47 52 48 47 47 48 18 7 1 3 3 32 SEQ 1003 x CAL 143 SEQ 1003 x NATAL SUGAR SEQ 1003 x PAN 127 SEQ 1003 x RC WONDER Total 2233 2071 2407 2041 2188 29 32 20 16 97 33 35 28 32 - 49 49 45 51 49 4 0 0 2 6 69 26 17 2 24 138 27 29 43 27 27 - 50 54 56 57 51 53 19 4 2 1 2 28 162 Cross AFR AFR AFR AFR AFR G G G G 298* x CAL 143 298 x SUG 131 298 x NATAL SUGAR 298 x PAN 127 298 x RC WONDER Total 4523 x CAL 143 4523 x NATAL SUGAR 4523 x PAN 127 4523 x RC WONDER Total SAB SAB SAB SAB SAB RAA RAA RAA RAA RAA 21 x CAL 143 21 x SUG 131 21 x NATAL SUGAR 21 x PAN 127 21 x RC WONDER Total Yield (kg ha-1) 2819 2389 2640 3186 2627 2732 Total of DAB line (AXA) * AFR 298 = ICA Quimbaya 53 CV (%) Weight of 100 seeds (g) Number of DAB lines* 30 39 25 28 35 - 58 63 54 54 57 57 31 6 1 3 9 41 Table 27. Selections made from the Andean x Mesoamerican cross combinations. SER 8 x CAL 143 SER 8 x SUG 131 SER 8 x NATAL SUGAR SER 8 x PAN 127 SER 8 x R C WONDER Total 1402 1890 2062 1980 2420 1951 Number of Harvested Lines (June Sept 08) 5 26 9 33 7 80 SER SER SER SER SER 16 x CAL 143 16 x SUG 131 16 x NATAL SUGAR 16 x PAN 127 16 x R C WONDER Total 2095 1978 2402 2714 2533 2344 26 24 21 18 12 101 43 45 37 51 28 - 31 32 27 31 32 31 0 2 4 4 3 13 SER SER SER SER 22 x CAL 143 22 x SUG 131 22 x NATAL SUGAR 22 x PAN 127 Total 2822 2794 1950 2119 2421 23 51 59 36 169 29 32 35 22 - 33 36 31 30 32 1 16 6 3 26 SEC SEC SEC SEC SEC 16 x CAL 143 16 x SUG 131 16 x NATAL SUGAR 16 x PAN 127 16 x R C WONDER Total 1777 1863 1168 1744 1389 1588 2 11 17 8 14 52 9 20 32 55 31 - 24 29 30 28 29 28 0 0 0 0 0 0 SEQ SEQ SEQ SEQ SEQ 11 x CAL 143 11 x SUG 131 11 x NATAL SUGAR 11 x PAN 127 11 x R C WONDER Total 22 7 19 15 14 77 52 43 35 47 29 - 30 35 27 27 42 32 1 1 3 1 2 8 54 Cross Contributors: Yield (kg ha-1) 1692 1853 1905 1852 2114 1883 Total of DAB lines (AxM) CV (%) Weight of 100 seeds (g) Number of DAB lines* 22 34 28 27 31 - 35 32 32 29 36 33 0 4 0 1 2 7 M.W. Blair, V. M. Durán, F. Monserrate (SBA-1, CIAT), G. Makunde (DR4DZimbabwe), S. Beebe (CIAT-HQ), R. Chirwa (CIAT-Malawi), D. Lungu (Univ. of Zambia) 54 2.1.1.5 Field evaluation of a common bean reference collection for drought tolerance Rationale: Common bean improvement relies heavily on the genetic diversity of parental sources used for various purposes in a breeding program. The widest genetic diversity for cultivated germplasm is thought to exist in landraces and yet these have not been widely exploited in most common bean improvement projects, although they have been important for finding sources of insect or disease resistance (Singh, 2001). Screening of a core collection of mostly landraces has also been useful for finding new sources of low fertility tolerance. Broadening of the genetic base for cultivated common bean and for specific commercial classes of beans is very important given the narrow genetic based of some cultivar groups (Kelly, 1999), especially within the Andean genepool (Beaver et al., 1999). The objective of this work was to screen part of the CIAT core collection (termed a reference collection) for the identification of new sources of drought tolerance among bush bean landraces. In the process, a large number of phenotypic traits associated with drought tolerance were considered. The reference collection is a smaller more manageable subsample of the core collection and has been selected based on molecular marker fingerprinting to represent most of the diversity in cultivated common bean and in the initial collection. It also is a collection with a well-understood population structure based on the previous molecular analysis which allows it to be efficiently analyzed for marker associations, something that was not possible previously with quantitative traits and with a loosely understood collection. Given this, the reference collection was carefully stratified according to Andean and Mesoamerican genepools with known subdivisions according to each of the common bean races that we validated in previous studies. Materials and Methods: A total of 199 genotypes were evaluated in the July-September 2008 season across three location/treatment combinations: 1) Darién – rainfed; 2) Palmira – rainfed and 3) Palmira – irrigated. Of these genotypes, 169 belonged to the CIAT core collection and 30 were additional genotypes added to create the reference collection and considered as checks (12 Andean controls and 18 Mesoamerican controls). The genotypes were stratified according to gene pool with separate experiments for the Andean genotypes (64 entries in an 8 x 8 lattice) versus the Mesoamerican genotypes (121 entries in an 11 x 11 lattice). Soil conditions in Palmira (Aquic Haplustoll, pH= 7.7 and available P levels of 67 to 77 ppm) were fairly different than in Darién (Andic Dystrudept, pH 5.7 and 2 to 6 ppm available P) and fertilizer application was necessary in the latter location (400 kg ha-1 of diammonium phosphate, supplying 79 kg ha-1 P) but not in Palmira. In addition the altitude difference in Palmira (1000 masl) versus Darién (1500 masl) led to differences in adaptation to climate regimes. For example in Palmira minimum and maximum temperatures varied between 17 / 25 °C for lows and 25 / 33 °C for highs while in Darién average temperatures were 16°C (average low) to 25°C (average high). Rainfall also varied with 163 mm unevenly distributed during the growing season in Palmira compared to Darién where 280 mm rain fell but mostly towards the end of the season (R7 and R8 pod filling growth stage). Drought stress was therefore more intense but intermittent in Palmira with severe stress around early establishment (V2 and V3 growth stages) compared to Darién where it was moderate but early. For Palmira, two treatments were implemented, one with adequate irrigation and one with rainfed conditions (i.e. to induce drought stress after establishment). Both treatments were established with three early gravity irrigations of about 35 mm each at (-1, 15 and 34 days from/after planting). The irrigated treatment received three additional gravity irrigations of the same amount at approximately 1.5 to two week intervals (45, 55 and 72 days after planting). 55 Data collection: In all three environments data were collected on basic phenological and yield component traits. These included days to flowering (DF), days to maturity (DM), yield (kg ha-1), number of pods per plant, number of seed per pod and number of empty pods. Physiological measurements including chlorophyll content (SPAD), stomatal conductance (mmol m-2 s-1), photosynthetic efficiency (Fv′/Fm′), canopy temperature depression (°C) and leaf area index were also taken during the R6 to R7 growth stage. Scatterplots were made to graphically represent the rainfed (drought) versus irrigated (control) averages for each trait and each genotype. In addition, the data were used for a principal component analysis (PCA) to determine the relatedness of genotypes from each race within each gene pool based on the phenological, physiological and yield traits. Results and Discussion: Preliminary analysis was conducted separately for each experiment representing each gene pool and each growing environment (Table 28 and Figures 13 and 14). Average yields across all Andean genotypes were 1644 kg ha-1 in rainfed treatment in Palmira, 1614 kg ha-1 in rainfed treatment in Darién and 1016 kg ha-1 in irrigated control treatment in Palmira. Meanwhile, average yields across all Mesoamerican genotypes were 1724 kg ha-1in rainfed treatment in Palmira, 1941 kg ha-1 in rainfed treatment in Darién and 1596 kg ha-1 in irrigated control treatment in Palmira. The lower yields in the irrigated control treatment in Palmira were due to Sclerotium rolfsii infection. This root rot is favored in the warm, alternating humid-and-dry conditions of the irrigated treatment during the dry season and was more prevalent in the field used for irrigated treatment which is not tiled as compared to the field which is used for rainfed treatment which was tiled for additional drainage. The lower yields of the irrigated field made some comparisons of yield potential and drought effects difficult. For example, least significant differences were higher in the irrigated control treatment in Palmira than in the rainfed treatments in both sites, also reflecting the variability in disease incidence. However, the results of the two rainfed treatments in Palmira and Darién allowed us to analyze the effects of different stress treatments on the different genotypes while the comparison of the drought and irrigated treatments allowed us to analyze the effect of the irrigation treatment and disease pressure on these same genotypes. Below we summarize some results for the genotypes tested from each gene pool. Table 28. Average yields, days to flowering and maturity, 100 seed weight and % empty pods in Andean and Mesoamerican genepools, races and subgroups as compared to control genotypes used in the reference collection experiments. Race Andean genepool Nueva Granada 1 Nueva Granada 2 Perú Total Andean (1) (2) (3) (1) (2) (3) (1) (2) (3) Weight of 100 seeds (g) (1) (2) (3) 1717 1384 1011 1395 1213 1110 756 1053 1580 1541 1437 1527 37 30 36 34 41 37 40 39 47 43 47 46 40 42 41 41 43 36 45 41 43 37 38 39 33 35 38 35 34 36 39 36 39 40 43 40 67 69 78 71 68 71 76 71 78 80 83 80 Andean checks SAB 645 SEQ 1003 CAL 96 KAT B1 AFR 298 URUGEZI SEQ 1027 KAT B9 SELIAN97 CAL 143 AFR 619 SAB 258 2596 2566 2290 2285 2209 1962 1927 1900 1689 1586 1492 1459 463 2110 997 1170 1442 1396 1960 1919 2540 1568 2531 1451 1779 1486 1653 1703 1232 1500 1777 1525 2445 1969 1954 1365 34 40 53 38 46 46 36 39 22 32 36 33 45 45 59 39 51 32 44 43 28 44 65 34 57 48 65 35 57 44 55 48 28 50 45 39 35 25 34 30 35 36 30 33 32 34 42 41 20 41 36 51 44 16 7 38 26 31 25 24 62 54 53 39 44 49 30 34 17 44 29 42 29 34 33 28 32 36 38 28 37 34 36 28 34 35 33 28 33 36 36 29 36 34 37 27 36 43 38 38 38 42 46 36 40 38 42 35 62 67 67 60 68 70 69 58 70 68 70 60 60 69 70 63 72 68 73 62 67 70 77 60 79 82 81 81 80 79 82 75 74 80 79 73 Yield (kg ha-1) Days to Flowering Days to Maturity 56 Empty Pods (%) (1) (2) (3) Table 28. cont´d. Yield (kg ha-1) Race (1) Days to Flowering Days to Maturity (1) (2) (1) (2) (3) Empty Pods (%) (2) (3) Mesoamerican genepool Durango 1 1742 Durango 2 1678 Guatemala 1240 Mesoamerican 1 1764 Mesoamerican 2 1780 Total Mesoamerican 1702 1385 1743 1268 1573 1742 1597 1847 1986 1895 2116 2002 1997 31 35 35 38 37 36 31 35 34 37 36 35 36 39 39 40 40 39 68 69 71 69 70 69 64 69 70 69 69 68 72 76 78 76 75 75 33 21 24 21 20 23 33 23 26 23 25 25 37 24 28 23 23 26 51 40 46 41 42 43 62 40 46 31 34 39 54 35 41 30 28 35 Mesoamerican checks SEA 15 DOR 390 SXB 418 SER 16 SER 118 NCB 280 BAT 477 SER 109 SEA 5 VAX 6 A. MELKA T.CANELA75 VAX 3 SEQ 11 M. SOJA P. VILLA BAT 93 M. RED 2871 1812 2430 1789 2962 1926 1637 2124 2169 2194 2019 1447 2332 3052 1269 1497 1218 1439 2481 2439 2437 2864 2658 2582 2076 1528 1443 2194 2336 2065 2827 1890 2813 2036 2302 1790 32 39 38 33 37 32 38 31 33 38 41 39 37 34 38 28 38 37 31 42 36 32 36 31 37 33 32 38 39 37 37 33 38 30 37 36 37 43 39 37 39 37 40 37 37 41 43 42 40 37 43 36 42 39 68 71 70 66 69 66 69 67 67 69 73 69 69 70 73 65 69 73 62 77 67 62 67 63 74 67 66 71 74 69 69 70 69 60 69 77 76 80 74 74 75 77 77 75 61 77 66 78 79 75 81 72 77 77 34 17 28 25 25 26 21 26 26 22 16 21 25 29 21 37 18 22 33 20 29 24 27 24 22 27 29 24 20 22 30 30 21 34 19 24 35 20 28 25 30 31 22 26 32 23 14 23 27 29 23 36 20 28 28 19 27 31 19 43 46 60 38 25 27 33 31 23 38 74 50 32 39 21 22 5 13 28 32 28 17 26 17 32 20 30 36 59 42 46 48 23 44 24 19 41 27 21 24 14 20 31 26 33 15 45 31 24 3292 2917 2912 2863 2854 2835 2690 2597 2505 2494 2375 2357 2083 2049 1974 1715 1676 1472 (3) Weight of 100 seeds (g) (1) (2) (3) (1) (2) (3) Environments: Rainfed with intermittent drought stress (1) and Irrigated (2) in Palmira. Rainfed with moderate drought stress (3) in Darién * Andean Race: P- Perú, NG1 – Nueva Granada 1, NG2- Nueva Granada 2 ** Primary and Secondary Seed Color: 1 - White; 2 - Cream; 3 - Yellow; 4- Brown; 5 - Pink; 6 - Red; 7 - Purple; 8 - Black; 9 - Others Andean genotypes: The best yielding genotypes among the Andean reference collection genotypes were G11727 (2230 kg ha-1) in Darién –rainfed; G18255 (2455 kg ha-1) in Palmira –rainfed and LRK31 (2018 kg ha-1) in Palmira – irrigated control. Among the check genotypes the best were SELIAN97 (2445 kg ha-1) in both Darién – rainfed and Palmira – irrigated as well as SEQ1003 (2566 kg ha-1) and SAB645 (2596 kg ha-1) in Palmira – rainfed. On the other hand, a number of Andean genotypes from the reference/core collection were very poorly adapted, yielding nothing at all in either the Palmira – rainfed (G19831, G11787, G19841) or Palmira – irrigated (G19841) treatments. Darién was a more favorable location for Andean genotypes due to the cooler temperatures encountered there during the growing season. 57 Figure 13A – ScatterPlot for Yield: Drought in Palmira vs Non-Drought in Palmira 3000 70 Yield (Kg/Ha) in Drought - Palmira 2500 64 31 72 65 8 1500 24 38 45 2000 Means: 1644 46 LSD0.01 : 809 47 3 19 21 23 4 53 500 12 13 16 9 0 55 75 73 57 54 49 27 10 39 35 59 74 71 66 11 69 43 17 60 63 2 62 15 Race Control genotypes Nueva Granada 1 Nueva Granada 2 Means: 1213 LSD0.01 : 1016 7 52 4814 0 18 22 51 40 58 56 28 67 37 29 26 61 32 41 30 50 34 42 5 36 44 1 25 20 6 1000 68 33 500 1000 Perú 1500 2000 2500 Yield (Kg/Ha) in Non-Drought - Palmira 3000 Figure 13B – ScatterPlot for Yield: Drought in Palmira vs Moderate Drought in Darien g g g 3000 70 68 33 Yield (Kg/Ha) in Drought - Palmira 2500 2000 64 72 31 65 Means: 1644 LSD0.01 : 809 39 19 30 6 17 60 2 63 1000 15 26 61 5 58 13 500 9 52 0 400 48 800 14 32 24 40 45 75 36 54 35 47 49 44 25 23 46 74 34 55 66 37 29 43 42 4 62 53 22 51 57 38 67 20 1 73 27 21 41 10 3 50 69 11 18 59 1500 8 16 56 28 Means: 161412 LSD0.01 : 952 1200 1600 7 Race Controlsgonotypes Control NG1 Granada 1 Nueva NG2 Granada 2 Nueva P Perú 2000 Yield (Kg/Ha) in Moderate-Drought - Darién 58 71 2400 Figure 13C 8 g – PCA for Yield, Empty pods and Weight per 100 Seeds , pyp Yield: Drought in Palmira (A), Non-Drought in Palmira (B), Moderate- Drought in Darién (C) Empty Pods: Drought in Palmira (D), NonDrought in Palmira (E), Moderate- Drought in Darién (F) Weight per 100 seeds: Drought in Palmira (G), Non-Drought in Palmira (H), ModerateDrought in Darién (I) Numbers are Genotypes (see Append table) 4 PC2 (23,56 %) 61 0 12 D 60 50 58 34 15 42 47 57 32 20 38 10 24 21 27 31 43 39 69 4 29 74 67 3 2 49 72 73 25 59 22 41 44 51 55 45 37 19 I 8 H 35 65 70 64 G 1 54 33 68 66 A B C 75 40 18 Race 6 23 62 63 28 30 53 -4 11 5 13 56 F E 46 9 p 17 26 16 7 g 36 Perú Nueva Granada 1 71 Nueva Granada 2 Control Genotypes -8 -12 -7 Figure 13D PC 1 (38,74 %) 3 8 – PCA for Yield and Weight per 100 Seeds g 6 4 -2 g Yield: Drought in Palmira (A), NonDrought in Palmira (B), Moderate- Drought in Darién (C) Weight per 100 seeds: Drought in Palmira (G), Non-Drought in Palmira (H), Moderate- Drought in Darién (I) p Race Perú Nueva Granada 1 Nueva Granada 2 Control genotypes 71 PC2 (20,06 %) Numbers are Genotypes (see Append table) 2 57 53 0 12 56 9 48 -2 B 36 49C 40 68 66 74 22 32 A 75 33 25 46 44 55 27 72 73 31 54 41 45 50 67 37 51 42 69 34 10 43 29 24 39 3 70 60 4 2 G 15 59 19 20 62 16 26 H 65 21 61 30 23 63 11 I 13 28 58 17 38 7 5 18 47 35 8 64 1 6 -4 -10 -6 -2 PC 1 (51,43 %) 2 6 Figure 13. Results of rainfed and irrigation treatments in the Andean 8x8 lattice. 59 Figure 14A – ScatterPlot for Yield: Drought in Palmira vs Non-Dought in Palmira Figure A ScatterPlot for Yield: rought in Palmira Vs Non rought in Palmira 3400 123 120 2900 108 Yield (Kg/Ha) in Drought-Palmira 22 6 2400 79 73 113 112 111 121 110 109 122 115 21 99 96 103 77 119 60 29 70 74 71 82 116 46 8 11 32 51 Means: 1724 117 81 10 34 25 33 2 85 64 19 LSD0.01 : 964 5 75 97 50 98 106 53 62 45 93 65 87 90 95 16 20 91 31 78 114 57 107 9 69 18 40 63 49 48 7 41 14 26 104 47 105 12 80 55 1 28 84 58 44 67 118 83 59 101 76 43 54 4 89 37 86 100 94 30 61 36 35 102 52 15 88 3 42 27 38 56 72 1900 1400 900 124 92 Races Control genotypes Durango 1 Durango 2 Guatemala Mesoamerican 1 Mesoamerican 2 17 Means: 1596 LSD0.01 : 1205 23 24 68 13 66 39 400 400 900 1400 1900 2400 2900 3400 Yield (Kg/Ha) in Non Drought in Palmira Figure 14B – ScatterPlot for Yield: Drought in Palmira vs Moderate-Dought in Darien Figure ScatterPlot for Yield: rought in Palmira Vs Moderate rought in arién 3500 123 Yield (Kg/Ha) in Drought-Palmira 3000 109 122 2500 103 6 29 90 41 1500 1000 500 48 47 77 74 25 115 22 99 113 119 72 13 120 71 110 5 65 87 8 60 124 39 Means: 1941 LSD0.01 : 954 0 1000 1400 1800 2200 Yield(Kg/Ha) in Moderate- Drought - Darién 60 73 96 68 121 111 32 82 51 70 46 34 11 64 19 2 33 98 31 97 106 53 16 7 20 78 40 95 107 114 62 49 9 18 14 1 55 105 57 26 80 84 12 58 100 118 92 44 104 83 28 101 86 43 89 54 37 4 76 30 59 94 102 35 61 36 15 52 88 27 3 42 38 56 23 24 Means: 1724 LSD0.01 : 964 75 2000 112 108 79 2600 21 66 85 91 116 117 63 Races Control genotypes Durango 1 Durango 2 Guatemala Mesoamerican 1 Mesoamerican 2 3000 Figure 14C – PCA for Yield, Empty pods and Weight per 100 Seeds g 8 , pyp 123 110 PC 2 (21,09 %) 4 A 121 68 120 119 103 0 -4 115 112 B 96 73 C 124 116 21 22 g p Yield: Drought in Palmira(A), Non-Drought in Palmira (B), Moderate- Drought in Darién (C) Empty Pods: Drought in Palmira (D), Non-Drought in Palmira (E), Moderate- Drought in Darién (F) Weight per 100 seeds: Drought in Palmira (G), NonDrought in Palmira (H), Moderate- Drought in Darién (I) Numbers areGenotypes (see Append table) 111 122 33 H 60 66 13 85 51 71 109 26 65 97 6479 50 72 34 19 70 108 117 30 77 25 87 49 91 93 62 9532 98 31 78 17 86 37 29 12 94 105 53 55 63 104 118 28 81 40 18 82 844676 136 20 47 69 59 35 107 61 67 75 100102 83 57 54 42 101 D 58 56 38 27 88 23 52 80 G 11 I F 43 8 2 44 45 92 99 6 114 7 E -8 -4 0 Figure 14D – Figure 6 PC 1 (42,12 %) A 68 103 99 120 21 C Races Durango 1 Durango 2 Guatemala Mesoamerican 1 Mesoamerican 2 Control genotypes 39 PC 2 (27,89 %) 0 -2 12 Yield: Drought in Palmira(A), Non-Drought in Palmira (B), Moderate- Drought in Darién (C) Weight per 100 seeds: Drought in Palmira (G, Non-Drought in Palmira (H), ModerateDrought in Darién (I) Numbers areGenotypes (see Append table) 121 116 111 73 124 13 51 85 60 66 92 119 109 108 22 33 122 H 70 19 82 64 63 79 2 I 34 45 62 32 113 25 117 72 95 17 98 G 29 74 65 7 46 114 94 77 31 55 12 106 50 6 28 100 81 93 26 97 86 59 89 78 49 40 53 76 30 44 35 105 87 118 18 83 20 69 84 37 3 47 67 36 75 101 5 15 57 90 102 104 1 27 61 43 88 56 54 42 38 58 52 39 23 2 8 PCA for Yield and Weight per 100 Seeds 112 96 115 41 : PCA for Yield and Weight per 00 Seeds 123 B 9 4 3 4 110 4 14 16 5 15 24 -8 10 80 10 11 8 9 16 14 4 41 Races Durango 1 Durango 2 Guatemala Mesoamerican 1 Mesoamerican 2 Control genotypes -4 24 -6 -4 Figure 14. -2 0 2 4 PC 1 (40,75 %) 6 8 10 Results of rainfed and irrigation treatments in Mesoamerican 12 x 12 lattice. 61 Scatterplot comparisons were made between the drought and irrigated treatments in Palmira (Figure 13A) and between the Palmira and Darién rainfed treatments (Figure 13B) to detect the stability of each genotype and to compare the different races/groups. These comparisons showed that check genotypes were more resistant to this stress and higher yielding generally than the majority of the reference collection genotypes. This was to be expected since the checks were selected to be drought tolerant and high-yielding while the reference collection represents diverse landraces which have not been improved through breeding. Meanwhile, the Nueva Granada group NG1 was higher yielding generally than group NG2 or than race Peru, which shows the adaptation of some Nueva Granada race bush beans to hotter and drier conditions than race Peru bush beans. Mesoamerican genotypes: The best yielding genotypes among the Mesoamerican reference collection genotypes were G4017 (3055 kg ha-1) and G5694 (3025 kg ha-1) in Darién – rainfed; G3142 (2660 kg ha1 ) and G21212 (with 2551 kg ha-1) in Palmira – rainfed; and G278 (2799 kg ha-1) and G2199 (2864 kg ha1 ) in Palmira – irrigated control. It was evident that the Mesoamerican genotypes had a higher yield potential than the Andean genotypes in all three environments. Furthermore, there were no Mesoamerican genotype that failed to produce seed in any of the environments and the lowest yields were 1164, 475 and 497 kg ha-1, respectively; with Darién under moderate level of water stress being more productive generally for Mesoamericans than Palmira with irrigation or under drought stress. This may be the result of cooler temperatures in Darién and heat stress being a factor in Palmira as well as disease pressure from Sclerotium rolfsii as mentioned above. Using the same Scatterplot comparisons as described for Andean genotypes, the Mesoamerican control genotypes were found to generally outperform the reference collection genotypes, although some Mesoamerica race and Durango-Jalisco group 2 genotypes were also high yielding. Durango-Jalisco group 1 genotypes were found to perform lower as did Guatemala race genotypes. The race Guatemala genotypes are normally climbing beans, therefore the production system of non-trellised field grown plants was not favorable for them, a factor that probably was also important for some of the DurangoJalisco genotypes. Principal component analysis: Principal component analysis was also used to determine the relationships between the genotypes and corresponding groups within each gene pool considering the variables for yield, 100 seed weight, and percent empty pods in the three environments (subfigures C and D of Figures 13 and 14). Biplots showed the separation of the subgroups and races discussed above, especially the separation of Mesoamerica and check genotypes from Durango-Jalisco group 1 genotypes within the Mesoamericans and Nueva Granada versus Peru race genotypes in the Andeans. This confirmed something observed in the field which was the poor adaptation of “Durango” type cultivars to the tropical environments of Palmira and Darién, both in terms of overall vegetative growth and disease pressure (mainly powdery mildew and Sclerotium rolfsii) and the high levels of flower abortion in the case of Peru race genotypes. Biplot analysis also showed that high yield in the three environments (letters A, B and C in the PCA) was associated with the check genotypes while empty pods (letters D, E and F) were associated with the less adapted groups (race Peru for the Andeans and the Durango-Jalisco group 1 genotypes for the Mesoamericans) and that these two characteristics were located in opposite quadrants as would be expected. Seed size (letters G, H and I) was associated with some genotypes of races Peru and Nueva Granada as well with certain race Durango and Guatemala accessions. Empty podding (lack of seed filling) was negatively correlated with yield potential, a reflection of heat and drought stress symptoms. Meanwhile, seed size was somewhat correlated with yield potential since drought tolerant genotypes filled seed better than drought susceptible genotypes. 62 Conclusion and Future Plans: This project was useful because we were able to establish a rainfed season planting in Darién to simulate moderate level of water stress, something which had not been done before, and we were able to obtain reliable data from that location for moderate drought stress tolerance in Andean beans. Conditions at our new site were equivalent of moderate drought stress throughout the season which would be typical of many growing environments where beans face this stress. Likewise, Palmira presented good intermittent drought stress conditions but heat stress may be a confounding factor in some cases for Andean or Durango race beans. Conditions in Palmira presented moderate stress with intermittent drought consisting in only 23 mm of rainfall between V2 and V3 stages and 47 mm between R6 and R8 stages. In terms of the genotypes evaluated from the reference collection and the check genotypes, interesting results were obtained differentiating the best accessions within each race and obtaining an understanding of differences between each group (races Durango Jalisco 1 and 2, Guatemala, Mesoamerica, Nueva Granada 1 and 2, Peru). It was notable that better adaptation was found in the Nueva Granada group 1 than in NG2 or in race Peru and that within the Mesoamerican genotypes, race Mesoamerica and Durango Jalisco group 2 were better adapted than Durango Jalisco group 1 or race Guatemala. This may be due to photoperiod, relative humidity and prevalent diseases at the altitudes the two sites used compared to the region to which these genotypes are better adapted and highlights the challenge of using Durango genotypes in cultivar development for the sub-humid tropics. Apart from the 8 x 8 and 12 x 12 lattices another small lattice of 36 genotypes (6 x 6 lattice) was also planted and will be analyzed soon. The genotypes have also been re-arranged into a larger Andean lattice design experiment as well as the Durango-Jalisco-Mesoamerica experiment, for testing in Eastern and Southern Africa as part of the TL1 drought project. From this study, the Nueva Granada genotypes G18255 (33), G16115 (31), G4001 (22), G17070 (32), G1688 (18), G5625 (24), G22247 (36), G6639 (25), G7945 (27), G11957 (29) are of interest for further studies or use in breeding of drought tolerance. Among the Mesoamerican genotypes, a large number of lines could be of interest but the G21212 genotype stands out as was found previously by CIAT researchers. Further analysis will be conducted to evaluate drought sensitivity indices and comparisons of drought versus control values for a number of phenotypic traits in Palmira and severe versus moderate drought across the two sites to determine the relative effect of drought on trait expression in the genotypes. In addition, further statistical analysis as well as the analysis of variance (ANOVA) are ongoing. Finally, the new lattices will probably be planted in the June-September drought season in Palmira and Darién to test the trends identified so far. 2.1.1.6 Evaluation of Andean breeding lines for adaptation to drought stress Rationale: As part of the drought screening activities described in the previous section, we have evaluated additional breeding lines for drought tolerance at a mid-elevation site useful for selecting Andean beans and instituted a dry season planting. The breeding lines (SAB series) are the product of recurrent selection for drought stress tolerance and earliness in CIAT headquarters and were tested in a new field in Darien in replicated yield trials during the July-August dry season. The SAB lines have Andean grain types and are of red mottled, cream mottled, large-red and large white commercial classes. These genotypes were developed from triple crosses between the drought-resistant genotype ICA QUIMBAYA crossed with drought susceptible but commercial type genotypes ABA36, ABA58 and COS16 and drought sources from the first cycle of selection (such as SAB258 and SAB259) that were derived from multiple crosses including a Durango source. Since ICA QUIMBAYA has wide adaptation we decided to test the resulting lines under mid-elevation conditions but with an off-season planting to coincide with drought stress. Materials and Methods: A total of 5 yield trials were planted in Darién (Valle del Cauca, Colombia) during the July-August dry season in 2008. The soils in Darién are Andic Dystrudept and the site is located at 1500 masl elevation with average temperature of 19°C. A new field was used for the 63 experiment to reduce the chances of root rot against which ICA Quimbaya and many SAB lines are highly susceptible. The soil pH was 5.7 and available P levels were 2 to 6 ppm, which were supplemented with 90 kg ha-1 of P fertilizer. The yield trials were organized as lattice design experiments with each trial containing SAB lines of a different commercial seed color class and 3 repetitions each. The first two experiments consisted in separate 6x6 lattices for red mottled and cream mottled grain types, respectively. The second two experiments consisted in separate 5x5 lattices for large-red and large-white grain types, respectively while a fifth lattice consisted on a 4x4 lattice with additional red mottled, cream mottled and large red lines. In addition the following check genotypes were included in the experiments: AFR298 (=ICA Qumbaya; large-red), COS16 (cream mottled), ABA36 (large-white), SAB560 (large red) and the local commercial variety Calima (red mottled). Results and Discussion: Rainfall pattern: Growing conditions during the season were appropriate for drought testing since total rainfall was 280 mm. Soil texture was heavier than in our previous fields in Darién, so residual water from the rainy season probably supplemented this moisture level. Drought stress was highest early in the season and especially soon after flowering since 198 mm of the rain during the season fell during pod filling (R7-8). Meanwhile, temperatures varied from 16 to 25 °C during the growth cycle. Yield data: Genotype yields for the five experiments are summarized in Figure 15 which compares the average yield in kg ha-1 for the check genotypes and advanced lines from each commercial class. In that figure each lattice is represented as a column of datapoints with the least significant difference (LSD) given at the base of the column and the check genotypes highlighted as diamonds. Comparisons of the trials, shows that the average yields of the large-red lines (column C) were higher than for the red mottled and cream mottled lines (columns B and D), while the lowest yields were in the lattices with the large-white beans (column E) and the mixed group (column A). Meanwhile, comparisons of the SAB lines with the check genotypes shows that for all of the trials (columns A through E), some of the advanced lines were significantly superior to the controls (AFR298, CALIMA, COS16 and ABA36) based on the LSD for each trial. For example, SAB 663 with an average yield of 2047 kg ha-1 was significantly higher yielding than the check genotype CALIMA with 1472 kg ha-1 within column B for the 6x6 red mottled trial (LSD = 560 kg ha-1). As expected in all the lattices except the 4 x 4 lattice (column A), AFR298, the moderately drought tolerant parent, had higher yields than COS16 or ABA36, which were susceptible parents although again these differences were not significant. Surprisingly AFR298 and Calima performed about equally well. Conclusions and Future Plans. In general, the results confirm the genetic gain for drought tolerance and yield potential that has occurred in the breeding of the SAB lines compared to both their droughttolerant and susceptible parents. In addition, certain SAB lines can be selected with greater stability across mid-elevation and lower-elevation sites based on this analysis. The most promising series of lines appear to be those of the large-red commercial class followed by some of the cream-mottled genotypes. However, differences between the trials could have been due to the innate drought tolerance and yield potential of the genotypes in the trials or to their location in the field, since available P levels were found to vary along a gradient from the upper part to the lower part of the field (from 2.4 to 6.2 ppm), which also reflected differences in organic matter and clay content. Future plans are to repeat these yield trials in other years or locations as part of the drought projects we are developing and to evaluate if selection for drought tolerance in the advanced lines improves adaptation to low fertility conditions. Collaborators: M.W. Blair, F. Monserrate, S. E. Beebe, M. Grajales, Y. Viera, A. Hoyos (SBA-1) 64 3000 SAB lines A - Red mottled and other beans B - Red mottled beans C - Red beans D - Cream striped beans E - White beans Control Genotypes Calima - Red Mottled bean AFR298 - Red bean COS16 - Cream Striped bean ABA36 - White bean 2700 2400 B Yield (Kg/Ha) A 2100 730 SAB560 728 720 1800 729 1500 723 724 726 727 725 721 COS16 1200 686 D C 677 669 679 670 680 622 671 623 678 665 672 668 673 675 620 621 676 674 732 667 CALIMA 666 731 661 642 655 663 643 619 617 654 640 645 657 652 637 638 664 644 650 CALIMA 646 658 656 662 660 647 653 649 648 618 639 651 616 687 685 733 682 684 683 630 624 631 626 625 627 701 629 691 693 689 690 702 688 694 692 696 695 628 681 AFR298 COS16 722 633 CALIMA AFR298 712 E 703 710 716 711 737 738 736 AFR298 714 713 707 704 706 718 735 719 717 708 709 734 715 ABA36 CALIMA COS16 659 AFR298 AFR298 COS16 632 700 699 641 697 698 705 739 900 LSD*: 534 634 636 LSD*: 560 LSD*: 498 LSD*: 402 LSD*: 512 * Least Significant Difference Kg/Ha (p=0.05) Figure 15. Comparison of average yield (kg ha-1) for SAB lines planted in 5 lattice design experiments in Darién in the 2008 dry season. 2.1.1.7 Physiological evaluation of drought resistance in elite lines under field conditions Rationale: Development of drought adapted bean varieties is an important strategy to minimize crop failure and improve food security in bean growing regions. Previous research indicated that the superior performance of common bean genotypes under drought was associated with their ability to mobilize photosynthates to developing grain and to utilize the acquired N and P more efficiently for grain production. Among the plant traits evaluated using elite lines and recombinant inbred lines, higher values of pod partitioning index, pod harvest index and stem biomass reduction were identified as useful traits to consider in the breeding program in addition to grain yield for identifying bean genotypes that are better adapted to both terminal and intermittent drought stress conditions. We evaluated drought adaptation of 36 elite lines including checks from the on-going breeding program on improving drought resistance to 65 quantify phenotypic differences in drought resistance under field conditions and to define the physiological basis for improved drought adaptation. Materials and Methods: A field trial was conducted at Palmira in 2007 (June to September). The trial included 36 genotypes A 686, A 774, BAT 477, Carioca, Cowpea Mouride, DOR 390, EAP 9503-32B, EAP 9653-16B-1, G19902, G24390, G40001, NCB 226, NCB 280, Perola, RCB 273, San Cristóbal 83, SEA 15, SEA 5, SEN 36, SEN 56, SER 109, SER 113, SER 118, SER 119, SER 125, SER 128, SER 16, SER 48, SER 78, SER 90, SXB 405, SXB 409, SXB 412, SXB 415, SXB 418 and Tio Canela (Table 29) to determine genotypic differences in tolerance to drought stress conditions. A 6 x 6 balanced lattice design with 3 replicates was used. Two levels of water supply (irrigated and rainfed) were applied. For the irrigated treatment, a total of 4 gravity irrigations (approximately 35 mm each) were applied while for the rainfed treatment only 2 irrigations were applied to assure good crop establishment (one before planting and another 24 days after planting). Details on planting and management of the trial were similar to those reported before. Experimental units consisted of 4 rows, 3.72 m long by 0.6 m wide. A number of plant attributes were measured at mid-podfilling under rainfed conditions in order to determine genotypic variation in drought resistance. These plant traits included leaf chlorophyll content (SPAD), canopy temperature, canopy temperature depression (CTD), leaf area index, canopy dry weight per plant and shoot TNC content (total nonstructural carbohydrates). Canopy temperature was measured with a Telatemp model AG-42D infrared thermometer. The instrument was held at a 45o angle at 50 cm from the canopy surface to measure canopy temperature and canopy temperature depression. At the time of harvest, grain yield and yield components (number of pods per plant, number of seeds per pod, and 100 seed weight) were determined. Stem biomass reduction (mobilization of photosynthate reserves) was determined based on difference in stem dry weight at harvest from the stem dry weight at mid-pod filling. Pod partitioning index (dry wt of pods at harvest/dry wt of total biomass at mid-podfill x 100), pod number per area, seed number per area, pod harvest index (dry wt of seed/dry wt of pod at harvest x 100), yield production efficiency (seed biomass dry weight at harvest/total shoot biomass dry weight at mid-pod filling), seed production efficiency (seed number per area/ total shoot biomass dry weight at mid-pod filling per area), pod production efficiency (pod number per area/ total shoot biomass dry weight at mid-pod filling per area) and grain filling index (100 seed weight of rainfed/100 seed weight of irrigated) were also determined. Eight genotypes (DOR 390, SER 16, SER 109, G40001, G24390, SXB 418, SEA 5 and Tio Canela 75) were selected to determine differences in root growth and distribution across soil depth. Root samples were taken at 53 days after planting at both levels of water supply. Samples were taken at 5 soil depths (05, 5-10, 10-20, 20-40 and 40-60 cm), using a 5 cm diameter soil corer. Five soil cores were taken, three cores between rows and two within rows. To facilitate washing, samples were first soaked for 30 minutes in 5% sodium hexametaphosphate solution. Soil and roots were separated by hand washing, cleaning and scanning. Root length and diameter were determined by image analysis system (WinRHIZO V. 2007b). Results and Discussion: Palmira – Soil, temperature, rainfall and evaporation: The soil is a Mollisol (Aquic Hapludoll) with no major fertility problems (pH = 7.7), and is estimated to permit storage of 130 mm of available water (assuming 1.0 m of effective root growth with –0.03 MPa and –1.5 MPa as upper and lower limits for soil matric potential). During the crop-growing season, maximum and minimum air temperatures were 30.2 and 18.6o C (Figure 16). The incident solar radiation ranged from 11.2 to 25.1 MJ m-2 d-1. The total rainfall during the active crop growth was 235.6 mm. The potential pan evaporation was of 316.2 mm. These data on total rainfall and pan evaporation together with rainfall distribution indicated that the crop suffered intermittent drought stress during active growth and development. The mean yield under rainfed conditions was 1560 kg ha-1 compared with the mean irrigated yield of 2063 kg ha-1 (Figure 17), although the difference in drought versus irrigated yield was wider for the commercial checks. 66 Table 29. Origin, growth habit, seed color, days to flowering, days to maturity and 100 seed weight of 36 bean genotypes tested in a Mollisol at Palmira. Genotype Origin A 686 Colombia Growth habit 2 A 774 BAT 477 Carioca Colombia Colombia Brazil 3 2 3 Cowpea Mouride DOR 390 EAP 9503-32B EAP 9653-16B-1 G19902* Senegal Colombia Honduras Honduras Argentina 2 2 2 2 4 G24390* Mexico 4 G40001** NCB 226 NCB 280 Perola Mexico Colombia Colombia Brazil 4 2B 2A 3 RCB 273 San Cristobal 83 2B 2 SEA 15 SEA 5 SEN 36 SEN 56 SER 109 SER 113 SER 118 SER 119 SER 125 SER 128 SER 16 SER 48 SER 78 SER 90 SXB 405 SXB 409 Colombia Dom. Republic Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia Colombia 2 2 2 2 2A 2 2A 2A 2A 2A 2 2 2A 2 2B 2B SXB 412 SXB 415 Colombia Colombia 2A 2A SXB 418 Colombia 2B Tio Canela 75 Honduras 2 Seed color Creambrown Cream Cream Creambrown Cream Black Red Red Creambrown Creamblack White Black Black Creambrown Red Redcream Roxo Cream Black Black Red Red Red Red Red Red Red Red Red Red Cream Creambrown Cream Creambrown Creambrown Red Days to flowering Irrigated Rainfed 39 38 Days to maturity Irrigated Rainfed 70 73 100 seed weight Irrigated Rainfed 29 28 37 38 39 38 38 38 68 70 68 66 64 67 28 24 28 25 21 24 48 38 36 37 37 45 38 37 38 37 70 69 68 67 69 79 72 65 71 67 15 22 24 24 5 14 22 22 21 7 44 43 80 78 4 4 34 35 33 39 32 34 31 40 63 68 65 70 60 68 61 73 12 33 29 30 12 33 27 27 35 38 33 38 66 70 62 65 24 30 24 27 36 33 37 35 35 37 36 35 35 35 34 35 36 35 39 38 31 33 37 32 33 35 36 33 32 32 32 34 35 32 37 38 68 63 67 66 65 66 65 67 66 67 65 65 65 66 67 68 60 65 69 63 63 64 65 63 62 63 61 63 64 62 65 66 32 30 26 28 29 29 29 28 28 30 27 34 24 32 30 31 34 28 24 28 25 28 28 27 29 29 25 32 23 31 27 28 37 37 37 37 67 69 65 66 28 29 25 28 37 38 70 67 32 28 37 37 68 68 24 22 * Wild common bean ** Phaseolus acutifolius 67 80 Palmira Rainfall Pan evaporation Maximum temperature Minimum temperature 60 60 40 40 20 20 0 Temperature (°C) Rainfall and pan evaporation (mm day-1) 80 0 0 10 20 30 40 50 60 70 80 Days after sown Figure 16. Rainfall distribution, pan evaporation, maximum and minimum temperatures during crop growing period at Palmira in 2007. Rainfed grain yield (kg ha-1) 2500 SEN 56 PALMIRA NCB 226 SER 125 SER 113 SER 16 SER 119 SXB 412 SER 118 SER 109 SEA 5 RCB 273 Cowpea M. SER 90 Tio Canela SXB 405 EAP 9653-16B-1 G 40001 BAT 477 Perola Carioca 2000 1500 Mean: 1560 LSD0.05: 449 EAP 9503-32B 1000 500 G 19902 Mean: 2063 LSD0.05: 565 G 24390 0 0 500 1000 1500 2000 2500 3000 3500 Irrigated grain yield (kg ha-1) Figure 17. Identification of genotypes that are adapted to rainfed conditions and are responsive to irrigation in a Mollisol at Palmira. Genotypes that yielded superior with drought and were also responsive to irrigation were identified in the upper, right hand quadrant. 68 Under drought stress conditions in the field, the seed yield of 36 genotypes ranged from 200 to 2374 kg ha-1. Among the lines tested, the lines NCB 226, SEN 56, SER 113, SER 125 and SER 16 were outstanding in their adaptation to rainfed (drought stress) conditions. These lines were also responsive to irrigation (Figure 17). Among the 36 lines tested, G19902 (Andean wild bean germplasm accession) and G24390 (MesoAmerican wild bean germplasm accession) were the most poorly adapted lines under both irrigated and rainfed conditions (Figure 17). Under rainfed conditions, significant genotypic differences were observed in canopy biomass production at mid-pod filling growth stage (Figure 18). Cowpea cv. Mouride showed greater vigor than the common bean lines. However this line showed lower value of harvest index indicating a limitation on mobilization of photosynthates to pod development. Among genotypes of P. vulgaris tested, SEN 56 and NCB 226 were outstanding in canopy biomass production (Figure 18); also the line SEN 56 yielded well under rainfed conditions due to greater ability to partition photosynthetically assimilated carbon to seeds as reflected by higher values of harvest index (Figure 19). The line SER 118 was outstanding in its harvest index value but it had lower canopy biomass under rainfed conditions indicating the need for adequate vegetative vigor to achieve higher values of grain yield. Among the 36 genotypes evaluated, G19902 and G24390 presented the lower values of canopy biomass and grain yield (Figure 18). (a) 3000 SXB 418 SER 16 Carioca SER 109 SXB 415 BAT 477 Perola SER 125 Tio Canela SER 119 SER 118 RCB 273 G 40001 2000 1500 1000 (b) NCB 226 SEN 56 2500 Grain yield (kg ha-1) Rainfed Irrigated 3500 Cowpea M Mean: 2063 LSD0.05: 565 G 24390 500 Mean: 4677 LSD0.05: 2121 G 19902 0 0 2000 SEN 56 NCB 226 SER 125 SXB 415 SER 118 SEA 5 Cowpea SXB 405 BAT 477 G 40001 Carioca EAP 9503-32B 4000 6000 Mean: 1560 LSD0.05: 449 G 19902 Mean: 3530 G 24390 LSD0.05: 1115 8000 10000 12000 0 2000 4000 6000 8000 10000 12000 Canopy biomass (kg ha-1) 3500 (c) SXB 418 Carioca SXB 412 SER 109 LSD0.05: 565 SXB 415 SEA 15 Cowpea SER 113 SER 125 SER 119 SER 118 Tio Canela RCB 273 G 40001 2500 Mean: 2063 2000 1500 (d) NCB 226 SEN 56 3000 SER 90 SEN 56 NCB 226 SER 125 SER 113 SER 75 SER 109 SER 118 Tio Canela SEN 36 Mean: 1560 Perola BAT 477 G 40001 LSD0.05: 449 EAP 9503-32B 1000 G 24390 500 G 19902 0 0 20 40 Mean: 58 LSD0.05: 42 60 G 24390 80 100 0 20 G 19902 Mean: 48.8 LSD0.05: 42 40 60 80 100 Harvest index (%) Figure 18. The relationship between grain yield and irrigated and rainfed canopy biomass (a, b) and grain yield and irrigated and rainfed harvest index (c, d) when grown in a Mollisol at Palmira. 69 Results on the relationship between rainfed pod harvest index (PHI) and rainfed grain yield indicated that SER 125, NCB 226 and SEN 56 were superior in mobilizing photosynthates from pod wall to seeds (Figure 19). The PHI values of G19902 and G24390 were markedly lower than that of other bean genotypes. The superior performance of lines SER 125, NCB 226 and SEN 56 was associated with greater values of pod harvest index, higher values of stem biomass reduction (Figure 19), higher values of seed and pod number per area, and 100 seed weight (Figure 20). Higher stem biomass reduction values indicate higher photosynthate mobilization from the stem to other plant structures such as pods. Irrigated 3500 (a) NCB 226 Mean: 2063 SEN 56 LSD0.05: 565 SXB 418 Carioca SER 16 EAP 9653-16B-1 SXB 415 SER 128 Perola SXB 409 SER 113 Tio Canela SER 118 EAP 9503-32B SER 119 SXB 405 RCB 273 G 40001 3000 2500 2000 1500 Grain yield (kg ha-1) Rainfed 1000 (b) SEN 56 NCB 226 SER 125 SXB 415 SER 123 SER 16 SER 78 SER 109 Tio CanelaSXB 409 SEA 5 SEN 36 EAP 9653-16B-1 Perola BAT 477 EAP 9503-32B Mean: 1560 LSD0.05: 449 Cowpea G 24390 500 Mean: 15.9 LSD0.05: NS G 19902 0 -40 -20 0 20 40 80 -40 60 G 19902 G 24390 Mean: 42.7 LSD0.05: 31 -20 20 0 40 60 80 Stem biomass reduction (%) 3500 (c) 2500 (d) NCB 226 SEN 56 3000 SXB 418 SER 16 SER 109 Carioca SER 128 Cowpea SER 119 SER 118 RCB 273 G 40001 Mean: 2063 LSD0.05: 565 2000 1500 1000 SEN 56 NCB 226 SER 125 SER 16 SXB 412 SXB 409 Cowpea SXB 405 BAT 477 G 40001 Carioca EAP 9503-32B Mean: 1560 LSD0.05: 449 G 24390 500 G 19902 Mean: 80.6 LSD0.05: 2.7 G 24390 G 19902 Mean: 76.8 LSD0.05: 3.4 0 30 40 50 60 70 80 90 30 40 50 60 70 80 90 Pod harvest index (%) Figure 19. The relationship between grain yield and irrigated and rainfed stem biomass reduction (a, b), grain yield and irrigated and rainfed pod harvest index (c, d) when grown in a Mollisol at Palmira. Yield production efficiency of SER 118 under rainfed conditions was outstanding and this was because of its greater value of harvest index while its canopy biomass production was only above average value (Figures 21, 18). Yield production efficiency is an integrated measure for photosynthate mobilization and SER 118 could be an excellent parent to improve yield under rainfed conditions. 70 Irrigated 3500 (a) 3000 Rainfed NCB 226 (b) SEN 56 SXB 418 SER 16 Carioca SXB 412 SER 109 Cowpea SER 118 Tio Canela SER 119 RCB 273 G 40001 2500 2000 1500 Cowpea Mean: 2063 LSD0.05: 565 1000 G 19902 500 0 0 SEN 56 NCB 226 SER 125 SER 118 SXB 412 DOR 390 SXB 405 EAP 9653-16B-1 Perola BAT 477 G 40001 EAP 9503-32B SER 113 G 24390 Mean: 1093 LSD0.05: 543 Cowpea Mean: 1560 LSD0.05: 449 Mean: 860 G 19902 G 24390 LSD0.05: 667 500 1000 1500 2000 2500 3000 3500 0 500 1000 1500 2000 2500 3000 3500 Grain yield (kg ha-1) Seed number per area (no. m-2) 3500 3000 2500 2000 1500 (d) (c) SEN 56 NCB 226 SXB 418 Carioca SER 109 SER 16 DOR 390 SEA 5 SER 48 BAT 477 SER 119 SXB 405 RCB 273 G 40001 Mean: 2063 LSD0.05: 565 Cowpea 1000 SEN 56 NCB 226 SER 113 SER 125 SXB 415 SER 16 SXB 409 SER 109 SXB 405 EAP 9653-16B-1 Perola G 40001 EAP 9503-32B G 24390 500 G 19902 Mean: 239 LSD0.05: 109 Mean: 200 LSD0.05: 112 G 19902 Mean: 1560 LSD0.05: 449 Cowpea G 24390 0 100 150 200 250 300 350 400 450 500100 150 200 250 300 350 400 450 500 Pod number per area (no. m-2) 3500 3000 2500 2000 1500 (e) (f) NCB 226 SEN 56 SXB 418 SER 16 SER 90 DOR 390 Carioca SEN 36 SER 128 Cowpea BAT 477 SER 48 Mean: 2063 Tio Canela SXB 405 LSD0.05: 565 RCB 273 G 40001 1000 G 24390 500 0 5 10 G 19902 Mean: 26.1 LSD0.05: 1.9 G 19902 0 SEN 56 NCB 226 SER 113 SER 125 SER 16 Mean: 1560 SER 48SEA 15 LSD0.05: 449 Cowpea SER 109 SER 90 Tio Canela SXB 405 G 40001 BAT 477 Perola EAP 9503-32B 15 20 25 30 Mean: 24.6 LSD0.05: 1.9 G 24390 35 40 0 5 10 15 20 25 30 35 40 100 seed weight (g) Figure 20. The relationship between grain yield and irrigated and rainfed seed number per area (a, b), grain yield and irrigated and rainfed pod number per area (c, d), and grain yield and irrigated and rainfed 100 seed weight (e, f) when grown in a Mollisol at Palmira. 71 Irrigated 3500 (a) 3000 Rainfed (b) NCB 226 SER 90 SER 16 SER 90 Carioca SXB 418 SXB 412 SER 109 SXB 415 SER 113 SEA 15 Cowpea BAT 477 Mean: 2063 SER 119 Tio Canela LSD0.05: 565 RCB 273 G 40001 2500 2000 1500 SEN 56 NCB 226 SER 125 SER 113 SER 119 A 774 SER 109 Tio CanelaSXBCowpea 405 Perola BAT 477G 40001 EAP 9530-32B 1000 Mean: 1560 LSD0.05: 449 G 24390 500 Mean: 0.59 LSD0.05: 0.41 G 19902 0 0.0 Grain yield (kg ha-1) SER 118 2500 2000 1500 0.4 0.6 0.8 1.0 1.2 0.0 (c) SEN 56 418 SER 16 SXB Carioca SXB 412 SER 109 SEA 15 Cowpea SER 125 SER 118 Tio Canela RCB 273 G 40001 Mean: 2063 LSD0.05: 565 G 19902 Mean: 2.61 LSD0.05: 2.35 500 0 0 3000 2500 2000 1500 0.4 0.6 0.8 1.0 1.2 (d) NCB 226 1000 3500 0.2 -1 Yield production efficiency (g g ) 3500 3000 0.2 G 24390 G 19902 Mean: 0.49 LSD0.05: 0.29 2 4 SEN 56 NCB 226 SER 113 SER 125 SER 118 SER 128 SER 109 Cowpea SXB 405 Perola BAT 477 G 40001 EAP 9503-32B G 24390 6 8 10 G 19902 Mean: 2.65 LSD0.05: 1.83 12 0 2 4 6 -1 Seed production efficiency (no. g ) (f) (e) Mean: 1560 LSD0.05: 449 G 24390 8 10 12 NCB 226 SEN 56 SER 16 SXB 418 SXB 412 SXB 415 SER 109 SEA 15 SER 125 Tio Canela SER 118 RAC 273 G 40001 Mean: 2063 LSD0.05: 565 1000 G 24390 500 G 19902 0 0.0 0.5 1.0 SEN 56 NCB 226 SER 113 SER 125 SXB 415 SER 118 SEA 5 Cowpea SXB 405 BAT 477 G 40001 EAP 9503-32B Mean: 0.58 LSD0.05: 0.60 1.5 G 19902 Mean: 0.64 LSD0.05: 0.46 2.0 3.0 0.0 2.5 0.5 1.0 Mean: 1560 LSD0.05: 449 1.5 2.0 G 24390 2.5 3.0 -1 Pod production efficiency (no. g ) Figure 21. The relationship between grain yield and irrigated and rainfed yield production efficiency (a, b), grain yield and irrigated and rainfed seed production efficiency (c, d), and grain yield and irrigated and rainfed pod production efficiency (e, f) when grown in a Mollisol at Palmira. 72 Since a major role of transpiration is leaf cooling, canopy temperature depression (CTD) relative to ambient air temperature is an indication of how capable is transpiration in cooling the leaves under a demanding environmental load. Relatively lower canopy temperature under drought stress could indicate a relatively better capacity for taking up soil moisture and for maintaining a relatively better plant water status. But CTD can be a poor indicator of resistance to drought if grain yield is highly dependent on limited amounts of soil-stored water. Relationship between CTD (canopy and ambient temperature difference at 1 pm) with rainfed grain yield indicated that the lines SEN 56, NCB 226, SEA 5, Cowpea, Carioca, G19902 and G24390 showed higher values of CTD that indicate greater rates of transpirational water loss (Figure 22). However SEN 56 and NCB 226 combined higher values of CTD with higher grain yield under rainfed conditions while Carioca, G19902 and G24390 yielded less. Thus it is important to identify which genotypes are resisting drought by combining mechanisms such as photosynthate mobilization with efficient water use (e.g., SER lines including SER 118), and which genotypes are using deeper roots to maintain higher values of CTD but were not efficient in using water to produce greater seed yield under rainfed conditions (e.g., Carioca, G19902 and G24390). Rainfed grain yield (kg ha-1) 3000 SEN 56 PALMIRA NCB 226 2500 SXB 415 SER 16 2000 1500 Cowpea M SEA 5 SER 125 SER 113 SER 78 SER 119 SER 118 SER 109 Tio Canela SXB 409 SER 90 EAP 9653-16B-1 G 40001 BAT 477 Perola SXB 418 Mean: 1812 LSD0.05: 518 Carioca EAP 9503-32B 1000 500 G 19902 Mean: -2.9 LSD0.05: 2.6 G 24390 0 -8 -7 -6 -5 -4 -3 -2 -1 0 Canopy temperature depression (oC) Figure 22. Identification of genotypes that combine superior seed yield with lower values of canopy temperature depression (CTD) when grown under rainfed conditions in a Mollisol at Palmira. Genotypes with greater seed yield and minimum differences were identified in the upper, right hand quadrant. Among the 8 genotypes evaluated for root attributes, the line SER 16 and G24390 (Table 30) presented the higher values of total root production under rainfed conditions in terms of length while SEA 5 and G40001 presented the lowest production. Results on root distribution through soil profile showed that G24390, SEA 5 and Tio Canela 75 had developed deeper root system as revealed by the length and biomass of roots at 40-60 cm soil depth (Table 30). By comparing the root distribution data with CTD values, genotypes such as G24390, Tio Canela 75 and SEA 5 used deeper roots to cool the leaves, i.e., higher CTD values probably due to higher rates of transpiration. But lines such as SER 16 presented relatively less amount of roots and lower values of CTD with higher grain yield under drought conditions compared with the other genotypes. These results indicate that SER 16 was more water use efficient than G24390, Tio Canela 75 and SEA 5 due to its greater ability to mobilize photosynthates to grain. Further work is needed to verify these observations. 73 Table 30. Root attributes evaluated across soil depth of 8 bean elite lines grown under irrigated and rainfed conditions in a Mollisol in Palmira. Genotype DOR 390 G24390 G40001 SEA 5 SER 16 SER 109 SXB 418 Tío Canela 75 Soil depth (cm) 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 0-5 5-10 10-20 20-40 40-60 Root length (m m-3) Root biomass (g m-3) Root diameter (mm) Specific root length (m g-1) Irrigated Rainfed Irrigated Rainfed Irrigated Rainfed Irrigated Rainfed 3842 4799 2907 1669 1168 4898 5912 3587 1532 1478 2870 6037 4571 1445 736 7145 7071 3059 2007 2162 3239 8679 5003 866 690 4854 7222 4266 716 817 2523 7391 4728 1626 1599 6551 8067 6400 2371 935 9658 9896 4894 805 660 4564 12148 4436 2975 1947 4265 10012 3841 1272 541 9056 5691 2912 2383 2134 8361 13573 3400 1414 1342 6342 9740 4554 2413 1645 4234 9871 4588 2454 1565 5424 13356 3866 1629 1771 33 51 25 21 16 31 47 25 10 12 35 57 46 15 8 72 77 37 23 21 34 72 39 7 6 47 69 37 7 7 27 61 50 15 14 51 63 50 21 7 136 119 48 12 7 28 91 41 30 18 28 71 40 14 5 98 89 39 34 26 78 132 32 15 14 60 86 49 29 17 40 151 64 25 16 60 137 40 15 15 0.25 0.27 0.25 0.30 0.29 0.25 0.25 0.25 0.26 0.29 0.27 0.26 0.27 0.29 0.32 0.28 0.27 0.30 0.34 0.33 0.25 0.25 0.25 0.28 0.27 0.26 0.26 0.26 0.29 0.30 0.32 0.25 0.27 0.29 0.28 0.25 0.24 0.25 0.30 0.30 0.28 0.26 0.26 0.33 0.31 0.26 0.26 0.29 0.30 0.30 0.26 0.25 0.27 0.29 0.30 0.29 0.35 0.29 0.35 0.33 0.27 0.26 0.28 0.29 0.30 0.26 0.25 0.27 0.29 0.29 0.27 0.29 0.29 0.31 0.31 0.28 0.27 0.29 0.28 0.30 110 100 114 80 75 156 129 142 147 101 77 101 101 93 91 99 100 77 96 105 100 121 130 116 96 102 102 111 104 105 92 122 89 111 124 127 134 128 113 135 77 93 102 69 100 143 139 105 99 118 169 148 96 96 101 86 56 82 70 89 96 104 104 96 97 101 113 96 84 89 106 84 78 95 93 98 101 97 115 117 74 Correlation coefficients between final grain yield and other shoot attributes under rainfed conditions indicated that greater seed yield was positively related to leaf area index, canopy biomass, harvest index, pod harvest index, stem biomass reduction, shoot and seed TNC content (Table 31). Significant negative correlations were observed between rainfed grain yield and seed production efficiency, pod production efficiency, days to flowering and days to maturity. These significant negative associations indicate that while earliness has contributed to superior performance under rainfed conditions, the formation of pods and seeds was not the factor limiting the grain yield. It was rather the ability to fill seeds as reflected by the significant positive associations between grain yield and harvest index, pod harvest index and 100 seed weight under rainfed conditions. Alternatively, the correlations were markedly influenced by including the two wild bean accessions that had greater values of pod and seed production efficiency per unit dry weight of shoot biomass. Table 31. Correlation coefficients (r) between final grain yield (kg ha-1) and other shoot attributes of elite lines grown under irrigated and rainfed conditions in a Mollisol in Palmira. Plant traits Leaf area index (m2/m2) Total chlorophyll content (SPAD) Canopy biomass (kg ha-1) Canopy temperature depression (oC) Shoot TNC content (mg g-1) Seed TNC content (mg g-1) Pod partitioning index (%) Harvest index (%) Pod harvest index (%) Yield production efficiency (g g-1) Seed production efficiency (no. g-1) Pod production efficiency (no. g-1) Stem biomass reduction (%) Days to flowering Pod number per area (no. m-2) Seed number per area (no. m-2) Days to maturity 100 seed weight (g) Grain filling index (%) Irrigated 0.45*** -0.01 0.48*** -0.46*** 0.07 0.27** 0.18 0.23* 0.63*** 0.23* -0.37*** -0.39*** -0.01 -0.26** 0.006 -0.004 -0.29** 0.67*** Rainfed 0.25** 0.01 0.47*** -0.10 0.19* 0.23* 0.14 0.32** 0.71*** 0.32** -0.38*** -0.43*** 0.30** -0.46*** -0.007 0.04 -0.34** 0.68*** -0.18* *, **, *** Significant at the 0.05, 0.01 and 0.001 probability levels, respectively. Conclusions: Field evaluation of elite lines at Palmira resulted in identification of five lines NCB 226, SEN 56, SER 113, SER 125 and SER 16 that were outstanding in their adaptation to drought stress conditions. The superior performance of these lines under drought stress was associated with higher values of harvest index, pod harvest index, leaf area index and canopy biomass. In contrast, results on root growth and distribution under field conditions indicated that G24390, Tio Canela 75 and SEA 5 were more deep rooted than the drought adapted SER 16, suggesting that root growth in G24390 and Tio Canela 75 occurred at the expense of photosynthate mobilization to seed under drought stress. G19902 and G24390 were identified as the most poorly adapted to drought, suggesting that wild beans do not have inherent drought resistance, and that domestication has improved this trait. The SER lines that were developed in the last few years seem to combine the desirable traits for drought adaptation such as greater mobilization of photosynthates to seed with efficient use of water through stomatal control. Contributors: J. Polanía, M. Grajales, C. Cajiao, R. García, J. Ricaurte, S. Beebe and I. M. Rao 75 2.1.1.8 Physiological evaluation of drought resistance of 33 recombinant inbred lines (RILs) of DOR 364 x BAT 477 under terminal drought stress over two seasons Rationale: The bred line BAT 477 is very well adapted to drought while DOR 364 is a commercial variety in Central America and is less adapted to drought stress. We evaluated drought adaptation of 33 RILs of the cross DOR 364 x BAT 477 over two seasons to obtain phenotypic data for eventual gene tagging for drought resistance. Materials and Methods: Two field trials were conducted at Palmira in 2005 (June to September) and 2006 (June to September). The two trials included 97 RILs of DOR 364 x BAT 477 along with 1 check (SEA 5) and 2 parents (DOR 364, BAT 477) but only 33 RILs were selected for intensive characterization to determine genotypic differences in tolerance to drought stress conditions. A 10 x 10 balanced lattice design with 3 replicates was used. Two levels of water supply (irrigated and rainfed) were applied. For the irrigated treatment, a total of 5 gravity irrigations were applied while for the rainfed treatment only 2 irrigations (approximately 35 mm each) in 2005 and 3 in 2006 were applied to assure good crop establishment. Details on planting and management of the trial were similar to those reported before. Experimental units consisted of 2 rows, 3.72 m long by 0.6 m wide. A number of plant attributes were measured at mid-pod filling only under rainfed conditions in 2005 and under both conditions in 2006 in order to determine genotypic variation in drought resistance. These plant traits included leaf chlorophyll content (SPAD); leaf area index; canopy dry weight per plant; shoot nutrient (N, P) uptake; shoot and seed ash content; and shoot and seed TNC (total nonstructural carbohydrates). At the time of harvest, grain yield and yield components (number of pods per plant, number of seeds per pod, and 100 seed weight) were determined. Stem biomass reduction (mobilization of photosynthate reserves) was determined based on difference in stem dry weight at harvest from the stem dry weight at mid-pod filling. Pod partitioning index (dry wt of pods at harvest/dry wt of total biomass at mid-podfill x 100), pod number per area, seed number per area, pod harvest index (dry wt of seed/dry wt of pod at harvest x 100), yield production efficiency (seed biomass dry weight at harvest/total shoot biomass dry weight at midpod filling), seed production efficiency (seed number per area/ total shoot biomass dry weight at mid-pod filling per area), pod production efficiency (pod number per area/ total shoot biomass dry weight at midpod filling per area) and grain filling index (100 seed weight of rainfed/100 seed weight of irrigated) were determined. Seed P content, ash content and TNC (total nonstructural carbohydrate) content were also measured. Results and Discussion: Palmira – Soil, temperature, rainfall and evaporation: The soil is a Mollisol (Aquic Hapludoll) with no major fertility problems (pH = 7.7), and is estimated to permit storage of 130 mm of available water (assuming 1.0 m of effective root growth with –0.03 MPa and –1.5 MPa upper and lower limits for soil matric potential). During the crop-growing season, maximum and minimum air temperatures were 34.5 and 15.8 °C in 2005 and 34.2 and 16 oC in 2006 (Figure 23). The incident solar radiation ranged from 10.2 to 22.8 MJ m-2 d-1 in 2005 and 9.2 to 23.9 MJ m-2 d-1 in 2006. The total rainfall during the active crop growth was 126.4 mm in 2005 (most of which fell at the end of the crop growing season) and 33.2 in 2006. The potential pan evaporation was of 400.5 mm in 2005 and 410.7 in 2006. These data on rainfall and pan evaporation together with rainfall distribution indicated that the crop suffered significant terminal drought stress during active growth and development in both years. The mean yield under rainfed conditions was 723 kg ha-1 compared with the mean irrigated yield of 1655 kg ha-1 (56% decrease in grain yield due to drought stress). Under drought stress conditions in the field, the seed yield of 33 RILs ranged from 537 to 1029 kg ha-1 (Figure 24). Among the RILs tested, two RILs BT 21138-17-1-1 and BT 21138-6-1-1 were outstanding in their adaptation to rainfed (water stress) conditions. The relationship between grain yield of rainfed and irrigated treatments indicated that several RILs were superior to the best parent, BAT 477 and the check 76 genotype, SEA 5. Among the 33 lines tested, BT 21138-31-1-1 and BT 21138-28-1-1 were found to be very poorly adapted RILs under rainfed conditions. Significant genotypic differences were observed in canopy biomass production at mid-pod filling growth stage (Figure 24). The RILs BT 21138-30-1-1, BT 21138-3-1-1 and BT 21138-83-1-1 showed greater vigor than the rest of the RILs; however these RILs showed lower value of harvest index indicating a limitation on mobilization of photosynthates to pod and seed development. The two RILs BT 21138-17-11 and BT 21138-6-1-1 with moderate values of canopy biomass (Figure 24) yielded well under rainfed conditions due to greater ability to partition photosynthetically assimilated carbon to pods as reflected by higher values of harvest index (Figure 24). The relationship between rainfed seed yield and other plant attributes indicated that the outstanding performance of lines BT 21138-17-1-1 and BT 21138-6-1-1 was associated with higher values of pod harvest index (Figure 24) indicating greater mobilization of photosynthates to the grain. 60 60 50 40 40 30 30 20 20 10 10 0 60 0 60 Rainfall Pan evaporation Maximum temperature Minimum temperature Palmira - 2006 50 50 40 40 30 30 20 20 10 10 0 Temperature (°C) Rainfall and pan evaporation (mm day-1) Palmira - 2005 50 0 0 10 20 30 40 50 60 70 80 Days after sown Figure 23. Rainfall distribution, pan evaporation, maximum and minimum temperatures during crop growing period at Palmira in 2005 and 2006. 77 1100 Mean: 1655 LSD0.05: 347 (a) 1000 900 Mean: 723 LSD0.05: 241 BT 36-1-1 Mean: 723 LSD0.05: 241 BT 96-1-1 BT 57-1-1 BT 124-1-3 BT 142-1-2 BT 73-1-1 BT 97-1-1 BT 7-1-1 BAT 477 BT 124-1-1 BT 69-1-1 BT 16-1-1 BT 3-1-1 BT 147-3 BT 50-1-1 DOR 364 BT 30-1-1 BT 51-1-1 BT 28-1-1 BT 31-1-1 BT 57-1-1 700 600 500 Rainfed grain yield (kg ha-1) BT 6-1-1 BT 83-1-1 BT 4-1-1 BT 64-1-1 BT 96-1-1 BT 124-1-2 BT 124-1-3 BT 7-1-1 BT 16-1-1 BT 84-1-1 BAT 477 BT 98-1-1 BT 69-1-1 BT 124-1-1 BT 30-1-1 BT 83-1-3 BT 50-1-1 DOR 364 BT 104-3 BT 25-1-1 BT 51-1-1 BT 28-1-1 BT 31-1-1 800 13 SEA 5 00 14 00 15 900 600 500 00 17 00 18 00 19 00 00 20 20 00 25 Mean: 723 LSD0.05: 241 30 Mean: 33 LSD0.05: 22 (e) 1000 900 800 700 600 500 60 70 80 BT 6-1-1 90 64 40 66 (f) BT 17-1-1 00 BT 64-1-1 BT 96-1-1 BT 124-1-3 BT 69-1-1 BT 98-1-1 68 70 Mean: 873 LSD0.05: 312 SEA 5 50 BAT 477 BT 13-1-1 72 74 76 78 BT 36-1-1 BT 17-1-1 BT 83-1-1 BT 6-1-1 BT 4-1-1 Mean: 723 LSD0.05: 241 BT 96-1-1 BT 64-1-1 BT 124-1-2 BT 57-1-1 BT 131-1-1BT 7-1-1 BAT 477 BT 97-1-1 BT 23-1-4 BT 84-1-1 BT 124-1-1 BT 98-1-1 BT 30-1-1 BT 83-1-3 BT 50-1-1 DOR 364 BT 51-1-1 BT 104-3 BT 28-1-1 BT 31-1-1 60 500 600 700 800 900 1000110012001300 Stem biomass reduction (%) Figure 24. 45 Pod harvest index (%) BT 4-1-1 BT 36-1-1 Mean: 723 LSD0.05: 241 BT 64-1-1 BT 96-1-1 BT 57-1-1 BT 131-1-1 BT 124-1-2 BT 124-1-3 BT 7-1-1 BAT 477 BT 124-1-1 BT 84-1-1 BT 16-1-1 BT 98-1-1 BT 30-1-1 BT 50-1-1 BT 147-3 DOR 364 BT 25-1-1 BT 28-1-1 BT 31-1-1 30 00 BT 83-1-3 BT 30-1-1 DOR 364 BT 28-1-1 BT 51-1-1 BT 104-3 BT 31-1-1 BT 83-1-1 20 40 BT 17-1-1 BT 6-1-1 SEA 5 BT 83-1-1 BT 4-1-1 BT 36-1-1 BT 7-1-1 BT 23-1-4 SEA 5 10 00 BT 7-1-1 BT 1-1-1 Harvest index (%) 1100 35 Mean: 72 LSD0.05: 3.8 Mean: 723 LSD0.05: 241 BT 96-1-1 BT 64-1-1 BT 124-1-2 50 00 BT 4-1-1 BT 36-1-1 40 30 (d) BT 131-1-1 BT 57-1-1 BT 7-1-1 BT 97-1-1 BT 23-1-4 BT 84-1-1 BAT 477 BT 69-1-1 BT 124-1-1 BT 96-1-1 BT 147-3 BT 30-1-1 DOR 364 BT 50-1-1 BT 51-1-1 BT 28-1-1 BT 31-1-1 20 00 Canopy biomass (kg ha-1) Mean: 52 LSD0.05: 22 BT 17-1-1 BT 6-1-1 SEA 5 BT 83-1-1 (c) 1000 700 16 Irrigated grain yield (kg ha-1) 1100 800 00 Mean: 3465 LSD0.05: 901 BT 17-1-1 BT 6-1-1 SEA 5 BT 83-1-1 BT 4-1-1 BT 36-1-1 (b) BT 17-1-1 Seed number per area (no. m-2) Identification of genotypes that are (a) adapted to rainfed conditions and are responsive to irrigation; and combine higher values of rainfed grain yield with superior values of (b) canopy biomass, (c) harvest index, (d) pod harvest index, (e) stem biomass reduction, and (f) seed number per area 78 Correlation coefficients between final grain yield and other shoot attributes under rainfed conditions indicated significant negative relationship between seed yield and total chlorophyll content and seed N and P content under rainfed conditions (Table 32). This observation indicates that genotypes that mobilized greater amounts of N and P together with photosynthates yielded better under rainfed conditions. It is important to note that the pod harvest index was significantly associated with seed yield under rainfed conditions. This indicates that the genotypes that mobilized a greater proportion of photosyntates from pod to seed performed better under rainfed conditions (Figure 24). Significant positive associations were also observed between rainfed grain yield and seed number per area, pod number per area, yield production efficiency, seed production efficiency and pod production efficiency indicating the contribution of improved plant efficiency under drought stress to produce grain. However reduction in stem biomass showed no significant correlation with rainfed grain yield and this may be due to the late rains in 2005 season that might have allowed some lines to grow during seed filling. Table 32. Correlation coefficients (r) between final grain yield (kg ha-1) and other plant attributes of RILs of common bean grown under rainfed conditions in a Mollisol in Palmira. Plant traits Leaf area index (m2 m-2) Total chlorophyll content (SPAD) Canopy biomass (kg ha-1) Seed TNC Content (mg g-1) Seed N Content (%) Seed P Content (%) Pod partitioning index (%) Pod harvest index (%) Harvest index (%) Seed number per area (no. m-2) Pod number per area (no. m-2) Yield production efficiency (g g-1) Seed production efficiency (no. g-1) Pod production efficiency (no. g-1) Stem biomass reduction (%) Rainfed 0.16* -0.41*** 0.26*** -0.11 -0.42*** -0.31*** 0.40*** 0.43*** 0.41*** 0.56*** 0.45*** 0.41*** 0.41*** 0.27*** -0.12 *, **, *** Significant at the 0.05, 0.01 and 0.001 probability levels, respectively. Conclusions: Field evaluation of 33 RILs of the cross DOR 364 x BAT 477 at Palmira over two seasons under terminal drought stress conditions resulted in identification of two lines (BT 21138-17-1-1 and BT 21138-6-1-1) that were superior in their adaptation to drought stress conditions. The superior performance of these lines under drought stress was associated with higher values of harvest index, pod harvest index and seed and pod number per area indicating the importance of greater mobilization of photosyntates to pods and to seeds under rainfed conditions. Contributors: J. Polanía, M. Grajales, C. Cajiao, R. García, J. Ricaurte, S. Beebe and I. M. Rao 79 2.1.1.9 Physiological evaluation of drought resistance in recombinant inbred lines (RILs) of DOR 364 x BAT 477 under intermittent drought stress Rationale: We evaluated drought adaptation of 33 RILs of the cross DOR 364 x BAT 477 over two seasons (2005 and 2006) under terminal drought stress. In 2007, we evaluated 97 RILs of the cross DOR 364 x BAT 477 under intermittent drought stress to obtain phenotypic data for eventual gene tagging for drought resistance. Materials and Methods: A field trial was conducted at Palmira in 2007 (June to September). The soil is a Mollisol (Aquic Hapludoll) with no major fertility problems (pH = 7.7), and is estimated to permit storage of 130 mm of available water (assuming 1.0 m of effective root growth with –0.03 MPa and –1.5 MPa upper and lower limits for soil matric potential). The trial included 97 RILs of DOR 364 x BAT 477 along with 1 check (SEA 5) and 2 parents (DOR 364, BAT 477) to determine genotypic differences in tolerance to drought stress conditions. A 10 x 10 balanced lattice design with 3 replicates was used. Two levels of water supply (irrigated and rainfed) were applied. For the irrigated treatment, a total of 4 gravity irrigations (approximately 35 mm each) were applied while for the rainfed treatment only 2 irrigations were applied to assure good crop establishment. Experimental units consisted of 2 rows, 3.72 m long by 0.6 m wide. A number of plant attributes were measured at mid-podfilling under both rainfed and irrigated conditions in order to determine genotypic variation in drought resistance. These plant traits included leaf chlorophyll content (SPAD), canopy temperature, canopy temperature depression (CTD), leaf area index, canopy dry weight per plant and shoot TNC content (total nonstructural carbohydrates). Canopy temperature was measured with a Telatemp model AG-42D infrared thermometer. The instrument was held at a 45o angle at 50 cm from the canopy surface to measure canopy temperature and canopy temperature depression. At the time of harvest, grain yield and yield components (number of pods per plant, number of seeds per pod, and 100 seed weight) were determined. Stem biomass reduction (mobilization of photosynthate reserves) was determined based on difference in stem dry weight at harvest from the stem dry weight at mid-pod filling. Pod partitioning index (dry wt of pods at harvest/dry wt of total biomass at mid-podfill x 100), pod harvest index (dry wt of seed/dry wt of pod at harvest x 100), pod number per area, seed number per area, yield production efficiency (seed biomass dry weight at harvest/total shoot biomass dry weight at mid-pod filling), seed production efficiency (seed number per area/ total shoot biomass dry weight at mid-pod filling per area), pod production efficiency (pod number per area/ total shoot biomass dry weight at mid-pod filling per area) and grain filling index (100 seed weight of rainfed/100 seed weight of irrigated) were determined. Shoot and seed TNC (total nonstructural carbohydrate) contents were also measured. Results and Discussion: During the crop-growing season, maximum and minimum air temperatures were 30.56 and 18.61 oC (Figure 25). The incident solar radiation ranged from 11.2 to 25.1 MJ m-2 d-1. The total rainfall during the active crop growth was 243.1 mm (a significant portion falling during grainfilling). The potential pan evaporation was of 431 mm. These data on rainfall and pan evaporation together with rainfall distribution indicated that the crop suffered intermittent drought stress during active growth and development. The mean yield under rainfed conditions was 849 kg ha-1 compared with the mean irrigated yield of 1741 kg ha-1 showing 51% decrease in mean grain yield due to drought stress (Figure 26). Under drought stress conditions in the field, the seed yield of 97 RILs ranged from 603 to 1171 kg ha-1 (Figure 26). Among the RILs tested, two RILs BT 21138-68-1-1 and BT 21138-74-1-1 were outstanding in their adaptation to rainfed (water stress) conditions. The relationship between grain yield of rainfed and irrigated treatments indicated that several RILs were superior to the best parent, BAT 477 and the check genotype, SEA 5. Among the 97 lines tested, BT 21138-31-1-1 and BT 21138-34-1-1 were found to be very poorly adapted lines under rainfed conditions. 80 60 Rainfall Pan evaporation Maximum temperature Minimum temperature 50 50 40 40 30 30 20 20 10 10 0 Temperature (°C) Rainfall and pan evaporation (mm day-1) 60 0 0 10 20 30 40 50 60 70 80 Days after planting Figure 25. Rainfall distribution, pan evaporation, maximum and minimum temperatures during crop growing period at Palmira in 2007. Rainfed grain yield (kg ha-1) 1200 BT 68-1-1 PALMIRA 1100 BT 22-1-1 BT 77-1-1 BT 79-1-1 BT 96-1 BT 64-1-1 1000 BT 74-1-1 BT 35-1-1 BT 61-1-1 BT 39-1-1 BT 8-1-1 BT 62-1-1 BT 28-1-1 BT 9-1-1 BT 91-1-1 BT 36-1-1 BT 81-1-1 BT 21-1-1 DOR 364 BAT 477 BT 124-1-1 900 Mean: 849 LSD0.05: 238 800 BT 14-1-1 BT 15-1-1 BT 47-1-1 BT 59-1-1 700 600 500 1200 BT 128-1 SEA 5 BT 63-1-1 BT 104-3 BT 23-1-1 BT 69-1-1 1400 1600 BT 2-1-1 BT 66-1-1 BT 92-1-1 BT 50-1-1 BT 7-1-1 BT 134-1-1 BT 31-1-1 Mean: 1741 LSD0.05: 431 1800 2000 2200 2400 Irrigated grain yield (kg ha-1) Figure 26. Identification of genotypes that are adapted to rainfed conditions and are responsive to irrigation in a Mollisol at Palmira. Genotypes that yield superior with drought and were also responsive to irrigation were identified in the upper, right hand quadrant. Significant genotypic differences were observed in canopy biomass production at mid-pod filling growth stage (Figure 27). The RILs BT 21138-64-1-1 and BT 21138-69-1-1 showed greater vigor than the rest of the RILs; however these RILs showed lower value of harvest index indicating a limitation on mobilization of photosynthates to pod and seed development. The relationship between rainfed seed yield and other plant attributes showed that the outstanding performance of lines BT 21138-68-1-1 and BT 21138-74-1-1 was associated with higher values of stem biomass reduction indicating greater mobilization of photosynthates from stems to pods (Figure 28). 81 (a) 2000 Mean: 1741 LSD0.05: 431 1600 1200 Grain yield (kg ha-1) Rainfed Irrigated 2400 BT 9-1-1 BT 21-1-1 BT 39-1-1 BT 91-1-1 BT 28-1-1BT 54-1-1 BAT 477 BT 13-1-1 DOR 364BT 25-1-1 SEA 5 BT 88-1-1 BT 29-1-1 BT 7-1-1 BT 45-1-1 BT 104-1-1 BT 15-1-1 BT 14-1-1 800 400 1200 Mean: 5397 LSD0.05: 1683 2400 3600 4800 6000 Mean: 2227 LSD0.05: 805 (b) BT 22-1-1 BT 68-1-1 BT 74-1-1 BT 77-1-1 BT 64-1-1 BAT 477 BT 10 DOR 364 BT 58-1-1 SEA 5 BT 69-1-1 BT 33 BT 31-1-1 7200 1200 2400 3600 Mean: 849 LSD0.05: 238 4800 6000 7200 Canopy biomass (kg ha-1) 2400 (c) BT 9-1-1 BT 39-1-1 BT 2-1-1 BT 28-1-1 BT 81-1-1 BAT 477 DOR 364 SEA 5 BT 27-1-1 BT 22-1-1 BT 99-1-1 BT 15-1-1 BT 124-1-3 BT 104-3 BT 14-1-1 (d) Mean: 52 LSD0.05: NS BT 61-1-1 BT 48-1-1 2000 1600 1200 800 Mean: 1741 LSD0.05: 431 BT 68-1-1 BT 74-1-1 BT 22-1-1 BT 79-1-1 BT 77-1-1 BT 64-1-1 BT 128-1 BT 147-1-1 BAT 477 BT 10-1-1 BT 95-1-1 DOR 364 BT 25-1-1 SEA 5 BT 38-1-1 BT 57-1-1 BT 59-1-1 BT 20-1-1 BT 33-1-1 BT 31-1-1 Mean: 40 LSD0.05: NS 400 20 30 40 50 60 70 Mean: 849 LSD0.05: 238 80 90 100 20 30 40 50 60 70 80 90 100 Harvest index (%) Figure 27. The relationship between grain yield and irrigated and rainfed canopy biomass (a, b) and grain yield and irrigated and rainfed harvest index (c, d) when grown in a Mollisol at Palmira. The relationship between rainfed seed yield and seed number per area, pod number per area and 100 seed weight showed that the line BT 21138-74-1-1 was outstanding in producing greater number of seeds and pods and also in filling the grain (Figure 29). Correlation coefficients between final grain yield and other shoot attributes under rainfed conditions indicated significant relationship between seed yield and leaf area index and canopy biomass under irrigated and rainfed conditions (Table 33). It is important to note that the harvest index, pod partitioning index, pod harvest index, seed number per area and pod number per area showed significant positive association with seed yield under rainfed conditions. This indicates that the genotypes that mobilized a greater proportion of photosyntates from plant structures to pod and seed formation performed better under rainfed conditions (Figures 28 and 29). 82 Rainfed Irrigated 2400 (a) BT 9-1-1 BT 39-1-1 BT 21-1-1 BT 61-1-1 BT 76-1-1 BT 28-1-1 BT 48-1-1 BT 81-1-1 BAT 477 BT 12-1-1 BT 1-1-1 DOR 364 SEA 5 BT 29-1-1 BT 131-1-1 BT 45-1-1 BT 34-1-1 BT 57-1-1 BT 104-3 BT 59-1-1 BT 15-1-1 BT 14-1-1 Mean: 1741 LSD0.05: 431 2000 1600 1200 800 Mean: 28.4 LSD0.05: NS 400 -40 0 20 40 60 BT 22-1-1 BT 68-1-1 BT 74-1-1 BT 79-1-1 BT 16-1-1 BAT 477 BT 8-1-1 BT 10-1-1 DOR 364 BT 89-1-1 SEA 5 BT 88-1-1 Mean: 849 BT 104-3 BT 23-1-4 LSD0.05: 238 BT 31-1-1 80 -40 -20 0 20 40 60 80 Stem biomass reduction (%) 2400 Grain yield (kg ha-1) -20 Mean: 42.1 LSD0.05: NS (b) (c) BT 9-1-1 BT 39-1-1 BT 61-1-1 BT 21-1-1 BT 2-1-1 BT 76-1-1 BT 48-1-1 BT 81-1-1 BT 12-1-1 BAT 477 BT 27-1-1 DOR 364 BT 29-1-1 SEA 5 Mean: 1741 BT 70-1-1 LSD0.05: 431 BT 45-1-1 BT 124-1-3 BT 15-1-1 BT 104-3 BT 59-1-1 BT 14-1-1 2000 1600 1200 800 Mean: 51.3 LSD0.05: NS (d) Mean: 70 LSD0.05: NS Mean: 849 LSD0.05: 238 BT 68-1-1 BT 22 BT 74-1-1 BT 79 BT 77-1-1 BT 64-1-1 BT 128-1 BT 147-3 BAT 477 BT 53-1-1 BT 10 BT 95-1-1 BT 25-1-1 BT 58 DOR 364 SEA 5 BT 56 BT 89 BT 59-1-1 BT 57 BT 104-3 BT 20-1-1 BT 31-1-1 BT 7-1-1 BT 33-1-1 400 20 40 60 80 100 120 20 40 60 80 100 120 140 Pod partitioning index (%) 2400 (e) 2000 Mean: 1741 LSD0.05: 431 1600 1200 BT 9-1-1 BT 61-1-1 BT 39-1-1 BT 10 BT 48-1-1 BT 63-1-1 BT 65-1-1 BAT 477 BT 4 BT 93-1-1 DOR 364 BT 29-1-1 BT 50 SEA 5 BT 69-1-1 BT 33 BT 57-1-1 BT 47 BT 15-1-1 BT 14-1-1 800 Mean: 80 LSD0.05: 2 400 68 70 72 74 76 78 80 (f) Mean: 849 BT 22-1-1 BT 68-1-1 BT 74-1-1 LSD0.05: 238 BT 64-1-1 BT 77-1-1 BT 79-1-1 BT 27-1-1 BT 131-1-1 BAT 477 BT 28-1-1 DOR 364 BT 89-1-1 SEA 5 BT 45-1-1 BT 69-1-1 BT 33-1-1 BT 34-1-1 BT 31-1-1 84 68 82 Mean: 75.6 LSD0.05: 4 70 72 74 76 78 80 82 84 Pod harvest index (%) Figure 28. The relationship between grain yield and irrigated and rainfed stem biomass reduction (a, b), grain yield and irrigated and rainfed pod partitioning index (c, d), and grain yield and irrigated and rainfed pod partitioning index (e, f) when grown in a Mollisol at Palmira. 83 Rainfed Irrigated 2400 (a) BT 9-1-1 BT 39-1-1 BT 61-1-1 BT 21 BT 10 BT 48 BT 81-1-1 Mean: 1741 BT 56 BAT 477 BT 66 LSD0.05: 431 BT 4 DOR 364 BT 27 BT 29-1-1 SEA 5 BT 24 BT 98 BT 35-1-1 BT 47-1-1 BT 15-1-1 BT 59-1-1 BT 124-1-3 BT 14-1-1 2000 1600 1200 800 Mean: 966 LSD0.05: 491 (b) Mean: 544 LSD0.05: 238 BT 68-1-1 BT 74-1-1 BT 22-1-1 BT 64-1-1 BT 95 BAT 477 BT 19-1-1 BT 25-1-1 DOR 364 BT 80-1-1 BT 59 SEA 5 BT 43-1-1 BT 34 BT 31-1-1 Mean: 849 LSD0.05: 238 400 200 400 600 800 1000 1200 1400 1600 200 400 600 800 1000 1200 1400 1600 Seed number per area (no. m-2) Grain yield (kg ha-1) 2400 (c) BT 9-1-1 BT 2-1-1 BT 39-1-1 BT 48-1-1 BT 76-1-1 BT 28-1-1 BT 62-1-1 BAT 477 BT 17-1-1 BT 13-1-1 DOR 364 BT 27 SEA 5 BT 70 BT 45-1-1 BT 98 BT 47 Mean: 1741 BT 59-1-1 BT 124-1-3 BT 57-1-1 LSD0.05: 431 BT 15-1-1 BT 14-1-1 2000 1600 1200 BT 61-1-1 800 Mean: 192 LSD0.05: 88 Mean: 145 LSD0.05: 55 (d) BT 68-1-1 BT 79BT 64 BT 74-1-1 BT 77 BT 19-1-1 BT 54 BAT 477 BT 6-1-1 BT 95 DOR 364 BT 54 BT 25-1-1 Mean: 849 BT 59-1-1 SEA 5 BT 80-1-1 LSD0.05: 238 BT 92 BT 33-1-1 BT 31-1-1 400 60 120 180 300 60 240 120 180 240 300 Pod number per area (no. m-2) 2400 (e) BT 9-1-1 BT 39-1-1 BT 2-1-1 BT 10-1-1 BT 76-1-1 BT 61-1-1 BT 66-1-1 BT 68-1-1 BAT 477 BT 31-1-1 DOR 364 BT 75-1-1 BT 19 BT 131-1-1 SEA 5 BT 43 BT 34-1-1 BT 45-1-1 BT 15 BT 57-1-1 BT 104-3 BT 59-1-1 BT 14-1-1 Mean: 1741 LSD0.05: 431 2000 1600 1200 800 Mean: 24.5 LSD0.05: 1.7 400 18 20 22 24 26 28 30 (f) Mean: 23.5 LSD0.05: 1.9 BT 74-1-1 BT 68-1-1 Mean: 849 BT 64-1-1 BT 22-1-1 LSD0.05: 238 BT 16-1-1 BT 79-1-1 BT 75-1-1 BAT 477 BT 55 BT 83-1-1 DOR 364 BT 46 BT 94 SEA 5 BT 104-3 BT 43-1-1 BT 33 BT 7-1-1 BT 34-1-1 BT 31-1-1 32 18 20 22 24 26 28 30 32 100 seed weight Figure 29. The relationship between grain yield and irrigated and rainfed seed number per area (a, b), grain yield and irrigated and rainfed pod number per area (c, d), and grain yield and irrigated and rainfed 100 seed weight (e, f) when grown in a Mollisol at Palmira. 84 Table 33. Correlation coefficients (r) between final grain yield (kg/ha) and other plant attributes of RILs of common bean grown under irrigated and rainfed conditions in a Mollisol in Palmira. Plant traits Leaf area index (m2/m2) Total chlorophyll content (SPAD) Canopy biomass (kg ha-1) Canopy temperature (oC) Canopy temperature depression (oC) Shoot TNC content (mg g-1) Seed TNC content (mg g-1) Pod partitioning index (%) Harvest index (%) Pod harvest index (%) Stem biomass reduction (%) Seed number per area (No m2) Pod number per area (No m2) Seed number per pod Days to flowering Days to maturity 100 seed weight (g) Grain filling index (%) Irrigated 0.19*** -0.12* 0.38*** -0.13* -0.16** 0.10 -0.09 0.02 0.03 0.11* 0.04 0.23*** 0.12* 0.17** 0.07 0.14* 0.11* Rainfed 0.34** 0.12* 0.33*** -0.08 -0.09 -0.02 -0.02 0.11* 0.13* 0.19*** -0.10 0.35*** 0.31*** 0.13* -0.05 0.08 0.11* 0.12* *, **, *** Significant at the 0.05, 0.01 and 0.001 probability levels, respectively. Conclusions: Field evaluation of 97 RILs of the cross DOR 364 x BAT 477 under intermittent drought stress resulted in identification of two RILs BT 21138-68-1-1 and BT 21138-74-1-1 that were outstanding in adaptation to intermittent drought stress conditions. The superior performance of these lines under intermittent drought stress was associated with higher values of harvest index, pod partitioning index, stem biomass reduction, seed number per area and pod number per area indicating the importance of greater mobilization of photosyntates to pods and seeds under rainfed conditions. Contributors: J. Polanía, M. Grajales, C. Cajiao, R. García, J. Ricaurte, S. Beebe and I. M. Rao 2.1.1.10 Evaluation of drought resistance in recombinant inbred lines (RILs) of MD 23-24 x SEA 5 under intermittent drought stress Rationale: We evaluated drought adaptation of 121 RILs of the cross MD 23-24 x SEA 5 over three seasons to obtain phenotypic data for eventual gene tagging. The bred line SEA 5 is very well adapted to drought while MD 23-24 is superior in commercial grain quality. The mean results over three seasons are reported. Materials and Methods: Three field trials were conducted at Palmira in 2003, 2004 and 2007 (June to September). The soil is a Mollisol (Aquic Hapludoll) with no major fertility problems (pH = 7.7), and is estimated to permit storage of 130 mm of available water (assuming 1.0 m of effective root growth with – 0.03 MPa and –1.5 MPa upper and lower limits for soil matric potential). The trials included 121 RILs of MD 23-24 x SEA 5 along with 5 checks (Cowpea, Tio Canela 75, DOR 390, EAP 9510-77 and SEA 15) 85 and 2 parents (MD 23-24, SEA 5) to determine genotypic differences in tolerance to drought stress conditions. An 11 x 11 balanced lattice design with 3 replicates was used. Two levels of water supply (irrigated and rainfed) were applied. Details on planting and management of the trial were similar to those reported before. Experimental units consisted of 2 rows, 3.72 m long by 0.6 m wide. A number of plant attributes were measured at mid-podfilling in order to determine genotypic variation in drought resistance. These plant traits included leaf chlorophyll content (SPAD), leaf area index; canopy dry weight per plant; shoot and seed ash content; and shoot and seed TNC (total nonstructural carbohydrates). At the time of harvest, grain yield and yield components (number of pods per plant, number of seeds per pod, 100 seed weight) were determined. Seed ash content and TNC (total nonstructural carbohydrates) were measure. Pod harvest index (seed weight/pod weight x 100) and grain filling index (100 seed weight of rainfed/100 seed weight of irrigated x 100) were also measured. Results and Discussion: During the crop-growing season, maximum and minimum air temperatures in 2003 were 33 and 14.6 °C, in 2004 were, 34.4 and 15.6 °C and in 2007 were, 30.5 and 18.6 °C, respectively (Figure 30). The incident solar radiation ranged from 8.2 to 23.3 MJ m-2 d-1 in 2003, 10.3 to 22.7 MJ m-2 d-1 in 2004 and 11.2 to 25.1 MJ m-2 d-1 in 2007. The total rainfall during the active crop growth was 126.5 mm in 2003, 110.4 mm in 2004 and 243.1 mm (a significant proportion of which fell during seed filling) in 2007. The potential pan evaporation was of 363 mm in 2003, 390 mm in 2004 and 431 mm in 2007. These data on rainfall and pan evaporation together with rainfall distribution indicated that the crop suffered intermittent drought during active growth and development. The mean yield under rainfed conditions was 1182 kg ha-1 compared with the mean irrigated yield of 1846 kg ha-1 with about 36% reduction of mean grain yield under drought stress (Figure 31). Under drought stress conditions in the field, the seed yield of 121 RILs ranged from 690 to 1574 kg ha-1 (Figure 31). Among the lines tested, three RILs, MR 81, MR 112 and MR 25 were outstanding in their adaptation to rainfed (water stress) conditions. These three lines were also responsive to irrigation. The relationship between grain yield of rainfed and irrigated treatments indicated that several RILs lines were superior to the best parent, SEA 5 and the 4 common bean check genotypes. Among the 121 lines tested, MR 8 was the most poorly adapted line under rainfed conditions. High significant correlation was observed between canopy biomass and grain yield; the genotype Cowpea Mouride showed the highest vigor with the highest canopy biomass under stress conditions (Figure 32). But this genotype showed a lower harvest index, indicating its limitation to mobilize photosyntates to seeds. Results on the relationship between rainfed grain yield and harvest index (HI) indicated that MR 25 and MR 81 were superior in mobilizing photosyntates to seeds (Figure 32). The HI value of EAP 951077, MR 109 and MR 40 was markedly lower than that of other bean genotypes. Results on the relationship between rainfed grain yield and pod harvest index indicated that MR 81 and MR 25 were superior in mobilizing photosyntates from pod to seeds (Figure 33). The superior performance of RILs MR25 and MR 81 was associated with greater values of harvest index, higher values of pod harvest index and seed number per area, higher values of stem biomass reduction (Figure 33). Higher stem biomass reduction values indicate higher photosyntates mobilization from the stem to other plant structures such as pods. 86 60 60 50 50 40 40 30 30 20 20 10 10 0 60 0 60 Palmira - 2004 Rainfall Pan evaporation Maximum temperature Minimum temperature 50 50 40 40 30 30 20 20 10 10 0 60 0 60 Temperature (°C) Rainfall and pan evaporation (mm day-1) Palmira - 2003 Palmira - 2007 50 50 40 40 30 30 20 20 10 10 0 0 0 10 20 30 40 50 60 70 80 Days after planting Figure 30. Rainfall distribution, pan evaporation, maximum and minimum temperatures during crop growing period at Palmira during 2003, 2004 and 2007 crop growing seasons. 87 Rainfed grain yield (kg ha-1) 1800 1600 Cowpea M MR 81 MR 112 MR 25 MR 95 MR 93 MR 27 MR 120 MR 65 SEA 15 MR 2 MR 55 MR 63 Mean: 1182 MR 64 SEA 5 MR 117 MR 108 LSD0.05: 297 MR 83 MR 98 Tio Canela MR 115 MR 20 MR 104 MD 23-24 MR 85 MR 97 MR 59 MR 1 MR 114 MR 6 MR 119 MR 33 MR 29 DOR 390 MR 49 MR 54 MR 116 MR 42 MR 109 MR 8 Mean: 1846 EAP 9510-77 LSD0.05: 248 MR 12 1400 1200 1000 800 600 1000 1200 1400 1600 1800 2000 2200 2400 2600 Irrigated grain yield (kg ha-1) Figure 31. Identification of genotypes that are adapted to rainfed conditions and are responsive to irrigation in a Mollisol at Palmira. Genotypes that yield superior with drought and were also responsive to irrigation were identified in the upper, right hand quadrant (a) 2400 (b) 1800 1500 1200 Mean: 1846 LSD0.05: 248 MR 8 900 Mean: 4364 LSD0.05: 867 600 1000 3000 5000 Mean: 2810 LSD0.05: 655 Cowpea M MR 108 MR 115 MR 120 MD 23-24 Tio Canela DOR 390 EAP 9510-77 SEA 15 MR 37 SEA 5 MR 112 MR 116 MR 88 MR 98 MR 64 MR 29 MR 20 MR 59 MR 109 2100 Grain yield (kg ha-1) Rainfed Irrigated 2700 Cowpea M MR 112 MR 25 MR 12 SEA 15MR 68 SEA 5 MR 83 MR 115 MD 23-24 MR 13 MR 113 MR 119 MR 33 DOR 390 MR 82 MR 116 MR 109 MR 8 EAP 9510-77 9000 1000 7000 3000 Mean: 1182 LSD0.05: 297 5000 7000 9000 Canopy biomass (kg ha-1) 2700 (c) (d) Cowpea M 2400 MR 108 MR 115 MD 23-24 Mean: 1846 MR 28 MR 3 DOR 390 EAP 9510-77SEA 15 LSD0.05: 248 MR 23 MR 21 MR 57 SEA 5 MR 116 MR 53 MR 98 MR 64 MR 20 MR 65 MR 29 MR 59 MR 109 MR 8 2100 1800 1500 1200 900 Mean: 55.3 LSD0.05: 17 600 30 40 50 60 70 80 90 Mean: 59.4 LSD0.05: 18 Cowpea M Mean: 1182 MR 25 MR 81 LSD0.05: 297 MR 12 MR 112 MR 106 SEA 15 MR 83 MR 32 MR 9 MR 101 SEA 5 MR 39 MR 115 MD 23-24 Tio Canela MR 40 MR 44 MR 119 MR 54 DOR 390 MR 114 MR 102 MR 109 MR 116 MR 42 MR 3 EAP 9510-77 MR 8 30 40 50 60 70 80 90 Harvest index (%) Figure 32. The relationship between grain yield and irrigated and rainfed canopy biomass (a, b) and grain yield and irrigated and rainfed harvest index (c, d) when grown in a Mollisol at Palmira 88 Irrigated 2700 Rainfed (a) 2400 Cowpea M MR 108 MR 56 MR 25 MR 120 DOR 390 MD Mean: 1846 Canela23-24MR 97 MR 28 MRTio 82 SEA 15 EAP 9510-77 LSD0.05: 248 MR 79 MR 24 SEA 5 MR 21 MR 11 MR 53 MR 98 MR 60 MR 6 MR 20 MR 65 MR 29 MR 59 MR 109 2100 1800 1500 1200 MR 112 Cowpea M MR 25 MR 24 MR 81 MR 120 SEA 15 MR 65 MR 106 MR 76 SEA MR 5 46 MR 105 Tio Canela MR 40 MR 102 MD 23-24 MR 36 MR 6 MR 114 DOR 390 MR 82 MR 33 MR 29 MR 37 MR 3 MR 109 EAP 9510-77 MR 8 Mean: 1182 LSD0.05: 297 MR 8 900 Mean: 19.7 LSD0.05: 23 600 -40 2700 Grain yield (kg ha-1) Mean: 27.2 LSD0.05: 20 (b) 2400 2100 1800 1500 1200 -20 0 20 60 -40 40 -20 0 Stem biomass reduction (%) (d) Cowpea M (c) MR 108 MR 115MR 25 MR 120 MR 3 MD 23-24DOR 390 EAP 9510-77 MR 118 Tio CanelaSEA MR 51 15 MR 101 MR 37 MR 26 MR 19 MR 73 Mean: 1846 SEA 5 MR 17 MR 96 LSD0.05: 248 MR 6 MR 85 MR 29 MR 20 MR 59 MR 109 MR 8 900 40 60 Mean: 75.4 LSD0.05: 3.9 MR 81 MR 112 MR 25 MR 95 Mean: 1182 Cowpea M MR 12 MR 27 MR 120 LSD0.05: 297 SEA 15 MR 47 MR 26 SEA 5 MR 88 MR 87 Tio Canela MD 23-24 MR 96 MR 113 MR 104 MR 49 DOR 390 MR 30 MR 116 MR 109 MR 42 EAP 9510-77 MR 8 Mean: 78.7 LSD0.05: 3.6 600 20 64 66 68 70 72 74 76 78 80 82 84 86 64 66 68 70 72 74 76 78 80 82 84 86 Pod harvest index (%) 2700 (e) 2400 MR 108 MR 120 MD 23-24 Tio CanelaDOR 390 EAP 9510-77 SEA 15 MR 111 MR 4 SEA 5 MR 66 MR 84 MR 6 MR 20 MR 59 MR 109 2100 1800 1500 1200 Mean: 1846 LSD0.05: 248 MR 8 900 Mean: 1168 LSD0.05: 329 600 0 (f) Cowpea M 1000 2000 3000 4000 Mean: 891 LSD0.05: 236 81 MR 112 MR MR 23 MR 120 MR 93 SEA 15 MR 65 SEA 5 MR 117 MD 23-24 MR 92 MR 85 MR 114 MR 119 DOR 390 MR 116 EAP 9510-77 5000 0 1000 Cowpea M 2000 3000 Mean: 1182 LSD0.05: 297 4000 5000 Seed number per area (no. m-2) Figure 33. The relationship between grain yield and irrigated and rainfed stem biomass reduction (a, b), grain yield and irrigated and rainfed pod harvest index (c, d), and grain yield and irrigated and rainfed seed number per area (e, f) when grown in a Mollisol at Palmira. 89 Correlation coefficients between final grain yield and other shoot attributes under rainfed conditions indicated significant positive relationship between final grain yield and leaf area index and canopy biomass under rainfed conditions (Table 34). Total chlorophyll content showed positive relationship with grain yield under irrigated conditions. It is important to note that the harvest index, pod partitioning index and pod harvest index were significantly associated with grain yield under rainfed conditions. This indicates that the genotypes that mobilized a greater proportion of photosyntates from vegetative organs to developing grains performed better under rainfed conditions (Figure 33). Seed number per area and pod number per area also showed significant positive association with grain yield only under rainfed conditions. Pod production efficiency showed significant negative association with grain yield under both irrigated and rainfed conditions. Days to maturity also showed significant negative association with grain yield indicating some contribution of earliness to superior performance under rainfed conditions. Table 34. Correlation coefficients (r) between final grain yield (kg ha-1) and other plant attributes of RILs of common bean grown under irrigated and rainfed conditions in a Mollisol in Palmira Plant traits Leaf area index (m2/m2) Total chlorophyll content (SPAD) Canopy biomass (kg ha-1) Shoot TNC content (mg g-1) Seed TNC content (mg g-1) Pod partitioning index (%) Pod harvest index (%) Stem biomass reduction (%) Seed number per area (no. m2) Pod number per area (no. m2) Harvest index (%) Yield production efficiency (g g-1) Seed production efficiency (no. g-1) Pod production efficiency (no. g-1) Days to maturity 100 seed weight (g) Irrigated -0.10** 0.15*** 0.38*** 0.08 0.18*** -0.14*** 0.08 0.03 0.05 -0.03 -0.12*** -0.12*** -0.25*** -0.32*** 0.45*** 0.17*** Rainfed 0.39*** -0.001 0.45*** -0.11** 0.02 0.07 0.37*** 0.11*** 0.34*** 0.15*** 0.13*** 0.13*** -0.03 -0.26*** -0.16*** 0.11** *, **, *** Significant at the 0.05, 0.01 and 0.001 probability levels, respectively. Conclusions: Field evaluation of 121 RILs of the cross MD 23-24 x SEA 5 over 3 seasons resulted in identification of the lines MR 81 and MR 25 that were superior in adaptation to drought stress conditions. The superior performance of these lines was associated with higher vigor, higher values of pod harvest index, harvest index and seed number per area, highlighting the importance of the photosyntate mobilization to pods and seeds under intermittent drought stress. Contributors: J. Polanía, M. Grajales, C. Cajiao, R. García, J. Ricaurte, S. Beebe and I. M. Rao 90 2.1.1.11 Evaluation of drought resistance and yield of 7 genotypes of Phaseolus vulgaris inoculated with and without Rhizobium etli strain CIAT 632 under greenhouse conditions Rationale: Published research from Mexico indicated that the inoculation of common bean genotype, Negro Jamapa, with over expressed trehalose-6-phosphate synthase gene in recombinant strain of Rhizobium etli CE3, increased grain production of common bean that was subjected to drought under greenhouse conditions. Before considering bean inoculation with mutant or recombinant strain of Rhizobium etli CE3, it is necessary to validate the efficacy of inoculation to improve bean growth under greenhouse conditions. We evaluated the response of 7 common genotypes to inoculation under terminal drought stress or maintenance of 80% of field capacity (well watered) in sterilized soil inoculated with or without Rhizobium etli CIAT 632 using greenhouse soil tube method. Materials and methods: A greenhouse study was conducted at CIAT - Palmira using a mix of an Andisol (from Darien of Colombia) with river sand (2:1 w/w), sterilized with steam for 2 h each day for 2 days and packed in soil cylinders (80 cm of height with 7.5 cm of diameter). Soil was fertilized with nutrients (kg ha-1 of 80 N, 50 P, 100 K, 101 Ca, 29.4 Mg, 20 S, 2 Zn, 2 Cu, 0.1 B and 0.1 Mo). The trial included 7 bean genotypes: BAT 477, BAT 477NN, DOR 364, DOR 364NN, Negro Jamapa, Pinto Villa and Alubia Cerrillos. The trial was planted as a randomized complete block arrangement with two levels of water supply: 80% field capacity (well-watered), and withholding of watering (to simulate terminal drought stress conditions); and 3 replications. For the treatment with inoculation, we added 1 ml of inoculum with the strain of Rhizobium etli CIAT 632, cultivated in YMA (yeast mannitol agar) liquid. Treatments of terminal drought were imposed at 14 days after planting. Plants with well-watered treatment were maintained by weighing each cylinder every two days and applying water to the soil at the top of the cylinder and plants with terminal drought were monitored for water stress by weighing each cylinder every two days for determination of decrease in soil moisture. Plants were harvested at the age of 35 days after establishment, i.e., 21 days of withholding of water application. After 2 weeks of establishment a number of shoot physiological characteristics were measured at weekly intervals: total chlorophyll content (SPAD) by chlorophyll meter SPAD-502; chlorophyll fluorescence FV’/FM’ by Fluorpen FP-100 (PSI – Photon Systems Instruments); stomatal conductance by porometer SC1 (Decagon Devices) and leaf temperature depression by the infrared thermometer (Telatemp model AG-42D). At the time of harvest, the following plant attributes were measured: leaf area; dry weight of stems, leaves, and pods; and root parameters including nodulation (nodules number and dry weight), root length and biomass distribution at different soil depths (0-5; 5-10; 10-20; 20-40; 40-60; 60-75 cm). Roots in each soil layer were washed free of soil and sand and root length, mean root diameter, specific root length, and root dry weight were measured. Root length and mean root diameter were measured using an image analysis system (WinRHIZO, Regent Instruments INC.). Results and Discussion: The average of soil moisture at 35 days after planting in the drought treatment was at 50% of field capacity and the maximum temperature in the greenhouse was between 35 to 40oC and the minimum temperature was between 19 to 20oC. Terminal drought condition decreased the nodulation, biomass production, and leaf area of all bean genotypes studied (Table 35). But there was no response to inoculation with the strain Rhizobium etli CIAT 632 either in well watered or with terminal drought treatment in terms of shoot biomass and leaf area. The inoculation only increased the nodule number and their dry weight under terminal drought (Table 36). The steam sterilization of soil for 2 h each day for 2 days was not adequate to eliminate native Rhizobia. Alubia Cerillos was found to be more sensitive to drought in terms of shoot biomass and leaf area reduction compared with the well watered condition (Table 35). 91 Table 35. Effect of inoculation and drought on nodulation, shoot biomass and leaf area. Nodules number per plant Well watered Terminal drought + Inoc - Inoc + Inoc - Inoc Genotypes Pinto Villa Alubia Cerrillos Negro Jamapa BAT 477 BAT 477NN DOR 364 DOR 364NN Pinto Villa Alubia Cerrillos Negro Jamapa BAT 477 BAT 477NN DOR 364 DOR 364NN Table 36. 111 82 21 16 0.030 0.030 0.003 0.003 189 241 123 0 138 0 168 362 157 0 131 0 37 45 8 0 12 0 5 30 5 0 2 0 0.088 0.116 0.035 0 0.061 0 0.074 0.047 0.046 0 0.081 0 0.011 0.008 0.0008 0 0.003 0 0.007 0.003 0.001 0 0.0003 0 Leaf area (cm2 plant-1) Well watered Terminal drought + Inoc - Inoc + Inoc - Inoc Genotypes Nodules dry weight per plant (g) Well watered Terminal drought + Inoc - Inoc + Inoc - Inoc Shoot biomass (g plant-1) Well watered Terminal drought + Inoc - Inoc + Inoc - Inoc 517.2 551.8 195.5 165.5 5.28 5.10 1.58 1.62 724.8 752.7 538.0 704.4 756.9 958.5 643.0 836.4 617.5 589.3 770.2 757.2 89.4 187.0 79.4 123.7 192.5 133.9 120.1 178.8 87.5 146.0 185.0 200.0 5.90 4.38 3.67 6.18 4.69 7.03 5.30 4.50 3.96 4.06 5.12 5.74 0.73 0.97 0.90 1.02 1.09 1.04 0.68 1.07 1.01 1.04 0.94 1.20 Effect of inoculation under terminal drought on nodule number and weight at 35 days after planting. Number CIAT 632 Well watered Weight (g) Terminal drought Well watered Terminal drought + Inoc 120 17 0.05 0.0037 - Inoc 129 8 0.04 0.0012 LSD 0.05 28 7 0.023 0.0017 All genotypes decreased the stomatal conductance after two weeks of drought stress but the decrease was more pronounced with Pinto Villa than the other genotypes (Figure 34). This ability to reduce stomatal conductance contributed to its superior performance under drought stress. BAT 477 and its nonnodulating line BAT 477NN maintained their photosynthetic efficiency (yield of quantum efficiency) during drought stress (Figure 34). There were no significant differences between the nodulating and nonnodulating lines because the N supply from soil was adequate. 92 Figure 34. Stomatal conductance and photosynthetic efficiency of 7 genotypes under well watered and drought stress conditions at different days after planting (dap). Conclusions: The greenhouse soil tube method was found to be very effective in simulating drought stress and the genotype Pinto Villa was better adapted to drought due to its ability to decrease stomatal conductance while Alubia Cerrillos was more affected due to drought stress. Although there was no response to inoculation with the strain Rhizobium etli CIAT 632, the effect of terminal drought stress on nodulation was very marked. Soil sterilization with steam was not effective in eliminating native Rhizobia. We need to use a soil type with low nitrogen availability and improve the efficiency of sterilization of soil to test the response to inoculation. Contributors: N. Asakawa, E. Melo, and I. M. Rao 93 2.1.2 Aluminum resistance Highlights: • • • • • • Lines derived from an interspecific cross of SER 16, a drought resistant line, by Phaseolus coccineus (G35346) yielded well under both rainfed and aluminum toxic conditions. Some actually yielded more in intermittent drought than SER 16. SER 16 proved to be an excellent common bean parent to cross with P. coccineus, perhaps due to its characteristic of excellent remobilization to grain. A filter paper-styrofoam sandwich germination method was developed to improve the phenotyping capacity for Al resistance in common bean and a primary root marking method was developed to evaluate short-term effects of Al on root elongation process. A method was adapted and validated for screening for aluminum resistance in common bean based on qualitative determination of aluminum-complexing compounds including citrate released from the roots. Phenotypic evaluation of 20 common bean genotypes for aluminium resistance confirmed the higher level of Al resistance of three Andean genotypes (ICA Quimbaya, BRB 198 and G5273) and identified one Mesoamerican genotype (G24601) also with higher level of Al resistance. Phenotypic evaluation of 97 RILs of DOR 364 x BAT 477 for aluminium resistance resulted in identification of a few RILs with low inhibition of root growth under high Al in solution. Two RILs (BT 21138-128-1-M-M-M and BT 21138- 2-1-1-M-M-M) were found to be outstanding in root growth both with and without aluminum in solution. 2.1.2.1 Evaluation in two environments, of interspecific lines selected under aluminum toxicity Rationale: Aluminum toxicity is a primary constraint of crops in tropical soils, limiting root development and nutrient and water capture. Phaseolus vulgaris is considered to be relatively sensitive to aluminum, while previous field observations of Phaseolus coccineus and subsequent greenhouse evaluations demonstrated that this latter species has superior aluminum tolerance. Aluminum toxicity is thought to exacerbate drought stress by limiting access to soil moisture. Thus, combining resistance to drought and tolerance to aluminum is an important objective. Interspecific crosses were created for this purpose. Materials and Methods: Two drought resistant common bean parents (Mesoamerican SER 16; Andean ICA Quimbaya) were crossed to several accessions of P. coccineus (G35346; G35066; G35464) that had been selected in the field based on vegetative vigor in an aluminum toxic soil. The F1 plants were backcrossed to the common bean parent, and pedigree selection for vigor was practiced over several generations in the field in an aluminum toxic soil in Santander de Quilichao, from the F2 to the F4 generation when selection was performed under drought pressure. These families had reached the F5 generation at the time of this evaluation, while other selections that were not planted in the drought season and had only been subjected to selection for aluminum tolerance were in the F4. Ninety F4 and F5 families were planted in Palmira under rainfed conditions in the dry season (JulySeptember, 2008). Two-row plots 3.75 m long were used in three replications. Ten check lines were included to complete a 10 x 10 lattice design. Checks included elite drought selections as well as intraspecific selections for aluminum tolerance from previous breeding work. The same trial was planted in Santander de Quilichao in an aluminum toxic soil (66% saturation) in a lattice design with 4 replications in the October-January planting season. Data on yield and days to maturity were taken, and yield per day was calculated as an indicator of crop efficiency in remobilization of biomass to grain. 94 Results and Discussion: In the rainfed treatment, the crop received an estimated 170 mm of moisture derived from one irrigation with sprinklers, one furrow irrigation, and 115 mm of rainfall up to 68 days from planting. However, drought stress was only moderate, due to excellent soil structure and cloudy days with low evapotranspiration. Based on common checks in another irrigated trial, yield reduction was estimated to be 25-30%. Under these conditions several lines yielded exceptionally well, and two lines (ALB’s 205 and 167) significantly outyielded the drought tolerant parent, SER 16 (Table 37). This yield advantage was in part due to a longer growth cycle, these lines being 5-6 days later to mature than SER 16. Nonetheless, these lines maintained their efficiency, yielding marginally better per day than SER 16. Other elite lines from previous years (e.g., BAT 477; A774; Tio Canela) presented comparable days to maturity as the ALB lines but yielded less. Thus it appears that better yield potential with the ability to resist at least moderate drought stress has been obtained in some lines. This may reflect improved rooting derived from G35346 combined with good remobilization of biomass to grain, derived from SER 16. Table 37. Best yielding lines and checks of interspecific progeny under intermittent drought, Palmira, and aluminum toxicity, Santander de Quilichao, Colombia. 2008. Rainfed Tr 83 61 90 54 15 67 62 20 100 31 60 21 76 87 17 96 22 94 19 48 66 85 89 92 65 33 74 99 88 16 98 63 91 97 93 95 Parents / identification SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) Line ALB 205 ALB 167 ALB 213 SER 16 X (SER 16 x G35066-1Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) ((VAX 1 x BRB191) x G21212) x (RAB 655 x G22041) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) G 21212 SER 16 X (SER 16 x G35346-3Q) ALB ALB ALB ALB ALB 159 188 180 168 214 ALB ALB ALB ALB ALB ALB ALB 252 204 166 215 197 210 190 VAX 1 SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35066-1Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) TIO CANELA 75 SER 16 X (SER 16 x G35346-3Q) ICA QUIMBAYA X (ICA QUIMBAYA x G35464-5Q) SER 16 X (SER 16 x G35346-3Q) ((G24601 x (MAM 38 x BRB 198)) x G11015) X (MAM 38 x G21212) SER 16 X (SER 16 x G35346-3Q) SER 16 X (SER 16 x G35346-3Q) ((VAX 1 x BRB 191) x G21212) x (RAB 655 x G22041) SER 16 X (SER 16 x G35346-3Q) SER 16 A 774 ICA QUIMBAYA BAT 477 ALB 216 ALB ALB ALB ALB ALB 208 149 179 207 212 ALB 178 ALB 130 ALB 195 ALB 229 ALB 211 ALB 189 ALB 253 ALB 169 COL . rd rd rd br, cr rd rd rd rd rd rd rd rd rd rd bl rd Cr str rd rd rd rd rd rd rd pk rd Cr br rd rd v rd rd Cr rd Cr R s n k 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 38 68 Aluminum toxicity kg ha-1 -1 d D T F D T M kg ha-1 35 37 33 68 69 67 3199 3174 3029 46,9 45,7 45,3 33 33 32 33 35 68 67 64 68 71 2974 2891 2887 2883 2879 43,4 43,1 45,0 42,4 40,4 38 36 37 34 33 38 34 38 38 70 68 69 65 65 68 71 68 71 2824 2781 2761 2722 2720 2714 2695 2682 2674 40,3 40,8 40,1 41,7 41,9 39,4 38,1 39,1 37,3 38 33 33 33 36 33 38 32 68 66 66 64 70 67 68 64 2653 2625 2620 2614 2594 2592 2581 2572 38,9 39,8 39,4 40,5 36,8 38,9 38,0 40,5 32 33 64 67 2569 2568 40,1 38,2 38 33 32 71 65 65 2567 2563 2554 35,7 39,5 39,1 38 34 32 36 33 38 1.8 70 67 63 68 68 68 2550 2528 2520 2517 2453 2165 568 36,3 37,6 40,3 37,3 36,1 31,8 8.1 R s n k D T M kg ha-1 46 27 19 60 35 35 34 35 63 64 61 63 656 716 755 591 10.4 11.2 12.5 9.5 13 63 37 9 5 37 34 34 36 38 66 61 62 64 64 779 582 684 846 882 11.9 9.6 11.4 13.3 13.7 1 93 7 39 79 21 4 55 71 34 37 33 34 39 36 35 38 39 63 63 59 60 63 63 63 65 65 906 399 859 681 525 749 883 610 560 14.4 6.4 14.5 11.4 8.3 11.9 13.8 9.3 8.7 19 54 25 36 11 31 17 89 33 33 33 36 34 38 33 33 62 62 59 64 61 64 59 61 877 611 720 684 826 704 758 471 14.2 9.8 12.1 10.8 13.4 10.9 12.6 7.9 14 28 34 39 62 67 773 715 12.4 10.6 3 34 40 33 34 39 60 64 66 896 698 676 14.9 10.9 10.3 82 23 26 99 24 35 32 37 32 40 63 58 63 65 63 514 742 719 294 733 231 8.2 12.7 11.3 4.5 11.5 3.7 2.4 1.8 LSD (0.05) COL=color; bl=black; rd=red; cr str=cream striped; pk=pink; v=variable; DTF=Days to flower; DTM=Days to maturity. 95 kg ha-1 -1 d D T F 2.0 In the aluminum toxicity treatment, no line significantly outyielded the SER 16 recurrent parent, which in fact resulted to be intermediate in its reaction to aluminum. Thus, based on this data it cannot be stated that we successfully transferred improved aluminum tolerance from P. coccineus to SER 16. However, several lines outyielded the tolerant check, VAX 1, by as much as 60%. What was more important is that a few lines performed relatively well in both treatments (Table 37). In the rainfed and aluminum treatments, respectively, ALB 213 ranked 3rd and 19th, ALB 188 ranked 5th and 13th, ALB 214 was 8th and 9th, ALB 252 was 9th and 5th, and ALB 204 was 10th and 1st. The correlation in yield across the two treatments was 0.54. These lines merit wider evaluation, given their positive performance and the fact they represent unusual genetic variability that has not previously been exploited in cultivated common bean. We speculate that it was possible to obtain better progenies from this cross as compared to other interspecific crosses in the past, due to SER 16’s excellent remobilization of biomass to grain. We hypothesize that that this trait serves to overcome the tendency for such crosses to produce lines with excessive vegetative growth and poor harvest index. Collaborators: S. Beebe, I. Rao, C. Cajiao, M. Grajales, M.L. Cortés, A.F. Guerrero 2.1.2.2 Improving phenotyping capacity to evaluate for aluminum resistance Rationale: Screening for Al resistance using hydroponic-based methodology normally requires that large amounts of genotypes or accessions should be tested at the same time under the same climatic and experimental conditions. Independently of the amount of materials to be tested, obtaining healthy and homogenous plantlets or seedlings to reduce the variability originating in the germination process is of great importance. Previously, germination of bean seedlings involved use of growing mediums such as peat, sand or a mix of both substrates. However, this method is time consuming, special skills are required to clean the peat from the roots in order to reduce the interference with the Al in the nutrient solution and in many cases the roots are not uniform. Normally, it was required up to 16 h to establish, germinate and transplant a trial with 60 genotypes. In spite of careful manipulation, in the end high standard deviations did not allow clear differentiation between genotypes for their level of Al resistance. Materials and Methods: After reviewing the previous method used in the screening of bean for Al resistance, some key modifications were made to increase the capacity to screen larger number of genotypes while reducing the time needed for the establishment and the sources of variability between replicates. These key modifications in order of importance were: 1. Replacing the sand-based germination by filter-paper/styrofoam sandwiches; 2. Manipulation of plantlets during the transplant to nutrient solution to avoid damage of basal roots (very important in the determination of root architecture parameters); 3. Measuring root length after 24 or 48 h of Al treatment using marked roots (important to correlate with physiological studies); 4. Controlling the pH of the nutrient solution, especially during the preparation of the Al treatments; 5. Controlling the aeration system to avoid root damage during the treatment time. Results: After the implementation of the sandwich system to germinate the plants, healthy (straight tap and basal roots) and homogenous plantlets were obtained (Figure 35 b and c), allowing quick selection of pairs of seedlings to be transplanted to containers with or without Al. The time to arrange and establish the whole germination system is less than 2 h, and after germination the transplanting of 120 genotypes (15 seeds x 2 treatments x 4 replicates) corresponding to circa 14,400 seeds could be completed in less than 4 h. 96 Figure 35. Comparison between seedlings germinated in sand (a) or in filter-paper/Styrofoam sandwich system after 3 (b) or 4 days (c) of germination. Physiological studies of Al resistance normally require measurements of root elongation of tap and basal roots during short periods of time (few hours) after Al treatment. Marking roots (Figure 36) 3 cm above the root tip (root zone already differentiated) just prior to the Al treatment, allows measuring the direct effect of Al on root elongation at different Al treatment times. After obtaining homogenous seedlings from the germination system, results from Figure 37 shows that no noticeable differences were observed between marked and complete root measurement methods using an Al resistant Quimbaya and Al sensitive VAX 1 genotypes. Therefore, for routine screening of Al resistance the complete root measurement could be used. However, to measure the Al effect during short or consecutive treatment times, the marking method is recommended. 97 Figure 36. Root elongation measured with two methods: a) complete root and b) marking 3 cm above the root tip. Control plants Al treated plants 3.0 3.0 24 h 48 h 24 h VAX-1 Quimbaya 2.0 1.5 1.0 48 h 2.5 Root growth rate -1 [mm h ] Root growth rate -1 [mm h ] 2.5 VAX-1 Quimbaya 2.0 1.5 1.0 0.5 0.5 0.0 0.0 marking complete marking marking complete complete marking complete Root measurement Root measurement Figure 37. Influence of two different methods (marking and complete root) to measure root growth rates of two common bean genotypes (Al-resistant, Quimbaya; Al-sensitive VAX-1) after germination using filter-paper/styrofoam germination system. Conclusions: A filter paper-styrofoam sandwich germination method was developed to improve the phenotyping capacity for Al resistance in common bean and a primary root marking method was developed to evaluate short-term effects of Al on root elongation process. Contributors: A.F. Rangel, J. Ricaurte, S. Beebe, I. M. Rao (CIAT); W. Horst (Leibniz University of Hannover, Germany) 98 2.1.2.3 Qualitative indication of Al-induced citrate exudation in different Phaseolus species using an Agarose-Aluminon method Rationale: Previous studies have shown that citrate exudation contributes to Al resistance in common bean. Two protocols were implemented in order to check the contribution of citrate exudation to Al resistance in different Phaseolus species using 1 cm excised root tips in nutrient solution (quantitative) or intact plants in agarose gels containing aluminon as color indicator (qualitative). The agarose gel technique shows that organic acid exudation occurs along the entire root system but mainly to the first 1 to 2 cm of the root tip, the most Al sensitive root zone. The amount of organic acids secreted to the agarose media of the three Phaseolus species tested decreased in the order of P. coccineus > P. vulgaris > P. acutifolius. The presence of Al-complexing chelators in root exudates and rhizosphere soil solution can be visualized using agarose gels, containing red-colored Al-aluminon complexes on the root surface. The presence of Al chelators with higher affinity to Al, compared to aluminon (e.g. organic acid anions, phenols) is indicated by discoloration zones. Materials and methods: Gels of agarose (low gelling point) of 3 mm (1 % w/v) were used as carrier matrix for the Al-aluminon complex. Agarose gels were prepared in nutrient solution containing 5 mM CaCl2, 0.5 mM KCl and 8 µM H3BO3. Aluminum (250 µM) was added to the agarose containing nutrient solution after cooling down to 50oC and adjusting the pH to 4.5. Aluminon stock solution was prepared by mixing NaOH (24g), Aluminon (175 mg) dissolved in acetic acid (120 ml) and adjusted to 500 ml with bi-distilled water and adjusting the pH to 4.2. Different bean accessions with contrasting levels of Al-resistance were pretreated for 24 h in nutrient solution containing 20 µM of Al. This treatment time is enough to detect significant differences in Al resistance among common bean genotypes based on inhibition of root elongation. Also, it has been demonstrated that these differences corresponded with the capacity to exude organic acid anions, mainly citrate. After Al pretreatment four plants of each accession were carefully placed into flat transparent acrylic cuvettes, the root system was dispersed and a mix of 25 ml aluminon, 70 ml of agarose containing nutrient solution and Al, and 5 ml ascorbic acid (0.5%) at 40oC, was carefully poured to cover the whole root system (3 to 4 mm thickness). Thereafter, the cuvettes were covered with plastic sheet to avoid desiccation of the agarose. Depending on the amount of Al-complexing compounds released from the root or accumulated in the rhizosphere, discoloration was visible after 6 to 8 h. Results: In general terms, the appearance of bleached zones were observed in the complete root system but mainly in the first 1 to 2 cm root apices. Similarly, all accessions showed the capacity to exude Alchelating substances from the roots. However, the grade and magnitude of discolored areas were in the order of P. acutifolius < P. vulgaris < P. coccineus. Additionally, the extent of discolored areas were in agreement with the known Al resistance rating of all six accessions used, so less blanched areas were observed in G40159 and VAX-1, both rated as Al-sensitive. SER16, an Al-intermediate accession, showed discolored areas mainly in the first 1 to 2 cm from the root tips and in areas where more root tips appeared. G19833, Quimbaya and G35346-3Q, all classified as Al-resistant, showed a greater capacity to exude Al-chelating substances along the whole root system (Figure 38). However, the capacity to exude these substances by the coccineus accession was outstanding. The validation of this result requires quantification of the Al-chelating substances exuded from the roots. According to previous studies, this should correspond mainly to citrate exudation. Thus, this method can be used as a fast test to check the existence of citrate exudation as a mechanism of Al resistance in different bean accessions. 99 Conclusions: A method was adapted and validated for screening for aluminum resistance in common bean based on qualitative determination of aluminum-complexing compounds including citrate released from the roots. Contributors: A.F. Rangel, J. Ricaurte, S. Beebe, I. M. Rao (CIAT); W. Horst (Leibniz University of Hannover, Germany) Figure 38. Qualitative determination of Al-complexing compounds (e.g. citrate) released from the roots of six bean genotypes differing in Al-resistance. G40159 (P. acutifolius); VAX-1, SER16, G19833 and Quimbaya (P. vulgaris); and G35346-3Q (P. coccineus). 100 2.1.2.4 Phenotypic differences in aluminum resistance of landraces and bred lines Rationale: Toxicity of Al in acid soils in the tropics is a major problem to resource-poor farmers for producing beans. Amending soils with lime is not economically feasible for most farmers. Improving the level of Al resistance in common bean could help farmers to produce beans locally in acid soils with high aluminum. Field screening of 5000 germplasm accessions and breeding lines in Al-toxic soils with and without lime (65% Al saturation) indicated significant genotypic variation in seed yield. Likewise, significant genotypic differences for Al resistance have been found in nutrient-solution based screenings using inhibition of root elongation after 48 h at 20 µM Al supply as a parameter for Al injury. The present work aimed to identify bean genotypes with Al resistance as determined by Al-induced inhibition of principal root elongation and total root length per plant. Genotypic differences in root vigor in the absence of Al in nutrient solution were also evaluated to identify genotypes that combine Al resistance with high root vigor. Materials and Methods: Three days old seedlings of 20 common bean genotypes listed in Table 38 were grown in nutrient solution containing 5 mM CaCl2.2H2O, 0.5 mM KCl and 8 µM H3BO3 under glasshouse environmental conditions at CIAT-Palmira. After 2 days of pH adjustment from 5.5 to 4.5 with a step wise decrease, plants were exposed to either 0 or 20 µM AlCl3.6H2O for up to 2 days (Figure 39). The experiment was repeated twice with 4 replications and the mean values are reported. Primary root elongation rate (mm h-1) was measured through marking of the primary root 3 cm above the root tip at the beginning and at the end of the treatments. The difference between the two markings was used to measure primary root elongation rate and percent inhibition of root elongation rate due to Al treatment. At 48 h of treatment with or without Al, total root length (cm plant-1), mean root diameter (mm), root volume (cm3 plant-1) and surface root area (cm² plant-1) were measured using Winrhizo software (WinRHIZO 2003, Regent Instruments INC). Root weight (g plant-1) was determined after drying roots until constant weight in an oven at 60 oC for 48 h. Specific root length (m g-1) was calculated dividing root length by root weight. Results: Root growth of all genotypes (Al-resistant and Al-sensitive) was inhibited by Al treatment (Figures 40 and 41). Three Andean bred lines (ICA Quimbaya, BRB 198 and G5273) and one Mesoamerican accession (G24601) showed resistance to Al in solution. Three Mesoamerican bred lines (VAX 1, ALB 220, BAT 477) were found to be more sensitive to Al in solution. Two bred lines (ALB 230, ALB 218) that combined Mesoamerican, Jalisco and Andean races showed higher values of root growth both without and with Al stress (Table 38). BAT 477 showed greater root vigor in the absence of Al in solution. Among the six INB lines tested, INB 606 showed intermediate level of Al resistance (Figures 40 and 41). C onclusions: Phenotypic evaluation of 20 common bean genotypes for aluminum resistance resulted in identification of three Andean genotypes (ICA Quimbaya, BRB 198 and G5273) and one Mesoamerican genotype (G24601) with higher level of Al resistance. Contributors: J. Ricaurte, R. García, A. F. Rangel, S. Beebe, I. M. Rao (CIAT); W. Horst (Leibniz University of Hannover, Germany) 101 Table 38. Description of 20 common bean genotypes (ordered by inhibition of primary root elongation, Figure 40) grown in a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with or without 20 µM Al for 48 h, pH 4.5. Genotype Origin Race Species Pedigree ICA Quimbaya CIAT N Canadian Wonder x A 487 G 6592 x A 487 BRB 198 CIAT N CAL 192 x MCR 2515 G 5273 MEXICO N CIAS 72 P. vulgaris x P. acutifolius INB 606 CIAT M ICA Pijao COLOMBIA M BKI 11 X DOR 390/-1C-1C-1C-1C-MC-MC-MC ICA Pijao G 5773 G 19833 RAB 655 PERU CIAT N M Chaucha Chuga (VAX 3xMAM 38)-1/-(NN)Q-115Q-(NN)Q-(NN)Q-(NN)C-(NN)C-(NN)Q INB 108 CIAT M MAM 38 CIAT J ALB 218 INB 604 CIAT CIAT M x J xN = M M ALB 230 CIAT MxJxN=M G40001 MEXICO INB 605 CIAT M INB 603 CIAT M INB 109 CIAT M BAT 477 CIAT M ALB 220 VAX 1 CIAT CIAT M M P.acutifolius P. vulgaris x P. acutifolius ICA Pijao x G40001 A 409 x (BAT1670 x G4000 x XAN112) P.vulgaris x P.acutifolius G 24601 x (G 22041 x BRB 198)F2/-MQ-7Q-2Q-MQ-MQ-7Q-MC DCBC-V x DCBC-V/-F5-MC-14C-MC-MC-MC ((G 24601x(MAM 38xBRB 198)F1)F1xG 11015)F1 X (MAM 38xG 21212)F1/-MQ-15Q-MQ-MQ-12Q-MC P. acutifolius P. vulgaris x P. acutifolius P. vulgaris x P. acutifolius P. vulgaris x P. acutifolius DCBC-V x DCBC-V/-F5-MC-4C-MC-MC-MC DCBC-V x DCBC-V/-F5-MC-4C-MC-MC-MC ICA Pijao x G40001 (51051 x ICA Bunsi)F1 x (51052 x Cornell 49-242)F1/-CM(11-C)-M-CM(8-C) (G 3834 x G 4493)F1 x (G 4792 x G 5694)F1/-CM(11-C)-M-CM(8-C) RAB 655 X (MAM 49 X RIB 66)F1/-MQ-MQ-MQ-8Q-MQ-MQ-14Q-MC A 769 x(A 775 x (G 5773 x G40001-B1) Race: M = Mesoamerica; J = Jalisco; N = Nueva Granada Pedigree: Local names at country of origen; CIAT codes 102 Figure 39. Refined methodology to evaluate aluminum resistance in common bean genotypes using nutrient solution under greenhouse conditions. A and B) seedlings germinated using filter paper-styrofoam sandwich germination paper; C) adapted to pH changes from 5.5 to 4.5; D) aluminum test ; E and F) acquisition and analysis of root images. 103 . Root elongation rate with Al (mm h-1) 2.2 A Mean = 1.74 LSD0.05 = 0.41 2.0 ICA Quimbaya 1.8 G 5273 BRB 198 1.6 G 19833 INB 108 1.4 INB 606 1.2 G 40001 MAM 48 G 24601 ALB 230 Mean = 1.28 LSD0.05 = 0.20 BAT 477 1.0 INB 605 ALB 220 0.8 VAX 1 0.6 1.2 1.4 1.6 1.8 2.0 2.2 Al-inhibited primary root elongation (%) Root elongation rate without Al (mm h-1) 80 B 60 40 20 IC A Q U IM B B AY R A B G 19 24 8 6 G 01 52 IN 7 IC B 3 A 60 PI 6 J G AO 19 R 83 AB 3 IN 655 B M 108 AM AL 3 B 8 IN 218 B AL 60 B 4 G 23 40 0 IN 001 B IN 605 B IN 603 B B 10 AT 9 AL 47 B 7 22 VA 0 X 1 0 Genotypes Figure 40. Root elongation rate (A, mm h-1) and inhibition of principal root elongation (B, %) of 20 common bean genotypes grown in a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with or without 20 µM Al for up to 48 h, pH 4.5. Bars are means ± SD of eight replicates from 2 experiments. Horizontal line represents the genotypic mean value. 104 Total root length with Al (cm plant-1) 300 A Mean = 303 LSD0.05 = 115 ALB 230 250 ICA Quimbaya BRB 198 200 ALB 218 G 19833 G 5273 INB 108 INB 606 150 BAT 477 INB 603 MAM 38 ALB 220 100 Mean = 163 LSD0.05 = 47 VAX 1 INB 604 INB 109 50 0 0 100 200 300 400 500 600 Total root length without Al (cm plant-1) Al-inhibited total root length (%) 80 B 60 40 20 G IC A Q U IM B AY 19 A B 83 R 3 B 1 G 98 52 IN 73 B R 60 AB 5 IN 655 B IN 606 B AL 109 B G 21 24 8 G 60 40 1 00 VA 1 IN X IC B 1 A 10 PI 8 AL JAO B IN 230 B M 603 AM B AT 38 IN 477 B AL 60 B 4 22 0 0 Genotypes Figure 41. Total root length (A, cm plant-1), inhibition of total root length (B, %) of 20 common bean genotypes grown in a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with or without 20 µM Al for up to 48 h, pH 4.5. Bars are means ± SD of eight replicates from 2 experiments. Horizontal line represents the genotypic mean value. 105 2.1.2.5 Phenotyping for aluminum resistance in recombinant inbred lines (RILs) of DOR364 x BAT 477 Rationale: The present work is aimed to determine differences in resistance to Al-induced inhibition of principal root elongation and total root length of 97 RILs of the cross DOR364 x BAT477. In addition, root vigor without Al in solution is also determined. These phenotypic data will be used to identify QTLs involved in Al resistance in common bean. Materials and Methods: Three day old seedlings of 100 common bean genotypes (97 RILs of the cross DOR364 x BAT477, two parents and one check, SEA 5; Table 39) were evaluated in nutrient solutions under greenhouse conditions at CIAT Palmira. Seedlings were obtained with germination in 1-2 mm diameter of cleaned sand. Nutrient solutions contained 5 mM CaCl2.6H2O, 0.5 mM KCl and 8 µM H3BO3. After 2 days of pH adjustment from 5.5 to 4.5 with a step wise decrease, plants were exposed to either 0 or 20 µM AlCl3.6H2O for up to 2 days. The experiment was repeated three times with 3 replications at each time and the mean values are reported. Primary root elongation rate (mm h-1) was measured through marking of the primary root 3 cm above the root tip at the beginning and at the end of the treatments. The difference between the two markings was used to measure primary root elongation rate and percent inhibition of root elongation rate due to Al treatment. At 48 h of treatment with or without Al, total root length (cm plant-1), mean root diameter (mm), root volume (cm3 plant-1) and surface root area (cm² plant-1) were measured using Winrhizo software (WinRHIZO 2003, Regent Instruments INC). Root weight (g plant-1) was determined after drying roots until constant weight in an oven at 60 oC for 48 h. Specific root length (m g-1) was calculated dividing root length by root weight. Results: Root growth of all genotypes (RILs, parents, check) was inhibited by Al treatment (Figures 42 and 43). Al treatment inhibited primary root elongation of DOR 364 and BAT 477 by 57% and 64%, respectively. The 6 outstanding RILs with lower percent inhibition of primary root elongation showed values from 29 (R100=BT 21138_128-1--M-M-M) to 36% (R71=BT 21138_71-1-1-M-M-M). In ascending order they were: R100 (BT 21138_128-1--M-M-M), R39 (BT 21138_39-1-1-M-M-M), R70 (BT 21138_70-1-1-M-M-M), R8 (BT 21138_ 8-1-1-M-M-M), R44 (BT 21138_124-1-1-M-M-M-M-M) and R71 (BT 21138_71-1-1-M-M-M). Total root length inhibition in DOR 364 and BAT 477 was 48% and 36%, respectively. The RILs R84 (BT 21138_84-1-1-M-M-M) , R72 (BT 21138_72-1-1-M-M-M), R61 (BT 21138_61-1-1-M-M-M), R56 (BT 21138_56-1-1-M-M-M), R73 (BT 21138_73-1-1-M-M-M), R41 (BT 21138_41-1-1-M-M-M) and R30 (BT 21138_30-1-1-M-M-M) showed lower values of total root length inhibition (5.0, 23.8, 23.9, 24.2, 24.3, 24.8 and 26.4 respectively). Two RILs (R100 = BT 21138_128-1--M-M-M and R2 = BT 21138_ 2-1-1-M-M-M) were found to be outstanding in root production both with and without Al in solution (Figure 44). 106 Table 39. RIL Pedigree of 98 RILs of DOR364 x BAT 477 evaluated in a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with 0 and 20 µM Al for up to 48 h, pH 4.5. Pedigree RIL 1 BT 21138_ 1-1-1-M-M-M 35 2 BT 21138_ 2-1-1-M-M-M 3 BT 21138_ 3-1-1-M-M-M 4 BT 21138_ 4-1-1-M-M-M 5 Pedigree RIL Pedigree BT 21138_35-1-1-M-M-M 68 36 BT 21138_36-1-1-M-M-M 69 BT 21138_69-1-1-M-M-M 37 BT 21138_104-3-M-M-M-M-M 70 BT 21138_70-1-1-M-M-M 38 BT 21138_38-1-1-M-M-M 71 BT 21138_71-1-1-M-M-M DOR 364 39 72 BT 21138_72-1-1-M-M-M 6 BT 21138_ 6-1-1-M-M-M 40 BT 21138_39-1-1-M-M-M BT 21138_115-1-1-M-M-M-MM 73 BT 21138_73-1-1-M-M-M 7 BT 21138_ 7-1-1-M-M-M 41 BT 21138_41-1-1-M-M-M 74 BT 21138_74-1-1-M-M-M 8 BT 21138_ 8-1-1-M-M-M 42 BT 21138_42-1-1-M-M-M 75 BT 21138_75-1-1-M-M-M 9 BT 21138_ 9-1-1-M-M-M 43 76 BT 21138_76-1-1-M-M-M 10 BT 21138_10-1-1-M-M-M 44 BT 21138_43-1-1-M-M-M BT 21138_124-1-1-M-M-M-MM 77 11 BAT 477 45 BT 21138_45-1-1-M-M-M 78 BT 21138_77-1-1-M-M-M BT 21138_131-1-1-M-M-MM-M 12 BT 21138_12-1-1-M-M-M 46 BT 21138_46-1-1-M-M-M 79 BT 21138_79-1-1-M-M-M 13 BT 21138_13-1-1-M-M-M 47 BT 21138_47-1-1-M-M-M 80 BT 21138_80-1-1-M-M-M 14 BT 21138_14-1-1-M-M-M 48 BT 21138_48-1-1-M-M-M 81 BT 21138_81-1-1-M-M-M 15 BT 21138_15-1-1-M-M-M 49 BT 21138_124-1-2-M-M-M-M 82 BT 21138_82-1-1-M-M-M 16 BT 21138_16-1-1-M-M-M 50 BT 21138_50-1-1-M-M-M 83 BT 21138_83-1-1-M-M-M 17 51 BT 21138_51-1-1-M-M-M 84 BT 21138_84-1-1-M-M-M 18 BT 21138_17-1-1-M-M-M BT 21138_ 23-1-4-M-M-MM 52 BT 21138_124-1-3-M-M-M-M 85 BT 21138_85-1-1-M-M-M 19 BT 21138_19-1-1-M-M-M 53 BT 21138_53-1-1-M-M-M 86 20 BT 21138_20-1-1-M-M-M 54 BT 21138_54-1-1-M-M-M 87 BT 21138_35-1-4-M-M-M BT 21138_147-3-M-M-MM-M 21 BT 21138_21-1-1-M-M-M 55 BT 21138_55-1-1-M-M-M 88 BT 21138_88-1-1-M-M-M 22 BT 21138_22-1-1-M-M-M 56 BT 21138_56-1-1-M-M-M 89 BT 21138_89-1-1-M-M-M 23 SEA 5 57 BT 21138_57-1-1-M-M-M 90 BT 21138_90-1-1-M-M-M 24 BT 21138_24-1-1-M-M-M 58 BT 21138_58-1-1-M-M-M 91 BT 21138_91-1-1-M-M-M 25 BT 21138_25-1-1-M-M-M 59 BT 21138_59-1-1-M-M-M 92 BT 21138_92-1-1-M-M-M 26 BT 21138_26-1-1-M-M-M 60 BT 21138_60-1-1-M-M-M 93 BT 21138_93-1-1-M-M-M 27 BT 21138_27-1-1-M-M-M 61 BT 21138_61-1-1-M-M-M 94 BT 21138_94-1-1-M-M-M 28 BT 21138_28-1-1-M-M-M 62 BT 21138_62-1-1-M-M-M 95 BT 21138_95-1-1-M-M-M 29 BT 21138_29-1-1-M-M-M 63 BT 21138_63-1-1-M-M-M 96 BT 21138_96-1-1-M-M-M 30 BT 21138_30-1-1-M-M-M 64 BT 21138_64-1-1-M-M-M 97 BT 21138_97-1-1-M-M-M 31 65 BT 21138_65-1-1-M-M-M 98 BT 21138_98-1-1-M-M-M 32 BT 21138_31-1-1-M-M-M BT 21138_ 83-1-3-M-M-M-MM 66 BT 21138_66-1-1-M-M-M 99 BT 21138_99-1-1-M-M-M 33 BT 21138_33-1-1-M-M-M 67 BT 21138-67-1-1-M-M-M 100 BT 21138_128-1--M-M-M 34 BT 21138_34-1-1-M-M-M 107 BT 21138_68-1-1-M-M-M Primary root elongation with Al (mm h-1) 1.6 A Mean = 1.59 LSD0.05= 0.42 1.4 R100 1.2 R39 1.0 R70 R82 R2 R98 R56 R44R36 R31 R47 R25 R71 SEA 5 R61 R68 R38 R40 R32 R95 R3 R49 R60 R20 R54 R34R80 R30 R45 R1 R52 R86 R67 R69 R42 R7 R63 R53 R18 R8 R96 R13 R14 R15 R78 R84 R16 R55R26 R17 DOR 364 R85R46 R92 R91 R22 R81 R58 R79 R19 R24R88 R33 R51 R43R93 R76 R94 R50 R6 R48 R41 R65 R29 R37 R74 R99R57 R89 R4 R12 R83 R72 R27 R9 R35 R28 R62 R75 0.05 R64 R87 R97 R90 R21 R73 BAT 477 R59 0.8 Mean = 0.79 LSD = 0.30 0.6 R66 R10 0.4 1.0 1.2 1.4 1.6 1.8 2.0 100 B 80 60 40 20 0 R100 R39 R70 R8 R44 R71 R98 R82 R45 R36 R31 R61 R59 R49 R S E A2 5 R68 R25 R84 R56 R47 R7 R72 R54 R22 R92 R20 R3 R38 R40 R80 R34 R13 R26 R86 R1 R74 R60 R67 R96 R46 R42 R66 R58 R69 R78 R63 R85 R91 R55 R30 R95 R79 R81 R18 R89 R6 R99 R32 R65 R15 R16 R62 R64 R17 R52 R43 R53 R24 R29 R51 R33 R48 R57 R87 R9 R19 R12 R27 R93 35 DORR 364 R4 R14 R37 R94 R10 R50 R76 R88 R41 R21 R83 R75 R28 73 BAT R 477 R97 R90 Al-inhibited primary root elongation (%) Primary root elongation without Al (mm h-1) Genotypes Figure 42. Root elongation rate (A, mm h-1) and Inhibition of principal root elongation (B, %) of 100 common bean genotypes (98 RILs of DOR364xBAT 477 and 3 checks)evaluated in a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with or without 20 µM Al for up to 48 h, pH 4.5. Bars are means ± SD of nine replicates from 3 experiments. Horizontal line represents the genotypic mean value. 108 Root length with Al (cm plant-1) 200 A Mean = 230 LSD0.05= 91 180 R3 R100 R21 R24 R41 160 R15 120 100 R84 R36 R95 R18 R30 R44 R19 R31 R78 R72 R52 R89 R91 R4 R7 R65 R88 R86 R97 R45 R29R32 R51 R61 R47 R58 R83 R53 R98 R62 R63 SEA 5 R92R20 R85 R79 R38 R69 R49 R55 R87 R66 R70 R96 R50 R33 R57 R27 R6 R26 R73 R9R68 R14 R39 R93 R59 R94 R12 R42 R76 R16 R67 R60 R64 R8 R10 R75 R17 R48 R22 R13 R71 R25 R99 R34 R82R74 R37 R28 R81 R1 Mean = 131 LSD0.05= 54 BAT 477 80 100 DOR 364 R35 R54 R46 R56 140 R90 R43R2 R40 150 R80 200 250 300 350 Root length without Al (cm plant-1) B 80 60 40 20 R53 R80 R39 R60 R1 R17 R88 R14 R42 R34 R64 0 R84 R72 R61 R56 R73 R41 R30 R24 R89 R81 R55 R92 R29 R66 R44 R3 R96 R45 R74 R20 R15 R70 R21 R38 R69 R4 R48 R49 R43 BAT 47 7 R18 R28 R76 R82 R91 R22 R19 R79 R95 R9 R62 R7 R36 R93 R90 R68 R32 R86 R98 R94 R27 R26 R16 R33 R52 R8 R100 R2 R58 R71 R65 R47 R99 R97 R51 R63 R78 R46 R75 SEA 5 R25 R37 R12 R13 R6 R59 R87 R54 R10 R40 R31 R57 R67 R83 R85 R35 DOR 36 4 R50 Al-inhibited total root length (%) 100 Genotypes Figure 43. Total root length (A, cm plant-1), and inhibition of total root length (B, %) of 100 common bean genotypes (98 RILs of DOR364 x BAT 477 and 3 checks) evaluated in a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with or without 20 µM Al for up to 48 h, pH 4.5. Bars are means ± SD of nine replicates from 3 experiments. Horizontal line represents the genotypic mean value. 109 Figure 44. Root system of 2 selected RILs of DOR364 x BAT 477 and their parents after 48 h of exposure to a solution containing 0.5 mM Ca, 0.5 mM K and 8 µM B with 0 and 20 µM Al, pH 4.5. Conclusions: Significant genotypic variation in resistance to aluminum in RILs of DOR 364 x BAT 477 was found. A few RILs with low inhibition of root growth under high Al in solution were identified. Two RILs (BT 21138_128-1-M-M-M and BT 21138_ 2-1-1-M-M-M) were found to be outstanding in root growth both with and without Al in solution. Contributors: J. Ricaurte, R. García, F. O. Ibáñez, A. F. Rangel, S. Beebe, I. M. Rao (CIAT); W. Horst (Leibniz University of Hannover, Germany) 110 Activity 2.2 Developing germplasm with resistance to insect pests: Bruchids and leafhopper Highlights : • New accessions from the gene bank were evaluated for insect resistance • Resistance to Zabrotes subfasciatus was reconfirmed • New breeding lines that have resistance to the leafhopper (Empoasca kraemeri) were identified • Some Andean bean lines presented high level of tolerance to Empoasca and less yield loss 2.2.1 Screening for sources of resistance to major insect pests Rationale: Identification of sources of resistance to major insect pests of beans is a continuous activity. Additional work is conducted to identify and characterize the mechanisms of resistance to major specific pests. Materials and Methods: Germplasm accessions and breeding lines are planted in the field under high levels of natural leafhopper infestation, usually with 4-5 replicates per genotype in randomized complete block designs. Evaluations for resistance include damage score and bean productivity ratings, insect counts, damage counts and in some cases, yield components and yields. In the case of Zabrotes, the combination of biochemical tests to confirm the presence of arcelin and insect feeding bioassays has given excellent results. Leafhopper (Empoasca kraemeri) No useful sources of resistance to the leafhopper were found among about 150 accessions of bean germplasm evaluated in 2008. In trials conducted under field conditions at CIAT headquarters, Mesoamerican parents with different resistance sources were evaluated with 240 lines in replicated nurseries. 26 of these lines were selected as resistant to be evaluated again in 2009 to confirm their resistance. Storage weevil (Zabrotes subfasciatus) In 2008, special emphasis was placed upon the reconfirmation of resistance in previously selected RAZ lines. These materials were later tested in nurseries with 3 repetitions with six pairs of Z. subfasciatus per 30 seeds. The genetic improvement method using backcrosses that combine biochemical tests to confirm the presence of arcelin and insect feeding bioassays have had satisfactory results whenever the resistance to the Mexican bean weevil (Z. subfasciatus) is to be incorporated into bean cultivars. It is necessary to reconfirm resistance in the RAZ lines with the objective of assuring purity of the obtained lines. Table 40 shows that the first lines generated maintain a high level of antibiosis in its control of Z. subfasciatus. Contributors: J. M. Bueno, O. Díaz, J.F. Valor 111 Table 40. Levels of resistance identified to Zabrotes subfasciatus in RAZ lines Line RAZ 1 RAZ 4 RAZ 4-1 RAZ 4-2 RAZ 4-3 RAZ 4-5 RAZ 6 RAZ 7 RAZ 8 RAZ 9 RAZ 9-1 RAZ 9-4 RAZ 11-1 RAZ 12 RAZ 13-3 RAZ 13-4 RAZ 13-6 RAZ 14 RAZ 15 RAZ 16 RAZ 17-4 RAZ 17-5 RAZ 17-10 RAZ 18-1 RAZ 20-1 RAZ 20-2 RAZ 24-4 RAZ 24-6 RAZ 24-7 RAZ 25-1 RAZ 25-8 RAZ 29 RAZ 31 RAZ 32 RAZ 34 RAZ 36 RAZ 45 RAZ 53 RAZ 59 RAZ 62 RAZ 65 Percentage of adult emergence 7.4 6.8 8.0 9.6 4.6 9.5 7.7 6.7 5.2 8.3 7.9 8.6 8.8 5.4 6.8 6.9 5.9 12.0 3.3 16.7 12.7 6.4 13.3 7.4 16.8 19.1 14.2 10.2 12.2 25.3 14.2 16.4 24.2 25.0 1.2 1.2 5.8 5.4 6.3 3.5 2.3 Days to adult emergence 46.5 44.2 39.2 43.4 44.4 45.0 42.5 47.2 55.4 46.7 46.2 49.6 48.2 48.8 45.9 44.9 46.1 42.4 48.8 42.8 45.0 45.1 47.2 49.9 44.9 45.1 45.9 48.9 46.1 41.5 42.9 46.3 45.2 42.0 47.3 51.8 52.5 47.6 46.6 48.8 52.7 112 Isa Rating 1.1 0.9 1.8 2.0 0.7 2.0 1.2 1.0 0.7 2.0 1.6 1.5 1.6 0.8 1.6 1.3 1.1 2.8 -0.1 3.4 2.3 1.1 2.3 1.1 3.5 3.8 3.0 1.7 2.8 4.6 3.2 3.0 4.0 4.6 -2.3 -2.5 0.7 0.6 0.8 -0.5 -1.5 R R R R R R R R R R R R R R R R R R R I R R R R I I R R R I I R I I R R R R R R R Table 40. cont´d. Line RAZ 68 RAZ 74 RAZ 82 RAZ 84 RAZ 86 RAZ 87 RAZ 89 RAZ 92 RAZ 98 RAZ 109 RAZ 110 RAZ 114 RAZ 118 RAZ 119 RAZ 124 RAZ 126 RAZ 137 RAZ 143 RAZ 167 RAZ 168 RAZ 169 RAZ 173 RAZ 193 RAZ 136b RAZ 51 ICA Pijaoc a b Percentage of adult emergence 2.5 8.0 2.3 7.9 4.5 5.8 4.1 6.6 4.7 8.3 4.6 13.3 7.8 6.4 3.8 1.7 3.2 6.4 15.6 8.6 16.0 10.0 2.0 9.7 7.2 96.7 Days to adult emergence 49.2 46.5 49.5 48.1 46.9 50.7 48.4 48.9 46.9 47.4 47.5 48.4 50.8 49.1 47.6 55.2 50.5 45.4 49.7 45.5 46.6 57.9 63.1 47.1 47.9 33.8 Isa Rating -0.8 1.0 -1.0 0.9 1.0 0.6 0.2 1.2 0.3 1.6 0.6 2.0 0.9 1.2 -0.4 -1.7 -0.4 0.8 2.6 2.0 3.3 1.3 -1.3 2.3 1.6 9.5 R R R R R R R R R R R R R R R R R R R R I R R TR TR TS Index of susceptibility: (ln progeny per female/days to adult emergence) x 100; Resistant P. vulgaris line; c Susceptible P. vulgaris cultivar. 2.2.2 Developing germplasm resistant to insects 2.2.2.1 Storage weevils (Zabrotes subfasciatus) Rationale: The identification of arcelin as a biochemical marker for the selection of progenies with resistance to Z. subfasciatus has greatly facilitated the incorporation of this trait into a range of cultivated beans. Using arcelin-1 donor parents in simple crosses or backcrosses to the cultivated parents we have been able to generate additional progenies (RAZ lines) that have shown high levels of resistance to Zabrotes. The purpose of this project was to evaluate the segregation of arcelin-based bruchid resistance in crosses of two RAZ lines with a red-mottled, drought tolerant Andean parent. Materials and Methods: The work on the development of a molecular DNA microsatellite marker for arcelin presence and resistance to the Mexican bean weevil was as described in the 2003 Annual Report. The crosses were made between the drought tolerant Andean breeding line SEQ1006 and RAZ 105 and 113 RAZ 106 as arcelin-donor parents in 2006 and advanced to the F2 generation where all individuals were tested for the arcelin marker and the F3 seed harvested from each single plant selection. The resulting 247 F3 progenies from two different crosses were tested for resistance to Z. subfasciatus in replicated nurseries in the laboratory. The resulting F3 progenies were used to reconfirm marker selection. Results and Discussion: As shown in Tables 41 and 42, very high levels of resistance to the Mexican bean weevil were identified with absolute correspondence between the presence of arcelin and resistance to the insect. Table 41. Levels of resistance to Zabrotes subfasciatus in F3 seeds of progenies derived from crosses with RAZ 105 Cross Percentage of emergence Days to adult emergence Isa Best lines selected for % emergence 0.0 79.0 -12.5 0.0 79.0 -12.5 0.0 79.0 -12.5 0.0 79.0 -12.5 1.2 60.3 -9.9 1.2 75.7 -8.7 1.3 69.0 -8.8 1.4 71.7 -8.8 1.8 63.0 -5.0 2.1 59.7 -5.1 2.8 57.0 -4.7 2.9 70.1 -8.1 3.2 59.0 -4.7 4.2 56.7 -4.7 4.8 56.7 -4.2 5.0 65.7 -4.2 5.5 59.7 -4.3 5.5 57.2 -3.9 5.6 57.7 -4.3 5.6 47.0 -1.0 6.2 55.4 -3.9 6.3 57.5 -4.1 7.4 60.8 -4.3 8.1 51.7 -3.8 8.6 43.7 -0.7 9.6 50.2 0.6 Checks RAZ 44b 9.7 48.4 1.5 RAZ 36b 0.0 79.0 -12.5 RAZ 193b 0.0 79.0 -12.5 RAZ 105c 10.4 42.9 2.8 ICA Pijaod 97.2 32.3 7.4 LSD 24.7 14.4 6.0 a Susceptibility Index: (ln progeny per female/days to adult emergence) x 100; b Resistant P. vulgaris line; c donor parents; d Susceptible P. vulgaris cultivar. SEQ1006 x RAZ 105-32 SEQ1006 x RAZ 105-49 SEQ1006 x RAZ 105-66 SEQ1006 x RAZ 105-94 SEQ1006 x RAZ 105-18 SEQ1006 x RAZ 105-116 SEQ1006 x RAZ 105-21 SEQ1006 x RAZ 105-77 SEQ1006 x RAZ 105-50 SEQ1006 x RAZ 105-38 SEQ1006 x RAZ 105-54 SEQ1006 x RAZ 105-24 SEQ1006 x RAZ 105-104 SEQ1006 x RAZ 105-46 SEQ1006 x RAZ 105-80 SEQ1006 x RAZ 105-20 SEQ1006 x RAZ 105-44 SEQ1006 x RAZ 105-30 SEQ1006 x RAZ 105-4 SEQ1006 x RAZ 105-45 SEQ1006 x RAZ 105-37 SEQ1006 x RAZ 105-65 SEQ1006 x RAZ 105-61 SEQ1006 x RAZ 105-85 SEQ1006 x RAZ 105-10 SEQ1006 x RAZ 105-23 114 Table 42. Levels of resistance to Zabrotes subfasciatus in F3 seeds of progenies derived from crosses with RAZ 106 Cross SEQ1006 x RAZ 106-132 SEQ1006 x RAZ 106-35 SEQ1006 x RAZ 106-137 SEQ1006 x RAZ 106-87 SEQ1006 x RAZ 106-120 SEQ1006 x RAZ 106-148 SEQ1006 x RAZ 106-146 SEQ1006 x RAZ 106-18 SEQ1006 x RAZ 106-54 SEQ1006 x RAZ 106-124 SEQ1006 x RAZ 106-83 SEQ1006 x RAZ 106-119 SEQ1006 x RAZ 106-8 SEQ1006 x RAZ 106-44 SEQ1006 x RAZ 106-106 SEQ1006 x RAZ 106-75 SEQ1006 x RAZ 106-74 SEQ1006 x RAZ 106-31 SEQ1006 x RAZ 106-122 SEQ1006 x RAZ 106-23 SEQ1006 x RAZ 106-133 SEQ1006 x RAZ 106-149 SEQ1006 x RAZ 106-88 SEQ1006 x RAZ 106-67 SEQ1006 x RAZ 106-32 SEQ1006 x RAZ 106-82 SEQ1006 x RAZ 106-80 SEQ1006 x RAZ 106-28 SEQ1006 x RAZ 106-76 RAZ 193b RAZ 36b RAZ 44b RAZ 106c ICA Pijaoc LSD Emergence Days to adult Percentage emergence Best lines selected for % emergence 0.0 71.0 1.0 63.0 1.5 68.0 1.8 54.3 2.6 57.3 2.7 58.0 3.1 56.3 3.2 64.8 3.3 62.3 3.3 50.7 3.7 54.3 3.9 56.0 4.0 56.0 4.1 59.2 4.1 57.3 4.4 62.7 5.4 50.0 5.7 49.2 6.4 47.8 6.7 51.4 7.2 50.9 7.4 47.3 7.6 54.2 8.0 47.5 8.8 51.3 8.8 51.2 8.9 51.1 9.2 49.8 9.2 59.1 Checks 0.0 71.0 4.3 46.0 7.1 45.6 4.0 56.4 94.6 33.3 47 22 a Isa -14.5 -10.5 -9.9 -6.5 -5.8 -5.9 -5.9 -9.7 -5.5 -1.8 -1.6 -1.5 -5.4 -5.5 -5.4 -9.5 -0.8 -1.4 -1.2 -0.8 -0.6 -0.4 -0.7 -0.3 -0.1 -0.1 -4.6 -4.4 -4.7 -14.5 2.0 0.1 -0.4 6.7 14 Susceptibility Index: (ln progeny per female/days to adult emergence) x 100; b Resistant P. vulgaris lines; c donor parents; d Susceptible P. vulgaris cultivar. Contributors: J. M. Bueno, M. Blair, J.F. Valor, C. Muñoz 115 2.2.2.2 Crosses to incorporate arcelin-based bruchid resistance into Andean beans Rationale: The Arcelin resistance gene is the most effective resistance factor for the most common storage pests of common bean, namely the Mexican bean weevil, Zabrotes subfasciatus (Boheman). We have been making crosses between arcelin containing RAZ lines and a series of Andean and Mesoamerican beans with drought tolerance useful for Eastern and Southern Africa (Ethiopia, Kenya, Malawi, Tanzania and Zimbabwe). In addition we have been conducting marker assisted selection for the arcelin gene. The long-term objective of this work is to increase the efficiency of breeding for multiple constraint resistance and facilitate the pyramiding of bruchid resistance with other biotic and abiotic stress resistances. Materials and Methods: Crosses were generated to incorporate Arcelin-based bruchid resistance into a drought tolerant background and then transfer that resistance/tolerance to the small white, Ethiopian variety Awash Melka and to various Andean bean types. The bruchid resistance sources used in the crosses were RAZ44, RAZ105, RAZ106, RAZ 107, RAZ168, RAZ169 and RAZ170, the first of these with small white grain type and the remainder all with large red mottled grain type. The drought tolerance sources were the DRK, RAA, SAB, SEA and SEQ lines described in other sections of the report as well as RCB588, RCB591, SER16, SER119, SXB405, SXB412 and SXB418 used in crosses with Awash Melka. In addition some triple crosses were generated for the nutrition breeding program using NUA35 and NUA56, and some double crosses were generated with the RMA lines discussed previously, the Malawian release CIM9314-34, the Kenyan releases KAT B1 and KAT B9 and other African released varieties, such as Canadian Wonder, CAL96 and CAL143. Marker assisted selection was carried out as described previously using microprep DNA. Results and Discussion: For Andeans, a total of 251 F1 plants segregated for the arcelin locus in Palmira 2008a (with 141 of these segregating for the arcelin locus alone and 110 segregating for both arcelin and the bc3 gene). Of these 236 amplified with the arcelin marker and the proportion of resistant and susceptible genotypes are shown in Table 43. For Mesoamericans (Awash Melka), a total of 498 F1 plants segregated for the arcelin locus in seven different pedigrees and the resulting resistant, susceptible and heterozygous selection are shown in Table 44. Pedigrees of crosses with arcelin genes that have been advanced to the F2:4 and F1:3 generations appear in Table 45, including 16 simple crosses, 32 triple crosses and 60 double crosses. Collaborators: M.W. Blair, H.F. Buendia, F. Monserrate, S. Beebe (SBA-1, CIAT) T. Assefa (EARO, Ethiopia) J.M. Bueno, C. Cardona (Entomology) 116 Table 43. Selections made in 2008 b for the arcelin gene in triple and double crosses Cross type Triple crosses Double crosses Table 44. Segregation 61 101 20 54 Triple crosses used for marker assisted selection in the small white (navy) commercial class Pedigree AWASH MELKA x (RCB588 x RAZ44) AWASH MELKA x (RCB591 x RAZ44) AWASH MELKA x (SER119 x RAZ44) AWAHS MELKA x (SER16 x RAZ44) AWASH MELKA x (SXB405 x RAZ44) AWASH MELKA x (SXB412 x RAZ44) AWASH MELKA x (SXB416 x RAZ44) Totals Table 45. Arcelin + + - No. of rows 2 4 4 3 5 4 5 R (+) 11 24 27 4 42 29 44 181 H 6 11 9 17 14 14 4 75 Simple, triple and double crosses with RAZ lines Simple crosses BRB211 x RAZ170 BRB211 x RAZ169 BRB263 x RAZ170 BRB 264 x RAZ 104 RMA68 x RAZ168 RMA69 x RAZ170 RMA70 x RAZ167 RMA71 x RAZ105 RAZ107 x SAB576 SAB 568 x RAZ103 SAB 575 x RAZ105 SAB 581 x RAZ168 SEQ 1006 x RAZ 107 SEQ11 x RAZ170 SEQ1003 x RAZ170 SEQ1027 x RAZ104 Triple crosses RMA44 x (BRB264 x RAZ104) RMA52 x (BRB215 x RAZ103) RMA58 x (BRB264 x RAZ104) (BRB264 x RAZ105) x SAB630 (BRB215 x RAZ 103) x SAB630 (BRB264 x RAZ105) X SAB711 (BRB263 x RAZ169) X SAB 711 (BRB285 x RAZ103) X SAB 711 (BRB285 x RAZ103) X SAB 711 (BRB285 x RAZ103) X SAB 711 (RMA69 x RAZ169) X SAB 712 (RMA69 x RAZ169) X SAB 712 (RMA70 x RAZ169) X SAB 712 (RMA 70 x RAZ 168) X SAB 712 (BRB 285 x RAZ 103) X SAB 712 (BRB 215 x RAZ 104) X NUA 35 (RMA 69 x RAZ 169) X NUA 35 NUA35 x (RMA70 x RAZ167) NUA 35 x (BRB 264 x RAZ 105) NUA 35 x (BRB264 x RAZ104) NUA 35 x (BRB 215 x RAZ 103) NUA56 x (RMA70 x RAZ167) NUA56 X (SEQ1006 X RMX20) NUA56 x (RMA70 x RAZ168) (BRB215 x RAZ103) X SAB650 RAA19 X (BRB264 X RAZ105) RAA20 x (BRB264 x RAZ105) RAA21 x (BRB264 x RAZ105) DRK149 x (BRB264 x RAZ104) DRK149 x (BRB264 x RAZ105) (BRB263 x RAZ169) X DRK 149 DRK156 x (BRB264 x RAZ105) 117 S (-) 14 38 47 16 43 50 31 239 Table 45 cont’d. Double crosses (CAL143 x SAB620) x (RMA70 x RAZ167) (CAL 143 x SAB 620) x (BRB 264 x RAZ 104) (CAL143 x SAB620) x (RMA70 x RAZ168) (SAB628 x CAL143) x (BRB285 x RAZ103) (SAB628 x CAL143) x (CMB106 x RAZ103) (SAB628 x CAL143) x (DRK149 x RAZ103) (KATB1 x SAB625) x (BRB215 x RAZ103) (KATB1 x SAB625) x (BRB215 x RAZ104) (KATB1 x SAB618) x (BRB215 x RAZ103) (KATB1 x SAB618) x (BRB215 x RAZ104) (KATB1 x SAB618) x (BRB263 x RAZ169) (KATB1 x SAB618) x (BRB263 x RAZ105) (KATB1 x SAB618) x (BRB263 x RAZ104) (KATB9 x SAB617) x (BRB215 x RAZ104) (KATB9 x SAB617) x (BRB263 x RAZ169) (KATB9 x SAB617) x (BRB264 x RAZ105) (KATB9 x SAB617) x (BRB264 X RAZ104) (KATB9 x SAB617) x (BRB215 x RAZ103) (KATB9 x SAB622) x (BRB285 x RAZ103) (KATB9 x SAB622) x (CMB106 x RAZ103) (KATB9 x SAB622) x (BRB264 x RAZ104) (SAB628 x KATB1) x (BRB285 x RAZ103) (SEQ1003 x BRB264) x (SEQ1027 x RAZ105) (BRB285 x RAZ103) x (SEQ1036 x RMX20) (SEQ1036 x RMX20) x (BRB285 x RAZ103) (RMA71 x RAZ104) x (BRB215 x VAX6) (BRB215 x RAZ104) x (SEQ11 x RAZ169) (BRB264 x RAZ104) x (SEQ1003 x RAZ169) (SEQ1003 x RAZ169) x (BRB264 x RAZ104) (BRB266 x RMA60) x (SAB568 x RAZ103) (BRB266 x RMX19) x (SEQ1004 x RAZ167) (BRB268 x RMX19) x (SAB575 x RAZ104) (BRB215 x VAX6) x (RMA70 x RAZ168) (BRB215 x VAX3) x (RMA70 x RAZ167) (SAB617 x SAB621) x (SEQ11 x RAZ169) (CWONDER x SAB623) x (BRB285 x RAZ103) (CAL96 x SAB621) x (RMA69 x RAZ169) (CAL96 x SAB621) x (BRB264 x RAZ104) (BRB211 x VAX3) x (RMA69 x RAZ169) (SEQ11 X RAZ169) X (SAB621 X SAB635) (RMA70 x RAZ168) x (SAB621 x SAB635) (RMA71 x RAZ104) x (SAB621 x SAB635) (RMA71 X RAZ104) X (SAB617 X SAB621) (RMA70 x RAZ168) x (SAB620 x SAB631) (SAB620 x SAB631) x (RMA70 x RAZ167) (RMA69 x RAZ169) x (SAB620 x SAB631) (RMA68 x RAZ167) x (SAB620 x SAB631) (SAB617 x SAB621) x (BRB264 x RAZ105) (SAB617 x SAB621) x (BRB264 x RAZ104) (SAB620 x SAB634) x (BRB285 x RAZ103) (SAB617 x SAB621) x (BRB263 x RAZ169) (SAB617 x SAB621) x (BRB215 x RAZ104) (SAB616 x SAB629) x (BRB215 x RAZ103) (CIM9314-34 x SAB622) x (BRB215 x RAZ103) (CIM9314-34 x SAB622) x (BRB285 x RAZ103) (CIM9314-34 x SAB622) x (BRB264 x RAZ105) (CIM9314-34 x SAB622) x (BRB263 x RAZ169) (CIM9314-34 x SAB622) x (SEQ11 x RAZ169) (CWONDER X SAB623) X (RMA68 X RAZ167) (CWONDER x SAB623) x (SEQ11 x RAZ169) 2.2.2.3 Leafhopper (Empoasca kraemeri) Rationale: Although there is little direct breeding for Empoasca tolerance today, routine evaluations of breeding materials is a service to the breeding programs to complement data on other traits. In 2008 support was given to the breeding program with the evaluation of resistance to the leafhopper in a large nursery of red, black, carioca, cream and white colored beans, with different resistances and agronomic characteristics (anthracnose, angular leaf spot, BCMV, BGYMV, CBB, drought, low fertility and high concentrations of grain iron). Materials and Methods: Evaluations for resistance to E. kraemeri were done in the field under high level conditions of natural infestation. A randomized complete block design was used for this evaluation with 5 repetitions per genotype. Evaluations for resistance include a damage score and bean production rating, insect counts, damage counts and in some cases, yield and yield components. 118 Results and Discussion: Of the 232 evaluated entries, 1 was rated as resistant (damage scale of 6.5 or less in a 1 to 9 scale) and 25 as intermediate (damage scale of 6.6 to 7.0 in a 1 to 9 scale), with high infestation of 14.2 leafhopper nymphs per leaf at 50 days after planting (Table 46). The resistant line, SEN 62, is a black seeded line selected in the drought breeding project. Among the other lines with intermediate resistance, the AQB lines and the INB lines are interspecific progeny, derived respectively from P. coccineus and P. acutifolius. The SAB lines are Andean genotypes, also selected in the drought project. The selected lines will be reconfirmed in a field evaluation in year 2009. Table 46. Leafhopper damage scores and reproductive adaptation score of best white, carioca, cream, black and red lines with high agronomic value in the VEF 2008. Leafhopper Reproductive damage score c adaptation scored 6 AQB 608 CrSt S 6.6 4.0 7 AQB 609 CrSt S 6.8 3.8 154 INB 604 Bl S 7.0 4.3 155 INB 605 St S 7.0 4.0 156 INB 606 Bl S 6.8 3.8 609 SAB 515 RSt M 7.0 3.8 676 SAB 582 Wh M 7.0 3.3 678 SAB 617 RSt L 7.0 3.5 679 SAB 618 RSt M 7.0 3.3 683 SAB 622 R M 6.6 4.0 690 SAB 629 CrSt M 7.0 3.5 698 SAB 637 RSt M 7.0 3.3 704 SAB 643 RSt M 7.0 3.8 707 SAB 646 RSt M 7.0 3.5 842 SEC 18 RSt M 6.9 5.8 843 SEC 19 Wh S 6.9 4.3 846 SEC 22 Wh S 7.0 5.3 853 SEC 29 Wh S 7.0 4.8 911 SEN 57 Bl S 7.0 4.3 912 SEN 58 Bl S 7.0 5.3 913 SEN 59 Bl S 7.0 5.0 914 SEN 60 Bl S 7.0 4.5 916 SEN 62 Bl M 6.4 5.8 919 SEN 65 Bl M 7.0 5.3 920 SEN 66 Bl M 7.0 5.0 922 SEN 68 Bl S 7.0 4.8 1393 EMP 250 CrSt M 5.5 6.8 1394 EMP 486 R M 5.6 6.0 1395 EMP 512 CrSt M 5.4 6.8 1396 EMP 547 Wh M 5.8 6.3 1397 EMP 567 Wh M 5.5 6.8 1398 EMP 576 Bl M 6.1 5.5 1399 EMP 584 CrSt M 6.0 5.8 1400 EMP 595 R M 5.8 6.3 1401 BAT 41e R M 8.3 3.8 1402 ICA PIJAOf Bl M 6.5 6.0 a Wh= White; Bl = Black; Cr = Cream; St = Striped; R = Red; b S= small; M= medium; L= large; cOn a 1-9 score scale (1 = no damage: 9 = severe damage); d On a 1-9 visual scale (1, no yield, no pod formation; 9, excellent pod formation and filling, excellent yield); e Susceptible check ; fTolerant check. Entry No. Line Seed colora Seed sizeb 119 Continuing with the search for resistance to leafhopper in 2008, 73 lines characterized by their resistance to drought and with the bc-3 gene, low fertility and/or high iron with codes MAB, SQX, SBX, NEB, MIB, SER, SEN, INB, and AQB were evaluated for leafhopper damage. Nine selections were made under high levels of leafhopper infestation (12.4 nymphs per leaf at 50 days after planting) (Figure 45). The selected lines will be reconfirmed in a field evaluation in 2009. These reconfirmations will include a damage score and yield production rating, insect counts, damage counts and in some cases, yield and yield components. The selected lines are: NEB 31, MAB 84, SXB 119, AQB 149, MAB 159, SXB 175, SXB 184, SXB 192, MAB 335. Lines Select Lines EMP lines Cheks 9 Reproductive adaptatio 8 EMP 486 7 MAB 335 MAB 84 EMP 600 EMP 250 EMP 512 6 ICA PIJAO 5 4 EMP 500 Mean 3.8 ± 1.3 3 2 BAT 41 1 Mean 7.4 ± 0.9 0 0 1 2 3 4 5 6 7 8 9 Damage score Figure 45. The relationship between damage scores and reproductive adaptation scores in 73 lines coded MAB, SQX, SBX, NEB, MIB, SER, SEN, INB, and AQB tested for resistance to Empoasca kraemeri Contributors: 2.2.2.4 J. M. Bueno, S. Beebe Evaluation of Andean beans for resistance to leafhoppers Rationale: Most progress in breeding for resistance to leafhopper has been performed with Mesoamerican beans. Special effort is needed to improve the resistance of Andean types that tend to be highly susceptible. In 2008, efforts were aimed at the development of Andean type beans with tolerance to the leafhopper, Empoasca kraemeri. In particular, Empoasca is a problem in warm, dry climates where bean golden yellow mosaic virus (BGYMV) is also prevalent, and thus combinations of these two resistances are desirable. 120 Materials and Methods: Red-seeded lines Andean genotypes with the bgm-1 recessive allele for BGYMV resistance were evaluated for their resistance to the leafhopper. Evaluations for resistance to E. kraemeri were made in the field under conditions of high levels of natural infestation. A randomized complete block design was used for this evaluation with 5 repetitions per genotype. Evaluations for resistance include a damage score and bean productivity rating, insect counts, damage counts and in some cases, yield and yield components Results: Table 47 shows lines that are better than ICA Pijao, the standard tolerant check. Five of these genotypes are more tolerant than their donor parents SEL 1428, SEL 1433, EMP 122, EMP 277 and EMP 320. Table 47. Reconfirmed resistance to Empoasca kraemeri and yield of red lines with Andean genotypes crosses and corresponding parents. Damage scorea Code Reproductive adaptation scoreb Yield (kg ha-1) Protected Percentage yield loss Susceptibility indexc Nonprotected Crosses 1990.1 1457.9 27.8 1.3 1828.0 1118.3 39.6 1.7 1751.4 1620.9 7.9 0.7 1694.6 1598.7 6.1 0.6 1580.0 1222.2 23.1 1.2 1572.2 1614.7 0.0 0.4 2086.1 1904.9 9.1 0.6 1670.5 1223.2 27.9 1.3 1727.0 1325.6 22.6 1.2 1414.0 1431.0 0.0 0.5 EMP lines EMP122d 6.8 4.3 1290.9 1029.1 21.0 1.2 EMP277d 6.8 4.3 1937.9 1451.6 24.8 1.2 EMP320d 6.8 5.3 1731.5 1516.2 13.9 0.9 EMP597 5.4 7.0 2099.1 2204.9 0.0 0.1 Checks SEL1428d 7.2 4.0 1326.2 1007.5 24.9 1.3 SEL1433d 7.8 3.0 1309.5 706.7 46.7 2.1 ICA PIJAO e 6.9 5.7 2264.1 1838.6 17.9 0.9 BAT 41f 8.6 3.0 1590.3 1086.6 30.8 1.5 LSD 0.5 0.72 393.9 309.1 a On a 1-9 visual scale (1, no damage; 9, severe damage); b On a 1-9 visual scale (1, no yield, no pod formation; 9, excellent pod formation and filling, excellent yield); c Calculated with respect to the mean of the trial and the mean of the tolerant check; d Donor parents; e Tolerant check ; f Susceptible check. 3438-39 3397-98-1 3397-98-4 3397-98-6 3397-98-8 3397-98-12 218759F2(M)W3 218757F2(M)W4 218757F2(M)W10 218757F2(M)W15 Contributors: 6.7 6.7 6.7 6.2 6.8 6.4 6.3 7.0 6.2 6.7 4.3 4.3 5.3 5.7 5.3 6.9 5.8 4.7 4.0 4.3 J. M. Bueno, M. Blair 121 Activity 2.3 Developing germplasm resistant to diseases Highlights: • A large number of crosses were made to pyramid insect (bruchid) and disease (BCMNV and CBB) resistance with drought tolerance or mid-elevation adaptation in Andean bush beans. The arcelin gene was used as the source of bruchid resistance and was effectively selected for in 115 different cross combinations. Meanwhile, 187 cross combinations were generated for disease resistance. These crosses are being advanced at our drought stress and mid-elevation sites in Colombia. • Twelve new, large seeded red mottled bean varieties with multiple resistance to diseases and up to 30% better yield compared to commercial varieties released in six countries in eastern Africa. • Thirteen new red kidney varieties combining multiple stress resistance with high yield potential and marketable grain characteristics are released in seven countries in east and central Africa. • Eight new speckled sugar varieties with multiple disease resistance, marketable grain types and high yield potential (up to 24% over commercial checks) released for smallholder production in four countries in eastern Africa. • Eight new small and medium red bean varieties combining multiple disease resistance, high yield potential and marketable grain characteristics released for smallholder production in three countries of eastern Africa. • Eighteen new tan, brown and yellow seed varieties with multiple resistance to diseases and high yield potential released for production by smallholder farmers in four countries in eastern Africa • Twenty-six new climbing bean varieties combining multiple resistances to diseases with high yield potential and marketable grain characteristics released for smallholder production in seven countries in east and central Africa. • Several segregating populations and fixed lines in different market classes from South Africa, Malawi and Tanzania are available for distribution to interested NARS partners within SABRN and others in Africa • From the regional breeding program 24 (brown/khaki), 73 (sugar) and 213 (red mottled) developed for resistance to angular leaf spot, or common bacterial blight or low soil fertility or a combination of these stresses were distributed to various NARS programs. • Twenty-nine lines in various market classes which were developed for rust resistance or a combination of rust and angular leaf spot or rust and halo blight resistance by the NARS in South Africa were sent to the SABRN coordinator for seed increase and onward distribution to other interested NARS for the next planting season. • Twenty four new lines with a resistance gene to Pythium root rot were identified and included in the Pythium root rot nursery. • Forty lines combining resistance to Pythium root rot and angular leaf spot were identified and available for sharing with partners 2.3.1 Crosses to incorporate BCMV and CBB resistance into Andean beans Rationale: Bean common mosaic necrosis virus (BCMNV) is an aphid and seed transmitted Potyvirus that is very important in Eastern and Southern Africa where it causes necrotic symptoms on I gene containing genotypes. The related virus BCMV causes typical potyvirus symptoms on susceptible genotypes that do not contain the I gene. Both BCMV and BCMNV resistance is very important in dryland areas of Africa were aphid vectors are common. The most appropriate resistance combination is the I gene with a recessive resistance gene bc-3 which protects I gene containing genotypes from necrosis or the bc-3 gene alone due to its wide spectrum of resistance. Another important disease in the region is common bacterial blight (CBB) which is a foliar and seed-borne disease of beans grown in most tropical 122 lowland and subtropical areas of production especially during hot, humid summer weather. The disease is caused by the pathogen Xanthomonas axonopodis pv. phaseoli (Xap), which is widespread and part of a complex of Xanthomonad bacterial pathogens attacking many broadleaf and vegetable crops. Resistant bush beans were developed to both of these diseases in CIAT in the 1990s but have not been widely deployed in Andean breeding lines. As both diseases are important constraints in the same areas where drought occurs, it has been our goal to pyramid resistance to both diseases together with drought tolerance. This project describes the crosses made so far for this goal and outlines the initial attempts at pyramiding through marker assisted selection. Materials and Methods: Hybridizations were made in CIAT greenhouses between commercial Andean seed types and drought tolerance genotypes (SEA, SEQ, SAB lines) with sources of bean common mosaic necrosis virus resistance (BRB lines), common bacterial blight resistance (VAX, RMX lines) and arcelinbased bruchid resistance (RAZ lines). F1 seed was then planted in the Darien 2007b for triple and double crosses. These F1 in turn were planted in Palmira 2008a so as to make gamete selections that were used for marker validation using the markers ROC11 for BCMNV resistance, SU91 for CBB resistance and two new microsatellites associated with bruchid resistance. The Andean parents were from the red mottled, cream mottled, yellow and large red commercial classes, namely AND277, AND279, Canadian Wonder, CIM 9314-3, CMB106, CMB107, KATB1, KATB9, RMA44, RMA46, RMA52, RMA58, RMA60, RMA63, RMA66, RMA68, RMA69, RMA70, RMA71, RMA72, RMC57, RMC58, RMC65. The specific CBB resistant lines were VAX3, VAX6, RMX2, RMX8, RMX19, RMX20; while the BCMNV resistant lines were BRB198, BRB211, BRB263, BRB264, BRB265, BRB 266, BRB267, BRB268. The drought tolerant parents were DRK149, DRK156, RAA21, RAA34, SAB514, SAB516, SAB560, SAB568, SAB575, SAB576, SAB581, SAB630, SEA5, SEQ11, SEQ1003, SEQ1004, SEQ1006, SEQ1027, SEQ1032. Finally the arcelin containing parents were RAZ105, RAZ106, RAZ 107, RAZ168, RAZ169 and RAZ170. Results and Discussion: A total of 196 multiple F1 combinations were generated (88 from triple crosses and 108 from double crosses). Examples of these are given in Tables 48 and 49. In addition, 91 other F1 combinations from simple crosses (28 with BCMNV genes, 31 with CBB genes and 32 combining drought tolerance from SAB lines with larger seed size) were made and advanced to the F2:3 generation. Most of the combinations target Andean seed classes (red mottled, large red). Table 48. Examples of triple crosses combining BCMV, CBB or insect resistance genes together with drought tolerant and non-drought tolerant Andean parents. For mid-altitude adaptation RMA44 x (BRB264 x RAZ104) RMA44 x (BRB215 x VAX6) RMA44 x (BRB264 x VAX3) RMA44 x (BRB214 x VAX3) RMA44 x (BRB265 x VAX6) RMA52 x (BRB266 x RMX19) RMA52 x (BRB215 x RAZ103) RMA58 x (BRB266 x RMX19) RMA58 x (BRB264 x RAZ104) RMA58 x (BRB268 x RMX19) RMA58 x (BRB211 x VAX3) RMA60 x (BRB214 x VAX3) RMA60 x (BRB266 x RMA60) For drought tolerance RAA20 x (BRB264 x RAZ105) RAA20 X (BRB211 X VAX3) RAA21 x (BRB264 x RAZ105) RAA21 X (BRB211 X VAX3) DRK149 x (BRB264 x RAZ104) DRK149 x (BRB264 x RAZ105) DRK156 X (BRB211 X VAX3) DRK156 x (BRB264 x RAZ105) (BRB264 x RAZ105) x SAB630 (BRB215 x VAX6) x SAB630 (BRB215 x RAZ 103) x SAB630 (BRB214 X VAX3) X SAB630 (BRB215 X VAX6) x SAB645 123 Table 49. Examples of double crosses combining BCMV, CBB or insect resistance genes together with drought tolerant and non-drought tolerant Andean parents. (SAB619 X CAL143) X (BRB214 X VAX3) (SAB619 X CAL143) X (CMB106 X VAX3) (SAB619 X CAL143) X (BRB215 X VAX6) (SAB628 x CAL143) x (BRB285 x RAZ103) (SAB628 x CAL143) x (CMB106 x RAZ103) (SAB628 x CAL143) x (DRK149 x RAZ103) (CAL143 x SAB620) x (BRB264 x RAZ105) (CAL143 x SAB620) x (RMA70 x RAZ167) (CAL143 x SAB 620) x (BRB264 x RAZ104) (CAL143 x SAB620) X (BRB264 X VAX3) (CAL143 x SAB620) x (RMA70 x RAZ168) Marker assisted selection was used on a total of 383 gametes developed from BRB lines containing the bc-3 resistance gene, and 79 gametes developed from VAX lines containing a CBB resistance QTL. Some of the gametes for BCMV resistance (110) also segregated for arcelin-based insect resistance effective against the bruchid, Zabrotes subfasciatus. The ROC11 based selection was carried out with alkaline extraction DNA and agarose gel multiplexing as described previously, while the SU91 evaluation was performed with a mixture of different dilutions of alkaline extraction or miniprep DNA without multiplexing which made it more time consuming and expensive to carry out. Collaborators: M.W. Blair, F. Monserrate, A. Hincapie, S. Beebe 2.3.2 Development and release of new red mottled bean varieties with multiple constraint resistance in eastern Africa Rationale. The large seeded red mottled bean is probably the most important market class in East and Central Africa and in major bean producing countries in southern Africa such as Malawi. Red mottled beans are an important grain type in west and central African countries such as Cameroon. Red mottled beans command an estimated 22% market share of bean markets in East, Central and Southern Africa. An estimated 740,000 ha are sown with red mottled bean in Africa annually. Despite the rapid growth in demand for red mottled bean, supply has not kept pace in some countries. For example, Kenya is net importer of red mottled bean. Attempts to increase productivity have been severely constrained by biotic and abiotic stresses. Major biotic stresses include angular leaf spot, anthracnose, root rots and common bacterial blight. The most important abiotic stresses in red mottled bean growing areas are low soil nitrogen, phosphorus, soil acidity and frequent droughts. Kenya, Uganda, Tanzania and Malawi are not only the leading producers of red mottled beans but are also major consumers. Red mottled bean is gaining popularity in other countries in the region. For example, red mottled is now a popular grain type in southern Ethiopia since its introduction in the area in 1999/2000. In Awassa, the red mottled bean was selling at a higher price in local markets (20 to 30 Birr per kg, equivalent to US$ 2 to 3) five years after introduction, compared to the traditionally popular small red (CIAT, 2004). However, most of the regionally important red mottled commercial cultivars such as GLP 2 (Rosecoco), K20, K132 and Lyamungu 85 are susceptible to diseases, pests and low soil fertility (CIAT, 2005). This has contributed to decline in national production and low yields in farmers fields, adversely affecting food security, nutrition and incomes of bean growers in the region. A regional program was started in 2001 to develop improved red mottled bean lines with resistance to diseases (especially angular leaf spot, anthracnose, common bacterial blight and root rots) and tolerance to abiotic stress factors especially low soil fertility and drought. Development of the new varieties involved assembling a working collection, identification of sources of resistance, development of breeding populations, identification of lines combining resistance to two or more biotic and abiotic stresses, regional evaluation of promising lines and release of varieties. Since 2000, the regional program adopted a decentralized market-led breeding strategy with responsibilities shared among the nine network member countries (CIAT, 2001; Kimani, 2005). Regional 124 breeding activities for red mottled grain type were based at Namulonge in Uganda and Kabete in Kenya. This report highlights milestones realized by this program towards development of a new generation of red mottled bean varieties in eastern Africa. Materials and Methods: Working Collection. A breeding collection, which comprised of segregating populations, advanced breeding lines and other sources of resistance to angular leaf spot, anthracnose, root rots and tolerance to low soil phosphorus and nitrogen was created at Kabete. The collection included materials from CIATColombia, Kenya and CIAT-Uganda. Sources of resistance. Potential parents for the crossing program were identified from old and current commercial cultivars contributed by national programs. Sources of resistance originated from previous germplasm evaluations for biotic and abiotic stresses in Africa and Colombia. Mexico 54, G5686 and BAT 332 were used as donors for resistance to angular leaf spot, G2333 and NB 123 for anthracnose, POA 2 and FEB 190 for common bacterial blight, GLP 92 (Mwitemania) for resistance to halo blight, SCAM 80 CM/5, L226-10 and RWR 719 for root rots, and RWR 2075 and RWR 1946 for tolerance to low soil fertility, and L226-10 for bean common mosaic virus. L226-10 is resistant to rust, bean common mosaic virus (BCMV), angular leaf spot and several root rots (Freytag et al., 1985). RWR 2075 and RWR 1946 are also resistant to root rots (Namayanja et al., 2003). Characteristics of other parental lines are shown in Table 50. Population development. A multiple stress breeding approach based on multi-parent crosses was used to create breeding populations (Singh, 1994). Backcrosses and simple crosses to correct specific deficiencies for simply inherited resistances also were made at Kabete and Namulonge. Complex crosses were made to combine single resistances from selected parental lines. Three sets of populations were created: KP 01, KP 86 and KP04. KP01 was developed from bi-parental and multi-parent crosses among 51 genetically diverse lines from Andean and Middle-American gene pools. The parents included lines with known resistance to angular leaf spot, anthracnose, common bacterial and other diseases and tolerance to low soil fertility. KP 86 was derived from a seven parent complete diallel cross at Kabete. It was used to generate 33 F2 populations. KP04 was derived from bi-parental crosses among recent sources of resistance and popular commercial varieties. This population was designed to correct deficiencies of the commercial parents. Selection for resistance from KP01 populations. Selections were made for resistance to angular leaf spot, anthracnose, common bacterial blight, bean common mosaic and necrotic viruses (BCMV/BCMNV), rust and plant type in the early generations (F2 to F4). Both artificial inoculations and natural epiphytotics were used to identify resistant genotypes. Fifty-two segregating populations from KP 01 and lines from other crosses were advanced to F4 generation at Kabete Field Station and subjected to natural epiphytotics of angular leaf spot, root rots, anthracnose and rust. F2 plants were advanced as progeny rows in F3 and F4. Selection criteria included growth habit, reaction to diseases, vegetative vigour, pod load and seed characteristics. Seed harvested from F4 plants was separated into seven market classes (red mottled, dark red kidneys, small reds, yellow and brown, sugars, carioca and pintos). Seed of each market class was divided into four parts. The first part was screened for tolerance to angular leaf spot and Pythium root rot under artificial inoculation at Kawanda; the second part was screened in a field plot heavily infested with root rots in Sabatia. A third part was grown in low soil phosphorus test site in Kakamega, in Western Kenya, for soil acidity at Mulungu Research Station in Kivu province (D. R. Congo), and for tolerance to low soil nitrogen at Selian Agricultural Research Institute (Tanzania). These low soil fertility screening sites were selected by the Bean Improvement for Low soil Fertility (BILFA) working group (Lunze et al., 2007). 125 Table 50. Line Characteristics of some parental lines used in multi-parent crosses. Seed color Seed size *Growth habit Anthracnose Common bacterial blight Angular leaf spot PVA 800A Red mottled medium IIB 2 7 4 ICA Quimbaya red large I 2 7 7 XR-12307-1 red small III 1 Catrachita red small III 2 8 7 AND 277 red medium I 3 5 4 A222 black small II 4 9 3 G16140 Gray stripped large III 1 7 3 AFR 188 red medium I 2 4 5 G10613 white small III 2 7 5 XAN 309 red small IIB 7 3 7 G5686 Yellow mottled large I 2 8 2 A193 Red mottled large II 2 3 3 MAR 3 pinto small III 2 5 2 G5653 pink small III 2 8 4 PVA 1111 red large I 2 7 4 AFR 612 Red mottled medium I 2 5 4 BRB 190 Red mottled medium I 2 7 3 MAM 48 pinto medium III 7 5 3 CAL 167 Red mottled medium II 7 6 3 ICA Tunduma Red large II 7 3 AND 279 Red mottled medium I 2 7 3 CAL 143 Red mottled medium I 2 6 4 Calima Red mottled large I 5 5 4 * Growth habit : I=determinate, upright; II= semi-determinate, III= semi-climber. Disease scores: 1-3= resistant, 46= intermediate and 7-9=susceptible. Selection from KP86 populations. The segregating populations were selected for eight generations for resistance to six diseases (rust, angular leaf spot, anthracnose, halo blight, BCMV and common bacterial blight), yield and seed characteristics both in the greenhouse and in the field. Disease assessment was based on artificial inoculations and natural epiphytotics in the field. Disease assessment was done 21 days after inoculation (R6) and also at mid-pod filling (R8). Susceptible plants were discarded. To determine the yield potential of early generation lines, more than 200 F4 and F5 lines were evaluated at five locations for two seasons. Selection from KP04 Populations. Eleven F2 populations segregating for red mottled grain types and resistance to diseases were developed from the KP04 crosses (KAB02, KAB 03, KAB 04, KAB05, KAB06, KAB 07, KAB10, KAB11, KAB12, KAB 13 and KAB 14) (CIAT, 2005). Single plants were selected from F2 populations grown at Thika and Kabete during long rain (LR) season (April to August). The selected plants were used to establish F2.3 progeny rows at Kabete and Thika during short rain (SR). Plants were rated for diseases, phenology, grain yield and other agronomic traits using the CIAT standard scale (Schoonhoven and Pastor-Corrales, 1987). Susceptible plants were discarded. Resistant plants within a progeny row were bulk-harvested and evaluated in replicated trials at the two locations during the LR (F2.4) and SR (F2.5) seasons. F2.5 generations were screened for angular leaf spot at Kabete and root rots in an infested field in Sabatia (western Kenya), and at a disease hotspot in Laikipia (Rift Valley region). 126 Line development. One hundred red mottled lines selected from KP01 were evaluated in major agroecological zones in Uganda, Ethiopia, D. R. Congo, Cameroon, Kenya, Rwanda and Tanzania between 2004 and 2008 to identify lines with broad and country specific adaptation both on-station and in farmers’ fields. Regionally important red mottled commercial cultivars were included as checks. The trials were laid out in lattice design with three or four replicates. Each plot had four, 5 m rows. Agronomic data was collected from the inner two rows. Intra-row spacing was 10 cm and 45 between rows. Rating for biotic and abiotic stresses followed CIAT (1987) standard system. Collaborators added other promising red mottled lines to the standard set (25 entries) for comparison. Fifty-one lines were selected from KP 86 populations and evaluated in advanced yield trials at 10 locations for three seasons. Four commercial varieties (GLP2, GLP 24, GLP 92 and GLP 1004) were included as checks. Twenty-one lines were selected for participatory on-farm evaluations in eight locations for two seasons. They were distributed as a regional nursery. Two hundred eighty one F2.6 lines selected from seven KP04 populations were evaluated in preliminary and intermediate yield trials at Kabete, Laikipia and Ol Jorok (Kenya). The 281 F2.6 lines were originated from seven of the 11 populations: KAB 02 (67 lines), KAB 05 (16 lines), KAB 06 (70 lines), KAB 12 (25 lines), KAB 10 (47 lines), KAB 11 (30 lines) and KAB 13 (26 lines). These lines were previously selected for resistance to diseases, plant type and other agronomic traits for five generations at Kabete, Sabatia, Ol Jorok and Laikipia field sites in Kenya. Variety release. Each country selected candidate varieties from the regional nurseries for final evaluations and variety released based on the national variety release procedures. Results and Discussion Reaction to Diseases Natural epiphytotics facilitated elimination of plants and progeny rows susceptible to angular leaf spot, anthracnose, rust, and common bacterial blight in F2, F3 and F4 at Kabete (1860 masl). However, the severity of diseases varied with seasons. Root rots occurred sporadically especially after the heavy rains. In Kawanda, 453 lines were inoculated with Mesoamerican and Andean races of Phaeoisariopsis griseola in a screenhouse. One hundred seventy-two lines were resistant to Mesoamerican races; 173 showed intermediate reactions and 86 were susceptible. When inoculated with Andean races, 184 lines showed resistant reactions, 99 were intermediate and 148 were susceptible. Both Andean and Mesoamerican races and their intermediates occur in many bean growing areas in East and Central Africa. However, Andean races are more widespread. Results showed that 143 lines were resistant to races from the two gene pools. Table 51 shows some of the genotypes with combining resistance to races from the two gene-pools. Two lines NM 12667-4A-1 and UBR (93)22-5-1 had no disease after inoculation with the two race groups. NM 12667-4A-1 was selected from the cross (PVA 773 x ICA Tunduma)F1 x(PVA 800A x((XAN 309 x A193)F1 x(MAR3 x G5653)F1) . ICA Tunduma, PVA 800A, A193, MAR 3 and G5653 are resistant to angular leaf spot (Table 50). This suggested that NM 12667-4A-1 may have several alleles conditioning resistance to races of Mesoamerican and Andean Phaeoisariopsis griseola. 127 Table 51. Disease scores for the best lines selected from the segregating populations and other nurseries at Kawanda under artificial inoculation with Mesoamerican and Andean races of Phaeoisariopisis griseola. Lines NM 12667-4A-1 DOR 676-1 L R K 32 NR 12632-3D-1 FEB 191-1 K28/13A I-1-1 VTTT 915/11-2 OBA 3-1 NR 12638-7C-1 NR 12638-3B-1 NR 12636-4A-2 NR 12634-13 NR 12631-12 NR12793-6 B UBR(93)22-21-1 VTTT 915/7 BOA 5-8/2 Mesoamerican 1 1 1 1 1 1 2 1 2 1 2 1 1 1 1 1 2 Andean 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 9 9 9 9 9 9 9 9 9 9 Checks VTTT 918/11-1 VTTT 919/8-1 VTTT 919/10-2 VTTT 919/16-2 VTTT 920 /10-1 A total of 715 lines were artificially inoculated with Pythium spp at Kawanda. Results showed that three lines were resistant, 239 intermediate and 473 were susceptible. Promisnig lines are presented in Table 52. Pythium spp. are frequently the initial incitant of root rots in bean growing areas of East and Central Africa. Outstanding lines resistant to Pythium were NR 12631-7-1, NM 12657-5C-2 and NM 12646-3-1. All were derived from multi-parent crosses. Three hundred lines were screened for resistance to root rots in a field plot heavily infested with the disease in Sabatia. Twenty-five lines succumbed to the disease and produced no seed. All the other lines produced seeds which varied from 0.5 to nearly 400 g m-2. Table 53 shows that the best lines produced significant amounts of seed despite heavy disease pressure. Outstanding lines included selections from RAB 475, DFA 62, NR 12797-8-1, NR 12633A-6-1, NR 12633-5-1 and NM 12805-7C-1. These lines produced over 300 g m-2. The results showed that some of the outstanding lines in greenhouse and field screening originated from the same crosses: NR 12631, NM 12657, NM 12646 and NR12633 (Tables 52 and 53). Mortality among the best lines ranged from 0% (DFA 62-1 and NM12646-3-1) to 48% (NR 12656-8A-1). A total of 275 lines were selected for further evaluation. These represented not only the red mottled class but another four commercial classes as well. The methodology described here is relevant for those classes, especially the red kidney class. 128 Table 52. Disease scores of selected red mottled and red kidney lines artificially inoculated with Pythium spp at Kawanda Agricultural Research Institute, Uganda. Line NR 12631-7-1 NM 12657-5C-2 NM 12646-3-1 NR 12632-3G-2 NR12633-11ª NR 12793-8-1 AND 1056-1 DOR 703 NM 12803-11 NR 12 638-7B RWR1873 RWR 719 NR 12633-5-1 RWK 10 NR12633-4E-1 P 94056 NR 12797-8C-1 NR 12638-10-1 NR 12634 -1B-1 NR 12633-5B-1 AND 1055-1 POA 8-1 RWR 10 Mean 2.8 2.9 3.0 3.1 3.1 3.2 3.3 3.3 3.3 3.3 3.3 3.4 3.4 3.4 3.5 3.6 3.7 3.7 3.8 3.8 3.8 8.8 9.0 Table 53. Seed yield and percent morality of bean lines selected in a field plot infested with root rots in Sabatia, Western Kenya. Line RAB 475-1 DFA 62-1 NR 12797-8-1 NR 12633 A-6-1 NR 12633-5-1 NM 12805-7C-1 NR 12631-3D-1 NM 12652-9-1 NM 12806-2A-1 AND 1055-1 NR 12657-12 NR 12634-1C-1 NR 12649-B-1 NM 12651-9-1 NR 12631-9 NM 12651-116-1 RWR 1742-1 NM 12651-14-1 NM 12646-3-1 NR 12656-8A-1 Percent mortality 15 0 10 36.8 27.7 25 26.3 10 16 10 20 35 31.5 30 16 15 17.6 20 0 48 129 Seed yield (g m-2) 399 385 336 319 305 300 299 293 286 282 274 262 253 250 250 246 244 242 231 224 Adaptation to low soil P, N and pH Red mottled lines with tolerance to low P and low pH were AFR709-1, NM 12650-4C, AND 932-1 and NR 12631-9. The lines AND 932-1, VTTT 919-1, AFR 709-1 and NM 12650-4C combined tolerance to low soil acidity and low N. Seven red mottled lines combined tolerance to low soil P and N. These were AFR 709-1, NM 12656-6-1, AND 932-1, NM 12650-4C, NM 126-2A-1, NM 12805-7C-1 and NM 12655-3-1. Three red mottled lines were tolerant to all three stresses: AFR 709-1, AND 932-1 and NM 12650-4C. Evaluation of advanced lines In Uganda, seven lines were selected. These were ECAB 0060, ECAB 0070, ECAB 0090, AFR 623, POA8, F7MG/1 and POA 4. POA 4 was released as NABE 4. Based on on-station and on-farm evaluations for two seasons, 15 lines were selected in D. R. Congo from the regional multiple constraint nursery. These included CAL 143, AND 907, AND 897, AND 1060, AFR735, AFR 699, UBR93/4, AND 1005, POA 2, CAL 175, CAL 172, VAC 49, POA 8, AFR 623 and CAL 176. The regional nurseries were also distributed for further evaluation and selection in M'vuazi and Equator regions in western DRC. Selections from western D. R. Congo were further distributed to countries in West and Central Africa Bean Research Network (WECABREN) in 2006, 2007 and 2008. In southern Ethiopia, ten lines produced higher yields compared regional commercial varieties (K132 from Uganda, CAL 143 from Malawi, GLP 2 from Kenya and Lyamungu 85 from Tanzania). Yields varied from 2018 kg ha-1 for the local check to 3321 kg ha-1 for ECAB 0027 (Table 54). Growing conditions were favourable and no major disease incidence was recorded. The best yielding lines were ECAB 0027, ECAB 0008, ECAB 0081, ECAB 0063, ECAB 0042, ECAB 0043, ECAB 0098 and ECAB 0019. In Rift Valley region of Ethiopia, selected lines were evaluated in multi-location trials and are in the final stages of release. In Madagascar, two red mottled lines were selected. Yields were lower in Madagascar compared to other sites. The best performing lines were ECAB 0034 (890 kg ha-1) and ECAB 0063 (866 kg ha-1). Based on performance at three locations over two seasons, the best performing red mottled lines in Kenya were: ECAB 0019, ECAB 0027, E8, ECAB 0063, ECAB 0041, ECAB 0060, ECAB 0023, ECAB 0081, ECAB 0098, ECAB 0013, ECAB 0097, ECAB 0056, ECAB 0043, ECAB 0008, ECAB OO20, ECAB 0047 and ECAB 0068 (Table 55). These results indicate that most of the lines performed well in more than one country, indicating broad adaptation. Two outstanding lines were selected at sites in four countries (ECAB 0060 and ECAB 0063). Six lines showed good performance in three countries. These were ECAB 0020, ECAB 0019, ECAB 0023, ECAB 0081 and ECAB 0013. These results indicate that it was possible to identify new red mottled lines with better grain yield and resistance to the major diseases with broad adaptation. Lines with broader adaptation are preferred by seed producers because they justify seed production for a larger market. In Rwanda, five red mottled varieties were selected in 2008 after evaluations in on-station and on-farm trials for the last four years (Table 56). 130 Table 54. Plant height, 100-seed mass and grain yield of red mottled bean lines selected at Awassa, southern Ethiopia. Line ECAB 0056 ECAB 0034 ECAB 0042 ECAB 0008 ECAB 0063 ECAB 0060 ECAB 0068 ECAB 0041 ECAB 0043 ECAB 0047 ECAB 0020 ECAB 0098 ECAB 0019 ECAB 0023 ECAB 0081 ECAB 0013 ECAB 0050 ECAB 0027 ECAB 0097 ECAB 0082 K132 CAL 143 GLP 2 Lyamungu 85 Local check Mean Plant height (cm) 39 39 100-seed mass 41.5 42.6 Grain yield (kg ha-1) 2555 2581 34 36 43 33 40 38 34 39 39 34 35 38 35 37 36 42 48 40 36 35 45 37 38 37.6 45.5 51.6 46.7 42.7 43.0 41.8 50.8 44.7 43.2 41.6 41.5 45.8 47.9 43.7 45.1 49.1 47.0 42.5 43.8 38.2 40.6 49.7 50.2 44.8 2991 3261 3163 2389 2576 2789 3121 2706 2574 2990 2981 2956 3173 2329 2783 3331 2780 2039 2156 2398 2455 2217 2018 2692 131 Table 55. Days to 50% flowering, maturity, seed mass and grain yield of advanced generation red mottled bean lines grown at two locations in Kenya. Genotype ECAB 0007 ECAB 0006 ECAB 0042 ECAB 0023 ECAB 0038 ECAB 0012 ECAB 0010 ECAB 0043 ECAB 0033 Grain yield (kg ha-1) Days to flowering Days to maturity 100-seed mass (g) 43 40 41 42 41 41 42 41 40 84 82 81 82 83 83 83 81 81 45.5 47.1 41.5 43.0 51.4 47.9 46.3 49.7 56.8 Kabete 2352 2415 2054 2015 1897 2199 2286 2011 2001 Thika 1884 1755 1974 1980 2014 1619 1456 1717 1719 Mean 2118 2085 2014 1998 1955 1909 1871 1864 1860 43 44 43 42 42 44 43 43 41 41 42 40.3 42.8 41.5 3.5 ** ** ** 84 84 84 82 83 85 83 83 83 83 84 81 84 83 2.0 ** ** ** 47.5 47.2 47.2 38.2 46.8 45.4 47.4 48.1 38.1 32.5 47.3 45.4 49.9 47.6 1714 1752 1834 1446 1745 1522 1470 1560 1623 1773 1830 1927 1927 1303 1134 1365 1348 1152 1294 1045 1208 1315 1390 1535 1372 1372 1508 1443 1600 1397 1448 1408 1257 1384 1469 1581 1683 1372 1927 1650 22.1 ** * NS Checks GLP 2 Goberasha K132 KK 8 Lyamungu 85 Lyamungu 90 Melke PVA 8 SCAM 80CM/ 15 Selian 94 Simama Mean (Thika) Mean (Kabete) Trial Mean CV (%) Locations (L) Genotypes (G) GxL Error ** ** NS 132 Table 56. Disease score and grain yield of four red mottled lines selected for multiple disease in Rwanda, 2008 Line* BCMV ECAB 026 ECAB 001 ECAB 019 ECAB 0037 ECAB 0019 Ascochyta 2 2 2 2 3 2 3 3 3 4 Anthracnose 1 1 1 1 1 Angular leaf spot 3 5 6 3 3 Rust Grain Yield (kg ha-1) 3333 2833 3267 2333 1833 2 2 2 1 1 * ECAB 026, ECAB 001 and ECAB 019 were selected at Rubona and ECAB 0037 and ECAB 0019 at Rukira. ECAB 0037 and ECAB 019 had resistant reaction to common bacterial blight at Rukira. Source: Félicité Nsanzebara, 2008 Performance of Advanced lines from KP 86 and MCN nurseries. Twenty-one lines from KP86 population were distributed to bean programs in Uganda, Ethiopia, D. R. Congo, Tanzania and Kenya. Two lines KS 65-2 and KS 47-1 were selected in western D. R. Congo and pre-released (Kimani, 2006). In Kenya, five lines were recommended for full release after evaluation in national performance trials in December 2007 (Table 57). The results showed that the candidate varieties were more resistant to angular leaf spot, BCMV, halo blight and common bacterial blight compared to the susceptible checks. All candidate varieties, except KK22, were more tolerant to anthracnose compared with GLP 2. KK22 also was more susceptible to bean common mosaic necrotic virus (BCMNV) compared with other varieties and checks. Table 57. Disease score and grain yield of five red mottled lines selected for multiple disease in Kenya and evaluated in national performance trials at nine sites for three seasons, 2008 Genotype Market class Angular* Leaf spot Anth. BCMNV BCMV Halo blight CBB E8 M22 AFR 708 KK8 Lyamungu 85 Checks GLP 2 GLP 92 LSD 0.05 Red mottled Red mottled Red mottled Red mottled Red mottled 2.3 2.1 2.2 2.3 2.3 1.4 1.6 1.7 1.6 1.4 1.1 1.0 1.2 1.1 1.2 2.2 2.4 1.8 1.9 2.6 1.5 1.7 2.0 1.7 1.8 2.2 2.4 1.8 2.0 1.8 Grain Yield (kg ha-1) 1320 1140 1050 1020 1010 Red mottled Pinto 2.4 2.8 0.41 1.9 1.7 0.54 1.1 1.2 0.75 2.2 2.8 0.76 2.1 1.6 0.32 2.2 2.9 0.68 1060 800 260 Source: NPT Reports 2006-2007, KEPHIS; * Disease severity scores, anth= anthracnose. Release of new varieties: Table 58 shows some of the bush red mottled varieties released in east and central Africa since 2003. Contributors: Paul Kimani, Robin Buruchara (CIAT), Annet Namayanja (Uganda), Félicité Nzansebara, (Rwanda), Asrat Asfaw (Ethiopia) and Nkonko Mbikayi (D. R. Congo). Collaborators: Bean Teams of Madagascar, Uganda, D. R. Congo, Rwanda, Ethiopia, Kenya and Tanzania 133 Table 58. Red mottled varieties released in eastern Africa between 2003 and 2008. Variety New Rosecoco Chelalang Kenya Umoja Super Rosecoco CAL 98 RWR 2355 RWR1180 Shyorongi Ibado RWR 2142 Lyamungu 90 NABE Line Code E8 Lyamungu 85 AFR 708 M22 CAL 98 RWR 2355 RWR 1180 RWR 2245 AFR 722 RWR 2142 Lyamungu 90 AFR721 Year of Release 2008 2008 2008 2008 2007 2007 2007 2007 2006 2006 2005 2003 Countries of release Kenya Kenya Kenya Kenya Madagascar Rwanda Rwanda Rwanda Ethiopia Rwanda DRC -West Uganda References CIAT. 2001. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2005. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2006. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia Schoonhoven A van and Pastor Corrales, M.A. 1987. Standard system for the evaluation of bean germplasm. CIAT, Cali, Colombia. Freytag, G. F., Kelly, J. D. and Lopez, R. J. 1985. Registration of two navy bean germplasm lines L226-10 and L227-1. Crop Sci. 25: 714. Kimani, P.M. 2005. Bean research for development strategy in central and eastern Africa. Internacional de Agricultura Tropical (CIAT), Kampala, Uganda. 2p. Highlights: CIAT in Africa Number 14. Lunze L., P.M. Kimani, R. Ngatoluwa, B. Rabary, G.O. Rachier, M. M. Ugen, V. Ruganzu and E. Awad Elkarim. 2007. Bean improvement for low soil fertility adaptation in Eastern and Central Africa, pages 325-332. In: Bationo et al (eds.). Advances in Integrated Soil Fertility Management in sub-Saharan Africa: Challenges and Opportunities. Springer, Dordrecht, The Netherlands. 1094pp. Namayanja, A., P. Tukamuhabwa, F. Opio, M.A. Ugen, P.M. Kimani, A. Babirye, X. Kitinda, P. Kabayi and R. Takusewanya. 2003. Selection for low soil fertility bean tolerant to root rot. Bean Improvement Cooperative J. 43: 95-96. Singh, S.P. 1994. Gamete selection for simultaneous improvement of multiple traits in common bean. Crop Sci. 34:352-355. 2.3.3 Breeding red kidney bean varieties with multiple constraint resistance in eastern Africa Rationale: Red kidney bean is one of the most important grain types grown by smallholders and consumed in rural and urban centres in East and Central Africa. An estimated 350,000 ha are grown annually in Africa (Wortmann et al., 1998). Red kidneys are highly marketable in many areas and have potential for export. They account for about 11% of beans marketed in this region. A market survey conducted in 2000 by the East and Central Africa Bean Research Network (ECABREN) showed that red kidney bean was important in Kenya, Tanzania, Burundi, Rwanda, Cameroon, southern Ethiopia, southern Sudan and Madagascar. However, these seed types are relatively low yielding under the lowinput management. They are susceptible to biotic and abiotic stresses. Their productivity is severely constrained by angular leaf spot, anthracnose, root rots, bean stem maggot, low soil phosphorus and nitrogen, and drought. Production is dominated by a single variety, Canadian Wonder which was introduced in the region more than 50 years ago (CIAT, 2004). However the productivity of this variety has been on the decline because of susceptibility to diseases, especially angular leaf spot, anthracnose, 134 common bacterial blight, root rots, drought and declining soil fertility. More recent releases such as Selian 97 have yet to meet consumer acceptability comparable to Canadian Wonder. Red kidney bean is predominantly produced by smallholder farmers, with limited options for inputs to reduce effects of biotic and abiotic stresses (CIAT, 2003). Because most of the resource poor smallholder farmers can hardly afford purchased inputs, breeding cultivars tolerant to the major stresses is considered as the most effective strategy for developing improved cultivars. A regional breeding program led by Selian Agricultural Research Institute (SARI) in Arusha, Tanzania and University of Nairobi, Kabete (Kenya) was started in 2001 to develop new high yielding red kidney cultivars with resistance to two or more biotic and abiotic stresses and with desirable grain characteristics for smallholder farmers in East and Central Africa ( Kimani, 2005). Selection was carried out concurrently from existing and new populations at Arusha and Kabete. Promising lines with resistance to two or more or biotic and abiotic and improved yields were constituted into a regional nursery in 2003. The regional red kidney nursery was distributed to other national programs for participatory evaluation by farmers and researchers. This report highlights progress made in development of red kidney varieties fro smallholder farmers in eastern Africa between 2003 and 2008. Materials and Methods: Red kidney program started with development of segregating populations from simple and multiple crosses, and introduction of red kidney grain type from CIAT, Colombia. During the first phase of this work, the segregating populations were selected for six to eight generations under natural and artificial disease epiphytotics, low soil fertility and drought stress. In second phase, red kidney lines with acceptable grain and resistance to major stresses were entered into regional evaluations to expose them to a wider range of pathogen diversity and production environments in East and Central Africa and to identify candidates for national and regional releases. Population development. Germplasm from multiple constraint nurseries, segregating populations, advanced lines, regional disease nurseries and the regional program at Kabete (Kenya) and Selian Agricultural Research Institute, Arusha (Tanzania). Sources of resistance to anthracnose, angular leaf spot and root rots were identified and used in crosses with susceptible commercial cultivars. At Selian, G 5686, BAT 332 and Mexico 54 were the used as sources of resistance to angular leaf spot, common bacterial blight, powdery mildew and rust based on local validation trials. UBR (92)25 was the source of resistance to low soil N, P and drought. G 22501, G22258 and G11746 were used as sources of resistance to bean stem maggot and drought. A total of 11 single, 21 three-way and 11 double cross populations were developed (Table 59). At Kabete, breeding started with creation of segregating populations from bi-parent and multi-parent crosses. Parents were selected for known resistances to angular leaf spot, anthracnose, drought and tolerance to low soil fertility. Selection for multiple stress resistance Selection at Selian. Selection for priority stresses was performed from eight multiple constraint nurseries (MCN) constituted at Kabete in 2000, and also from segregating populations. MCN nurseries had 294 red kidney lines distributed as follows: MCN I (12 lines), MCN II (157 lines), MCN III (20 lines), MCN IV (47 lines), MCN V (12 lines), MCN VI (21 lines), Kabete PSP (18 lines) and KS-PSP (7 lines). These lines were evaluated for most important diseases in northern Tanzania which include angular leaf spot, common bacterial blight, anthracnose, rust, bean common mosaic virus and bean stem maggot at Selian and Olmotony field sites. Starting in 2001, a regional nursery of 100 red kidney advanced lines from the regional program at Kabete was evaluated for resistance to angular leaf spot, common bacterial blight, anthracnose, bean common mosaic virus and tolerance to manganese and iron toxicity at Selian. 135 Table 59. Populations generated at Selian Agricultural Research Institute, Tanzania to combine marketable grain characteristics of commercial cultivars with resistance to major biotic and abiotic stresses. Single cross Populations CW* x G 11746 Three way cross populations Double cross populations (CW x G 22258) x Mex 54 (CW x G22258) x ( MASAI RED x Mex 54) CW x G22258 (CW x G 22258) x G 5686 (CW x G 22258) x ( MASAI RED x G 5686) CW x G22501 (CW x G 22258) x BAT 332 (CW x G22258) x ( MASAI RED x BAT 332) CW x Mex 54 (CW x G 22258) x UBR (92) 25 (CW x G22258) x ( MASAI RED x BAT 332) CW x UBR (92)25 (CW x Mex 54) x UBR (92) 25 CW x G 5686 (CW x Mex 54) x G 11746 CW x BAT 332 (CW x Mex 54) x G 22501 (CW X Mex 54) X ( MASAI RED X UBR(92) 25) (CW x MEXICO 54) x ( MASAI RED x G 11746) (CW x G5686) x ( MASAI RED x (UBR92) 25) Selian 97 x G11746 (CW x G5686) x UBR(92) 25 (CW x G 5686) x ( MASAI RED x BAT 332) Selian 97 x G22258 (CW x G 5686) x BAT 332 (CW x G 11746) x ( MASAI RED x G 5686) Selian 97 x Mex 54 (CW x G 5686) x G 11746 (CW x G 11746) x ( MASAI RED x BAT 332) Selian 97 x G5686 (CW x G 5686) x G 22501 (CW x G 11746) x ( MASAI RED x UBR (92) 25) (CW x G 5686) x G 22258 (CW x G 11746) x UBR (92) 25 (CW x G 11746) x BAT 332 (CW x G 11746) x Mex 54 (CW x G 11746) x G 5686 (SELIAN 97 x G 5686 )x G 22258 (SELIAN 97 x G 5686) x G 22501 (SELIAN 97 x G 5686 )x G 11746 (SELIAN 97 X G 5686) X BAT 332 (SELIAN 97 X G 5686) X UBR (92)25 * CW= Canadian Wonder (syn GLP 24 in Kenya) Selection at Kabete. More than 60 F2 populations were initially grown Kabete Field station and selected for tolerance to diseases, seed type, type I or II growth habit over two generations. Selected plants were advanced in single plant progeny rows in F3 and F4 generations. F4 bulks were screened for resistance to Pythium root rot, angular leaf spot and anthracnose using artificial inoculation at Kawanda Agricultural Research Institute (Uganda). They were concurrently tested for tolerance to low soil phosphorus in Kakamega and in a root rot infested field plot in Sabatia (Western Kenya). To select for yield potential, F5 and F6 lines were grown in preliminary and intermediate yield trials at four locations in Kenya. About 102 selected lines were evaluated in advanced yield trials at three locations. Two regionally important red kidney cultivars, Canadian Wonder and Selian 97, were included as checks. The 102 were constituted into a regional red kidney nursery which was distributed to collaborating countries interested in this grain type (CIAT, 2004). 136 Results and Discussion: Selection for multiple resistance. More than 140 populations from single, three-way and double crosses were generated at Selian and Kabete. F2 populations were screened for resistance to diseases at Kabete Field station and selected for seed type, type I or II growth habit over two generations. Selected plants were advanced in single plant progeny rows in F3 and F4 generations. Populations were managed as described for the red mottled class, including disease inoculations and evaluations. Red kidney lines found to combine tolerance to low soil P included NM 12806-2A-1, AND 1055-1, RWR 1742-1, NR 12634-6-1 and NR 12634-13C-1. AND 1055-1 was resistant to root rots in Kakamega and under artificial inoculation in Kawanda. Lines with combined tolerance to low P and low N included RWR 1742-1 and full sibs from the crosses NM 12806-2A and NR 12634-13C, NR 12638-1B, NR 12639-5 and NR 126349B-1, RWK 10, VTTT 920-26, NM 12806-2A and NR 12634-13C. Two lines showed high levels of tolerance to the three stresses. These were NM 12806-2A-1 and NR 12634-13C. Selections were also made from KP86 populations. Canadian Wonder (GLP 24) was one of the parents from which these populations were derived (Table 60). Table 60. F4 and F5 lines with multiple resistance selected from KP 86 populations inoculated with disease isolates at Kabete. Cross/population Number of lines Combinations of resistance* GLP 288 x M535 25 CBB, BCMV, angular leaf spot, bean stem maggot L226-10 x NB 123 M535 x L226-10 GLP 2 x NB 123 GLP 2 x M535 GLP2 x GLP 288 GLP 2 x L226-10 GLP 92 x NB 123 GLP 288 x GLP 24 19 10 5 3 6 18 3 19 BCMV, rust, angular leaf spot, halo blight, bean stem maggot BCMV, halo blight, CBB BCMV, rust, anthracnose and halo blight BCMV, anthracnose, CBB, rust and angular leaf spot Rust, angular leaf spot, BCMV, anthracnose, CBB, BSM Rust, angular leaf spot, anthracnose, CBB Rust, angular leaf spot, halo blight, BSM Rust, angular leaf spot, CBB and BCMV * BCMV= bean common mosaic virus, CBB= common bacterial blight, BSM= bean stem maggot. Source: CIAT, 2007 Evaluation of Advanced lines. At Selian, 43 lines selected from the MCN nurseries were subsequently distributed for further evaluation in Madagascar, Uganda and Rwanda (Table 61). In Tanzania, 23 lines with multiple resistance to diseases, low soil fertility and drought were selected from 294 MCN lines and evaluated in preliminary and advanced yield trials, uniformity cultivar trials (UCT) and finally the national bean yield trials (NBYT). However, selection from the segregating populations was delayed following of departure of the breeder for further studies. Low yields were recorded in Madagascar. Yields of the best 12 lines varied from 710 to 1693 kg ha-1. However, only five lines had better yields than Canadian Wonder (GLP 24) with a mean yield of 825 kg ha-1. These were ECAB 0240 (889 kg ha-1), ECAB 0247 (875 kg ha-1), AND 931-B1 (982 kg ha-1), TZ 201-439-3 (777 kg ha-1), EMP 250-51 (1142 kg ha-1), VTT 926/2-4 (1693 kg ha-1) and UBR (91)45-1 (1266 kg ha-1). ECAB 0240 and UBR (91)45-1 flowered in 40 days. TZ 201-439-3 was the last to reach 50% flowering (48 days). 137 Table 61. Red kidney lines selected at Selian and distributed to Uganda, Madagascar and Rwanda. Source Nursery MCN I Lines AND 932B –1, FEB 147-1, EMP 263A-1, RAA 28-1, TZ 201439-3, RWR 1742-1,RWR 1946-2, RWR 1873, RWR 1896-1 MCN II NR 1263–7-1, NR 12634 –2A-1, NR 12634 –6A-1, NR 12634-9B-1, NR12634 –11B-1, NR12634 -11C, NR 12634 –12B –1, NR 12635 –2B –2, NR 12638 –2 -1, NM 12531 –9 –1, NM 12657 – 10B –1, NR 12672 – 5 –1, EMP250 –5-1 MCN III MCN IV LRK 32, AND 1064-1, AND 1063-1, FEB 195-1, POA 8-1, RAA 31-2 AND OBA 3-1 VTT 920/26, VTT 921/11, VTT 921/11-1, VTT 926/2-4 and VTT 926/12-2 MCN IV Kabete PSP KS PSP UBR (91) 15 –1, UBR (92)17-2 and UBR (91)45-1 K13/4A –C – 1 and K13/41 AF 12-12 KS 7 –1, KS 45-1, KS 45-3-2 and KS 56-1 In contrast, relatively high yields were recorded in Ethiopia. Grain yield varied from 2300 kg ha-1 (ECAB 0281) to 3452 kg ha-1 (ECAB 0270). However, 21 lines (including Canadian Wonder) yielded better than Selian 97. In Kenya, the best yielding lines were (in decreasing order) ECAB 0296 , ECAB 0224 , ECAB 0240, ECAB 0234, ECAB 0219, ECAB 0246, ECAB 0228, ECAB 0290, ECAB 0262, ECAB 0288, ECAB 0251, ECAB 0231, ECAB 0232, ECAB 0248, ECAB 0282, and ECAB 0292. Mean yields at Kabete and Thika varied from 1908 kg ha-1 for ECAB 0292, to 2218 kg ha-1for ECAB 0296. Mean yields were 1322 kg ha-1for Canadian Wonder and 1326 kg ha-1 for Selian 97. Twenty-one lines selected at Awassa (Ethiopia) were also selected in previous trials in Kenya. Among the lines selected in four countries were: ECAB 0201, ECAB 0252, ECAB 0240, ECAB 0267 and ECAB 0247. Thirteen lines performed well and were selected in three countries. Only four lines were selected in two countries (ECAB 0295 and ECAB 0224). Variety Release. Three red kidney (M18, L36 and L41) were among the 16 candidate varieties validated in national performance trials conducted over three seasons in 2005 and 2006 at nine locations in Kenya. Table 62 shows some characteristics of these lines. Table 62. Genotype Mean performance of red kidney lines in the national performance trials conducted at nine sites for three seasons in Kenya. Market class Angular* Leaf spot Anth. BCMNV BCMV Halo blight CBB Grain Yield (kg ha-1) 1130 1090 1050 L41 Red kidney 2.1 1.6 1.1 2.7 1.9 2.2 M18 Red kidney 2.0 1.4 1.0 2.7 1.7 2.0 L36 Red kidney 2.3 1.6 1.0 2.4 1.7 2.2 Checks GLP 2 Red mottled 2.4 1.9 1.1 2.2 2.1 2.2 1060 GLP 92 Pinto 2.8 1.7 1.2 2.8 1.6 2.9 800 LSD 0.05 0.41 0.54 0.75 0.76 0.32 0.68 260 Source: NPT Report 2006, KEPHIS; * Disease severity scores, anth= anthracnose, BCMNV= bean common mosaic necrotic virus, BCMV=bean common mosaic virus, CBB= common bacterial blight. 138 Several lines of red kidney have been released in eastern Africa. Table 63 shows some of released lines between 2003 and 2008. NABE 13 (RWR 1946) is high yielding and is resistant to bean root rots, angular leaf spot, anthracnose and is adapted to low soil fertility. NABE 14 (RWR 2075) is a high yielding variety with multiple resistances to low fertility acid soils, root rot, anthracnose and angular leaf spot. Table 63. Red kidney and large red bush varieties released in eastern Africa between 2003 and 2008. Variety Line Code Kenya Red Kidney M18 Kabete Super L36 Kenya Wonder L41 Nitu G16157 DRK 64 DRK 64 UBR (91)45-1 UBR (91)45-1 ODR ODR RWR 2091 RWR 2091 Selian 06 Flor de Mayo NABE 14 RWR 2075 NABE 13 RWR 1946 ACOS Red Source: National Bean program reports, 2006-2008. Contributors: Year of Release 2008 2008 2008 2005 2008(pre-release) 2008 (pre-release) 2008 (pre-release) 2006 2008 2006 2006 2007 Country of release Kenya Kenya Kenya D. R. Congo Madagascar Madagascar Madagascar Rwanda Tanzania Uganda Uganda Ethiopia Paul Kimani, Sostenes Kweka (Tanzania), Annet Namayanja (Uganda), Hery (Madagascar), Lodi Lama (D. R. Congo) and Robin Buruchara (CIAT) Collaborators: Asrat Asfaw, Bean Teams at Selian, Kabete, Namulonge, Kawanda, Antananarivo, Awassa, Rubona and Melkassa. References CIAT. 2003. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2004. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2005. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2007. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia. Kimani, P.M. 2005. Bean research for development strategy in central and eastern Africa. Internacional de Agricultura Tropical (CIAT), Kampala, Uganda. 2p. Highlights: CIAT in Africa Number 14. Wortmann, C.S, R.A. Kirkby, C. A. Eledu and D. J. Allan. 1998. Bean Atlas. CIAT, Colombia. 2.3.4 Development and release of new Speckled Sugar bean varieties with multiple disease resistance in eastern Africa Rationale: Speckled sugars belong to the cream colored market class, which accounts for 10% of Africa’s annual production. They are produced on 240,000 ha annually in eastern and central Africa, and on 120,000 ha in southern Africa. Wortmann et al (1998) reported that speckled sugars are of high to moderate importance in Kenya, South Africa, Angola, Uganda, Zambia, Zimbabwe, Lesotho and eastern Congo. Speckled sugars are high in demand in South Africa and Zimbabwe. South Africa imports speckled sugars from China and other countries to meet its domestic requirements. Southern Africa countries are therefore a potential market for ECABREN countries. In the Great lakes region, sugar bean 139 is grown in eastern D. R. Congo for export to urban centers in the western part of the country. Brown and speckled sugars are also important for domestic markets in Ethiopia, Uganda, Tanzania and Rwanda. Speckled sugars are exported to regional markets by D. R. Congo and to international markets by Madagascar. However, productivity of speckled sugars is constrained by rust, angular leaf spot, common bacterial blight, halo blight and poor tolerance to low soil nitrogen and phosphorus. Varieties with multiple resistance to diseases is probably the most effective and efficient strategy for managing these diseases, especially in low input systems common in eastern Africa. The East and Central Africa Bean Research Network (ECABREN) in collaboration with Southern Africa Bean Research (SABRN) network started a collaborative program to develop high yielding speckled sugar varieties with resistance to two or more biotic and abiotic constraints and marketable grain characteristics. The national bean program of D. R. Congo (INERA) has been leading a regional initiative to develop and disseminate improved varieties of speckled sugars for the last eight years, with a back-up regional program in Kenya, focusing on smallholder production. Objective of the program is to develop speckled sugar cultivars with improved grain yield and resistance to angular leaf spot, common bacterial blight, rust and tolerance to low soil fertility (Kimani, 2005). Selection activities were conducted collaboratively in Kenya and D. R. Congo countries with promising candidate lines constituting a regional nursery for evaluation by interested member countries. This report highlights progress made in this program since its inception. Materials and Methods: A regional germplasm collection was assembled at Kabete and screened for sugar grain type. The collection comprised of segregating populations (KP01) derived from 52 parents of diverse genetic backgrounds and known resistance to specific stress factors, especially angular leaf spot, common bacterial blight and halo blight. Included in this collection were six multiple constraint nurseries (MCN) constituted from advanced lines developed at CIAT, Colombia and from regional breeding programs in East and Central Africa. The materials were initially screened for adaptation and tolerance to biotic stress factors at Kabete Field Station, University of Nairobi and at INERA- Mulungu Research Station in D. R. Congo. At Mulungu, lines selected from F2 and F3 populations were evaluated in participatory trial sites in eastern D. R. Congo (Mbikayi and Kimani, 2004). In Kenya, single plant selections were selected from KP01 population and used to establish F3 and F4 progeny rows at Kabete Field Station. Progeny rows were selected on basis of reaction to angular leaf spot, rust, root rot, common bacterial blight and plant characteristics. F5 lines were grown in preliminary yield trials. F6 lines were grown in intermediate yield trials at three locations. Twenty-seven selected lines were finally selected for advanced regional yield trials in Uganda, D. R. Congo and Kenya. ‘Brown Speckled’ released in Ethiopia and Sugar 73 (Uganda) were included as checks. Disease scoring followed the standard CIAT scale (Schoonhoven and Pastor-Corrales, 1987). In Kenya, progeny from KP 86 F2 populations were selected for eight generations for resistance to six diseases (rust, angular leaf spot, anthracnose, halo blight, BCMV and common bacterial blight), yield and seed characteristics both in the greenhouse and in the field. Disease assessment was based on artificial inoculations and natural epiphytotics in the field. Disease assessment was done 21 days after inoculation (R6) and also at mid-pod filling (R8). Susceptible plants were discarded. To determine the yield potential of early generation lines, more than 200 F4 and F5 lines were evaluated at five locations for two seasons. Fifty-one lines were selected and evaluated in advanced yield trials at 10 locations for three seasons and also distributed to countries interested in this grain type (Tanzania, Uganda, D. R. Congo and Kenya). Four commercial varieties (GLP2, GLP 24, GLP 92 and GLP 1004) were included as checks. Twenty-one lines were selected for participatory on-farm evaluation in eight districts for two seasons. Yield data for 51 lines and the four check cultivars in 24 environments were analyzed for type 1 and type 4 stability parameters (Francis and Kannenberg, 1978; Lin et al 1986; Lin and Binns, 1988, 1991). Finally, nine lines were selected for multi-location national performance trials conducted at nine sites for three seasons. 140 Results and Discussion: Performance of F8 lines selected from the KP01 populations in Kenya is shown in Table 64. Results showed that there were significant genotypic differences for duration to flowering, maturity, pod m-2, 100-seed mass and grain yield (Table 64). Environmental effects were significant for phenology, angular leaf spot, anthracnose, rust, root rot, 100- seed mass and grain yield (P>0.01). Significant genotype x environment interaction was detected for days to flowering, root rot, days to maturity, 100-seed mass and grain yield. Angular leaf spot incidence was higher at Thika. Anthracnose incidence was highest at Juja. Rust incidence was highest at Juja during short rain season. Root rot was most severe at Thika. ECAB 0811 and ECAB 0822 showed intermediate reactions to root rot at this site. All other lines were rated resistant. The results indicated that some of the new lines had considerable yield advantage compared to the check cultivars. The results showed that 21 new lines produced higher grain yield compared to ‘Brown Speckled’ (Table 64). Seven lines produced higher grain yield than Sugar 73. These results were subsequently confirmed in advanced yield trials. Ten lines showed better yield that Sugar 73 and Brown Speckled. These were ECAB 0806, ECAB 0822, ECAB 0810, ECAB0807, ECAB 0805, ECAB 0808, ECAB 0817, ECAB 0826 and ECAB 0802. Mean yield of test lines over two environments varied from 1484 to 1675 kg ha-1 compared with 1455 kg ha-1 for Sugar 73 and 1411 kg ha-1 for Speckled Sugar. The test lines showed resistant reactions to angular leaf spot, anthracnose, rust and root rots in two environments. Results from INERA-Mulungu are presented in Table 65. Seven sugar lines were selected from 40 test lines. Five of the selected lines flowered and matured earlier than M’Mafutala, the check variety. Five lines showed higher 100-seed mass and grain yield than the check. These lines showed either resistant or intermediate reactions to angular leaf spot, anthracnose, ascochyta blight and rust at Mulungu. Two lines had more than 500 kg ha-1 yield advantage over the check. Advanced yield trials were conducted at INERA-Mulungu for the 27 lines received from the regional speckled sugar nursery in Kenya. These were separated into bush growth habit (Type I and II) and climbers (Type III and IV). Results showed that lines with bush growth habit (Types I and II) flowered in 39 to 48 days, and matured in 82 to 94 days. As expected, climbing bean lines flowered and matured later. There was high disease pressure due to angular leaf spot at Mulungu. However, all selected lines had intermediate reactions to angular leafspot. Most of the lines showed resistant scores to anthracnose, aschochyta and rust. Grain yield varied from 1009 kg ha-1 for RWV 1128-2 to 2004 kg ha-1 for KS 1513F11-1, and 2024 kg ha-1 for P94056. The climbing sugar bean lines did not show the expected superiority for grain yield compared to the bush lines. Ten bush lines and three climbers were selected for further evaluation. The selected bush lines were P94056, KS 65-2, NM 12652/9A-1, NM 12650/4A-1, NM 12633/9A, VTTT 926/3-5, NM 12656/14-1, MX 875-3T, NM 12647/A-1 and KS 151-3F11-1. The selected climbers were MAC 70-2, RWV 1134 and RWV1128-2. In Western Congo, two lines KS 65-2 and KS 47-1 which performed well in lowland conditions were recommended for pre-release. In Uganda, MAC 31 was selected. This line became very popular in local markets because of its large seeds and pods. It is now widely grown in eastern Uganda (Mbale district) for domestic and export markets. 141 Table 64. Mean duration to flowering, maturity, pods m-2, 100-seed mass and grain yield of F8 speckled sugar bean lines selected from KP01 populations and grown at four environments in Kenya. Genotype Days to 50% flower (d) Days to maturity (d) Pod m-2 100seed mass (g) Grain yield (kg ha-1) Kabete ECAB0811 ECAB0807 ECAB0801 ECAB0822 ECAB0813 ECAB0815 ECAB0823 SUGAR 73 ECAB0810 ECAB0808 ECAB0827 ECAB0809 ECAB0821 ECAB0802 ECAB0805 ECAB0806 ECAB0817 ECAB0814 ECAB0804 ECAB0826 ECAB0818 ECAB0824 Brown Speckled 2263 1567 1684 1955 1678 1437 1373 1597 1149 1756 1889 900 1373 1356 1040 1785 1390 1449 1026 1364 806 684 831 Juja (SR) Juja (LR) Mean 82.1 81.1 83.5 82.5 83.0 80.6 80.3 82.9 83.3 80.9 82.8 82.2 80.3 82.2 81.3 83.2 80.8 81.0 82.7 81.4 81.9 81.1 81.9 174.8 170.0 188.9 146.3 192.4 150.9 170.7 197.8 179.1 168.2 154.8 185.3 120.4 179.8 201.5 186.5 156.9 147.2 197.2 163.7 154.3 168.1 171.1 50.7 45.6 37.1 41.7 45.8 45.7 42.0 52.3 46.8 42.3 39.8 47.1 44.3 45.8 42.6 44.0 43.8 44.4 37.8 43.1 40.6 41.5 34.7 41.5 3.9 81.0 2.1 165.9 25.1 43.0 4.5 Genotypes (G) ** ** ** ** ** Environments (E) ** ** ** ** ** Mean CV(%) 42.8 40.6 42.2 43.0 42.4 39.7 39.4 42.8 43.3 40.4 43.2 43.0 38.2 41.7 40.5 42.9 40.2 39.5 43.1 40.2 40.9 41.3 41.8 Thika 1330 1862 1768 1784 1238 1179 1686 898 1675 1602 1500 1071 1755 1373 1711 1463 1566 1126 1314 1810 807 1859 1592 926 2105 1986 1858 1910 2167 1598 2231 1276 1684 2001 1579 1812 1757 1399 1617 876 1574 1879 1260 1257 1135 1775 1578 2185 2079 1925 2067 2059 2208 2297 2115 2212 1386 1930 1899 1857 1846 2093 1538 1496 762 1226 1879 1252 809 1421 2104 1850 1813 1793 1771 1732 1700 1666 1662 1661 1617 1592 1590 1578 1553 1441 1397 1351 1331 1327 1263 1215 1189 1413 1521 1602 1476 21.0 GxE ** ** NS ** ** *, **: Significant at 5 and 1 % probability levels, respectively; NS= not significant; SR= short rain and LR=long rain seasons. 142 Table 65. Days to 50% flowering, maturity, and seed mass and grain yield of sugar grain type bean lines selected, Mulungu, D. R. Congo. Genotype P94056 KS 65-2 MCD 2519-1 NM 12652/9A-1 NM 12633/9A VTTT 926/3-5 NM 12656/14-1 M’Mafutala (Check) Trial Mean LSD (0.05) CV (%) Days to flowering 42 48 41 41 39 41 39 48 Days to maturity 92 92 85 85 82 88 82 92 100-seed mass (g) 27.5 34.8 33.5 16.0 44.2 40.2 42.5 23.1 Grain yield (kg ha-1) 2024 1858 1707 1418 1411 1327 1246 1216 1191 495 23.7 Participatory Variety Selection. A total of 113 on-farm trials were conducted in Kenya; 47 in the long rain season and 66 during the short rain season. Sixty-six farmers were interviewed during the post-trial evaluation. Most farmers had grown the test lines for the two seasons. Over 95% of the participating farmers were women. Except for Nakuru, Embu and Kisii, farmers in other districts grew the lines in pure stands. In Nyeri, farmers selected line E2 because of high yield, taste, thick soup and fast cooking. In Taita-Taveta, farmers selected E7 based on grain color, yield, earliness, seed and pod size, disease resistance and tolerance to heavy rain. In Embu, farmers selected E4 and E7. Primary selection criteria included maturity, yield, seed traits, taste, reaction to diseases and pests, and tolerance to excess rain. Farmers in Kisii, Kakamega and Machakos listed similar criteria. Stability analysis. Plotting the grand mean yield against the coefficient of variation (CV%) divided the genotypes into four groups: Group I had high yield, small variation; Group II had high yield, large variation, Group III had low yield, small variation, and Group IV had low yield and large variation. E7 showed the showed the smallest variation across environments. GLP 585, a commercial check had the lowest grain yield and the largest CV across environments. Analyses based on type 1 stability parameter showed that E2 and E7 combined high yield and stability. These two lines were rated better than the commercial checks. E2 was rated as high yielding and stable by type 4 stability analyses. National performance trials and release. Based on farmer assessments and on-station evaluation, four sugar lines were registered for the nation performance trials (NPT). This trial with 16 bush entries lines and three checks was conducted at nine sites. The national trials were conducted over two long rain (March-August) and short rain seasons (November –December). The check varieties were GLP x 92 (‘Mwitemania’), GLP 2 (Rosecoco) and GLP 1127 (‘New Mwezi Moja’). Table 66 shows the performance of sugar lines in these evaluations. 143 Table 66. Genotype E2 E4 E7 Checks GLP 92 GLP 2 Mean performance of speckled sugar genotypes in the national performance trials conducted at nine sites for three seasons in Kenya. Angular Leaf spot* Anth. BCMNV BCMV Halo blight CBB 2.3 2.2 2.5 1.5 1.5 1.4 1.0 1.0 1.1 2.0 2.6 2.5 1.8 2.0 1.7 2.2 2.6 2.4 Grain Yield (kg ha-1) 1200 1070 1080 2.4 2.8 1.7 1.9 1.1 1.3 2.8 2.2 2.2 2.9 2.9 2.2 800 1060 Source: NPT Reports 2006 and 2007, KEPHIS ; * Disease severity scores. The results showed that E2 had 17.7% yield advantage over the best check and 24.8 % over the mean of the checks. E4 had a 4.2% yield advantage over the best check across the nine environments and 11.1% over the mean of the checks. E7 showed a 5.1% yield advantage over the best check and 12.1% over the mean of the checks. Disease severity rating showed that the test lines had favorable reactions to several diseases, and showed lower levels of susceptibility compared with the check varieties. For example GLP 92 had 6.63% incidence for angular leaf spot compared to 4.4% for E2, 4.6% for E4 and 5.3% for E7. For anthracnose, GLP 2 had a 3.5% incidence compared with 1.9% for E2, 1% for E7 and 1.9% for E4. BCMV incidence was 23.8% for GLP 92 compared with 12.5% for E2, 18.9% for E7 and 19% for E4. Incidence of common bacterial blight was 17.4% for GLP 92 compared with 9.7% for E2, 9.8% for E7 and 7.6% for E4. Mean incidence for halo blight was 3.6% for GLP 2, compared with 2.7 % for E2, 3.2 for E7 and 1.5% for E4. The three sugar lines (E2, E4 and E7) were recommended for full release the National Variety Release Committee. Variety Release and Conclusions: Several new speckled sugar varieties were released in eastern Africa between 2003 and 2008 (Table 67). In Uganda, Sugar 73 was released as NABE 5. MAC 31, a climber was released as NABE 12. In Ethiopia, ‘Kranskop’, which originated from South Africa was released. In Kenya, three bush and one climbing bean speckled sugar varieties were formally released in 2008. These results indicate that improved sugar bean varieties were identified from the working collection and locally developed populations. However, color retention in sugars remains a challenge. Most sugar varieties change from preferred wine red speckles on cream or white background to brown after storage, thus losing their consumer appeal. Table 67. Speckled sugar bush and climbing bean varieties released in eastern Africa between 2003 and 2008. Variety Line Code Miezi Mbili E2 Kenya Early E4 Kenya Sugar bean E7 Kenya Safi MAC 13 NABE 5 Sugar 73 NABE 12 MAC 13 Kranskop Kranskop KS 65-2 KS 65-2 Source: National program reports, 2006-2008 Year of Release 2008 2008 2008 2008 2006 2007 2007 2006 (pre-release) 144 Country of release Kenya Kenya Kenya Kenya Uganda Uganda Ethiopia D. R. Congo (west) Contributor: Paul Kimani (CIAT) and Agnes Mwangombe (Kenya). Collaborators: Nkonko Mbikayi (D. R. Congo), Annet Namayanja (Uganda), Teshale Assefa (Ethiopia) References Mbikayi, N and P.M. Kimani. 2004. Participatory selection of yellow, brown, sugar and tan bean market classes in Eastern Congo. Bean Improvement Cooperative 47: 305-306. Francis, T. R. and Kannenberg, L. W. 1978. Yield stability studies in short-season maize. I. A. descriptive method of grouping genotypes. Canadian Journal of Plant Science. 58:1029-1034. Lin, C. S. and Binns, M. R.. 1991. Genetic properties of four types of stability parameter. Theoretical and Applied Genetics 82 : 505-509. Lin, C. S. and Binns , M. R. .1988. A method of analyzing cultivar x location x year experiment: a new stability parameter. Theoretical and Applied Genet ics 76:425-430. Lin, C. S., Binns, M. R. and Lefkovich, L. P.. 1986. Stability analysis: where do we stand? Crop Science: 894-900. Schoonhoven A van and Pastor Corrales, M.A. 1987. Standard system for the evaluation of bean germplasm. CIAT, Cali, Colombia. 2.3.5 Breeding small and medium red bean varieties resistant to multiple stresses for smallholder producers in eastern Africa Rationale: Small and medium red bean cultivars are the second most important grain type grown by smallholder farmers in East and central Africa after the red mottled class. They account for more than 20% of bean grown and marketed in East, Central and Southern Africa. An estimated 670,000 ha are sown with small red bean each year. They are widely grown by smallholder farmers and consumed in rural and urban centres in the region. Small reds are of high to moderate importance in Ethiopia, Rwanda, Burundi, Kenya, Madagascar, D. R. Congo, Uganda and Tanzania (Wortmann et al., 1998). Although they are generally moderate to high yielding, their productivity is severely constrained by rust, angular leaf spot, root rots, low soil P and N. For example, a recently released and widely adopted climber ‘Umubano’ succumbed to Fusarium wilt in Rwanda and bean common mosaic virus in Kenya. Many farmers have stopped growing this variety despite its good yield potential. Red Wolaita, probably the most important variety in Ethiopia for domestic market, is susceptible to rust, angular leaf spot, anthracnose and common bacterial blight. GLP 585, released in 1984, is probably the most popular small red cultivar in Kenya. It is however very susceptible to rust, anthracnose, root rots and angular leaf spot. Masaai Red is a popular landrace in Tanzania but susceptible to rust and other diseases. A regional program led by Ethiopian national program in partnership with University of Nairobi, Selian Agricultural Research Institute at Arusha, Tanzania and CIAT was initiated in 2001 to develop high yielding, marketable small red bush bean cultivars with tolerance and/or resistance to a combination of three or more biotic and abiotic stresses and suitable for production in sole and intercrop systems in Africa. Our specific objectives were to: i) initiate crossing programs to develop new populations segregating for priority traits at Awassa, Selian and Kabete, ii) identify recombinants from the populations, iii) evaluate promising lines for resistance to biotic and abiotic stresses, iv) constitute a small and medium red bean nursery for regional evaluation, and v) conduct multi-location and participatory evaluations with farmers and release candidate varieties. Materials and Methods: A regional breeding nursery for small and medium red was constituted from advanced lines from CIAT and national programs of Kenya and Ethiopia in 1998/1999 (CIAT, 2002). Multi-parent crosses were used to create segregating populations. Parents included in the crossing block were selected based on known resistance to angular leaf spot, anthracnose, rust and tolerance to low soil fertility. Selected commercial varieties and donor parents for specific traits were used in crossing programs in Kenya, Ethiopia and Tanzania to combine resistances to biotic and abiotic stresses with 145 farmer preferred traits in single, three way and double crosses. The segregating populations were grown at Kabete Field Station, (Kenya) and at the Southern Agricultural Research Institute (SARI) in Awassa (Ethiopia) and selected for tolerance to diseases, plant and grain type for three generations. F4 and F5 bulks were screened for tolerance to root rots and tolerance to low soil fertility in Kakamega and Mulungu Station in Eastern Congo. Lines selected from segregating populations for disease resistance and other agronomic traits were evaluated in preliminary, intermediate and advanced trials in Ethiopia, Kenya, Tanzania, Uganda, Madagascar and Rwanda. Twenty-five advanced generation small red lines were evaluated at eight locations in Kenya, three in Cameroon, two in Ethiopia (Awassa and Melkassa), and one each in Madagascar and Tanzania (CIAT, 2003). The trials were laid out in a 6 x 6 lattice design with three replicates. Each entry was sown in four, 5 m rows. Regionally important commercial cultivars were included as checks. These were: Red Wolaita from Ethiopia, Maasai Red from Tanzania and GLP 585 (Wairimu) from Kenya. The entries were also included in participatory variety selection trials at two sites in Uganda, three in Rwanda and two in Kenya. Results and Discussion: Population development. In Kenya, crosses were made to transfer rust, anthracnose, root rots and angular leaf spot resistance to GLP 585 (locally known as ‘Red Haricot’) and Maasai Red. Roba-1 and Awash were used as sources of rust resistance in these crosses. G2333 and Vunikingi contributed resistance to root rots and anthracnose. Mexico 54 and G5686 were donors for angular leaf spot resistance genes. Four hundred twenty successful pollinations were made for the GLP 585 improvement program and 469 pollinations for Maasai Red program. The Ethiopian crossing program for small reds was started in 2001 at Awassa Regional Research Centre. It focused on improvement of Red Wolaita, the most popular and widely grown small red in Ethiopia. Red Wolaita is low yielding and susceptible to rust angular leaf spot (ALS), common bacterial blight (CBB), anthracnose, drought and bean stem maggot (BSM). It is widely adapted and has attractive and marketable seed color. Donor parents used in the crossing program included G6, EMP 376, G6450, DOR 794, DOR 716, EMP 375 and EMP 252. Additional donor parents selected for the expanded program included Mexico 54 (ALS), G5686 (ALS), XAN lines (CBB), G2333 (anthracnose), Beshbesh and Melka (BSM), Awash-1 and Roba (rust). In addition to simple crosses, the crossing program developed multiparent males with multiple resistances for backcrossing to the recurrent parents (Red Haricot, Maasai Red and Red Wolaita). Several populations were developed at Awassa in 2005 and 2006 (Table 68). These populations were advanced through single pod bulk and modified pedigree methods and also used for genetic studies (Asrat, 2006). Selection criteria include resistance to angular leaf spot, common bacterial blight, floury leaf spot, rust and drought. Table 68. Code SN 1 SN 2 SN 3 SN 4 SN 5 BSE-03-01 BSE-03-03 AWB-0401 AWB-0402 Populations developed at Southern Agricultural Research Institute, Awassa, 2005-2008. Pedigree Red WolaIta x Vax-6 SNNPR-1-20 (MA4 x Mex-235) RAB-589 x SNNPR-1-42 ETAW-01-L-5-19A x ETAW-01-L-3-12A AFR-702 x Mex-54 CIFAC-87100 / CIM9314-36 // HAL-5 / MEX-54 RWR-719 /// G1175-3/ Red Wolaita // RAB-585 / Mex-54 Red Wolaita///RAB-585/DOR-716//RWR-719/RAZ-54 Red Wolaita ///MUC-95/EMP-212//RAB-585/Mex-54 146 Remarks Small seeded progeny Small seeded progeny Small seeded progeny Medium white progeny Large seeded progeny Large seeded progeny Small seeded progeny >70 F2 plants selected >80 F2 plants selected At Selian, 6 F2 populations were generated from single crosses of Maasai Red with Mex 54 (angular leaf spot), BAT 332 (angular leaf spot), UBR (92)25 (low soil N), G5686 (angular leaf spot), G11746 and G22258 (bean stem maggot). Fourteen three-way and 11 double crosses were subsequently made with the same set of parents to generate combinations of resistance to angular leaf spot, bean stem maggot and adaptation to low soil fertility and small red grain type. Due to departure of the breeder for further studies, limited selection was conducted on these populations. Early generation selection. Five populations developed at Awassa were evaluated for different constraints including moisture stress. Five F1.2 populations were first evaluated at Awassa under early and late sown conditions during the main season (meher). The best performing F1.2's were advanced to F3. 169 F1.3 seeds of each family were divided into three equal parts and evaluated for stresses prevalent at each of the three contrasting test sites during the short rain season (belg); Awassa (1750 m) for common bacterial blight, Kokate (2161m) for tolerance to low soil P and N, and at Amaro (1426 m) for drought. Five checks Omo95(RWR-719), DOR-554, DICTA-105, Roba and Red Wolaita were included in the trial. The best populations were selected and advanced to F4 using single pod bulk method. Seventy-one F1.3 families combining high yield with biotic and abiotic stress resistance/tolerance under natural conditions were selected and advanced to F4. The moisture stress was very severe during the cropping season at Amaro and even cowpea which is drought tolerant totally failed at Amaro in this season. A majority of the families did not yield at all, but 12 families gave grain yield of more than 1000 kg ha-1 (Table 69). CAW02-03-10-7, CAW-02-04-7-7 and CAW-02-01-2-1 gave the best yields of 1549, 1520 and 1442 kg ha-1 under moisture stress, compared with yields of DOR-554 and Omo-95 that gave 678 and 661 kg ha-1, respectively. These families also expressed good yielding potential and resistance to diseases both Awassa and Kokate test locations (Tables 69 and 70). The 71 F1.4 families from five populations were further evaluated for two generations (F1.4 and F1.5) under drought stress at Amaro during the meher and belg seasons following population bulk method. From best performing families, five most vigorous plants were selected to create progeny rows for line selection. This resulted in 265 F1.6 lines. These lines were evaluated in a preliminary yield trial at Awassa during the meher season. The trial was laid in augmented randomized block design. Three checks (Omo-95, Red Wolaita and DOR-554) were included. The lines were planted in single, 2m row plots spaced at 80cm. The test lines expressed variability for grain yield. Grain yield of the 2 m row plots varied from 251 g for (CAW-02-04-7-7-2) to 967g for (CAW-02-03-8-1-1). Fifty-one lines out-yielded the best check variety. Two populations (RWR-719///Red Wolaita/ICTA JU-95-4//XAN-317/DOR-794 and ROBA///EMP445/DFA-64//Red Wolaita/RAB-589) accounted for more than 92 % of the best yielding lines. The best performing 95 lines were advanced to F7 and planted in advanced yield trials at six locations during the meher season. Results showed that lines developed from drought tolerant families expressed good performance in favorable environments. Advanced Yield Trials. In Kenya, 10 advanced lines were selected from the preliminary and intermediate yield trials at Kabete, Juja and Thika (Tables 71 and 72). These had a yield potential above 2 t ha-1 and matured in less than 95 days. CAL 170B-4, DFA 52-1, DFA 57, EMP250-3-1, FEB 200-1, ECAB 0408, ECAB 0416, ECAB 0424, ECAB 0428, ECAB 0426 and ECAB 0411 were susceptible to rust (score of 7 and above) at Thika (CIAT, 2001). BRB 71-1 and TLP 8-1 were discarded due to susceptibility to black root. FEB 200-1 showed moderate tolerance to low soil P at Kakamega. All lines showed intermediate or resistant reactions to rust, angular leaf spot and anthracnose at the three sites. 147 Table 69. Grain yield (kg ha-1) of 12 F1.3 and F1.2 families at three locations in southern Ethiopia. F1.2 (Meher ) F1.3 (Belg season) Families Pedigree Amaro (1426m) Mean Awassa (E) Awassa 2187.5 1259.1 1747.8 5615.0 3118.8 2271.3 2265.6 1307.2 1948.0 4207.5 3958.8 CAW-02-01-2-1 2064.4 1875.0 1441.9 1793.8 5566.3 2002.5 CAW-02-01-5-1 1591.9 1484.4 1032.2 1369.5 5737.5 2382.5 Red Wolaita /// XAN-314 / EMP375//MOC106 / DOR-548] 1552.8 1328.1 1408.4 1429.8 3966.3 3107.5 3272.2 1562.5 1198.8 2011.2 3536.3 2108.8 RWR-719 ///Red Wolaita / ICTAJU 95-4 // XAN-317 / DOR-794 2437.5 2656.3 1243.8 2112.5 3982.5 2138.8 2529.4 1875.0 1549.1 1984.5 4282.5 3565.0 2136.9 2500.0 1355.3 1997.4 4120.0 3032.5 1577.5 1250.0 1355.3 1333.7 4500.0 3398.8 2137.2 2109.4 1520.3 1922.3 4100.0 2413.8 CAW-02-04-3-5 1251.3 1250.0 1215.3 1238.9 4828.8 2855.0 Red Wolaita ** 1749.7 1953.1 317.8 1340.2 3986.1 - Omo-95(RWR-719)** 1431.6 2031.3 660.9 1374.6 3644.1 - Roba-1** 1555.9 1679.7 68.8 1101.5 - - DOR-554** 2169.1 1835.9 678.4 1561.2 - - DICTA-105** 1780.9 1796.9 211.6 1263.1 - - 3681.6 3671.9 1549.1 2300.4 - - CAW-02-01-1-1 CAW-02-01-1-2 CAW-02-02-5-1 CAW-02-02-6-3 CAW-02-03-10-6 CAW-02-03-10-7 Red Wolaita /// [DOR-716 / ICTAJU95-2//G-6 / DICTA-106] CAW-02-03-15-4 CAW-02-04-8-3 CAW-02-04-7-7 Roba / EMP-445 / DFA-64 //Red Wolaita / RAB-589 Highest yield obtained Awassa Kokate (1750m) (2161m) 1796.6 E = early sown , L = late sown, ** Checks 148 (L) Table 70. Disease, pod borer resistance scores, days to flowering and maturity of 12 F1.2 and F1.3 families at three locations in southern Ethiopia. Families F1.3 Awassa (Belg ) ALS* CBB HB F1.2 Awassa (Meher early sowing) Pod borer 3 3 3 2 1 1 3 4 2 2 1 2 ALS CBB Rust FLS DF Awassa (Meher late sowing) DM ALS CBB Rust FLS DF DM CAW-02-01-1-1 1 6 1 1 5 1 2 52 102 1 5 1 2 47 96 CAW-02-01-1-2 1 6 1 1 6 1 1 52 104 1 6 1 1 50 95 CAW-02-01-2-1 1 7 1 1 6 4 2 50 102 1 5 4 2 51 95 CAW-02-01-5-1 1 6 1 1 5 1 1 50 106 1 7 1 1 51 93 CAW-02-02-5-1 1 5 2 1 5 1 1 50 101 1 7 1 1 50 91 CAW-02-02-6-3 1 5 2 1 1 1 1 48 102 1 8 1 1 48 91 CAW-02-03-10-6 1 3 2 1 2 1 1 51 103 1 5 1 1 47 96 CAW-02-03-10-7 1 4 2 1 2 3 1 51 106 1 6 1 1 47 97 CAW-02-03-15-4 1 4 1 1 3 1 1 46 107 1 5 1 1 47 98 CAW-02-04-8-3 1 5 2 1 4 1 1 49 106 1 6 1 1 49 102 CAW-02-04-7-7 1 6 1 1 4 1 1 51 100 1 5 1 1 51 98 CAW-02-04-3-5 1 4 2 1 4 1 1 51 102 1 4 1 4 48 99 Checks Red Wolaita 1 2 2 2 4.5 6.5 2 Omo-95(RWR-719) 1 4 2 2 1 3 1 Roba-1 1 2 2 3 DOR-554 1 4 2 2 DICTA-105 1 3 1 2 Abbreviations: ALS= angular leaf spot, CBB= common bacterial blight, FLS=floury leaf spot, HB= halo blight, DF= days to 50% flowering, and DM=days to 50% pod maturity. Disease score using CIAT (1987) standard system. 149 Table 71. Days to flowering, maturity, seed mass and grain yield of top 10 small red F5/F6 bean lines selected from segregating populations and other nurseries. Yield (kg ha-1) PYT IYTb Mean (3 sites) (2 sites) ECAB 00421 45 87.9 27.6 2586 2393 2489 ECAB 00419 45 90 27.5 2362 2269 2316 ECAB 00413 43 87.3 26.5 2413 2182 2298 ECAB 00406 48.8 92.5 25.2 1860 2624 2242 ECAB 00426 45.9 89.7 27.4 2442 2028 2235 ECAB 00417 48 92.2 26.9 2148 2243 2196 ECAB 00420 44 86.7 26.3 2665 1631 2148 ECAB 00414 44 87.8 24.9 2247 2046 2147 ECAB 00422 44 90.7 26.9 2025 2250 2138 Trial mean 46.3 90.3 26.0 2067 1944 2005 Genotypes (G) ** ** ** ** NS Locations (L) ** ** NS ** ** G XL ** ** ** ** * LSD 0.05 2.8 3.0 1.8 290.2 665 CV(%) 5.4 2.9 6.0 19.9 34.3 *,** : Significant at 5 and 1% probability levels, respectively; NS= not significant. PYTa = preliminary yield trial of F5 ; IYTb= intermediate yield trial of F6 and other advanced lines. Genotype Table 72. Genotype ECAB 0429 ECAB 0426 ECAB 0420 ECAB 0428 ECAB 0424 ECAB 0408 ECAB 0410 ECAB 0402 Days to 50 % flower Days to 75% maturity 100-Seed mass (g) a Days to 50% flowering, maturity, seed mass and grain yield of advanced generation small red bean lines grown at two locations in Kenya. Days to flowering Days to maturity 100-seed mass (g) 43 43 44 43 45 43 45 45 85 88 86 85 86 85 86 87 26.9 27.7 28.5 28.3 28.1 26.9 27.4 32.3 Grain yield (kg ha-1) Kabete 2367 2111 1980 2052 2044 1782 1697 1992 Checks GLP 585 43 85 26.7 1741 Maasai Red 43 84 27.9 1787 Red Wolaita 44 85 25.9 1533 Mean (Thika) 43 85 25.0 Mean (Kabete) 44 86 30.9 1752 Trial Mean 43 85 27.9 1752 CV (%) 8.0 1.9 5.7 Locations (L) NS ** ** Genotypes (G) NS ** ** GxL NS NS ** Error *, **: Significant at 5 and 1% probability levels, respectively; NS= not significant 150 Thika 1482 1680 1646 1462 1308 1551 1590 1242 Mean 1925 1895 1813 1757 1676 1617 1644 1617 1352 1646 1408 1365 1365 1547 1717 1471 1365 1742 1554 31 ** NS NS In Ethiopia, advanced medium and small red lines from the first regional nursery constituted in 2001, were evaluated at 13 sites representing a wide range of production zones. The most promising entries from national variety trials were DOR 527, DOR 711 and DOR 811. Farmers also evaluated and selected advanced lines in on-farm trials conducted in Melkassa, Awassa and Alemaya in participatory variety selection program (Assefa et al., 2007). Two new regional small red nurseries were developed at Awassa between 2005 and 2008; the first nursery had 97 SNNPR lines, and second had 74 ETAW lines. The nurseries were distributed for further evaluation and identification of candidate varieties to programs in D. R. Congo, Kenya and Tanzania. Regional Trials Ninety-seven SNNPR lines and three checks were evaluated at Amaro in southern Ethiopia during the belg season. The 33 best yielding lines selected at Amaro were further evaluated in multi-location trials in nine environments in southern Ethiopia for two years. The test lines expressed highly significant variation for grain yield both at Awassa and Amaro. The mean yield ranged from 3531 (local) to 6355 (SNNPR-130) kg ha-1 at Awassa location and 178 (SNNPR-1-11) to 675 (SNNPR-1-29) kg ha-1 at Amaro test location. Sixteen test lines significantly out-yielded the best check DOR-554 (4556 kg ha-1) at Awassa. At Amaro under severe drought conditions, five lines out yielded the best local check. Among checks used in the trial, local farmer variety performed better under moisture stress at Amaro. Seventy 74 ETAW lines along with four checks were evaluated at Kokate (low soil P and N) in 2 m row plots using a lattice design with two replicates. The grain yield ranged from 1859 (ETAW-02-2-7) to 6537 kg ha-1 (ETAW-02-4-9). Thirty five lines performed better than the best check DOR-554. In Madagascar, seven small red lines were selected. Grain yield of the selected lines varied from 771 to 1278 kg ha-1. The selected lines were ECAB 0427 (771 kg ha-1), ECAB 0418 (707 kg ha-1), ECAB 0411 (1155 kg ha-1), ECAB 0417 (1278 kg ha-1), ECAB 0415 (1072 kg ha-1), ECAB 0422 (824 kg ha-1) and ECAB 0410 (780 kg ha-1). This compared with 870 kg ha-1for GLP 585. The selected lines flowered in 42 to 48 days in Madagascar. GLP 585, ECAB 0415 and ECAB 0410 flowered in 48 days. All other lines flowered in 42 to 43 days. In Tanzania, 20 small red ECAB lines were evaluated for two seasons in Madiira. Growing conditions were favourable for plant growth and disease development. Results showed that test lines showed had intermediate to resistant reactions to rust and common bacterial blight. ECAB0411, ECAB0417 and MCM 2001 showed intermediate reactions to anthracnose. All other lines showed resistant reactions. Disease pressure was high for anthracnose, bean common mosaic virus and angular leaf spot. Diseases with the highest mean score on this trial were angular leaf spot (mean 4) and BCMV (mean 5). Nevertheless all genotypes showed resistant to intermediate reactions to diseases (scores 1-6), except entry Red Wolaita which was susceptible to BCMV. Flowering duration was between 41 and 48 days. Plant vigor ranged from 2.0 to 5. Nine lines had better yields compared to the check varieties (GLP 585, Red Wolaita and Maasai Red). The best yielding entries were ECAB0420 (4717 kg ha-1), ECAB 0429 (4485 kg ha-1), ECAB 0421 (4366 kg ha-1), ECAB 0412 (4319 kg ha-1), ECAB 0426 (4286 kg ha-1) and ECAB0401 (4091 kg ha-1). Yield of checks varied from 2.9 to 3.3 t ha-1 in these trials. Regionally, four new lines were selected in the three countries. These were ECAB 0418, ECAB 0411, ECAB 0417 and ECAB 0410. Eight lines were selected in two of the three countries. Three lines were selected in one country. Variety Release: Several varieties have been released in countries where this grain type is important (Table 73). Melka Dima (XAN 310) was released in Ethiopia in response to demand for a cover crop that 151 reduces loss of soil moisture. It is high yielding and resistant to common bacteria blight. Nasser (DICTA 105) is high yielding, resistant to common bacterial blight and early maturing. Omo 95 (RWR 719) was selected during the participatory breeding program in southern Ethiopia. It has light red seeds that cook fast. Omo 95 is high yielding, resistant to common bacterial blight and root rots. It has been reported to be drought tolerant in southern Ethiopia. Batagonia is a red seeded, high yielding climbing bean variety, and probably the first formally released climbing bean in Ethiopia. Table 73. Small and medium red varieties released in eastern Africa between 2003 and 2008. Variety Melka Dima Dinknesh Wairimu Dwarf Batagonia Omo 95 Nasser Dimtu Menakely* Line Code XAN 310 RAB 484 RWV 482 RWR 719 DICTA 105 DOR 554 Local landrace Year of Release 2006 2006 2007 2004 2003 2003 2003 2007 (pre-release) Country of release Ethiopia Ethiopia Kenya Ethiopia Ethiopia Ethiopia Ethiopia Madagascar * Madagascar has no formal variety release system and is working with ECABREN to develop release procedures. Contributors: Paul Kimani, Asrat Asfaw and Teshale Assefa (Ethiopia) Collaborators: Sostenes Kweka (Tanzania), Hery (Madagascar), Beam Teams at Kabete (Kenya), Selian (Tanzania), Awassa (Southern Ethiopia), FOFIFA (Madagascar), Melkassa (Central Ethiopia) and Mulungu (D. R. Congo) References Asrat Asfaw and D. Dauro. 2006. Annual Bean Report. Awassa Agricultural Research Institute, Awassa, Ethiopia. CIAT. 2002. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia. CIAT. 2003. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia. Schoonhoven A van and Pastor Corrales, M.A. 1987. Standard system for the evaluation of bean germplasm. CIAT, Cali, Colombia. Teshale Assefa, H. Assefa and P.M. Kimani. 2007. Development of improved haricot bean germplasm for mid- and low altitude sub-humid ecologies of Ethiopia, pages 87-94. In: Food and Forage Legume of Ethiopia: Progress and Prospects. ICARDA, Allepo, Syria. 2.3.6 Development and release of brown and tan colored bean varieties with multiple stress resistance in eastern Africa Rationale: Brown, tan and yellow beans account for about 11% of Africa’s bean production. They are grown on 290,000 ha in eastern Africa and 90,000 ha in southern Africa. They are of high to moderate importance in southwest Kenya, along Lake Tanganyika region in Tanzania, eastern D. R. Congo, Rwanda, Burundi, Uganda, southern Sudan, Madagascar and southwest Cameroon. In the Great lakes region, brown and tan colored beans are part of popular mixtures. In southern Africa, brown, tan and yellow beans are important in Angola, Zambia, northern Mozambique, Swaziland and Lesotho. Some small seeded types such as ‘Ubososera’ are tolerant to soil acidity. Carioca types have high yield potential but average market potential in the region. However, they are a potential food security crop for smallholder, resource poor farmers because of their plasticity and good performance in marginal 152 environments. Carioca bean may have potential in niche markets. For example, carioca grain type has been in high demand in the Copper Belt of Zambia and sugar estates of Swaziland which are underexploited markets for ECABREN countries. However, available cultivars of brown, tan and yellow beans are susceptible to angular leaf spot, anthracnose, root rots, rust, and halo blight, low soil P and nitrogen and Al/Mn toxicity. These factors contribute to the low production and inability to meet demand in domestic and regional markets. Although these grain types are important to smallholder farmers, little effort has been devoted to their improvement in Africa. Regional programs were therefore started in 2001 to develop improved cultivars of these grain types with a high yield potential, a combination of tolerance to three or more biotic and abiotic stresses and grain characteristics acceptable to consumers in target markets. The main achievements of these programs are presented briefly in this report. Materials and Methods: Brown, tan, yellow and carioca bean grain types were selected from F2 and F3 segregating populations at Kabete. The populations were derived from multi-parent crossing program, which included lines with known tolerance to major biotic and abiotic stresses (Kimani et al., 2004; Lunze et al., 2007). The selections were grown in progeny rows at Kabete to increase uniformity within families and to expose them to disease and drought stress. F5 and F6 lines were grown in preliminary and intermediate yield trials at three locations in Kenya including Thika. Thirteen lines selected in advanced yield trials and lines from other multiple constraint nurseries were used to constitute a regional nursery. The regional nursery was distributed for further evaluation to member countries interested in these grain types between 2000 and 2005 (CIAT 2000, 2001, 2002, 2003, 2004 and 2005). Breeding populations were also generated at INERA -Mulungu Research Station in eastern D. R. Congo. This program started in 2001, and focused on improving resistance to angular leaf spot (ALS) in three locally popular but susceptible cultivars: Kirundo (yellow), Munyu (brown) and Nakaja (tan). Six hundred sixty-nine pollinations were made with A285, A235, Mex 54, G5686, MLB-36-89A and A339 as sources for resistance to ALS. Other resistance sources used were: G11727, ACC 714 and Besh-Besh for bean stem maggot. VEF(88), LPY6, CIM 9314, LSA 32 and PAN 150 were used a sources of resistance to low N, P and low pH complex. These sources were combined in complex crosses and backcrossed to the three recurrent parents. Early generation (F2 to F4) populations were evaluated for resistance to angular leaf spot, root rots, anthracnose and common bacterial blight in disease ‘hot spots’. F5 and F6 generations were evaluated for grain yield and other agronomic traits in replicated trials at 2 to 4 locations in Kenya and D. R. Congo. Selected lines with multiple constraint resistance were subjected to participatory selection with men and women farmers to identify lines that met breeder, farmer and consumer criteria in F7 and F8 generations. Gofta, a brown seeded variety released in Ethiopia, line A197, Kirundo, and other yellow and tan colored local varieties were included as checks. Standard agronomic practices were followed. Best performing lines were entered in multi-location national variety yield trials to identify candidates for release in collaborating countries. Results and Discussion: ECAB 0761 showed an intermediate reaction to root rot at Thika. All other genotypes showed resistant reaction to angular leaf spot, anthracnose and rust. Genotypes flowered and matured earliest at Thika and later at Kabete. Average duration to maturity was 87 days at Kabete compared to 82 days at Thika. Average pod yield was 161 pods m-2 at Thika compared with 162 pods m-2 at Thika. Mean seed mass was highest at Kabete (30.8 g per 100-seeds) and lowest at Thika (24.1 g per 100-seeds). Mean grain yield was highest at Juja. Only four lines produced higher grain yield than A197 (Table 74). Five new lines had higher grain yield than Gofta. However, the yield advantage was modest. In eastern D. R. Congo, two lines DRK 7 (1620 kg ha-1) and MAC 16 (997 kg ha-1) had higher yields than the best check, MLB 118/96B (943 kg ha-1) in preliminary yield trials. However, seven lines had better yields than the second check, Kirundo (623 kg ha-1). The selected lines showed better tolerance to biotic and abiotic stresses compared to the checks. In the confirmatory yield trial (CYT), nine lines retained their superior performance compared to Kirundo (check). These were MLB 118/94B (1693 kg ha-1), 153 CODMLB 001/03 (1690 kg ha-1), LSA 144 (1624 kg ha-1), COD MLB 007/03 (1605 kg ha-1), MLB 174/94B (1557 kg ha-1), COD MLB 033/03 (1489 kg ha-1), CODMLB 005/03 (1482 kg ha-1), M’Sole (1301 kg ha-1) and COD 037/03 (1210 kg ha-1). In these trials Kirundo had a yield of 1198 kg ha-1. Fifteen bush bean lines were tested for consumer and farmer acceptability at five sites in South Kivu. Seven were selected as potential candidates for release. In descending order of acceptability, they were: ZKA 9310M/95 (30% acceptability), SCAM 80 CM/2 (22%), KS 65-2 (18.5%), RWR 1873 (11%), LSA 60-1 (11%), G5686 (3.7%) and MCD 2518-1 (3.7%). Table 74. Mean duration to flowering, maturity, pods m-2, 100-seed mass and grain yield of selected F8 brown, yellow and tan colored bean lines grown at four environments in Kenya. Genotype ECAB0755 ECAB0753 ECAB0761 ECAB0758 A 197 ECAB0757 GOFTA ECAB0756 ECAB0752 ECAB0763 ECAB0760 Environmental mean CV(%) Days to 50% flower (d) Days to maturity (d) Pod m-2 100-seed mass (g) 44.0 44.0 42.8 42.7 41.7 43.8 41.9 43.5 44.2 45.5 43.7 43.8 84.6 83.7 83.6 83.0 82.7 85.5 82.8 84.9 85.3 85.8 84.9 84.6 207.9 208.4 211.6 204.4 205.2 219.8 184.2 219.0 198.5 199.3 201.7 201.7 23.0 21.8 22.3 28.6 48.8 25.9 29.4 21.6 25.5 24.1 25.6 26.6 3.7 2.2 16.1 5.7 Grain yield (kg ha-1) Kabete LR Thika SR Juja SR Mean 1646 1666 1302 650 1387 1320 1197 1206 890 922 1182 1117 2658 2078 2773 2322 1633 2247 2587 2510 2657 2512 2488 2373 1325 1763 1397 2076 1953 1400 1175 1206 1333 1417 1147 1395 1876 1836 1824 1683 1658 1656 1653 1641 1627 1617 1606 1628 17.4 Genotypes (G) ** ** NS ** ** Environments ** ** ** ** ** (E) GxE NS ** * ** ** *, **: Significant at 5 and 1 % probability levels, respectively; NS= not significant, LR= long rain (April-July), SR= short rain seasons (Nov-December). In western D. R. Congo, bean germplasm was introduced to INERA-M’vuazi in western D. R. Congo from INERA research stations at Mulungu, Gandanjika, FOFIFA (Madagascar) and University of Nairobi (Kenya) from 2000 (Kimani, 2006). The collection comprised of 80 sugar bean lines and 40 BILFA 5 nursery lines from Mulungu, 8 entries from FOFIFA bean program, more than 86 F2 and F3 segregating populations from the regional multiple constraint nurseries at University of Nairobi, and local collections. The collection was evaluated at M’vuazi, Kisantu and several on-farm sites in Bas Congo, Kinshasa and Bandu Provinces. All trial sites were below 1000 masl. The evaluations were conducted in collaboration with farmer groups, NGOs and community based organizations (CBOs). Variety Release: Table 75 shows some of the tan, yellow and cream colored varieties released in eastern Africa between 2003 and 2008. ‘Wedo’ is a medium seeded, high yielding cream/light tan colored variety with combined resistance to common bacterial blight and rust. ‘Haramaya’ is a large seeded variety with light tan seeds. It was released by Haramaya University (formerly ‘Alemaya University’) for production 154 in eastern Ethiopia. Inamunihire is yellow seeded determinate bush variety. It is high yielding and has good taste and other cooking characteristics. ‘Lumbua’ is a tan seeded bush (type 1) variety with combined resistance to common bacterial blight and web blight. It is tolerant to heat, low soil nitrogen, phosphorus and potassium. ‘Sepe’ has large tan colored seeds. It is a bush variety with combined tolerance to web blight, common bacterial blight, heat, low soil nitrogen, phosphorus and potassium. ‘Manseki’ is yellow seeded climbing bean variety with combined resistance to common bacterial blight, web blight, heat and low soil fertility. It is a large seeded tan colored bush (type 1) variety adapted to hot humid lowlands of western D. R. Congo. It is resistant to common bacterial blight, and tolerant to heat and low soil fertility. G22501, BF 10 and BF 12 are cream seeded varieties with resistance to common bacterial blight, tolerance to web blight, bean stem maggot, heat and/or low soil fertility. They have type 1 growth habit. RWR 2172 and RWR 2154 have medium, cream seeds, and determinate bush growth habit. Table 75. Brown, tan and yellow seeded varieties released in eastern Africa between 2003 and 2008. Variety Line Code RWR 2154 ISAR line MAM 48 MAM 48 Wedo MAM 41 Inamunihire IZ 0201245 IZ 0201513 IZ 0201513 Mbidi Local landrace Lumbua L4 Sepe G8047 Manseki Landrace I7 Landrace G22501 G22501 BF12 BF12 BF10 BF10 RWR 2172 ISAR line MAM 48 MAM 48 Wedo MAM 41 Inamunihire IZ 0201245 IZ 0201513 IZ 0201513 Source: National Bean program reports, 2003-2008 Contributors: Year of Release 2006 2003 2003 2003 2003 2005 2005 2005 2005 2005 2005 2005 2005 2004 2003 2003 2003 2003 Country of release Rwanda Ethiopia Ethiopia Burundi Burundi D. R. Congo (west) D. R. Congo (west) D. R. Congo (west) D. R. Congo (west) D. R. Congo (west) D. R. Congo (west) D. R. Congo (west) D. R. Congo (west) Rwanda Ethiopia Ethiopia Burundi Burundi Paul Kimani, Nkonko Mbikayi and Lodi Lama (INERA, D. R. Congo), Teshale Assefa (EAIR, Ethiopia), Capitoline Ruradama (ISABU, Burundi), Félicité Nzanzebara (ISAR, Rwanda). Collaborators : Bean Teams at Kabete (Kenya), Mvuasi and Mulungu (D. R. Congo), Melkassa (Ethiopia), Alemaya University, Bujumbura (Burundi) and Rubona (Rwanda). 155 References CIAT.2000. Annual Report, IP-2. Cali, Colombia. CIAT.2001. Annual Report, IP-1, Cali, Colombia CIAT. 2003. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2004. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia CIAT. 2005. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia Kimani, P.M. 2004. Germplasm issues in participatory bean breeding in Africa. Participatory breeding workshop, 17-25 May 2004, Kakamega, Kenya. Pan African Bean Research Alliance, Kampala, Uganda. Kimani, P.M. 2006. Bean varieties for humid tropic regions: Reality or fiction? (on line). [On line]. Internacional de Agricultura Tropical (CIAT), Kampala, Uganda. 2p. Highlights: CIAT in Africa No. 34. Lubanga, L., P.M. Kimani, R. Ngatoluwa, B. Rabary, G.O. Rachier, M. M. Ugen, V. Ruganzu and E. Awad Elkarim. 2007. Bean improvement for low soil fertility adaptation in Eastern and Central Africa, pages 325-332. In: Bationo et al (eds.). Advances in Integrated Soil Fertility Management in sub-Saharan Africa: Challenges and Opportunities. Springer, Dordrecht, The Netherlands. 1094pp 2.3.7 Development and release of improved climbing bean varieties in eastern Africa Rationale: Although climbing beans a relatively recent introduction in most of the countries in East and Central Africa, they have gained popularity in the last decade among smallholder farmers because of their conspicuous yield advantage over bush types, and more effective and efficient utilization of cultivated plots which are decreasing rapidly in size due to fast population growth. Climbing beans, which were introduced in Rwanda in mid 1980’s spread rapidly to Burundi, D. R. Congo, Uganda, Kenya, Tanzania and Ethiopia. However, continued expansion and adoption of climbing bean technology is constrained by susceptibility of available cultivars to anthracnose, angular leaf spot, root rots complex, aschochyta, common bacterial blight, halo blight, low soil N and P drought and lack of preferred grain types. Expanding cultivation of climbing beans from their traditional production high altitude zones with adequate rainfall, moderately fertile soils and high population pressure to the less densely populated and more expansive middle altitudes is constrained by poor adaptation of popular cultivars, moisture, heat and low soil fertility stresses. Spread of climbing beans to regions where traditional mixtures do not adequately meet new market demand for specific seed types has put pressure on national programs to develop cultivars that respond better to these demands. A regional program was therefore initiated in 2000 to develop high yielding, marketable climbing beans cultivars with resistance to two or more biotic stresses and wider adaptation. In this report, we highlight major milestones reached in development and release of improved climbing beans in east, central and west Africa between 2003 and 2008. Materials and Methods: A working collection was made from climbing bean genetic resources maintained by CIAT, Colombia and the national program of Rwanda, which has the regional mandate for development of climbing beans. Additional germplasm was received from national program of D. R. Congo. The collection included entries from CIAT’s core collection, advanced breeding lines, landraces and commercial cultivars. Parental lines were selected for the crossing programs at Kabete (Kenya) and Rubona (Rwanda). Crosses were made to incorporate root rot, fusarium wilt, angular leaf spot, and anthracnose resistance and red mottled seed type into popular and well-adapted climbers. Adapted parents included Umubano, Vunikingi, Ngwinurare, Puebla and Urugezi. Umubano is resistant to anthracnose but susceptible to fusarium wilt. Vunikingi is high yielding and resistant to root rots, fusarium wilt and anthracnose. However, its small pink seeds are not popular in major markets in the region. Ngwinurare was included in the crosses because of its large red seeds popular in certain market sectors. It is not resistant to priority constraints. Puebla is tolerant to low soil fertility. Mexico 54 provided resistance to angular leaf spot, and SCAM 80/CM 15 resistance to root rots. Rubona 5, PVA 8 and Urugezi are popular red mottled cultivars but susceptible to anthracnose, angular leaf spot, fusarium wilt, and root rots. These parental lines were combined in single, three way and double crosses (Table 76). More than 691 156 pollinations were made at Kabete to create populations segregating for red mottled, red kidney and other priority seed types and resistance to angular leaf spot, anthracnose, root rots and fusarium wilt. Screening for multiple disease resistance. Both advanced lines and segregating populations were screened for multiple disease resistance in Rwanda, Kenya and D. R. Congo. In Rwanda, 11 F2 populations were divided into four portions and their F3, F4, F5 and F6 progenies screened in a systematic rotation in disease hot spots at Rubona (angular leaf spot), Rwerere (anthracnose), Gikongoro (root rots) and Ntendezi (fusarium wilt and anthracnose). Disease pressure was moderate at Rwerere (2300m), high at Rubona (1650m), and very high at Ntendezi (1600m) and Gikongoro (2300m). At each planting cycle, pedigree selection method was used to obtain diseases resistant plants based on a severity score rating of 3 or less on a CIAT scale of 1-9. Sixty-six F7 lines resistant to 2 or more diseases and different market seed-types were planted in replicated yield trials at Rwerere and Rubona sites representing the high (2300 m) and mid altitude (1650 m) zones. Table 76. Multiple constraint resistance climbing bean populations created at ISAR, Rubona. Population Code MMCR-RW-1 MMCR-RW-2 MMCR-RW-3 MMCR-RW-4 MMCR-RW-5 MMCR-RW-6 MMCR-RW-7 MMCR-RW-8 MMCR-RW-9 MMCR-RW-10 MMCR-RW-11 Source: Musoni, 2007. Pedigree Ngwinurare/SCAM 80CM-15//Ngwinurare/Puebla Ngwinurare/Puebla// Mex 54 Umubano/SCAM 80 CM-15//Umubano/Ngwinurare Umubano/SCAM 80 CM-15//RWR13121 Umubano /Mex 54 Vuninkingi/Mex 54 Urugezi/Puebla Umubano/Urugezi//Mex 54 Vuninkingi/Urugezi//Umubano Umubano/Vuninkingi Umubano/Vuninkingi // Umubano In Kenya, 75 advanced lines and segregating populations were evaluated at Kabete, Thika, Ol Jorok and Embu. Ol Jorok (2350m) received heavy rainfall from planting to mid-pod filling, creating favourable conditions for disease development. At Embu, participatory selection was conducted among 25 F4 segregating populations of medium altitude climbers from CIAT, Colombia and from the regional climbing bean nursery. Preliminary evaluation for morphological traits (growth habit, pod clearance and plant height) and yield components was conducted on-station during the long rain season. During the following short rain season, representative farmer groups from Meru, Embu and Kirinyaga districts were invited to evaluate and select single plants (F4.6) with preferred traits. Farmers agreed on the most important selection criteria. Progeny rows of the F4.6 lines were established at the station during the following long rain season to produce adequate seed for on-farm testing. About 32 F4.7 lines were subsequently evaluated on-farm and on-station in the three districts. Five lines were selected for national performance trials which were conducted at seven sites (Katumani, Thika, Embu, Eldoret, Njoro, Kakamega and Kapsabet) over two years (CIAT, 2007). Between 2003 and 2007, the regional nursery was also distributed for further evaluation and selection by programs in D. R. Congo, Uganda, Tanzania, Madagascar, Burundi, Cameroon and Ethiopia. Additional crosses were generated at INERA- Mulungu (D. R. Congo) and selected for multiple resistance to common bacterial blight, angular leaf spot and tolerance low fertility acid soils (Mbikayi and Kimani, 2004). 157 Results and Discussion: The eleven bi-parental and multi-parent populations developed at Rubona showed considerable segregation for grain type, growth habit and disease resistance (Tables 77 and 78). Thirty-eight F2.6 lines combining resistance to two or more diseases and high yield potential were selected (Table 79). Fourteen lines showed adaptation to medium altitude zones, 15 to high altitude zones and nine to both medium and high altitude zones. Thirty-two lines combined multiple diseases with the preferred red and red mottled grain types. Results showed that Gikongoro and Ntendezi were good hot spots for the selection against root rots while Rubona was ideal for the selection against angular leaf spot. However, Rwerere was not as reliable as Ntendezi for the selection for anthracnose resistance. The new elite lines with a yield range of 2.5 ton ha-1 to 4.5 ton ha-1, or 101-141% of the yields of improved checks were selected at Rwerere and Rubona (Tables 80a, b, and c). The bush bean cultivar, SCAM 80 CM / 15 was the most effective donor of the red-mottled seed coat color compared with Urugezi and RWR 1312. Sixty-five per cent of the new red and red mottled climbing bean lines were large seeded. Table 77. Combinations seed color and disease resistance observed in 11 populations developed from simple and multiple parent crosses at ISAR Rubona station. Population Seed color Red Red mottled MMCRW-1 + + MMCRW -2 + MMCRW -3 + + MMCRW -4 + + MMCRW -5 + MMCRW -6 MMCRW -7 + MMCRW -8 + + MMCRW -9 + + MMCRW -10 + MMCRW -11 + + = trait observed, - = not observed. Anthracnose + + + + + + + + Disease resistance Angular Pythium root leaf spot rot + + + + + + + + + + + + + + + + Fusarium wilt + + + + + + + + + Table 78. Distribution of new climbing bean lines selected from 11 populations by market class. Population MMCRW-1 MMCRW-2 MMCRW-3 MMCRW-4 MMCRW-5 MMCRW-6 MMCRW-7 MMCRW-8 MMCRW-9 MMCRW-10 MMCRW-11 Total Red 11 0 0 0 1 0 0 0 0 0 0 13 158 Red-mottled 18 0 4 1 0 0 3 0 4 0 0 30 Purple 4 1 0 0 2 4 0 0 0 2 0 12 Cream 3 1 0 0 0 0 0 0 0 0 0 4 Black 2 0 0 0 3 0 1 0 0 1 0 7 Total 39 2 4 1 6 4 4 0 4 2 0 66 Table 79. Mean yield of new climbing bean lines selected for multiple disease resistance at Rubona and Rwerere ISAR stations. Grain yield (kg ha-1) by site Market class No. of lines Rubona Rwerere Mean Reds 13 2232 2771 2505 Red mottled 30 2416 2521 2468 Purple 12 1421 2799 2110 Cream 4 1386 2306 1847 Black 7 1771 2736 2254 Parents 7 1838 1938 1886 Checks 2 1678 2663 2171 Table 80a. Seed color Disease score, market class and yield advantage of new climbing bean lines at Rubona, Rwanda. Population Red MMCRW-1 Red mottled MMCRW-9 MMCRW-1 MMCRW-2 MMCRW-7 Black Table 80b. Seed color MMCRW-5 RWV 2573 RWV 2575 RWV 2581 RWV 2599 RWV 2606 RWV 2676 RWV 2680 RWV 2681 RWV 2682 RWV 2697 RWV 2698 RWV 2694 RWV 2695 RWV 2856 Anth. 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Disease severity ALS Root rot 3 2 3 2 4 3 4 2 4 4 4 3 3 4 4 4 3 4 4 4 4 4 2 2 4 3 3 4 Yield increase above check (%) 141 126 131 107 111 155 103 130 158 109 110 129 112 102 Disease score and yield advantage of new climbing bean lines selected for multiple disease resistance at Rwerere, Rwanda. Population Red MMCRW-1 Red-mottled MMCRW-1 MMCRW-7 Purple Black Variety MMCRW-10 MMCRW-7 MMCRW-5 MMCRW-5 Variety RWV 2572 RWV 2573 RWV 2575 RWV 2581 RWV 2594 RWV 2599 RWV 2680 RWV 2691 RWV 2697 RWV 2694 RWV 2695 RWV 2654 RWV 2851 RWV 2852 RWV 2856 Anth. 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 Disease severity ALS Root rot 3 4 4 2 3 2 4 3 4 4 4 2 3 4 3 4 4 4 3 2 4 3 4 4 3 4 3 4 3 4 159 Percent yield advantage over check (%) 151 125 105 108 107 100 115 101 122 111 109 134 107 106 118 Table 80c. Seed color Disease score, market class and yield advantage of new climbing bean lines selected for multiple disease resistance at Rubona and Rwerere trial sites, 2005-2007. Source population Variety Code Disease severity rating Anthracnose Red MMCRW-1 Red mottled MMCRW-1 MMCRW-7 Black MMCRW-5 RWV 2573 RWV 2575 RWV 2581 RWV 2599 RWV 2680 RWV 2697 RWV 2694 RWV 2695 RWV 2856 1 1 1 1 1 1 1 1 1 Angular leaf spot 3 4 3 4 3 4 3 4 3 Root rots 2 2 3 2 4 4 2 4 4 Yield advantage over check (%) Rubona Rwerere 141 126 131 107 103 109 129 112 102 125 105 108 100 115 122 111 109 118 At Ol Jorok (Kenya) test lines showed delayed flowering (64 to 94 days) compared with 40 to 49 days at Kabete and Thika (Table 81). Most of the lines were attacked severely by anthracnose, web blight, angular leaf spot, CBB, BCMV, ascochyta and halo blight or a combination of these diseases. However, a few lines were either completely free of the diseases or showed very low levels of infection. Eleven lines were selected because of their vigor and low infection. These were: MLV-76/97A, VCB 87012, AND 10, VCB 81012, MLV 198/97A, MLV 222/97A, MLV 227/97A, MLV 216/97A, G24517, Urugezi and G20751. Urugezi showed moderate infection by viruses. Umubano and Vunikingi were susceptible to BCMV. Lines apparently adapted to conditions prevailing at the three sites were: G24517, G20751 and Urugezi. At Embu, the F4 populations segregated for a wide range of grain types, which included red mottled, red kidney, small reds, pinto, sugars and yellows. The lines also showed considerable variation in duration to maturity, plant height, vigor, pod load and grain yield (Table 82). The broad variation observed in these populations suggested possible outcrossing, in addition to the expected segregation. Some lines showed instability with segregation persisting in F4.7 and F4.8. For example, selections from MAC 26 continued to segregate for grain type even in advanced generations. Pinto, grey (mwezi moja type), yellow pinto and red mottled types could be identified from progeny rows in F4.8 generation. Grain yield varied from 800 to 3800 kg ha-1. Duration to maturity varied from 80 to 108 days at Embu (about 1550 m). Twenty-two bean F4.6 lines combining preferred grain, yield and plant traits were selected. Five lines (Table 82) were registered for national performance trials. Results showed that Umubano and Vunikingi were very susceptible to BCMV (score of 9) at Thika and Kabete. MAC 34 showed intermediate reaction to rust and angular leaf spot. MAC 13, MAC 34 and MAC 64 showed outstanding performance across agroecological zones. Release of New Varieties: Considerable progress has been made in development and popularisation of climbing bean varieties in east and central Africa in the last five years. Nearly all countries are evaluating or have released improved climbing varieties. Table 83 shows some of the new climbing bean varieties released in eastern Africa between 2003 and 2008. 160 Table 81. Duration to 50% flowering, seed size, grain type and reaction to bean common mosaic virus, web blight and common bacterial blight of 26 climbing bean lines selected at Ol Jorok, Kenya. Line MLV 59/97A MLV 76/97A Kirundo VCB 87012 Nakaja AND 10 VCB 81012 M’Sole AFR 441 MLV 198/97A MLV 6/90B MLV 222/97A MLV 56/96B MLV 227/97A MLV 216/97A Cuarentino 0817 SEQ 1006 G59/1-2 Nain de Kyondo G50330 G24517 G20875 Gisenyi G20833 G31479 G20751 Days to flowering 72 63 76 Seed size M S L Grain type *CBB *BCMV sugar brown yellow 5 1 1 2 1 7 Web blight 4 2 1 70 57 85 61 76 71 71 71 69 62 63 85 63 M S L S S M S S S S S S S brown brown sugar brown brown zebra zebra brown white brown brown black white 1 1 1 1 1 1 2 1 1 1 1 1 1 1 5 1 7 2 1 1 1 1 5 1 1 1 1 1 1 1 2 3 3 9 1 1 1 1 1 70 71 76 L L S zebra brown white 1 2 1 3 7 2 4 1 1 94 76 86 91 85 76 84 M M L L S M M brown yellow brown sugar black black yellow 1 3 1 1 3 2 2 2 1 7 2 9 7 7 1 1 1 1 1 1 1 Other observations Susceptible to ALS vigorous vigorous vigorous Seed size: S= small (< 25g/100 seeds), M= medium (25-39 g/100 seeds) and L=large (> 40 g/100 seeds) * CBB= common bacterial blight, ALS= angular leaf spot and BCMV= bean common mosaic virus. Table 82. Genotype MAC 13 MAC 34 MAC 64 MAC 26 RWV 524B Checks Umubano Some characteristics of five climbing bean lines selected in advanced yield trials in Kenya. Duration to flowering 48 48 40 48 Duration to maturity 49 102 48 95 99 91 88 91 Grain type Medium, red round seeds Medium, red mottled Red mottled Segregating for pinto, red and yellow grain types Red kidney, large seeded, heavy podding Small, glossy red 161 Grain yield (kg ha-1) 3000 3600 4100 2667 3000 2933 Table 83. Climbing bean varieties released in eastern Africa between 2003 and 2008. Variety Line Code Kenya Mavuno MAC 64-1 Kenya Safi MAC 13-3 Kenya Tamu MAC 34-5 Cheupe CAB 19 M 211* M211 Kayana* Kayana VNB 81010* VNB 81010 RWV 1365* RWV 1365 MLV 198/97A* MLV 198/97A G 59/1-2* G59/1-2 VCB 81013* VCB 81013 LIB 1* LIB 1 Kiangara* MLV 59/97A RWV 1892 RWV 1892 RWV 2070 RWV 2070 RWV 1892 RWV 1892 CODMLV 052* CODMLV 052 CODMLV 056* CODMLV 056 MLV 224/97A* MLV 224/97A MLV 198/97A* MLV 198/97A MLV 59/97A* MLV 59/97A Batagonia-1 RWV 482 MAC 28 MAC 28 NABE 12C Sugar 131/MAC 31 Ndamirabashonji RWV 167 Amakwamirire RWV 296 Source: National Bean program reports, 2003-2008 * Pre-releases. Year of Release 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2008 2007 2007 2007 2007 2007 2007 2007 2007 2004 2004 2003 2003 2003 Country of release Kenya Kenya Kenya Tanzania Madagascar Madagascar Madagascar Madagascar Madagascar DRC (west) DRC (west) DRC (west) DRC (west) Rwanda Rwanda Rwanda DRC (east) DRC (east) DRC (east) DRC (east) DRC (east) Ethiopia Rwanda Uganda Rwanda Rwanda Contribution: Paul Kimani (CIAT), Matthew Blair (CIAT), Augustine Musoni (ISAR, Rubona), Nkonko Mbikayi (INERA, D. R. Congo), Lodi Lama (INERA, D. R. Congo), Teshale Assefa (EIAR, Ethiopia), Annet Namayanja (NARO-NACRRI, Uganda), Festo Ngulu (SARI, Tanzania) and Heri Andriamazaolo (FOFIFA, Madagascar). Collaborators: Bean Teams in Kenya, Rwanda, D. R. Congo, Madagascar, Ethiopia and Uganda References CIAT. 2007. Bean Improvement for the Tropics. Annual Report I-P1. CIAT, Cali, Colombia. Mbikayi, N and P.M. Kimani. 2004. Participatory selection of yellow, brown, sugar and tan bean market classes in Eastern Congo. Bean Improvement Cooperative 47: 305-306. Musoni, A. 2007. M.Sc. Thesis, University of Nairobi, Kenya. 162 2.3.8 Breeding for specific bean market classes within Southern Africa Bean Research Network (SABRN) Rationale: Within the SABRN, not all NARS programs have the capacity to generate segregating populations of their own. The countries that have active breeding programs include, Malawi, South Africa, southern highlands of Tanzania, Zambia and Zimbabwe. All these NARS bean breeding programs have embraced the market class breeding program initiative, where each country concentrates on a market class or two, usually focusing on market classes which are important to the nation, and improve them for specific biotic and or abiotic stresses of importance to the nation as well as to the region. The resulting germplasm is shared to all NARS programs with interest in the said market classes. In this reporting period, a number of countries in the region, including the back-up regional breeding program had developed segregating populations and lines which combined specific market classes and one or more biotic and or abiotic stresses: Progress in breeding for market classes • In Malawi, the focus is on red kidney, khaki, red mottled (calima) market classes, and important stresses are common bacterial blight (CBB), angular leaf spot (ALS), halo blight (HB), and low soil fertility (LSF). Many lines in these market classes developed by the national program were in preliminary trials (PYT) and some in advanced yield trials (AYT). In addition, there were some sugar lines, which were developed using molecular tools (marker assisted selection) in collaboration with Washington State University. • In South Africa, the focus is on sugar and navy bean, targeting such stresses as: ALS, CBB, Rust, bean common mosaic virus (BCMV) and halo blight (HB). During reporting period there were over 1800 lines in different generations (F3 - F6), and in different populations which combined sugar bean market class with various multiple stresses like: CBB/rust/ALS/BCMV; Rust/ALS/BCMV; Rust/CBB/BCMV and Rust/BCMV. Likewise there were several lines at different generations which combined navy bean market classes with different combination of biotic stresses. In addition some lines were in check-row trials, and many others in replicated yield trials. Some of the lines in different market classes, which were developed for rust and ALS or rust and halo blight (HB) or only rust resistance were sent to the SABRN regional coordinator for seed increase and distribution to other interested NARS programs within the region. The southern highlands of Tanzania (SHTZ) focus on such market classes as: red beans (small, medium and large) and yellow beans, targeting such traits as: ALS and anthracnose (ANTH). There are several lines in the red market class which were in F5, F6, F7 and F8 generation. Some of these lines are ready for distribution to various interested NARS partners in SABRN or beyond. The progenies in the yellow bean market class are at F4 generation. They also had some calima bean lines which were doing very well in the SHTZ environments and they were ready to share the lines with other national programs. • In Zambia, the focus is on yellow and brown/tan (khaki), targeted such traits as: ALS and CBB. They had over 300 lines at various F3, F4, BC1 and BC2 in different populations combining different parental sources for the market class and donor parents for ALS and CBB resistance. • In Zimbabwe, the target market classes were: red beans (large red kidney, medium and small red) and sugar bean, targeting such traits as: ALS and CBB for biotic and drought and low P for abiotic stresses. They had a total of 56 lines from 13 populations at F5 generation. • The regional network breeding program has a back-up breeding program to support the NARS breeding programs in various market classes: red mottled, brown/khaki, sugar, reds, and purples. 163 The targeted stresses include; ALS, CBB, bean stem maggot (BSM), low P and drought. During this reporting period, there were over 20 populations at F3-F5 generation combining red mottled market class with various stresses such as ALS, CBB, BSM and low P. In addition there were 300 lines at F7 and above. In the brown/khaki market class, there were 10 populations in F2-F3 generations and 70 lines at F7 generation combining grain type with ALS, CBB, BSM and drought. In addition there were over 100 lines in red kidney, 80 lines in sugar and 70 lines in purple, all at F7 generation and above. These lines are ready for distribution to NARS partners within SABRN and others. Results and Discussions: The active national breeding programs in SABRN are making progress in generating segregating populations in different bean market classes, to improve them for resistance to various biotic and abiotic constraints. The segregating populations were at different generations, ranging from F2 through F7 and above. In some countries like Malawi, South Africa and southern highlands of Tanzania, they also had some fixed lines which were already in preliminary and advanced yield trials. These countries were ready to share the segregating populations as well the fixed lines with others in the network, who are interested in the available market classes. This is particularly useful to national programs which have interest in several bean market classes, but they do not have the resources and capacity to run breeding programs in all market classes. Thus sharing breeding responsibility among various NARS programs, and exchange of resulting germplasm becomes a key to ensure that a number of bean market classes are included in the breeding programs across the region – unleashing the power of networks. Contributor: R. Chirwa Collaborators: A. Liebenberg, M. Liebenberg, D. Fourie, J. Bokosi, C. Madata, G. Makunde, K. Muimui, S. Beebe, M. Blair, R. Buruchara, P. Kimani 2.3.9 Developing bush and climbing bean lines with resistance to Pythium root rot, angular leaf spot, and bean common mosaic and necrotic viruses Rationale: Pythium root rots and angular leaf spot (ALS) are some of the important diseases affecting bean production in Africa, particularly, on major commercial and adapted bush and climbing bean cultivars. At the same time, some of the good sources of resistance to other diseases are either susceptible (e.g. CAB 19, G2333) to bean common mosaic (BCMV) and bean common mosaic necrotic virus (BCMNV) or have the dominant “I” gene (RWR 719, RAB 487, RWR 2075, RWR 1946) which confers resistance to a wide range of BCMV strains. However, BCMNV strains induces the lethal black root hypersensitive resistant (HR) reaction on germplasm with “I” gene and therefore limits their usefulness. To provide stable, broad-based resistance to the latter, a suitable strategy is to protect the “I” gene by combining it with race non-specific (bc-3) or race-specific resistance recessive genes (typically bc-22). Efforts have been on-going to improve resistance against these diseases by new introductions or pyramiding resistances. Several populations have been generated, and selection is done using a variety of methods, including marker assisted selection (MAS). Last year, we reported progress made in improving resistance against Pythium root rot ALS and efforts to introgress BCMV and BCMNV resistances in commercial and adapted bush and climbing bean cultivars. This year we continued selection from segregating populations and lines with a focus on resistance, yield and seed types. 164 Materials and Methods: i) Evaluation and advancement of backcross populations developed for Pythium resistance Since 2004, selections from 20 backcross populations developed as an effort to introgress Pythium root rot resistance to into key commercial bean varieties have been going on. Selections are based on resistance to Pythium root rot and ALS using both phenotypic and genetic markers. This year, phenotypic selections were done on three F4BCS4 families having RWR 719 as a Pythium root rot resistant donor parent, i.e., F4BCS4 GLP 585 x RWR 719, F4BCS4 CAL 96 x RWR 719 and F4BCS4 URUGEZI x RWR 719. Five plants per selected plot were selected based on plant architecture, pod load, disease and pest resistance, early maturity and physical similarity to the recurrent parent (seed size, seed color, growth habit etc.). The selected plants were assayed for the presence of the Pythium root rot resistance gene using the PYAA19800 SCAR marker. Three hundred and eleven back cross (F5BCS4) lines from populations developed from AND1062, AND1055, SCAM-80CM/15 and MLB-49-89A as Pythium root rot resistant donors were planted and fifty plants selected per population based on phenotypic similarity (growth habit, seed size and color etc) to the recurrent parent. ii) Marker assisted selection from families developed for combined resistance to Pythium and Angular leaf spot Efforts to combine resistance genes to Pythium root rot and angular leaf spot (ALS) resulted in a number of combined crosses. Seventy-eight F8 lines that were F6 derived progenies previously selected under screenhouse condition for resistance to angular leaf spot were assayed for the presence of PYAA19800 SCAR marker associated with Pythium root rot resistance gene in RWR 719 and OPE709 SCAR marker associated with the ALS resistance gene in MEX54. iii) Introgressing bc-3 gene into commercial root rot resistant cultivars with I-gene through backcrossing Commercial varieties that are resistant to Pythium root rots but susceptible to bean common mosaic necrosis virus (RWR 719, RWR 1946, RWR 2075) were crossed with nine bean lines, MCM 2001, MCM 5001, MCM 1015, UBR 92(25), USWK-6, TARS-VR- 7S, USCR-9, USCR-7, that have both the I and bc-3 gene that confers resistance to bean common mosaic necrosis virus. F1 populations were generated and advanced to F2 by selfing. The rationale was to introgress the bc-3 gene into the market class varieties and use both phenotypic selection and molecular markers to select and advance only those materials that had either bc-3 gene alone or a combination of both genes. This past season, 50 plants from each of the 12 crosses involving UBR (92) 25, MCM 1015, MCM 2001 and MCM 5001 (a total of 600 F2 plants) and the four susceptible parents MLB-49-89A, RWR 719, RWR 2075 and RWR 1946 were screened for the presence of the “I”, “bc-3” and Pythium root rot resistance genes using molecular markers. In addition a backcross program was set up for these crosses. At F2, plants were inoculated with BCMV inoculum and resistant plants backcrossed to the respective recurrent parents. BC1S1 seed was planted in the field to get BCS2 plants. Single plants selected from the F2BCS2 populations based on good plant architecture, seed characteristics and phenotypic background of the susceptible parents (RWR719, RWR 1946 and RWR 2075) from each population were inoculated BCMNV inoculum. Those not showing symptoms of black root (Plate 1) were assayed for resistance genes using molecular markers as described below. 165 Plate 1 a. F2 Plant showing black root symptoms Plate 1b. F2 plant showing typical BCMV symptoms DNA Extraction: Using a 2mm bore, 5 discs per leaf were cut out from 2-week old plants and placed on a Whatman plant saver card (taking precautions to prevent cross contamination) and spotted by applying gentle pressure using a pestle. The latter (saver card) were air dried at room temperature for 1 hour. Two millimeter plant saver card discs impregnated with leaf tissue were cut and each washed twice with 200µL Whatman purification reagent, incubating for 3 minutes at room temperature after each wash. The card discs were similarly washed twice with 200µL of isopropanol as described above and left to completely dry before being used as DNA template in a PCR reaction. DNA Amplification: Leaf discs were used as template in a 20µL PCR premix (Bioneer Corp, Korea) containing the following: 1U Top DNA polymerase, 250mM dNTPs, 1.5mM MgCl2, 10mM Tris-HCl (pH 9), 30mM KCl, stabilizer and tracking dye. 0.5µM of the primer and water were added to top up to the required volume. The mixture was vortexed gently and loaded in a Biorad Mycycler (BIO-RAD laboratories, Hercules, California) thermocycler. For the SW13 marker which is associated with “I” gene and PYAA19800 SCAR marker associated with Pythium root rot resistance gene in RWR 719, the mixtures were subjected to 35 amplification cycles which consisted of: an initial denaturation step at 95oC for 5 minutes and 34 cycles at 94oC for 15 seconds, annealing temperature 65oC for 40 seconds, extension 72oC for 1 minute, a final extension of 72oC for 10 minutes and a holding temperature of 4oC. For ROC 11 marker associated with “bc-3” gene, the amplification cycles were similar except that the annealing temperature of 54oC for 40 seconds. Amplicons were resolved on 1.2% agarose gel stained with 10mg ml1 Ethidium bromide and the gel subsequently immersed in 0.5XTBE. Electrophoresis was performed at 70V for 1 hour and bands visualized under UV light and the image captured on a digital camera mounted on a computer. Results and Discussion: i) Selection of backcross (BCs) populations for resistance to Pythium root rot: The RWR 719 SCAR marker identified 28 plants from three F4BCS4 families having the gene linked to Pythium root rot resistance in RWR 719 (Table 84). Three hundred and fifty six plants were selected from 11 backcross populations based on agronomic characteristics (Table 85). 166 Table 84. Selection of F4BCS4 lines using the PYAA800 SCAR marker Population Selected plots F4BCS4 GLP 585 x RWR 719 F4BCS4 CAL 96 x RWR 719 F4BCS4 URUGEZI x RWR 719 Table 85. 2 8 29 Total samples from a cross-code 10 40 145 Plants +ve for RWR719 marker 8 20 28 Phenotypic selections from 11 backcross populations developed for Pythium root rot resistance Population No. progenies per population Plants evaluated per population No. plants selected 39 49 24 19 19 42 16 21 21 27 31 50 50 50 50 50 50 50 50 50 50 50 32 36 40 36 31 34 35 34 25 28 25 308 550 356 F6BCS4 (GLP2 x MLB-49-89A) F6BCS4 (GLP2 x SCAM80 CM/15) F6BCS4 (GLP2 x AND 1062) F6BCS4 (GLP585 x MLB-49-89A) F6BCS4 (GLP585 x AND1055) F6BCS4 (CAL96 x MLB-49-89A) F6BCS4 (CAL96 x SACM 80 CM/15) F6BCS4 (CAL96 x AND1055) F6BCS4 (Urugezi x MLB-49-89A) F6BCS4 (Urugezi x AND1055) F6BCS4 (Urugezi x AND 1062) ii) Screening of combined crosses for resistance ALS and Pythium root rot: The SCAR markers identified 40 lines combining PRR and ALS resistance genes (Figure 46 and Table 86). Agronomic data is being assessed to characterize the materials and make them available to national program partners. iii) Introgressing bc-3 gene into commercial root rot resistant cultivars with I-gene through backcrossing. Locally adapted resistance sources were successfully used to transfer the bc-3 gene into commercial root rot resistant cultivars. Although a relatively higher number of plants had the bc-3 gene (Table 87, Figure 47), very few had a combination of I and bc-3 genes. Plants with at least bc-3 gene (alone or in combination with “I” gene) have been advanced. Furthermore other results show that RWR 2075, a newly released variety in Uganda for its root rot resistance, seems to have a similar marker as that associated with resistance in RWR 719. Results are underway to confirm this observation. RWR 1946 (also newly released for its root rot resistance in Uganda) seems not to have the marker. 167 Figure 46. Screening of combined crosses for the presence of the gene conditioning resistance to Pythium root rots using the marker PY AA19. Left 25/100 mixed DNA ladder. Sample 3 is positive control RWR 719, and sample 26 is variety CAL 96. Absence of a band indicates absence of the gene. Table 86. Lines with both angular leaf spot and Pythium resistance genes Cross Progenies evaluated F8 (CAL 96 x RWR 719) x (CAL 96 x MEX 54) F8 (CAL 96 x MLB-49-89A) x (CAL 96 x MEX 54) F8 (CAL 96 x SCAM 80CM /15 ) x (CAL 96 x MEX 54 Total Table 87 . 30 36 12 78 Plants selected/ progeny 150 180 60 390 No. plants with both PRR and ALS resistance genes 18 20 2 40 Plants with various combinations of genes using the SW13 and ROC11 markers. Cross codes RWR2075 x MCM 5001 RWR2075 x UBR (92) 25 RWR 2075 x MCM 1015 RWR 2975 x MCM 2001 RWR 1946 x UBR (92) 25 RWR 1946 x MCM 1015 RWR 1946 x MCM 2001 RWR 1946 X MCM 5001 RWR 719 X MCM 1015 RWR 719 X MCM 2001 Total No. evaluated 50 50 50 50 50 50 50 50 50 50 500 No. with bc-3 gene 3 8 4 5 4 4 3 11 3 5 50 168 No. with I and bc-3 genes 2 3 2 1 1 2 2 3 1 3 20 Figure 47. Screening of the cross RWR 2075 x MCM 5001 for the presence of the bc-3 gene using the ROC 11 marker. Left 25/100 mixed DNA ladder, sample 9 is positive control MCM 5001; Sample 18 is CAB 19. Absence of a band indicates presence of the gene and vice versa. A total of 95 plants (Table 88) were selected based on their phenotype and advanced to F3BCS2 where selections will be based on the presence of the bc-3 and I gene using the respective molecular markers. Table 88. Phenotypic selections on F3BCS2 plants Crosses RWR 719 x USWK 6 RWR 719 x TARS VR 1s RWR 719 x TARS VR 7s RWR 719 x USCR 7 RWR 719 x USCR 9 RWR 1946 x USWK 6 RWR 1946 x TARS VR 1s RWR 1946 x TARS VR 7s RWR 1946 x USCR 7 RWR 1946 x USCR 9 RWR 2075 x USWK 6 RWR 2075 x TARS VR 1s RWR 2075 x TARS VR 7s RWR 2075 x USCR 7 RWR 2075 x USCR 9 Selected plants No. plants with BCMV No. plants with black root 10 12 1 5 - 2 5 7 2 - 10 9 10 10 9 11 1 7 10 10 - 169 No. plants advanced to BC-S2 8 7 3 7 10 10 8 11 9 2 10 10 95 iv) Pythium Root rot nursery A number of lines derived from RWR 719 and identified (phenotypically and using molecular markers) to be resistant to Pythium root rot were incorporated into the Root Rot Nursery formed last year. This brings to 75 the number of entries in this nursery which in addition to resistance, have desirable seed and agronomic characteristics. Contributors: R. Buruchara, C. Mukankusi, S. Sebuliba, A. Male, C. Acam, F. Mururokwere, P. Kimani, (CIAT); G. Mukeshimana, A. Namayanja, and G. Tusiime Collaborators: S. Beebe and M. Blair References Vallejos C.E., Astua-Monge G., Jones V., Plyler T.R., Sakiyama N.S., Mackenzie S.A. 2006 Genetic and molecular characterization of the I locus of Phaseolus vulgaris. Genetics, 172: 1229-1242. USDA. Agricultural Research Services. Accomplishment Report. Plant disease national program, 2006. Mukeshimana. G., A. Paneda, C. Rodriguez, and J.D.Kelly. 2005. Markers linked to the bc-3 gene conditioning resistance to bean common mosaic potyviruses in common bean. Euphytica 144:291-299. Payne, R.W., Murray, D.A., Harding, S.A., Baird, D.B. and Sourtar, D.M. 2007. Introduction to GenStat for windows 10th edition. VSN International, Hemel Hempstead, UK. . 170 Activity 2.4 Yield potential: climbing beans Highlights • QTL for growth habit and climbing ability were identified on six chromosomes, although many were located on B04. This illustrates the complexity of growth habit, and implicitly, of crop domestication as growth habit was reduced from climbing to bush type. 2.4.1 Detection of QTL for climbing ability and component traits in common bean Rationale: Common bean varies in growth habit from aggressive climbing types to bush beans. While most production of common beans worldwide has been with bush beans, the highest yields are obtained with climbing types, which under favorable growing conditions, can produce as much as 4,500 kg ha–1. One inherent characteristic of climbing beans is their capacity to climb, which is closely related to growth habit. Growth habit is determined by a combination of factors including determinate versus indeterminate growth, total plant height, degree of branching and internode length. Together these factors make up climbing ability. The objective of this research was to determine the quantitative trait loci (QTL) controlling climbing ability in a F5:8 recombinant inbred line population derived from an inter-gene pool cross of an aggressive indeterminate climbing bean with type IV growth habit (G2333) by an indeterminate bush bean of type IIb growth habit (G19839). Materials and Methods: Plant materials: A total of 84 F5:8 recombinant inbred lines (RILs) were developed from the cross G2333 x G19839 by single-seed descent where G2333 is from Mexico and possesses type IVa growth habit, while G19839 is from Peru and possesses type IIb growth. Both are somewhat tolerant of low phosphorus and have been used to study phosphorus use efficiency. Planting sites: The population was planted in four experiments across environments that varied in altitude (from 1000 to 1750 masl) and soil fertility (low versus high phosphorus) over three sites: Darién (1,485 masl), Palmira (965 masl), both in the department of Valle de Cauca while a final experiment was in Popayán (1,730 masl). In Darién, the two trials were planted under conditions of high (64.3 ppm) and low phosphorus (1.7 ppm), respectively. Agronomic management was the same for the four trials, except for fertilizer applications for the low-phosphorus trial in Darién, where, to maintain the low phosphorus levels, only 7.5 kg ha–1 of phosphorus in the form of triple super-phosphate was applied. The other trials received high phosphorus treatment of 45 kg ha–1. Experimental design: At each of the four sites, an experimental design of randomized complete blocks was established, with 86 treatments and 2 replications. The size of the plot or experimental unit was one row, 3 m long, in which 30 seeds were sown at 10-cm intervals each, with a distance of 1.2 m between rows. Field data were collected for climbing ability, plant height, internode length and branch number. Climbing ability (CA) was measured according to a visual scale that ranged from 1 to 9, where 1 corresponded to the highest and most aggressive plants and 9 to the smallest and least aggressive plants. Two evaluations were made, one at 45 days and the second 75 days after planting for both climbing ability and plant height. Results and Discussion: QTL were identified by composite interval mapping for plant height, internode length and number of branches per plant on a genetic map covering all common bean linkage groups with a total length of 1175 cM. A total of 7 QTL were found for plant height, 9 for climbing ability, 6 for internode length and 1 for branch number (Table 89). LR values ranged from 14.8 to 25.3. 171 QTL number varied per location with the favorable environmental conditions in Darién HP and Popayán producing a higher proportion of significant marker association (with 7 and 9 QTL, respectively) than the less favorable environments of Palmira and Darién LP (with 6 and 1, respectively). Table 89. QTL for plant height (PH), internode length (IL), climbing ability (CA) and branch number (BN) in the G2333 x G19839 RIL population detected at either 45 or 75 days after planting in four field experiments (Darién under high phosphorus (HP) or low phosphorus (LP), Popayán (Pop) and Palmira (Pal). Trait - Evaluation Site PH-1 PH-1 PH-1 PH-1 PH-2 PH-2 PH-2 IL IL IL IL IL IL CA-1 CA-1 CA-1 CA-1 CA-1 CA-1 CA-1 CA-2 CA-2 BN HP Pop Pop Pop HP LP Pal HP HP Pal Pal Pal Pop HP HP Pal Pop Pop Pop Pop HP Pal Pop QTL name Plh1-2 Plh1-1 Plh1-3 Plh1-4 Plh2-1 Plh2-3 Plh2-2 Int2 Int3 Int4 Int2 Int3 Int1 Cab1-1 Cab1-4 Cab1-2 Cab1-1 Cab1-3 Cab1-5 Cab1-6 Cab2-1 Cab2-1 Brn1 Linkage group B04 B03 B04 B08 B04 B11 B04 B04 B04 B04 B04 B04 B03 B04 B07 B04 B04 B05 B10 B11 B04 B04 B04 Closest marker X010.85 BM189 PV-ctt001 M130.7 PV-ctt001 BMd033 BMd026 X010.85 PV-ctt001 L040.75 BMd026 PV-ctt001 Q170.46 PV-ctt001 BM046 BMd026 PV-ctt001 F081.0 W060.6 BMd033 PV-ctt001 PV-ctt001 BM161 R2 LR 22.82 15.11 16.14 15.41 20.53 15.35 16.90 23.69 15.97 22.52 19.95 19.48 19.72 20.74 18.84 19.19 16.36 14.81 15.53 14.79 22.01 23.54 25.34 0.1952 0.1399 0.1403 0.1055 0.2489 0.1637 0.1445 0.1839 0.2033 0.2836 0.1910 0.1574 0.1609 0.2026 0.1872 0.1759 0.1353 0.1152 0.1381 0.2161 0.2373 0.2542 0.2235 Total R2 Additivity 0.4812 0.5119 0.5276 0.4762 0.4158 0.3039 0.4624 0.5407 0.453 0.4067 0.3141 0.474 0.3926 0.4946 0.4884 0.4234 0.5432 0.4093 0.4279 0.5161 0.4771 0.4313 0.5321 0.1903 0.1358 0.1395 0.1233 0.2523 0.1887 0.1255 1.8164 1.9515 1.1886 0.9671 0.8533 1.1631 -0.5859 0.5846 -0.3575 -0.3522 -0.3356 -0.3833 -0.4408 -0.5622 -0.6004 0.6890 In addition more QTL were observed at the earlier evaluation stage of 45 DAP compared to the later evaluation stage of 75 DAP for both plant height (4 versus 3 QTL) and climbing ability (7 versus 2 QTL). The largest number and most significant QTL were found on the lower half of linkage group B04 suggesting a major pleiotropic locus for growth habit traits at this location of the genome that is distinct from previously characterized genes which control plant morphology of the crop. Contributors: M.W. Blair (SBA-1, CIAT), O.E. Checa (Univ. Nariño) Collaborators: I. Ochoa (CORPOICA), S. Beebe (CIAT) 172 Activity 2.5 Characterizing and monitoring pathogen and insect diversity Highlights: • 2.5.1 Sixteen Pythium species were found to be associated with beans root rots in Rwanda and the cultivars CAL 96, RWR 617-97A, Urugezi and RWR 1668 were susceptible to all these species. G2331, AND 1062, MLB-40-89A, Vuninkingi, AND 1064 and RWR 719 were resistant. Monitoring of whitefly populations in the Andean zone Rationale: Monitoring the changes that could occur in white fly populations and the composition of species in the target areas of the Andean Zone, where bean and snap bean crops are important, is one of the main objectives of the Project financed by the Regional Fund of Agricultural Technology (FONTAGRO). In addition, the Project financed by the Ministry of Agriculture and Rural Development (MADR), also has as important goal to periodically sample whitefly species on hot and sweet peppers in the Cauca Valley, in order to measure resistance to insecticides. Since the same species of white fly pass between bean and peppers, this activity is relevant for the bean program. This information is necessary to modify the existing management systems and to be prepared for new situations in the future. Materials and Methods: In 2008 a total of 20 white fly samples (adults and pupae) were processed. These were collected in 8 locations of the Antioquia, Cundinamarca and Tolima departments of Colombia, at altitudes ranging between 1730 and 2465 meters above sea level (masl). Samples were taken from beans and snap beans within the project financed by FONTAGRO. For the MADR project, a total of 12 whitefly samples (adults and pupae) were processed, having been collected in 4 locations of the Cauca Valley Department of Colombia, at altitudes ranging between 941 and 1437 meters above sea level (masl). Samples were taken from both hot pepper and sweet peppers. RAPD techniques (primer OPA-04) were used to identify species of pupae and adults. The identification was based on morphological characteristics of pupae and comparisons between RAPD patterns in samples brought from the field with those of existing mass-rearing colonies maintained at CIAT. Results and Discussion: The analysis of 20 samples taken in 8 locations in the Antioquia, Cundinamarca and Tolima departments of Colombia (Figure 48), showed that 100 % of the whiteflies collected belong to Trialeurodes vaporariorum, identified as the most important whitefly species on bean and snap beans crops in the hillside areas of Antioquia, Cundinamarca and Tolima. In order to determine the distribution of the whitefly in sweet pepper, other areas in Colombia continue to be monitored, where it is considered to be an important pest. Four areas located in the northern and central part of the Valle de Cauca department were sampled, with altitudes ranging between 941 and 1437 masl. As an example of the RAPD patterns obtained, Figure 49 shows that 17% of the collected samples were of T. vaporariorum and the other 83 %, found between 900 and 1400 masl, correspond to B. tabaci biotype B. Presence of biotype B of B. tabaci was evidenced in the field by proofs of physiological disorders (irregular ripening) and by morphological differentiations with T. vaporariorum. These identifications were confirmed by the means of RAPD type molecular trials. Since Bemisia spp. are vectors of Gemini viruses of Phaseolus, these results are directly relevant to common bean. 173 Figure 48. RAPD’s of white flies collected in Antioquia and Cundinamarca (Colombia) in common bean and snap bean. Amplification of the primer OPA-04: M, Marker 100 pb; 1, T. vaporariorum CIAT; 2, B. tabaci biotype A CIAT; 3, B. tabaci biotype B CIAT; 4-5, adults of T. vaporariorum collected in Aguasclaras (Carmen de Viboral, Antioquia, 2165 masl); 6, pupae (not determined) collected in La Aurora (C. de Viboral, Ant. 2189 masl); 7, pupae de T. vaporariorum collected in La Aurora (Carmen de Viboral, Antioquia. 2189 masl); 8-9, adults of T. vaporariorum collected in La Aldana (Carmen de Viboral, Antioquia, 2169 masl); 10-11, adults de T. vaporariorum collected in San Juan Bosco (Marinilla, Antioquia, 2200 masl); 12-13, adults of T. vaporariorum collected in La Floresta (Santuario, Antioquia, 2134 masl), 14-15, adults of T. vaporariorum collected in Laderas (Fómeque, Cundinamarca, 1982 masl); 16, adults of T. vaporariorum collected in La Unión (Fómeque, Cundinamarca, 1730 masl); 17, B. tabaci biotype B CIAT; 18, B. tabaci biotype A CIAT; 19, T. vaporariorum CIAT. Figure 49. RAPD’s of white flies collected in the northern and central Cauca Valley in hot pepper. Amplification ofthe primer OPA-04: M, Marker 100pb; 1, T. vaporariorum CIAT; 2, B. tabaci biotype A CIAT; 3, B. tabaci biotype B CIAT; 45, adults of B. tabaci biotype B collected in La Unión (941 masl), 6-7, adults of B. tabaci biotype B collected in La Cumbre (1437 masl); 8, pupae of T. vaporariorum collected in La Cumbre (1437 masl); 9, pupae of B. tabaci biotype B collected in La Cumbre (1437 masl); 10-11, adults of B. tabaci biotype B collected in La Cumbre (1398 masl); 12, adults of B. tabaci biotype B collected in Restrepo (1181 masl); 13, adults of T. vaporariorum collected in Restrepo (1182 masl); 14-15, pupae of B. tabaci biotype B collected in Restrepo (1181 masl); 16, B. tabaci biotype B CIAT; 17, B. tabaci biotype A CIAT; 18, T. vaporariorum CIAT; Bl, Reaction blank. 174 2.5.2 Diversity, distribution and pathogenicity of Pythium species in Rwanda Rationale: Beans (P. vulgaris L.) is a major source of protein in Eastern Africa and particularly in Rwanda. Produced under low input agriculture, beans are vulnerable to attack by diseases and other constraints. One of these diseases is Pythium root rots. The increase in severity and incidence of root rots have been associated with a relatively recent evolution of farming systems especially under high demographic pressure and the resulting decline in soil fertility (Rusuku et al., 1997). This is typical of Rwanda where the importance of root rots was recognized as far back as late 80s (CIAT, 1992). There are efforts in Rwanda to routinely incorporate resistance to Pythium root rots in improved bush and climbing beans varieties. However, as this is done, it is considered important to determine and monitor the variation, distribution and pathogenicity of Pythium species in Rwanda. The information will then be used to identify or verify sources of resistance and for phenotypic evaluations in varietal improvements. The aim of this study was to characterization Pythium species responsible for bean root rot in Rwanda. Materials and Methods: Soil and plant samples were collected from fields in 24 districts of Rwanda (Table 90) at predetermined positions and at different altitude levels [low (900 – 1400 m), medium (1400 – 1650 m) and high (1650 – 2300 m)]. At each field, information was taken on the physical location including: Province, District and Sector; GPS location. Attributes of the beans varieties in the fields were observed and noted, e.g. type of growth; as well as on the cropping systems. Table 90. Distribution of the samples by district in Rwanda Number of samples Huye 14 Gisagara 14 Nyanza 26 Karongi 27 Muhanga 4 Ruhango 8 Nyamasheke 12 Rusizi 18 Nyamagabe 5 Bugesera 5 Gasabo 6 Rwamagana 6 Kayonza 10 Ngoma 12 Kirehe 5 Gatsibo 4 Nyagatare 19 Rurindo 5 Gicumbi 14 Nyarugenge 6 Gakenke 1 Nyabihu 5 Rubavu 3 Musanze 2 Total of samples 231 LA: Low altitude, MA: Middle altitude, HA: High altitude. 175 Altitude MA MA MA MA MA MA MA LA MA LA MA LA LA MA MA MA LA HA HA MA HA HA HA HA Isolation and purification of Pythium species: Initial steps of isolation were done at ISAR’s laboratories at Rubona and Ruhengeri in Rwanda. Follow-up laboratory and screenhouse activities were carried out at Kawanda Agricultural Research Laboratories. Isolation of Pythium spp. from plant tissues was done as described by White (1988). Infected root pieces from the field samples (approximately 0.5- 2 cm long) were cut from expanding lesions and plated onto the selective medium of cornmeal agar (CMA) amended with 2ml/l and 3ml/l of antibiotics pimaricin and rifamycin respectively, to stop bacterial growth. Cultures were incubated at 20oC for 2-5 days. Hyphal tip method was used to sub-culture and transfer mycelia onto potato dextrose agar (PDA) slants. Molecular characterization of Pythium spp: DNA was extracted from harvested mycelia according to the procedure described by Mahuku (2004). Mycelia were ground to a fine paste in a mortar containing TES extraction buffer (0.2 M Tris-HCl [pH 8], 10 mM EDTA [pH 8], 0.5 M NaCl, 1% SDS) and sterilized acid-washed sea sand. Additional TES buffer containing Proteinase K was added and the mixture incubated at 65ºC for 30 min. DNA was precipitated using ice-cold isopropanol and the pellet was washed twice with 70% ethanol, dried and dissolved in TE buffer (10 mM Tris-HCl [pH 8], 1 mM EDTA). PCR analysis was performed using Oomycete ITS (Internal Transcribed Sequence) region primers to differentiate Pythium from other closely related fungi (White et al., 1990). The PCR reaction was performed in 50µl-reaction volumes containing 5µl of 10X PCR buffer, 8µl of 25mM MgCl2, 2.5 µl of 1.25mM dNTP, 0.2 µl of each primer (20µM) 18S (5’-TCC GTA GGT GAA CCT GCG G-3’) and 28S (5’-TCC TCC GCT TAT TGA TAT GC-3’), 20 ng of DNA, and 0.2µl Taq DNA polymerase (5U/µl). Amplification was performed in a BIORAD Mycycler Thermocycler programmed for initial denaturation at 94°C for 5 min, for 35 cycles at 94ºC for 1 min, annealing at 68°C for 1 min, and extension at 72°C for 1.5 min, followed by a final extension for 7 min at 72°C. The products were run on 2% agarose gels containing 5mg ml-1 of ethidium bromide at a voltage of 100V for 2 hours and visualized under UV light. rDNA sequence analysis. Residual primers and dNTPS in the PCR products were removed using QIAquickTM PCR purification spin columns following the manufacturer’s protocol (QIAGEN, Crawley, 1997). Direct sequencing of the PCR amplified products was carried out using ITS 2 primers (White et al., 1990). Sequences from ITS 1 region of the ribosomal gene were edited using the Editseq program (DNASTAR Inc., Madison, Wis). The ITS1 sequences of the test isolates were compared with ITS 1 sequences of known Pythium species available in the public databases using Seqman program (DNASTAR). Multiple alignment of the sequenced product of ITS 1 was performed for comparison. Pathogenicity of Pythium species: The pathogenicity of the characterized Pythium spp was evaluated on the susceptible bean cultivars CAL 96 and “Urugezi”, using 2 or 3 isolates of each species. Similar evaluations were carried out on a few Rwandan varieties (G2331, R617-97A, RWR 1668, Vuninkingi) and other sources of resistance (RWR 719 MLB-40-89A AND 1064 and AND 1062) in the screenhouse. Seed of these cultivars were planted in wooden trays each infested with a distinct Pythium species. The trial was set up in a completely randomized block design (CRBD). Three weeks after emergence, the surviving plants were uprooted, washed and severity of root rots assessed using the CIAT scale of 1-9 (Abawi and Pastor Corrales, 1990). Isolates that had a mean disease score of 1-2, were considered non pathogenic; those with a score of 3-5 were considered mildly pathogenic and those that gave a score of 69 were highly pathogenic. Results Distribution of Pythium spp: Results of this study showed that, P. vexans was the most widespread species (23 isolates), followed by P. indigoferae (9 isolates); P. rostratifingens (6 isolates), P. torulosum (5 isolates), P. ultimum and P spinosum (4 isolates each) (Table 91). 176 Table 91. Distribution of the Pythium species by district in Rwanda District P. indigoferae P. chamaehyph on P. torulosu m P. cucurbitacear um P. diclinu m P. conidiophor um P. P. P. arrhenoman pachycau ultimu es le m P. P. vexan folliculosu s m P. P. P. macrosporu rostratifinge spinosu m ns m P. P. dissotocu rostratu m m Total Huye - - 2 - - - - - - - - 1 - 1 - 1 5 Gisagara - - - - - - - - - - - - 2 - - - 2 7 Nyanza 2 - 1 - - - - - - 3 1 - - - - - Karongi 1 - - - - - - - 1 1 1 - - - 1 - 5 Muhanga - - 1 - - 1 - - - - - - - - - - 2 Ruhango - - - - 1 - - - - - - - - - - - 1 Nyamasheke - - - - - - - - - 1 - - - 1 - - 2 Rusizi - - - - - - 1 - - - - - 2 2 1 - 6 Nyamagabe 1 - - - - 1 - 1 - - - - 1 - - - 4 Bugesera - - - - - - - - 1 3 1 - - - - - 5 Gasabo - - - - - - - - - 4 - - - - - - 4 Rwamagana - - - - - - - - - - - - - - - - 0 Kayonza - - - - - - - - - 1 - - 1 - - - 2 Ngoma - - - - - - - - - 2 - - - - - - 2 Kirehe - - - - - - - - - 1 - - - - - - 1 Gatsibo - - - - - - - - - - - - - - - - 0 Ngoma 4 1 - - - - - - - - - - - - - - 5 Nyagatare 1 - - - - 1 - - 1 2 - - - - - - 5 Rurindo - - - - - - - - - 1 - - - - 1 - 2 Gicumbi - 1 - 1 1 - - - - 1 - - - - - - 4 Nyarugenge - - 1 - - - - - 1 3 - - - - - - 5 Gakenke - - - - - - - - - - - - - - - - 0 0 Nyabihu - - - - - - - - - - - - - - - - Rubavu - - - - - - - - - - - - - - - - 0 Musanze - - - - 1 - - - - - - - - - - - 1 Total 9 2 5 1 3 3 1 1 4 23 3 1 6 4 3 1 70 177 Molecular characterization: A total, of 231 isolates were collected. However, 96 representing different regions were sequenced. ITS sequences were compared using Blast searches with sequences deposited at the National Center for Biotechnology Information (NCBI). Identity of species with sequences having less than 100% similarity to published sequences in GenBank were further analysed using phylogenetic analysis using parsimony (PAUP) version 4·0b (Swofford, 2001). The criterion for the selection of closely related species was based on clades of Pythium spp. generated by Lévesque and de Cock (2004). As a result a total of 16 different Pythium species were identified (Table 91). These were P. indigoferae, P. chamaehyphon, P. torulosum, P. cucurbitacearum, P. diclinum, P. conidiophorum, P. arrhenomanes, P. pachycaule, P. ultimum, P. vexans, P. folliculosum, P. macrosporum, P. spinosum, P. rostratifingens, P. dissotocum and P. rostratum. Pathogenecity: All the Pythium species identified were pathogenic to the susceptible varieties [CAL 96 (Plate 2) and URUGEZI] and other varieties (RWR 617-97A and RWR 1668) evaluated (Table 92). Bean cultivars G2331, AND 1062, MLB-40-89A, Vuninkingi, AND 1064 and RWR 719 (Plate 3) gave resistant reactions. The study confirmed previous reports regarding resistance of some of the known sources of resistance and the value of this resistance (Otsyula et al., 1998; Buruchara et al., 1999; Buruchara and Kimani 2001, Buruchara et al., 2007, Gichuru et al., 2008). It was however interesting that the known susceptible varieties were affected by all the species identified indicating the potential impact of Pythium species under favorable disease conditions. Plate 2: Roots of the susceptible (CAL 96) bean variety to the Pythium spp. Plate3: Roots from resistant variety (RWR 719) to the Pythium spp.. 178 Table 92. Pathogenicity and disease severity following artificial inoculation by different Pythium species on bean varieties cultivated in Rwanda. Beans Varieties Pythium species severity1 P. P. P. P. P. P. disso P. follicu P. indigo P. pachy P. rostrati P. P. P. arrheno chamae conidio cucurbita diclinum tocum losum Ferae caule fingens spinosum torulosum ultimum P. vexans P. P. macrospor rostratum um manes Hyphon Phorum Cearum CAL 96 8.5AB 8.1A 8.3A 8.8A 8.3B 8.4A 8.0B 7.8B 7.1B 8.4A 8.7A 8.2A 8.4A 8.4A 8.2B 8.6A G2331 3.9D 2.1 DC 2.4B 2.0DE 1.7DC 1.6ED 2.1DC 2.0C 1.8ED 1.4DC 1.6E 2.0D 1.7C 2.0ED 2.8E 1.6DC RWR 617-97A 7.9BC 8.4A 8.0A 7.2C 8.3B 7.9B 7.8B 7.7B 8.2A 7.3B 6.0C 7.4B 7.3B 7.7B 6.8D 6.2B URUGEZI 8.7A 8.0A 8.3A 8.7A 8.7A 8.5A 8.5A 8.3A 8.2A 8.7A 8.7A 8.5A 8.8A 8.8A 9.0A 8.5A RWR 1668 7.7C 6.1B 7.7A 7.8B 8.3B 5.8C 8.2BA 7.4B 6.3C 8.5A 8.2B 8.1A 8.6A 6.0C 7.4C 6.5B AND 1062 2.0E 1.9CD 1.9BC 2.2D 1.4DE 1.5ED 1.7DE 1.5DCE 1.8ED 1.2DC 1.6E 1.8D 1.5DC 1.9ED 2.0FG 1.6DC MLB 40-89A 2.3E 1.6DE 1.7CD 2.1DE 1.3DE 1.5ED 1.6E 1.5DE 2.1D 1.3DC 1.4EF 1.7ED 1.6DC 1.8E 1.9G 1.9DC VUNINKINGI 1.8E 1.4E 1.2D 1.7E 1.2E 1.2ED 1.1F 1.2E 1.5E 1.1D 1.1F 1.3E 1.3D 1.2F 2.0FG 1.3D AND 1064 2.2E 2.2C 2.0BC 2.1DE 1.9C 1.9D 2.2C 1.9DC 2.3D 1.5C 2.0D 2.6C 1.9C 2.4D 2.5FE 2.1C RWR 719 2.2E 1.6DE 1.6CD 2.0DE 1.5DE 1.7D 1.5FE 1.4E 1.9ED 1.2DC 1.8ED 1.6ED 1.2D 1.6EF 1.9G 1.4D SE 0.25 0.16 0.22 0.17 0.13 0.17 0.18 0.18 0.20 0.13 0.13 0.17 0.16 0.18 0.19 0.21 F(9,288) < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 < 0.0001 1: Based on CIAT scale of 1 to 9 179 References Abawi, G.S., and Pastor–Corrales, M.A. 1990. Root rots of beans in Latin America and Africa: diagnosis, research, methodologies and management strategies. CIAT, Cali, Colombia.114 p. CIAT, (1992). Pathology in Africa. In: CIAT annual report 1992.CIAT Bean Program, Cali, Colombia. Mahuku, G. (2004). A Simple Extraction Method Suitable for PCR-Based Analysis of Plant, Fungal, and Bacterial DNA. Plant Molecular Biology Reporter 22: 71–81, March 2004. Miklas, N.P., Kelly, J.D., Beebe, S.E. and Blair, M.W. 2006. Common bean breeding for resistance against biotic and abiotic stresses: From classical to Marker assisted selection breeding. Euphytica147:105-131. Otsyula, R.M and Ajanga, S.I. 1998. Development of an intergrated bean root rot control for western Kenya. African Crop Science Journal 6:62-67. Rusuku, G., Buruchara, R.A., Gatabazi, M. and Pastor-Corrales, Schmitthenner, A.F. (1997). Effect of crop rotation on Pythium ultimum and other Pythium species in the soil. Phytopathology 52:27. White T.J., Bruns T., Lee S., Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Shinsky JJ, White TJ, editors. PCR Protocols: A Guide to Methods and Applications, 315–322. Academic Press, San Diego. Wortmann C.S., Kirkby, R.A., Eledu, C.K.A., Allen, D.J . 1998. Atlas of Common Bean (Phaseolus vulgaris L.) Production in Africa. CIAT, Cali, Colombia (Publication No. 297). Contributors: Nzungize R. J1., Buah S2. , Mukankusi2 C., Buruchara2 R., Baudoin J.P3. 1 = ISAR, 2 = CIAT- Africa 3 = Gembloux Agricultural University, Belgium 180 Activity 2.6 Developing integrated disease and pest management components Highlights: • • • • 2.6.1 All of the bean-planted areas in Colombia and Ecuador included in the Fontagro-financed project “Reduction in the Use of Pesticides and Development of Resistance in Rice and Common Bean Crops in Colombia, Venezuela and Ecuador” were monitored for white fly populations. Whitefly species and biotypes, thrips and leafminers were identified, patterns of pesticide use for whitefly, thrips and leafminers registered, and levels of pesticide resistance in whitefly, thrips, and leafminer populations in Colombia quantified. Psuedomonas sp were isolated and shown to have antagonistic effects on Pythium spp Genetic diversity of varietal mixtures from southwest Uganda was characterized with indications of its potential value in the region. Reduction of pesticide use in common bean and snap bean crops through the development and implementation of IPM strategies in Colombia and Ecuador: on-farm surveys and baseline studies of pest resistance Rationale: As indicated in the 2007 Annual Report, monitoring of insecticide resistance levels in adults and immature populations of whiteflies, thrips and leafminers is a major objective of the Fontagrofinanced project. The two main whitefly species, thrips and leafminers in the Andean zone are a target of excessive use of insecticides. This is reflected in ever increasing levels of resistance to insecticide and difficulties in control. The principal purpose of a continuous monitoring of insecticide resistance is to develop alternative management strategies that will help to overcome resistance or delay the onset of this phenomenon. Materials and Methods: The main areas sampled were the northern part of Ecuador, and Valle de Cauca, Antioquia, Cundinamarca and Tolima departments in Colombia. Tests for insecticide resistance measurements were carried out in 43 different sites: 30 in Colombia and 13 in Ecuador, at altitudes ranging between 940 and 2465 masl. Using previously established diagnostic dosages for nymphs and adults, populations were tested in target areas. Adult resistance levels were monitored under field conditions by means of the insecticide-coated glass vial technique. Resistance of immature stages was tested by using the foliage dipping technique. Systemic novel insecticides (mostly neonicotinoids) were tested using the petri dish technique. Results and Discussion: Trialeurodes vaporariorum Valle de Cauca and Antioquia regions: Important changes were detected in adults of T. vaporariorum. High levels of resistance to organophosphates (methamydophos) were registered in most of the evaluated sites. Intermediate levels of resistance were registered to pyrethroid (cypermethrin) in Antioquia. All the races showed susceptibility to diagnostic dosages of carbamates (methomyl), neonicotinoids (imidacloprid) and the neirextoxine (thioxiclam hydrogen oxalate) (Table 93). 181 Table 93. Response (percentage mortality) of Trialeurodes vaporariorum adults to five insecticides in 10 areas of the Valle de Cauca – Antioquia regions of Colombia. Diagnostic dosages in µg vial-1 were tested using insecticide-coated glass vials, Diagnostic dosages in µg vial-1 or ppm. thioxiclam hydrogen oxalate (1500 ppm) CIAT 100.0 ab 93.8 a 89.7 ab 88.2 bcd 87.2 cde Pradera 2 (V) 100.0 a 8.5 f 75.7 abcde 89.1 bcd 94.6 abcd La Cumbre (V) 100.0 a 42.1 cd 86.3 abc 84.2 cd 97.0 ab C.Viboral 1 (A) 100.0 a 21.7 de 66.3 bcde 91.8 bc 91.8 bcde C.Viboral 2 (A) 100.0 a 22.0 ef 62.6 de 92.9 bc 85.9 de Santuario 1 (A) 100.0 a 53.0 bc 53.7 e 98.0 a 89.0 cde Marinilla (A) 100.0 a 36.8 cde 63.2 cde 99.0 a 91.0 cde Santuario 2 (A) 100.0 a 58.1 bc 79.9 abcd 95.9 ab 97.9 a El Cerrito (V) 97.9 b 46.9 bc 88.8 a 91.6 bcd 81.1 e Pradera1 (V) 94.9 c 63.8 b 85.1 abc 84.9 cd 93.7 abc Tulua (V) 93.8 c 38.1 cde 93.8 a 82.3 d 92.6 abc a Departments (A)=Antioquia, (V)= Valle de Cauca; b Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. methomyl Site methamidophos (32 µg vial-1 ) (Department)a (2.5 µg vial-1 ) cypermethrin (500 µg vial-1 ) imidacloprid (40 ppm) Corrected percentage mortality Figure 50 presents results of periodic measurements (1997-2008) of resistance to methamidophos in T. vaporariorum adult populations collected in three zones of Valle de Cauca and one in Antioquia. A significant decrease in resistance levels was detected in Pradera and El Cerrito in the Cauca Valley. Increases in resistance levels were found in La Cumbre (Valle) and in Carmen de Viboral (Antioquia). 1997 100 ab a 2002 2001 2003 2005 2008 a c c bc b 80 a 60 c c 40 a c b bc 20 a a c bc b 0 CIAT Pradera (Valle) La Cumbre (Valle) El Cerrito (Valle) Carmen de Viboral (Ant) Races Figure 50. Changes in toxicological responses to methamidophos (32 µg vial-1) in adult populations of Trialeurodes vaporariorum. Columns labeled with the same letter are not significantly different (P = 0.05) using the LSD test. Each site analyzed separately. CIAT = reference strains. A reduced response to the growth regulator buprofezin was detected in first instar nymphs of T. vaporariorum collected in El Cerrito and Pradera in Valle de Cauca. The Pradera strain exhibited intermediate levels of resistance to the neonicotinoid imidacloprid (Table 94). 182 Table 94. Response (percentage mortality) of Trialeurodes vaporariorum nymphs to three insecticides in Valle de Cauca and Antioquia Departments of Colombia. Diagnostic dosages in ppm. buprofezin diafenthiuron imidacloprid (16 ppm) (300 ppm) (300 ppm) C.Viboral 1 (A) 100.0 ab 100.0 a 99.5 a Santuario 2 (A) 100.0 a 100.0 a 99.4 ab CIAT 100.0 a 100.0 a 100.0 a C.Viboral 2 (A) 100.0 a 97.6 b 82.3 cd La Cumbre (V) 100.0 a 100.0 a 97.9 ab Santuario 1 (A) 100.0 a 100.0 a 90.3 cd Marinilla (A) 100.0 a 100.0 a 97.6 ab Tulua (V) 98.4 a 100.0 a 88.9 c El Cerrito (V) 52.2 b 100.0 a 100.0 a Pradera 2 (V) 35.0 c 100.0 a 76.4 d Pradera 1 (V) 29.0 c 100.0 a 91.8 bc a Departments (A)=Antioquia, (V)= Cauca Valley; b Means within a column followed by the same letter are not significantly different at the P = 0.05 by LSD test. Site (Department)a Corrected percentage mortality Periodic resistance measurements were also made on nymphal populations of T. vaporariorum collected in Pradera and El Cerrito (Valle de Cauca). A significant increase in the level of resistance to buprofezin was detected in Pradera and El Cerrito, possibly due to excessive use of this insecticide in those areas of Valle de Cauca in Colombia (Figure 51). 2002 2001 100 a a a a a a 2006 2005 a a a a a b 80 2003 a 2008 a a b b b b 60 c 40 c 20 0 CIAT Pradera La Cumbre El Cerrito El Dovio Races Figure 51. Changes in toxicological responses to buprofezin (16 ppm) in nymphal populations of Trialeurodes vaporariorum. Columns labeled with the same letter are not significantly different (P = 0.05) using the LSD test. Each site analyzed separately. CIAT = reference strains. Cundinamarca-Tolima region: In this region greater changes were detected. In general, the response (mortality) of T. vaporariorum adults to methamidophos was low (Table 95); however, nymphs are still susceptible to the insecticides that were tested (Table 96). 183 Table 95. Response (percentage mortality) of Trialeurodes vaporariorum adults to five insecticides in the Cundinamarca and Tolima regions of Colombia. Diagnostic dosages in µg vial-1 were tested using insecticide-coated glass vials, Diagnostic dosages in µg vial-1 or ppm. thiocyclam hydrogen oxalate (1500 ppm) CIAT 100.0 ab 93.8 a 89.7 a 88.2 bc 87.2 b Cajamarca 1 (T) 100.0 a 43.2 b 82.1 ab 89.7 ab 77.3 b Cajamarca 2 (T) 100.0 a 12.9 b 83.9 ab 95.7 a 78.5 b Pasca (C) 100.0 a 31.2 b 81.7 a 85.4 bc 99.0 a Fómeque 2 (C) 100.0 a 27.0 b 57.9 c 86.2 bc 95.7 a Fómeque1 (C) 97.9 a 26.6 b 69.1 bc 81.7 c 95.7 a a Departments (C) = Cundinamarca, (T) = Tolima; b Means within a column followed by the same letter are not significantly different at the P = 0.05 level by LSD. Site (Department)a methomyl (2.5 µg vial-1) methamidophos (32 µg vial-1) cypermethrin (500 µg vial-1) imidacloprid (40 ppm) Table 96. Response (percentage mortality) of Trialeurodes vaporariorum nymphs to three insecticides in the Cundinamarca and Tolima regions of Colombia, using diagnostic dosages in ppm. Tests were done following the methodology suggested by Prabhaker et al. (1985)a. buprofezin diafenthiuron imidacloprid (16 ppm) (300 ppm) (300 ppm) Cajamarca 1 (T) 100.0 ac 100.0 a 100.0 a CIAT 100.0 a 100.0 a 100.0 a Fómeque 2 (C) 94.4 ab 88.3 b 83.5 b Cajamarca 2 (T) 94.3 b 100.0 a 100.0 a Fómeque1 (C) 92.7 b 81.1 b 74.0 b Pasca (C) 50.8 c 85.0 b 91.3 b a Prabhaker, N.; Coudriet, D.; Meyerdirk, D. 1985. Insecticide resistance in the sweetpotato whitefly Bemisia tabaci (Homoptera: Aleyrodidae). J. Econ. Entomol. 78: 748-752; b Departments (C) = Cundinamarca, (T) = Tolima; c Means within a column followed by the same letter are not significantly different at the P = 0.05 using the LSD test. Place (Department)b Ecuador zone: The mortality response of T. vaporariorum to methamidophos was low in 80% of the sampled sites in Ecuador. Although neonicotinoids (imidacloprid) are not widely used in the northern region of Ecuador, a slight decrease in the response to this insecticide was found in Chalguayacu (Table 97). The excessive use of organophosphates in Ecuador is responsible for the significant increases in resistance levels through time (Figure 52). It is possible that the use of neonicotinoids at dosages below recommended levels has led to the development of resistance to neonicotinoids (Figure 53). No major changes were detected in nymphal populations. Bemisia tabaci biotype B The B biotype of B. tabaci has been shown to be more aggressive and to have a major capacity to develop resistance to insecticides. As can be seen in Table 98, the insect presents high levels of resistance to organophosphates, and intermediate levels of resistance to pyrethroids and neonicotinoids in Roldanillo (Cauca Valley of Colombia). 184 Table 97. Response (percentage mortality) of Trialeurodes vaporariorum adults to five insecticides in Ecuadorian provinces. Diagnostic dosages in µg vial -1 were tested using insecticide-coated glass vialsa. Diagnostic dosages in ppm, tested using the Cahill et al. (1996)b technique. thiocyclam hydrogen oxalate (1500 ppm) CIAT 100.0 ac 93.8 a 89.7 b 88.2 bc 87.2 c San Vicente 100.0 a 36.5 bc 96.9 a 83.7 cd 100.0 a Chalguayacu 100.0 a 59.6 ab 100.0 a 72.0 d 100.0 a Carpuela 100.0 a 24.7 c 100.0 a 92.9 ab 98.0 ab Pusir 100.0 a 61.6 bc 83.8 b 97.0 a 95.7 b Ibarra 100.0 a 31.3 bc 86.9 b 89.6 bc 100.0 a Concepción 93.3 b 64.6 b 98.1 a 81.5 cd 100.0 a a Plapp, F. W.; Jackman, J. A.; Campanhola, C.; Frisbie, R. E.; Graves, J. B.; Lutrell, R. G.; Kitten, W. F.; Wall, M. 1990. Monitoring and management of pyrethroid resistance in the tobacco budworm (Lepidoptera: Noctuidae) in Texas, Mississippi, Louisiana, Arkansas and Oklahoma. J. Econ. Entomol. 78: 748-752; b Cahill, M., K. Gorman, S. Day & I. Denholm. 1996. Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae). Bull. Entol. Res. 86: 343-349; c Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. methomyl (2.5 µg vial-1 ) Corrected percentage mortality Race methamidophos (32 µg vial-1 ) 2002 1997 100 a a a cypermethrin (500 µg vial-1 ) 2003 imidacloprid (40 ppm) 2008 a 80 a a 60 a a ab a 40 a a a a ab b 20 b a a a b a 0 CIAT San Vicente Ibarra Pimampiro Pusir Carpuela Races Figure 52. Changes in toxicological responses to methamidophos (32 µg vial-1) in adult populations of Trialeurodes vaporariorum in Ecuador. Columns labeled by the same letter are not significantly different (P = 0.05) using the LSD test. Each site analyzed separately. CIAT = reference strains. 185 Corrected percentage mortality 2002 100 a a 2003 a a b b 80 a a 2008 a a b b ab a a a a b 60 40 20 0 Figure 53. Table 98. CIAT San Vicente Ibarra Pimampiro Pusir Carpuela Races Changes in toxicological responses to imidacloprid (40 ppm) in adult populations of Trialeurodes vaporariorum. Columns followed by the same letter are not significantly different (P = 0.05) using the LSD test. Each site analyzed separately. CIAT = reference strains. Response (percentage mortality) of Bemisia tabaci B Biotype adults to five insecticides in Colombia and Ecuador. Diagnostic dosages in µg vial-1 were tested using insecticide-coated glass vialsa, Diagnostic dosages in ppm, tested using the Cahill et al. (1996)b technique. thiocyclam hydrogen oxalate (1500 ppm) Palmira (C) 100.0 ad 7.3 c 79.2 b 67.7 c 62.6 b Cuambo(E) 99.0 a 47.9 b 96.9 a 94.8 a 100.0 a CIAT 95.1 b 95.9 a 95.1 a 88.7 ab 94.0 a Roldanillo (C) 89.9 b 25.0 bc 62.5 c 79.6 bc 94.7 a a Plapp, F. W.; Jackman, J. A.; Campanhola, C.; Frisbie, R. E.; Graves, J. B.; Lutrell, R. G.; Kitten, W. F.; Wall, M. 1990. Monitoring and management of pyrethroid resistance in the tobacco budworm (Lepidoptera: Noctuidae) in Texas, Mississippi, Louisiana, Arkansas, and Oklahoma. J. Econ. Entomol. 78: 748-752; b Cahill, M., K. Gorman, S. Day & I. Denholm. 1996. Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae). Bull. Entol. Res. 86: 343-349; c Country (C) = Colombia, (E) = Ecuador; d Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. Place (Country)c methomyl (2.5 µg vial-1 ) methamidophos (32 µg vial-1 ) cypermethrin (500 µg vial-1 ) imidacloprid (40 ppm) The evaluation of nymphs was done with three insecticides. No significant changes were detected. The Roldanillo race in Colombia showed intermediate resistance to imidacloprid. In general, the response of B. tabaci nymphs (Table 99) is still that of susceptibility to the insecticides that were tested. 186 Table 99. Response (percentage mortality) of Bemisia tabaci B biotype nymphs to three insecticides in Colombia and Ecuador. Diagnostic dosages in ppm, tested using the Prabhaker et al. (1985)a technique. buprofezin (16 ppm) 100.0 ac 99.0 a 98.3 a 93.4 a Place (Country)b Cuambo(E) Palmira (C) Roldanillo (C) CIAT diafenthiuron (100 ppm) 100.0 a 94.9 b 100.0 a 100.0 a imidacloprid (1000 ppm) 100.0 a 100.0 a 75.0 c 91.6 b a Prabhaker, N.; Coudriet, D.; Meyerdirk, D. 1985. Insecticide resistance in the sweetpotato whitefly Bemisia tabaci (Homoptera: Aleyrodidae). J. Econ. Entomol. 78: 748-752; b Country (C)= Colombia, (E)= Ecuador; c Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. Thrips palmi Karny T. palmi has become a major pest of vegetables and beans in Colombia and Ecuador. A survey showed that snap beans, dry beans, sweet pepper, melon, squash and cucumber are the crops most affected by this pest in the Antioquia Department of Colombia and in El Carchi province of Ecuador. Up to 60% of farmers surveyed in these countries use pesticides as the only method of control of thrips populations (see Annual Report 2007). Table 100 shows that in Colombia this insect has developed high levels of resistance to organophosphate and pyrethroid insecticides. In the Antioquia Department, novel products like pirazoles (fipronil) and, to a lesser extent, neonicotinoids, have lost effectiveness to control this insect. Table 100. Response (percentage mortality) of Thrips palmi adults to six insecticides in Colombia. Diagnostic dosages in ppm, tested using the Cahill et al. (1996)a technique. Place (Department)b imidacloprid (1000 ppm) spinosad (2000 ppm) fipronil (100 ppm) carbosulfan (1000 ppm) cypermethrin (16 ppm) methamidophos (16 ppm) CIAT 100.0 ac 100.0 a 100.0 a 100.0 a 100.0 a 100.0 a Pradera (V) 100.0 a 100.0 a 60.0 c 100.0 a 3.0 d 1.0 e Cajamarca1 (T) 100.0 a 100.0 a 100.0 a 100.0 a 13.3 c 8.9 de Cajamarca2 (T) 100.0 a 100.0 a 100.0 a 100.0 a 15.2 c 5.1 de C.Viboral1 (A) 83.5 b 95.1 c 78.6 b 86.4 c 12.6 bc 22.3 bc C.Viboral2 (A) 83.2 b 95.0 c 25.7 d 26.7 d 5.5 cd 9.9 cd C.Viboral3 (A) 83.0 b 92.0 c 64.0 c 82.0 c 10.0 c 24.0 b El Cerrito (V) 56.0 c 97.0 b 85.0 b 95.0 b 29.0 b 7.0 de Ubaque (C) ---100.0 a 100.0 a 100.0 a 14.4 c 11.2 bcd a Cahill, M., K. Gorman, S. Day & I. Denholm. 1996. Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae). Bull. Entol. Res. 86: 343-349; b Departments: (A)= Antioquia, (C)=Cundinamarca, (T)= Tolima,(V)= Valle de Cauca; c Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. In Ecuador, T. palmi showed a high incidence in the province of El Carchi. The levels of resistance evaluated with six products show the development of resistance to organophosphates, carbamate, and pyrethroid insecticides (Table 101). As in Colombia, resistance to products that had been recently introduced into the market were found to be high (imidacloprid and fipronil). 187 Table 101. Response (percentage mortality) of Thrips palmi adults to six insecticides in Ecuador. Diagnostic dosages in ppm, tested using the Cahill et al. (1996)a technique. imidacloprid spinosad fipronil carbosulfan cypermethrin methamidophos (1000 ppm) (2000 ppm) (100 ppm) (1000 ppm) (16 ppm) (16 ppm) CIAT 100.0 ab 100.0 a 100.0 a 100.0 a 100.0 a 100.0 a Chagualyacu 71.1 b 100.0 a 89.9 a 63.6 b 6.3 d 38.4 c Carpuela 67.3 b 100.0 a 63.3 b 41.8 c 37.8 b 61.2 b San Rafael 66.0 b 100.0 a 72.4 b 72.4 b 23.5 c 45.9 c San Vicente 55.6 c 100.0 a 39.4 c 69.7 b 16.2 c 20.2 d a Cahill, M., K. Gorman, S. Day & I. Denholm. 1996. Baseline determination and detection of resistance to imidacloprid in Bemisia tabaci (Homoptera: Aleyrodidae). Bull. Entol. Res. 86: 343-349; b Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. Place Leafminers (Liriomyza huidobrensis ) This insect is the cause of numerous applications in regions located above 1400 masl. Although it has very good biological control, the great amount of insecticides used in the zone affects populations of beneficial insects. This may be one of the reasons for periodic outbreaks of this pest in dry and snap bean crops. The continuous use of carbamates, pyrethroids and abamectines that in previous years were efficient to control this pest, has led to the development of intermediate resistance levels to these insecticides (Table 102). Table 102. Response (percentage mortality) of Liriomyza huidobrensis adults and larvae to five insecticides in Colombia and Ecuador. Diagnostic dosages in µg vial-1 were tested using insecticide-coated glass vialsa, Diagnostic dosages in ppm, tested using the Ferguson (2004) techniqueb. Place (Country)c Concepción (E) CIAT C.Viboral (C) San Vicente (C) Pradera (C) La Cumbre (C) cypermethrin (2000 µg/vial) 99.0 ad 98.9 a 97.8 a 95.8 a 58.7 b --- Adults a chlorpyrifos (1000 µg/vial) 97.9 a 98.9 a 96.8 a 100.0 a 85.9 a --- methomyl (3000 µg/vial) 57.7 bc 95.7 a 90.3 a 74.7 b 45.7 c --- Larvae b abamectin cyromazine (1000 ppm) (50 ppm) 100.0 a 85.9 a 96.3 ab 97.9 a ----100.0 a 100.0 a 67.9 c 98.5 a 73.5 bc 100.0 a a Mason, G. A., Johnson, M. W., Tabashnik, B. E. 1987. Susceptibility of Liriomyza sativae and L. trifolii (Diptera: Agromyzidae) to permethrin and fenvalerate. J. Econ. Entomol. 80 (6):1262-1266; bFerguson, J. S. 2004. Development and stability of insecticide resistance in the leafminer Liriomyza trifolii (Diptera: Agromyzidae) to cyromazine, abamectin, and spinosad. J. Econ. Entomol. 97 (1):112-119; c Country (C) = Colombia, (E) = Ecuador ; d Means within a column followed by the same letter are not significantly different at the P = 0.05 using LSD test. 2.6.2 Continued monitoring of resistance to insecticides in Bemisia tabaci, on pepper hosts in Valle de Cauca Rationale: Bemisia attacks multiple hosts, and the insect readily passes from one crop to the next. Mismanagement of the insect on one crop may lead to resistance to insecticides and thus increase problems on other crops. Knowledge of the behavior of Bemisia occurring on other crops is therefore relevant for beans and the potential for bean production. A study of the zones affected by Bemisia tabaci attacking hot and sweet pepper crops is one of our main responsibilities within the project “Development of a management system of Bemisia tabaci in hot pepper and sweet pepper in Valle de Cauca”, financed 188 by the Ministry of Agriculture and Rural Development (MADR). Materials and methods: Changes in the composition of white fly species and levels of resistance to insecticides in B. tabaci populations were monitored throughout the target area. The target area included the municipalities of La Unión, Roldanillo and Bolívar in the northern region of Valle de Cauca; La Cumbre, Yotoco, Trujillo and Restrepo in the central region; El Cerrito, Palmira and Pradera in the southern region (Figure 54). All these rural municipalities were selected because of their agricultural importance in terms of hot and sweet pepper production. Resistance levels to insecticides were measured on adult populations using the vial techniques adopted by the project. One carbamate (methomyl), one organophosphate (methamidophos), one pyrethroid (cypermethrine), one neonicotinoid (imidacloprid) and one nereistoxine (thiocyclam hydrogen oxalate) were chosen as most representative of the insecticides used by local farmers. To test nymphal populations, the growth regulators buprofezin and diafenthiuron and the neonicotinoid imidacloprid were used. CL50 and CL90 values were calculated for the novel insecticides pyriproxyfen and spiromesifen (Table 103). With this information, diagnostic dosages for these two products were calculated. La Unión El Dovio Roldanillo Bolívar Trujillo Darién Buga Yotoco Restrepo Vijes La Cumbre El Cerrito Yumbo Palmira Pradera Figure 54. Target areas sampled and surveyed within the whitefly management project. 189 Table 103. Toxicological responses of laboratory races of Trialeurodes vaporariorum and Bemisia tabaci B biotype nymphs to two insecticides. Tests with spiromesifen and pyriproxyfen were conducted using the methodology suggested by Prabhaker et al. (1985)a. Cl50 Cl90 b + SEM (LC 95%) (LC 95%) 0.2 5.4 spiromesifen 1064 0.9 ± 0.07 (0.2 - 0.3) (3.6 – 9.1) T. vaporariorum 0. 5 4.5 pyriproxyfen 1593 1.3 ± 0.07 (0.4 - 0.6) (3.5 – 6.1) 0.05 1.5 0.9 ± 0.05 spiromesifen 1841 (0.04 - 0.07) (1.1 - 2.3) B. tabaci B biotype 1.1 10.7 pyriproxyfen 2788 1.3 ±0.05 (0.9 – 1.3) (8.9 – 13.1) a Prabhaker, N.; Coudriet, D.; Meyerdirk, D. 1985. Insecticide resistance in the sweetpotato whitefly Bemisia tabaci (Homoptera: Aleyrodidae). J. Econ. Entomol. 78: 748-752. Species Insecticide n χ2 3.3 3.4 1.7 4.9 Figure 55 shows some of the results obtained in 2008. It is evident that adults of the B. tabaci biotype have developed high levels of resistance to methamidophos and cypermetrine, intermediate resistance to imidacloprid and thiocyclam hydrogen oxalate. Figure 56 shows that nymph populations of this biotype are still susceptible to buprofezin and diafenthiuron. Our results also show that in some areas of the Valle de Cauca (Roldanillo, Yotoco) the B biotype is in the process of developing intermediate resistance levels. This situation has to be reconfirmed because resistance to novel insecticides like the neonicotinoids would make management of the whitefly extremely difficult. 190 100.0 a b a b b 80.0 60.0 methomyl (2.5 µg/v) 40.0 20.0 0.0 a 100.0 80.0 methamidophos (32 µg/v) 60.0 Corrected percentage mortality b 40.0 bc c 20.0 c 0.0 a 100.0 b 80.0 c c 60.0 cypermethrin (500 µg/v) 40.0 20.0 d 0.0 100.0 80.0 a b bc c d 60.0 imidacloprid (40 ppm) 40.0 20.0 0.0 a a 100.0 b 80.0 c c 60.0 thiocyclam hydrogen oxalate (1000 ppm) 40.0 20.0 0.0 Yotoco CIAT Rozo (Palmira) Morelia Pavas (Roldanillo) (La Cumbre) Races Figure 55. Toxicological response of adult populations of Bemisia tabaci B biotype to diagnostic dosages of five selected insecticides. Tests conducted in 2008. ¨CIAT¨ is the susceptible reference strain. Diagnostic dosages in µg vial-1 were tested using insecticide-coated glass vials. Diagnostic dosages in ppm, were tested using the Cahill et al. (1996) technique. 191 100 a a a a a 80 buprofezin (16 ppm) 60 40 20 Corrected percentage mortality 0 100 a a ab ab b 80 60 diafenthiuron (100 ppm) 40 20 0 a 100 80 b c b c imidacloprid (1000 ppm) 60 40 20 0 Morelia (Roldanillo) Yotoco CIAT Rozo (Palmira) Pavas (La Cumbre) Races Figure 56. 2.6.3 Toxicological response of Bemisia tabaci B biotype nymphs to diagnostic dosage of three selected insecticides in the Valle de Cauca in Colombia. ¨CIAT¨ is the susceptible reference strain. Diagnostic dosages in ppm tested using the Prabhaker et al. (1985) technique. Isolation and assessment of 2,4-diacetylphloroglucinol (DAPG/Phl)-producing fluorescent Pseudomonas strains for biological control of Pythium root rots Rationale: The genus Pseudomonas comprises the relatively large and important group of gram-negative, non-spore forming, motile rod bacteria. They produce a wide variety of antibiotics, which confer a competitive advantage and microbial fitness to survive in most environments (Haas and Keel, 2003; Paulsen et al., 2005). This genus also comprises beneficial bacteria such as plant growth promoters and biocontrol agents (Raaijmakers et al., 2002). Antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) is a polyketide compound with broad-spectrum antiviral, antifungal, antibacterial, and antitumor activity and phytotoxic properties (Raaijmaker et al., 2002; Haas and Keel, 2003). It is synthesized by several plantassociated fluorescent pseudomonads, and it plays a key role in the suppression of a wide variety of soil192 borne diseases (Haas and Defago, 2005; Ramette et al., 2006). 2,4-DAPG inhibits zoospores produced by Pythium spp. and also damages the membrane of this Oomycete (de Souza, 2003b). The aim of this study was to isolate and assess the in vitro antagonistic potential of fluorescent Pseudomonas towards Pythium spp causing root rot in common beans. Materials and Methods: Plant samples including maize (5), sorghum (2), pigeon peas (2) and beans (1) were collected from different sites (suppressive and non suppressive) around Kawanda Agricultural Research Institute. Isolation was done according to Mazzola et al., 2004. Briefly, the bean plants in their third growth phase (V3) were harvested, separated from the loose soil by gentle shaking and 0.5 g of the root system were excised from the plants and placed in 10 mL sterile distilled water. The roots and associated rhizosphere soil were vortexed for 60 s and serial dilutions of the resulting roots washed and plated on King’s medium B (KB) agar amended with ampicillin (100µg mL-1), chloramphenicol (13 µg mL-1) and cycloheximide (75 µg mL-1). Plates were incubated at 25oC for 48 h and colonies of fluorescent pseudomonads were differentiated from non-fluorescent colonies under UV light (wavelength 366 nm). Fluorescent pseudomonads were selected and transferred to fresh KB+ agar plates. A Gram stain was performed to examine to cell types. Inhibition assays were performed using the central disk method (McSpadden Gardener et al., 2005). Agar plugs (6mm) containing the fungal mycelium, taken from the growing edge of each culture were placed in the centre of fresh potato dextrose agar (PDA) plates and incubated at room temperature for 24 h. A loopfull of fluorescent inoculum were inoculated at three equidistant positions along the perimeter of the assay plate. Two plates, one with no bacteria inoculum and the other with bacteria alone were included as controls. The assay plates were incubated at 24oC. Each bacterial isolate was replicated in three plates entire assay was repeated three times. Fungal growth was assayed when the growth on the Pythium control plates extended the full radius of the plate. Two measurements were made: distance from the edge of the plug to the growing edge of the fungus (X), and the distance from the edge of the bacterial growth to the growing edge of the fungus (Y). Inhibition index (I) was calculated as: I=Y/(X+Y) (McSpadden et al., 2005). Results and discussion After 48hrs of incubation, cream bacterial colonies could be observed growing at different intensities from different dilutions of all the 10 root washes. On examination under UV light, 9 isolates had mixtures of fluorescent and non fluorescent colonies. Of the 9 fluorescent isolates, one (from bean) was Gram positive and was excluded from further investigation. Eight Gram negative rod-shaped isolates found were tested for antagonism against Pythium ultimum isolate on Potato Dextrose Agar (PDA) plates according to McSpadden et al., 2005. One isolate, P4 from pigeon peas, showed the highest antagonism with an inhibition index of 0.38 (Table 104, Figure 57). Table 104. Inhibition indices of eight fluorescent Pseudomonas isolates from Kawanda Isolate P1 P2 P3 P4 P7 P8 P9 P10 X (mm) 26.5 70 40 15 44 66 47.5 50.5 193 Y (mm) 8.5 -26 -25 9 0 -23.5 -24.5 -22 I 0.24 -0.59 -1.67 0.38 0.00 -0.55 -1.07 -0.77 Figure 57. Petri dishes showing: a) Pythium ultimum alone, b) antagonism of Pseudomonas inoculated at 3 equidistant positions and c) Pseudomonas alone Detection of 2,4-DAPG-producing fluorescent pseudomonads in Uganda has previously been conducted through extraction and PCR amplification of DNA from individual isolates (Kamenya, 2008). This detection is via amplification of phlD, which is a key gene in the biosynthesis of 2,4- DAPG. We have obtained these primers and will use it to detect the presence of the phlD gene in Pseudomonas isolate P4 and further screen more isolates for inclusion into this trial. We will continue to explore the potential of antagonistic organisms to suppress Pythium spp. through further collections and screening. Our target will be suppressive soils that many authors propose are a reservoir of natural, effective and valuable microbial antagonists, and the chance of selecting effective biocontrol strains might be improved if isolations are made from such soils. Contributors: Buruchara R., Buah S., Acam C. Collaborators: Edema R. (Makerere University) References de Souza, J.T., Arnould, C., Deulvot, C., Lemanceau, P., Gianinazzi-Pearson, V., Raaijmakers, J. M. 2003b. Effect of 2,4-diacetylphloroglucinol on Pythium: 187 cellular responses and variation in sensitivity among propagules and species. Phytopathology 93: 966–975. Haas, D., Défago, G. 2005. Biological control of soil-borne pathogens by fluorescent pseudomonads. Nature Reviews Microbiology 3: 307-319. Haas, D., Keel, C. 2003. Regulation of antibiotic production in root colonizing Pseudomonas spp. and relevance for biological control of plant disease. Annual Review of Phytopathology 41: 117–153. Kamenya, N, S. 2007. Detection, isolation and baseline characterization of Indigenous phlD+ fluorescent pseudomonas species in the control of bean root rot and coffee wilt disease in Uganda. Makerere University, Kampala,Uganda. Mazzola, M., Funnell, D. L., Raaijmakers, J. M. 2004. Wheat Cultivar-Specific Selection of 2,4Diacetylphloroglucinol-Producing Fluorescent Pseudomonas Species from Resident Soil Populations. Papers in Plant Pathology, available online at http://digitalcommons.unl.edu/plantpathpapers/37, University of Nebraska - Lincoln McSpadden Gardener, B. B., Gutierrez, L. J., Joshi, R., Edema, R., and Lutton, E. 2005. Distribution and biocontrol potential of phlD+ pseudomonads in corn and soybean fields. Phytopathology 95:715-724. Paulsen, I.T., Press, C.M., Ravel, J., Kobayashi, D.Y., Myers, G.S.A., Mavrodi, D.V., DeBoy, R.T., Seshadri, R., Ren, Q., Madupu, R., Dodson, R. J., Durkin, A. S., Brinkac, L.M., Daugherty, S.C., Sullivan, S.A., Rosovitz, M.J., Gwinn, M.L., Zhou, L., Schneider, D.J. Cartinhour, S.W., Nelson, W.C., Weidman, J., Watkins, K., Tran, K., Khouri, H., Pierson, E.A., Pierson,L.S., Thomashow, L.S., Loper, J.E. 2005. Complete genome sequence of the plant commensally Pseudomonas fluorescens Pf-5. Nature Biotechnology 23: 873-878. 194 Raaijmakers, J.M., Vlami, M., de Souza, J.T. 2002. Antibiotic production by bacterial biocontrol agents. Antonie van Leeuwenhoek 81: 537–547. Ramette, A., Moënne-Loccoz, Y., Defago, G. 2006. Genetic diversity and biocontrol potential of fluorescent pseudomonads producing phloroglucinols and hydrogen cyanide from Swiss soils naturally suppressive or conducive to Thielaviopsis basicola-mediated black root rot of tobacco. FEMS Microbiol. Ecol. 55: 369-381. Weller, D.M. 1988. Biological control of soil-borne plant pathogens in the rhizosphere with bacteria. Annual Review of Phytopathology 26: 379-407. 2.6.4 Microsatellite analysis of common bean mixtures from SW Uganda Rationale: Despite most consumers’ preference for pure varieties in Uganda, small scale farmers in Southwester Uganda grow bean varietal mixtures. There are many reasons for growing varietal mixtures, but prominent among this is being risk averse and ensuring yield stability in the face of a variety of production constraints including Pythium root rots, one of the major diseases in the area. We previously reported farmer’s management strategies for bean root rots using the genetic diversity, morpho-agronomic and characterization and pathological reaction of components of bean mixtures against Pythium ultimum. The objective of this part of the study was to analyze the amount of genetic diversity associated with bean mixtures in Southwestern Uganda using Simple Sequence Repeats (SSR) markers. We expect that this information will be useful in developing strategies to manage, use and conserve the diversity in the region. Materials and Methods: Seed materials used: Representative seed samples (20%) of the different morpho-agronomically categories of the diversity of the bean mixtures (Franco et al., 2005) were used. Twenty seeds of each selected variety were planted in the field in a completely randomized block design with 2 replications. Plants in a population were tagged and sampled for DNA extraction. All tagged plants were harvested singly into paper bags and labeled accordingly. Microsatellite DNA analysis: DNA was extracted from 2-week-old seedlings of the test materials. Twenty-three microsatellite primer pairs developed by various authors (Lioi et al., 2005; Yu et al., 2000; Gaitan-Solis et al., 2002; de Campos et al., 2007) were screened. As a result six pairs (Table 105) were chosen for full screening across all genotypes including CAL 96 and RWR 719 controls. The choice was based on clear polymorphism and stability of amplification. Markers were amplified with a hot start of 94oC for 5 min; followed by 10 cycles of a touchdown of decreasing annealing temperature by 1oC/cycle until the optimal annealing temperature was reached; then 30 cycles of 94oC for 30 sec, XoC for 45 sec and 72oC for 45 sec. (X was dependent on primer combination); followed by a final extension at 72oC for 7 min and the reaction was held at 4oC. The PCR reaction was carried out in a final volume of 20µl containing 10ng of genomic DNA, 0.1 µM of each of the forward and reverse primers, 10 mM of Tris-HCl (pH 7.2), 50 mM of KCl, 1.5 to 2.5 mM of MgCl2, depending on the primer combination, 250 mM of total dNTP and 1 unit of Taq polymerase. The products were checked for amplification in 2% agarose gels and stained with ethidium bromide. Proper separation of both phaseolin and microsatellite alleles were achieved on 4% denaturing polyacrylamide gels. The gel was run in a vertical position at a constant voltage of 200V for 2.5 hours and bands were visualized by a silver staining procedure. 195 Table 105. Accession no. SSR primers selected for bean diversity study. J04555 Size (bp) 152 M75856 157 U18791 239 X80051 192 GATS91 229 BM143 143 Forward primer Reverse primer GAGGGTGTTTCACTAT TGTCACTGC CAATCCTCTCTCTCTC ATTTCCAATC GGGAGGGTAGGGAAG CAGTG AGTTAAATTATACGA GGTTAGCCTAAATC GAGTGCGGAAGCGAG TAGAG GGGAAATGAACAGAG GAAA TTCATGGATGGTGGA GGAACAG GACCTTGAAGTCGGT GTCGTTT GCGAACCACGTTCAT GAATGA CATTCCCTTCACACAT TCACCG TCCGTGTTCCTCTGTC TGTG ATGTTGGGAACTTTTA GTGTG Ta (oC) 51 Reference 51 Yu et al., 2000 49 Yu et al., 2000 49 Yu et al., 2000 47 Gaitan-Solis et al., 2002 42 Gaitan-Solis et al., 2002 Yu et al., 2000 Phaseolin analysis: Phaseolin, the major seed storage protein of common bean, was analyzed using primers developed by Kami et al., (1995) that differentiate S phaseolin gene family from the T phaseolin types. The S phaseolin genomic DNA, that is characteristic of the Mesoamerican gene pool, yields two homoduplex PCR products whereas T phaseolin DNA produce three homoduplex amplification bands that is characteristic of the Andean gene pool. Both phaseolin types produce identical smallest amplification products (Figure 58, g). The primer pair Phas1 (5’-AGCATATTCTAGAGGCCTCC-3’) and Phas2 (5’-GCTCAGTTCCTCAATCTGTTC-3’ was used under optimal conditions of 2mMMgCl2, 200µM dNTPs, 0.2µM primers, and 40 cycles of 94oC for 30 s, 55oC for 30 s, and 72oC, for 45 s, with 10ng of DNA. Each clearly separated SSR amplification band was considered as an allele and coded for presence (1) or absence (0). The amount of genetic diversity was quantified using various indices implemented in the program ARLEQUIN version 3.11 (Schneider et al., 2007). The relationships between selected varieties and gene pool groups were derived from calculated genetic distances and dendrograms constructed with the neighbor joining method using NTSYS computer software. Results and Discussion: A total of 39 alleles were obtained from the 5 loci amplified across all genotypes with an average of 7.8 alleles per locus ranging from 2 alleles for X80051 to 14 alleles for U18791 (Figures. 58, a-f). This observed average number of alleles is comparable to those obtained in other studies such as the average of 7.7 alleles reported by Maras et al., 2006. This finding suggests that a significant number of germplasm accessions have to be preserved in order to retain the original diversity. A total of 38 haplotypes were observed in the first 102 genotypes showing 36 polymorphic sites over the 39 alleles (94.7% polymorphism) and an average gene/allele diversity of 29.3% over the loci. Haplotype real differences were checked before tests (Ewens et al., 1972) and an observed and expected F values of 0.068 and 0.059 were obtained (p = 0.93). The microsatellite analysis uncovered two major groups (Figure 59) corresponding to Andean and Mesoamerican gene pools based on the position of the control genotypes CAL 96 and RWR 719 in a cluster analysis performed using the Neighbor Joining method according to Nei (1978). There was a significant difference between the haplotypes of the two groups (F = 0.11, p = 0.000). However members of the Andean gene pool were much fewer than those of the Mesoamerican types. What was more remarkable was that, despite the few numbers, the Andean group was more diverse in terms of both haplotype diversity and percentage polymorphism compared to their Mesoamerican counterparts. This finding is in congruence with those of Santalla et al. (2004) who established that the Andean gene pool in Argentina has a large genetic base on the basis of morphological and adaptive variability as well as biochemical (allozymes and phaseolin) analysis. This high diversity in a susceptible group could be a result of the presence of infrequent alleles in the Andean group. 196 a) Marker BM143 b) Marker GATS 91 c) Marker J04555 d) Marker M75865 e) Marker U18971 f) Marker X80051 Andean Mesoamerican g) Phaseolin PCR Figure 58. Microsatellite marker profiles on silver-stained polyacrylamide gels 197 Mesoamerican Andean 14 19 61 256 77 57 10 86 12 100 97 67 66 26 44 68 8 70 76 30 14 92 33 64 90 71 843 60 15 17 46 102 35 43 85 47 53 72 54 73 79 91 31 39 9 87 22 23 96 88 507 27 80 37 89 2 95 65 18 29 515 the four growth habit types were present in both Andean and Mesoamerican groups although All 98 62 20 growth type IV and III were more prevalent (54.3% and 35.9% respectively) in the 28 36 42 75 Mesoamerican group while there was a more even distribution of growth types in the Andean 81 11 38 41 group. 94 74 58 82 13 21 24 40 The phaseolin seed protein PCR analysis revealed the presence of the two phaseolin types i.e., 56 59 99 the S and T phaseolin types. The former is found among cultivars of Mesoamerican origin 69 78 16 45 whereas the latter is found among cultivars descended from the Andean origin (Kami et al., 83 34 49 63 1995). Phaseolin analysis generally agreed with microsatellite groups providing further strength 32 93 55 48 for the structure. The Andean group comprised of 55.6% genotypes which possessed the T-type 101 52 phaseolin characteristic of7.25Andean races 2.00 12.50 Coefficient Figure 59. Microsatellite cluster analysis at 5 loci using NJ method. 198 17.75 23.00 Fewer genotypes were found in the Andean group than in the Mesoamerican group. The Andean clade was composed of 37 genotypes made of 16 haplotypes (43.2%). The observed and expected F values for this group were 0.12 and 0.10 respectively (p = 0.78). 71.8% of the loci were polymorphic for this group with a 9.62 mean number of pairwise differences. The Mesoamerican group comprised of 65 genotypes and 22 haplotypes accounting for 33.8% haplotype frequency, almost 10 times less than the Andean haplotype frequency. Percentage polymorphism over loci was observed at 69.2% with a mean pairwise difference of 7.27. The observed and expected F values were the same as those of the Andean genotypes (p = 0.95). The dominant seed shape in the Andeans was elongate flat (69.4%) most of which presented primary coat colors of red (36.1%) and cream (25%). 58.3% of this group was monochromatic leaving only 15 genotypes which had secondary colors, 8 of which were cream, 5 purple and 2 brown. These secondary colors were mainly patterned as speckles or zebra stripes across the seeds. Mesoamerican genotypes, on the other hand, were principally comprised of round flat seeds (50%) bearing cream primary colors (54.7%). 67.2% of Mesoamerican seeds were monochromatic and the dominant secondary color occurring on 52.3% of the seeds in this group was black that were mainly mottled in pattern. Urban markets mainly prefer large red mottled-to-maroon colored seeds most of which probably belong to the Andean gene pool. Hence farmers’ sources of these seeds could have been markets or NGOs. Additionally, the patterns of secondary colors on seeds were mainly speckles or Zebra stripes for the Andean group and mottled in Mesoamericans. Diversity in phaseolin types has been very useful for the classification of beans into Andean and Mesoamerican gene pools since most of the cultivars from one center of domestication possess a certain set of phaseolin type which is not found in cultivars from the other center of domestication (Duran et al., 2005). However, in this study the Andean group comprised of only 55.6% genotypes which possessed the T-type phaseolin characteristic of Andean races. Meanwhile 75% of the Mesoamerican genotypes had Type S phaseolin which is mainly used to designate this group. Average yield per plant was slightly higher in the Mesoamerican group. In terms of pathogenicity, the overall average disease score for Pythium in the Andean group was 5.7 (standard deviation = 1.94) compared to 4.7 (standard deviation = 1.71) for the Mesoamerican group. This is consistent with previous findings that the genotypes of Mesoamerican origin are more resistant to the root rot disease (Buruchara, 2003). Since large-seeded Andeans have more market potential, we recommend crossing between the two gene pools to transfer original resistance genes from Mesoamerican genotypes into the Andeans with a major focus on those genotypes that have already been naturally introgressed. Contributors: R. Buruchara, S. Buah, P. Okori (Makerere University) Collaborators: Matthew Blair, Paul Gepts 199 References Buruchara, R.A. 2003. Integrated management strategies for bean root rots in Africa. Centro Internacional de Agricultura Tropical (CIAT), Kampala, UG. 2 p. (Highlights: CIAT in Africa no. 2). Chiorato, A.F., Carbonell, S.A., Benchimol, L.L., Chiavegato, M.B., Dias, L.A, Colombo, C.A. 2007. Genetic diversity on common bean accessions evaluated by means of morpho-agronomical and RAPD data. Sci. Agric., Piracicaba, Brazil; 64:256-262. de Campos, T., Benchimol, L.B., Carbonell, S.A.M., Chioratto, A.F., Formighieri, E.F., and de Souza A.P. 2007. Microsatellites for genetic studies and breeding programs in common bean, Pesq. agropec. bras., Brasília, 42: 589-592. Duran, L. A., Blair, M. W., Giraldo, M. C., Macchiavelli, R., Prophete, E., Nin, J. C., and Beaver, J. S. (2005). Morphological and Molecular Characterization of Common Bean Landraces and Cultivars from the Caribbean. Crop Science 45: 1320-1328 FAO, 2007. Food and Agriculture Organization of the United Nations. Faostat agriculture data. Website: http://faostat.fao.org/default.aspx , Accessed 02 Nov, 2007. Franco, J., Crossa, J., Taba, S., and Sanad, H. (2005). A sampling strategy for conserving genetic diversity when forming core subsets. Crop Science 45: 1035-1044. Gaitan-Solis, E., Duque, M. C., Edwards, K. J and Tohme, J. (2002). Microsatellite Repeats in Common Bean (Phaseolus vulgaris): Isolation, Characterization, and Cross-Species Amplification in Phaseolus ssp. Crop Science 42:2128-2136. Kami, J., Velasquez, V.B., Debouck, D.G., and Gepts, P. 1995. Identification of presumed ancestral DNA sequences of phaseolin in Phaseolus vulgaris. Proc. Natl. Acad. Sci., USA. 92: 1101-1104. Lioi, L., Piergiovanni, A. R., Pignone, D., Puglisi, S., Santantonio, M. and Sonnante G. (2005). Genetic diversity of some surviving on-farm Italian common bean (Phaseolus vulgaris L.) landraces. Plant Breeding 124: 576-581. Schneider S, Roessli D, Excoffier L (2007) ARLEQUIN: a Software for Population Genetics Data Analysis, Version 2.000. University of Geneva, Geneva, Switzerland. Santalla, M., Menendez-Sevillano, M.C., Monteagudo, A.B., and De Ron, A.M. 2004. Genetic diversity of Argentinean common bean and its evolution during domestication. Euphytica 135: 75-87. Yu, K., Park, S. J., Poysa, V., and Gepts, P. (2000b). Integration of simple sequence repeat (SSR) markers into a molecular linkage map of common bean (Phaseolus vulgaris L.). J. Hered. 91: 429-434. Zhang, X., Blair, M.W., and Wang, S. 2008. Genetic diversity of Chinese common bean (Phaseolus vulgaris L.) landraces assessed with simple sequence repeat markers. Theor Appl. Genet, 117:629-640. 200 Product 3: Beans that respond to market opportunities Activity 3.1 Development of large white beans for international markets Highlights: • Crosses have been developed to combine alubia grain type (uniform, milky white, long cylindrical seed) with drought resistance. Rationale: International bean markets in some cases demand the same grain types as local and regional markets. For example, the small red type is widely accepted in Central America and in East Africa, and also finds a modest market in Pakistan and in the United States. Other types are cultivated in East Africa primarily for export, with a relatively minor component of local consumption, such as navy beans. Although productivity is relevant in such market types, meeting stringent market criteria to obtain the best prices often takes priority in farmers’ choices of which varieties to plant. One of the highest value beans is the large white class, which in several parts of the world demands very high prices. In Spain for example, the very large fabada type sells for as much as US$10 per kilo. Alubia beans are one of the highest value types on the international market, and can sell for almost double the price of small black beans. The alubia type presents seed size and long cylindrical shape more typical of Andean beans, and with milky white grain. Argentina has been one of the largest producers of alubia beans, but East Africa has the opportunity to enter this market for sale to the mid-eastern countries. Some production of large white types already occurs in the region, but there is potential for much more. Materials and Methods: In section 2.1.1.3 we reported on the development of large white beans for drought resistance. This is a particularly valuable trait for white beans, since production in dry areas is conducive to avoiding grain spotting and maintaining grain quality, but implies risk of drought. While the lines reported in the above section represent gains in agronomic traits, grain size and shape are still not adequate for the most demanding markets. It has been a particular challenge to obtain large, long cylindrical grain type with the best drought resistance, in spite of the fact that our drought resistant check, ICA Quimbaya (AFR 298) has excellent grain size and shape. In 2008 F2 populations developed within the drought breeding project were evaluated under intermittent drought. These included populations with alubia parents, and that were segregating excellent white color (milky, uniform, without veins or hilum discoloration). Out of F2 families that expressed relatively good productivity under intermittent drought we selected the seed with the best grain color and to the extent possible, acceptable alubia size and shape, although grain type still fell short in this regard. These were crossed to ICA Quimbaya to recover the grain characteristics of Quimbaya with the alubia color. This represents a divergence from the standard drought breeding approach, in that it places primary importance on the grain characteristics. Additional crosses were planned to combine the sources of grain color with the best drought sources, as a back up plan to the crosses with Quimbaya. Crosses are presented in Table 106. Results and Discussion: F1 and F2 seed was obtained for all combinations. Simultaneously, individual F3 plant selections were taken within the lines that served as sources of alubia type to seek improved types in available families. All materials will be screened in the drought nursery in July, 2009 Collaborators: S. Beebe, M. Grajales 201 Table 106. Crosses to combine alubia grain color with improved grain size and shape and/or drought resistance. Sources of grain form and/or drought resistance SAB SAB SAB SAB Quimbaya 618 630 650 686 Sources of commercial white color Entry (G 7930 x SAB 621)F1 x SAB 686/-MC 155 (G 7930 x SAB 621)F1 x ICA QUIMBAYA/-MC 159 (G 7930 x SAB 621)F1 x ICA QUIMBAYA/-MC 160 (G 7930 x SAB 621)F1 x ICA QUIMBAYA/-MC 162 SAB 676 x(G 7930 x SAB 634)F1/-MC 164 X X (G 7930 x SAB 634)F1 x ICA QUIMBAYA/-MC 170 X X ICA QUIMBAYA x (SAB 620 x SAB 634)F1/-MC 226 X X Activity 3.2 X X X X X X X X X Breeding Navy and Large White bean varieties with multiple stress resistance in eastern Africa Highlights: • Fourteen new navy and large white bean varieties combining multiple resistance to diseases and abiotic stress factors, high yield potential and marketable grain characteristics released for smallholder production in four countries in eastern Africa Introduction: Smallholder farmers in East and Central Africa predominantly grow small white bean (also known as white pea or navy bean) for export and local canning industries. Navy beans are grown on an estimated 310,000 ha per year and account for 9.6% of total bean production in Africa. They are particularly important in Ethiopia and Sudan. Ethiopia is the leading producer of navy bean in East and Central Africa. Farmers in the Rift Valley region of Ethiopia export nearly 90% of their navy bean. Navy bean is also important in Nile Valley of Sudan, southwestern Uganda, northeastern Tanzania, Kenya, Madagascar, Cameroon and the Great lakes region where it is a component of the mixtures (Wortmann et al., 1998). They are in high demand for the canning industries of Kenya, South Africa and Zimbabwe and in urban areas where they are popular because of their taste, short cooking time and low levels of flatulence. However, yields of navy bean under low input systems common in eastern Africa are low, typically 400 to 700 kg ha-1 due to susceptibility to diseases such as rust, common bacterial blight, anthracnose, angular leaf spot, root rots and edaphic stresses such low soil nitrogen and phosphorus, and drought. Large white bean is an important commodity for domestic and export markets of Madagascar and Sudan. Large white is gaining popularity in Ethiopia. Large whites are mostly exported from Madagascar (CTHA and Masumin exporters), Sudan, Ethiopia, Rwanda and D. R. Congo. Uganda is evaluating two varieties for Saudi Arabian markets. They tend to be susceptible to rust, common bacterial blight and angular leaf spot. Most of the commercial varieties are susceptible to diseases. There is need to identify desired varieties for new and emerging markets. There is indication that the Spanish types are more 202 popular in the export markets. Development of improved cultivars of navy and large white beans has been one of the priorities of the market-led breeding program of the East and Central Africa Bean Research Network (ECABREN). A regional breeding program was started in 2001 to develop improved, marketable navy and large white bean cultivars with resistance and/or tolerance to two or more priority constraints. Selection for improved lines was conducted collaboratively at Melkassa (Ethiopia), FOFIFA in Madagascar, and University of Nairobi in Kenya from existing and new breeding populations. The navy bean program is led by Ethiopian National Bean program (EIAR), large white program by FOFIFA bean program in Madagascar with back up programs at the University of Nairobi, Kenya and CIAT, Colombia. This report highlights some of the main achievements of this collaborative research for development program. Materials and Methods: A working collection was made from segregating populations, advanced lines, commercial cultivars, breeding populations, and constraint nurseries held by CIAT programs in Uganda, Colombia, Kenya and the Ethiopian national program. Parental lines from this collection were used for crossing programs at Kabete (Kenya), Melkassa Agricultural Research Centre, Nazareth (Ethiopia) and FOFIFA (Madagascar). One hundred thirty three pollinations were made at Kabete to transfer anthracnose and rust resistance from Roba-1 to Mexican 142. Mexican 142 is probably the oldest canning variety in the region and one of the few varieties which adequately meets the stringent canning requirements. It is however very susceptible to rust and anthracnose. It has type III growth habit. Farmers prefer type I growth habit because of ease of harvesting and threshing. We also made 97 pollinations to combine the high canning quality, tolerance to rust, bean stem maggot resistance and type 1 growth habit of Awash 1 with high yield potential and resistance to anthracnose of Goberasha. The F1 was backcrossed to Mexican 142 to generate segregating populations. Navy bean grain type was selected from 52 F2 segregating populations at Kabete Field Station, and used to establish F3 progeny rows. Progeny rows were rated for disease reaction, plant type and tolerance to moisture stress at Kabete. F4 bulks were evaluated for tolerance to low soil P at Kakamega and root rots in Sabatia test sites. The F4 bulks, F5 and F6 lines were grown in preliminary and intermediate yield trials at three locations. Thirty-eight new lines were finally selected for regional distribution and advanced yield trials. In Melkassa, crosses were made to generate new breeding populations. The adapted parents in the new crosses were PAN 182, Dresden, Mex 142 and Awash 1. Donor parents for anthracnose were Roba and Goberasha. PAN 182 and Awash contributed resistance to rust. HAL–5 was used a source of resistance to halo blight. A262, A197 and TY 3396-3 were included in these crosses due to their high yield potential. Awash also contributed type 1 growth habit. Dresden has excellent canning quality but is susceptible to anthracnose, CBB, rust and angular leaf spot. The first single crosses were made in 2000 main season. Development of three way, double and backcrosses started during the 2001off-season. Included in these crosses was ‘Omar’, a medium sized white bean that was identified by exporters for its quality. It is however susceptible to rust and other diseases. In Ethiopia, advanced lines were evaluated in multi-location trials. In Madagascar, selection was conducted from 19 advanced lines from the regional program at Kabete, and from new populations generated from crosses between Ranjonoby (a commercial large white variety) and sources of resistance to angular leaf spot, anthracnose , rust, common bacterial blight and tolerance to low soil fertility (Ikinimba). Ikinimba has shown good levels of resistance to rust, angular leaf spot, anthracnose and low soil fertility under Madagascar conditions. Awash was used as source of resistance to rust and anthracnose. XAN 74 was source of resistance to common bacterial blight. F1 were backcrossed to Ranjonoby for three generations. 203 Results and Discussion: Ten high yielding small white lines were selected from the preliminary and intermediate trials in Kenya. Some of their characteristics are shown in Table 107. Mean yields across sites were above 2 t ha-1. DB 190-84-1, ECAB 0601, ECAB 0628, UBR(92)17-1 and UBR(92)25-27 succumbed to rust at Thika and were discarded. All other lines showed resistant or intermediate reactions (1 to 9 on CIAT rust scale). ARA 8-4-1 and BL 207562 were susceptible to black root. All lines showed resistance to angular leaf spot and anthracnose. Selected F8 lines were evaluated at Kabete and Thika over three seasons. Table 107. Days to flowering, maturity, seed mass and grain yield of best 10 navy F6 bean lines selected from segregating populations and other nurseries in eastern Africa. Genotype BRB 148-1 ECAB 00621 BL 207562-1 ECAB 00622 ECAB 00612 ECAB 00605 ECAB 00624 BRB 45-1 ECAB 00623 ECAB 00625 Trial mean Genotypes (G) Locations (L) G XL LSD 0.05 CV(%) Days to 50 % flower 45.5 54.5 46.9 50.7 50.5 53.3 52.7 47.8 55.0 49.8 49.9 ** ** ** 2.6 4.5 Days to 75% maturity 100-Seed mass (g) 89 100 90 96 94.7 95 97 91 99 95 94.7 ** NS ** 2.9 2.7 23.7 28.4 27.0 24.6 18.4 23.5 24.8 24.0 21.7 26.8 24.5 ** ** ** 1.9 7.0 Yield (kg ha-1) PYTa (3 sites) 3129 2690 2861 2210 2367 2936 2652 2713 2528 2777 2319 ** ** ** 423.2 18.2 IYTb (2 sites) 3002 2299 2084 2559 2348 1771 1972 1838 2015 1760 1911 * NS * 621 32.5 Mean 3066 2495 2473 2385 2358 2354 2312 2276 2272 2269 2115 *,** : Significant at 5 and 1% probability levels, respectively; NS= not significant. PYTa = preliminary yield trial of F5 ; IYTb= intermediate yield trial of F6 and other advanced lines. Thirty-eight advanced lines were evaluated in multi-location trials in Kenya and compared with regional check varieties. Results showed significant genotypic differences for days to flower, maturity, rust, pod load, 100-seed mass and grain yield (Table 108). Effects of environment were significant for all traits (P>0.01). A significant genotype x environment interaction was detected for phenology, rust, pods m-2, 100-seed mass and grain yield. Rust was most severe at Kabete. Sarrag was rated very susceptible in three of the four environments with scores of 9, 9 and 8.3, respectively (CIAT scale). HRS was rated susceptible at Kabete with a score of 7. Mexican 142 showed intermediate score of 3.7 at Kabete and 4.3 at Thika. Awash 1 showed intermediate score of 3.6 for the two seasons at Kabete. ECAB 0614, ECAB 0617 and ECAB 0619 had intermediate rust scores at Kabete. ECAB 0602 also had an intermediate rust score of 3.7 at Juja. All other test lines showed resistant reactions to rust. Root rot was most severe at Kabete. ECAB 0602, ECAB0631 and ECAB0632 had scores of 5, 4.3 and 3.7, respectively at this site. All other lines and checks showed resistant reactions. Mean grain yield was lowest at Kabete during the long rain season and highest at the same site during the short rain season (Table 108). This was partly attributed to effects of rust and moisture stress at Kabete. Results showed that 37 new lines produced more grain than the best check (Mexican 142) (Table 108). ECAB0601 was the best line among the new lines with a high yield advantage over the check varieties. The results indicated that new lines have 204 considerable yield potential compared to the commercial cultivars. However, these lines need to be evaluated for canning quality. In Ethiopia, advanced lines were evaluated in multi-location trials at Alemaya, Awassa, Bako, Jimma, Melkassa, Ziway, Ambo, Mekele, Asassa, Adet, Pawe and Sirinka. Four lines promising were identified. These were UTT 24-131, UTT 27-24, NZBR 2-5, BZBR 2-8 (Teshale et al., 2007). Several lines were selected at Melkassa and Alemaya for the national yield trials (NYT) and farmer verification trials (FVT) under the supervision of the National Variety Releasing Committee (NVRC). Similar evaluations were conducted in Madagascar and Sudan. Release of New Varieties: Table 109 shows some of the new navy and large white varieties released or pre-released in eastern Africa between 2003 and 2008. RJ 1, RI 5-1, RI 5-5, RI 5-3, IL 5-53 are large seeded. They were selected from populations developed in Madagascar. Ibarya and Mutwakil have medium sized seeds (29 to 37 g/100 seed) but considered as large seeded in Sudan. Most of the new releases have resistance to two or more biotic and abiotic stresses. CAB 19 is a small white climbing bean released in Tanzania but originated from Rwanda. Contributors: Paul Kimani, Teshale Assefa (Ethiopia), Amee Rakoriahanta (Madagascar), Bulti Tesso and Chemeda Fininsa (Alemaya University, Ethiopia), Festo Ngulu (Tanzania) and Ghamal Khalifa (Sudan) Collaborators: Hery Andrimazaolo ( Madagascar) and Bean Teams at Kabete (Kenya), Melkassa (Ethiopia), FOFIFA (Madagascar), Alemaya ( Ethiopia) and Hudeiba (Sudan) References Teshale Assefa, H. Assefa and P.M. Kimani. 2006. Development of improved haricot bean germplasm for mid- and low altitude sub-humid ecologies of Ethiopia, pages 87-94. In: Food and Forage Legume of Ethiopia: Progress and Prospects. ICARDA, Allepo, Syria. Wortmann, C.S, R.A. Kirkby, C. A. Eledu and D. J. Allan. 1998. Bean Atlas. CIAT, Colombia. 205 Table 108. Mean duration to flowering, maturity, pods m-2, 100-seed mass and grain yield of selected F8 navy bean lines grown at four environments in Kenya Genotype ECAB0601 ECAB0627 ECAB0612 ECAB0629 ECAB0608 ECAB0614 ECAB0603 ECAB0617 ECAB0604 ECAB0621 ECAB0615 ECAB0607 ECAB0611 ECAB0620 ECAB0616 ECAB0610 ECAB0630 ECAB0613 ECAB0619 ECAB0623 ECAB0606 ECAB0632 ECAB0622 ECAB0609 ECAB0628 ECAB0626 ECAB0624 ECAB0634 ECAB0637 ECAB0633 ECAB0635 ECAB0618 ECAB0605 ECAB0636 HRS 545 ECAB0602 MEXICAN 142 BASSABEER ECAB0625 ECAB0631 AWASH 1 SARRAG Environmental mean C.V.(%) Days to 50% flower (d) Days to maturity (d) Pod m-2 100seed mass (g) 47.0 46.2 46.9 46.4 45.9 45.8 46.2 47.8 46.9 48.5 47.4 46.9 45.6 47.1 44.7 47.2 44.7 47.3 47.6 47.9 47.5 45.9 47.8 46.7 44.2 46.1 37.8 44.8 45.0 44.2 34.6 46.2 38.0 44.4 43.3 45.2 43.3 43.2 46.6 46.0 41.5 44.1 46.0 89.5 87.3 87.4 88.7 86.9 87.8 87.8 88.9 88.3 88.5 89.2 87.9 88.1 88.5 85.7 88.2 85.8 87.4 88.3 89.7 88.0 86.7 88.8 86.6 85.4 87.1 88.3 85.7 86.8 86.3 86.0 87.7 88.3 85.6 83.4 86.4 85.3 84.6 87.9 87.0 83.9 86.3 87.2 239.7 234.3 268.9 247.6 225.8 234.6 236.1 232.1 233.6 218.9 239.3 226.5 222.2 237.2 225.8 225.4 225.0 233.4 236.5 218.2 208.0 223.9 218.2 229.1 204.8 198.3 228.0 214.6 220.7 239.7 241.5 220.6 238.9 235.2 218.7 203.7 214.1 231.1 228.5 232.6 208.3 200.4 227.0 24.5 23.5 20.5 24.4 22.2 22.3 20.2 19.5 20.0 20.5 22.6 22.7 22.3 22.0 22.8 21.2 18.2 21.4 21.7 21.1 20.0 20.9 21.5 22.7 25.3 23.2 21.6 25.9 25.9 23.5 26.2 20.7 21.6 23.3 21.6 26.2 18.4 22.6 24.1 23.1 19.5 23.8 22.4 3.9 2.3 20.9 7.4 Grain yield (kg ha-1) Kab ete SR 2329 2688 2389 2204 2283 2568 2824 2658 1860 2809 1975 2086 2622 2670 2050 2611 2562 2243 2365 2445 2326 1852 2411 2255 2389 2356 1985 2396 2271 2380 1888 2515 1976 1154 2032 939 1981 1415 2609 717 1311 2340 2184 Kabete LR Thika SR Juja LR Mean 2490 2501 1702 2470 1684 1307 1420 1621 1743 1669 2075 1252 1707 1314 1660 925 1692 983 816 1169 861 1268 1772 281 1539 1613 2275 1304 830 1908 1546 1203 1549 1112 1450 861 1127 1123 1817 1200 557 591 1452 2805 2001 2723 2262 2578 2542 2205 1730 2471 2472 1983 2539 2771 1905 1879 1829 2338 1823 1676 2456 1716 2259 260 1830 1427 988 1686 1056 1721 630 852 1318 2300 1549 1261 2683 825 1908 940 1274 1561 1086 1868 2686 2299 2538 2325 2571 2554 2181 2503 2421 1416 2170 2279 977 2180 2406 2415 918 2357 2548 1209 2220 1644 2508 2541 1536 1892 895 1889 1751 1635 2245 1432 622 2319 1295 1432 1792 1086 151 1916 1077 447 1873 2578 2372 2338 2315 2279 2243 2158 2128 2124 2092 2051 2039 2019 2017 1999 1945 1878 1852 1851 1820 1781 1756 1738 1727 1723 1712 1710 1661 1643 1638 1633 1617 1612 1534 1510 1479 1431 1383 1379 1277 1127 1116 1844 Replications NS NS NS NS Genotypes (G) ** ** NS ** Environments (E) ** ** ** ** GxE ** ** * ** *, **: Significant at 5 and 1 % probability levels, respectively; NS= not significant 206 18.2 NS ** ** ** Table 109. Navy and large white varieties released in eastern Africa between 2003 and 2008. Variety Line Code Cheupe CAB 19 RJ1 RJ1 RI 5-1 RI 5-1 RI 5-5 RI 5-5 RI 5-3 RI 5-3 IL 5-53 IL 5-53 Cranscope Kranskop Argane AR04GY TAO4 JI TAO4- JI Chercher STTT-165-96 Chore STTT-165-92 Hirna STTT-165-95 Mutwakil Berber Large Ibarya ABA 61 Source: National Bean program reports 2004-2008. Activity 3.3 Year of Release 2008 2008 2008 2008 2008 2008 2007 2005 2005 2006 2006 2006 2003 2003 Country of release Tanzania Madagascar Madagascar Madagascar Madagascar Madagascar Ethiopia Ethiopia Ethiopia Ethiopia Ethiopia Ethiopia Sudan Sudan Identification of a varietal candidate in Nicaragua with potential for international export Highlights: • The bean program of INTA-Nicaragua has selected a line for varietal release with the purpose of exporting grain to the USA. Rationale: Beans have become increasingly commercial and are an important income earner in both Latin America and Africa. Surveys in rural Nicaragua indicated that beans rank among the top three income generators for more that 90% of the population. With many million of Latins now living in the USA, a population with increased buying power is willing to pay for traditional foods imported from the home country. Thus Nicaragua has organized a value chain to link farmers producing traditional red beans with the “nostalgia” market in the United States. Landraces with light red color have serious phytosanitary problems that have been solved in improved varieties with darker color, but now there is a demand for lines with color comparable to the landraces that also have improved agronomic traits. Materiales and Methods: In August, 2005 lines were selected by the Nicaraguan breeder in CIAT’s drought breeding nurseries in Cali, Colombia based on drought performance and commercial grain color. Among these was the line SM15212-33-3, derived from the parents SER 38 x (MAB 87 x SER 31). MAB 87 was developed for resistance to angular leaf spot (ALS) and is derived from G10474, a climbing bean from Guatemala that presents the widest known resistance to races of Phaeoisariopsis griseola, causal agent of ALS. The line was dubbed “628” in Nicaragua in reference to its serial number in the original nursery. Line 628 stood out for its excellent grain type. It was shipped to Nicaragua and underwent reselection and systematic evaluation for agronomic traits and culinary characteristics. Results and Discussion: Line 628 has now advanced through regional and validation trials. In the process it has proven to have stable resistance to races of ALS in Nicaragua. ALS has been increasing in 207 importance over the past decade and has reached dangerous levels in most countries. If Line 628 is released, it will be the first variety in Central America to be bred specifically for ALS resistance. This is relevant for the commercial market, since ALS causes early defoliation and reduced grain size and quality. Anecdotally, Line 628 also has proven to be relatively tolerant to excess rainfall. One farmer reported that while the local variety was totally lost in the unusually rainy “postrera” season (September-December) of 2008, Line 628 produced nearly 300 kg ha-1. In the very dry north of Nicaragua in Somoto, Line 628 was one of the better yielding lines in the “primera” season (May-August), although in most environments it is not outstanding in yield. Its advantage, in addition to its resistance to ALS, is its commercial quality and color. In the 2008 “postrera” season, a farmer association planted 44 hectares for seed production, even though the line is not officially released. Unfortunately, given the unfavorable climatic conditions, little was harvested. It is expected that formal release with take place in 2009. Collaborators: S. Beebe, M. Grajales, C. Cajiao, A. Llanos, J. Molina, R. Urbina Activity 3.4 Progress in development of Snap and runner beans for smallholder production in East and Central Africa Highlights: • Twenty bush and climbing snap bean lines with consumer preferred pod characteristics and resistance to diseases selected in four countries in eastern Africa. • Nineteen new short-day runner bean lines with high pod yield potential selected making smallholder production under short-day conditions in eastern Africa feasible. Rationale: Snap bean (Phaseolus vulgaris L.) and runner beans (Phaseolus coccineus L.) are the probably the most important high value beans grown in East and Central Africa. They are mainly grown for export markets but the domestic markets especially in urban areas are growing rapidly. It is a major source of income for smallholder farmers especially in Kenya, Uganda, Sudan, Tanzania, Madagascar in eastern Africa and Senegal, Cameroon and other countries in West Africa. Snap beans, also known as French beans are grown in North Africa for export to Europe and Middle East. There is growing interest to increase snap bean production for domestic and export markets in Rwanda, Ethiopia, Burundi, and other countries in East and Central Africa. Snap bean is also grown by large commercial companies for export to overseas supermarkets and for canning industries. Yield of snap bean in smallholder farmers’ fields varies from 2 to 8 t ha-1 ( CIAT, 2004), compared to over 14 t ha-1 among large scale producers. Smallholder production is constrained by diseases especially rust, angular leaf spot, root rots, bean common mosaic virus and pests especially bean stem maggots, thrips and nematodes. The intensive nature of cultivation of this crop leads to high disease and insect pressure, and consequently excessive use of pesticides. Smallholder production is further constrained by high costs of seed because most of the varieties produced by private companies are protected by legislation. Thus seed produced by contract in the region is exported for processing and packaging, and re-imported for production. The few varieties developed by public institutions (especially in Kenya) are often susceptible to diseases and pests. Very little has been done to develop improved snap bean varieties freely accessible to smallholder farmers and informal seed producers (who supply over 90% of dry bean seed grown in the region) in the region. Due to the high quality demands, smallholder farmers rely on fungicides and insecticides to reduce production and post harvest losses associated with diseases and pests. This is no longer a viable option because recently instituted minimum residue levels, and preference by importers to source produce from large 208 scale producers threaten to push smallholder farmers out of business ( Kimani, 2005; CIAT, 2004). In East and Central Africa, production is based on determinate types. Unlike their counterparts in South America, East African farmers normally do not grow the indeterminate types, which are higher yielding and have longer harvest duration. Breeding for high yield, disease and pest resistance, tolerance to abiotic stresses, general adaptation to tropical conditions and acceptable market quality is a critical component of an integrated strategy to address constraints to snap bean production in the region. Unlike snap beans, the main constraint to snap runner bean production is requirement for extended light. Current varieties are long day. They require at least three hours of additional lighting to stimulate flowering and pod setting. This demands installation of expensive infrastructure in production fields. Consequently, only large commercial firms can produce and market this high value crop. Runner beans are generally more resistant to major bean diseases and have been proposed as possible source of resistance to common bean. Development of short day runner beans will facilitate smallholder farmers to gainfully participate in runner bean sub-sector and also reduce production costs and make local produce more competitive in international markets. A regional program was therefore started in 2006 to support the development of improved snap bean varieties with high yield potential, resistance to biotic stresses and consumer preferred pod quality, and short-day runner beans for smallholder production. This report highlights progress in this program. Materials and Methods: The regional snap bean program is based at six institutions in four countries, Uganda, Kenya, Rwanda and Tanzania. In Uganda, snap bean research is based at National Crops Resources Research Institute (NACRRI), which has been coordinating the program since 2006. In Rwanda activities are based at ISAR station in Rubona. In Kenya, the national snap bean program in coordinated by the University of Nairobi with activities at National Horticultural Research Centre at KARI-Thika and at Moi University in Eldoret. In Tanzania, snap bean research is based at Selian Agricultural Research Institute (SARI), Arusha. Work at Kawanda has focused on screening snap bean varieties with farmers and developing production packages. At Moi University, crosses were made to develop locally adapted snap bean cultivars with improved pod yield, resistance to anthracnose and rust, and marketable pod quality (van Rheenen et al., 2003). After six generations of selection, 23 lines were identified. Following preliminary evaluations, the number of lines was reduced to 12. The 12 lines were evaluated in national performance trials at six locations (Eldoret, Thika, Kakamega, Marigat, Lanet and Njoro) in collaboration with Kenya Plant Health Inspectorate (KEPHIS) which coordinates testing of candidate varieties from public and private breeders and formal release of the best varieties. The trial included three commercial cultivars as checks. At the University of Nairobi, activities have focused on development of regional nurseries of bush and climbing snap beans, development of crosses to transfer rust, anthracnose, angular leaf spot, root rots and anthracnose resistance to popular commercial varieties, and breeding short day snap runner beans. Snap bean is a relatively new sub-sector in Tanzania. Activities at Selian, therefore focused on a baseline survey to better understand the major constraints and the production and marketing environment, evaluation local bush lines and 35 advanced climbing bean lines from CIAT, development of agronomic and crop protection management practises. In Rwanda, focus has been on evaluation of 40 advanced climbing and 18 bush snap lines received from CIAT, Colombia and selection for pod characteristics, (colour, shape, texture and taste) and resistance to rust, angular leaf spot and anthracnose from local variety ‘Vunikingi’ and its crosses with bush snap beans. Results and Discussion: In Uganda, 11 lines introduced from CIAT and Kenya were evaluated for three seasons. Paulista and Helda were used as checks. Major constraints were rust, angular leaf spot and common bacterial blight. However, field surveys showed that rust was most limiting in farmers’ fields. Results showed that pod inoculation was most effective for screening for resistance to common bacterial blight. Six lines (HAB 433, BC 4.8, A 20, J 12, L1 and L 12) showed combined tolerance to the three 209 major diseases (Table 110). Line HAB 433 was selected on-farm for yield and quality (pod size and shape, length, snappiness and taste). Table 110. Line HAB 173 HAB 414 HAB 433 A20 K3 L1 L12 J12 BC4.5 BC 4.8 BC 7.5 Paulista Helda Disease scores of 11 snap bean lines at Namulonge, Uganda. Common bacterial blight Rust Angular leaf spot 4 7 2 3 4 4 4 4 6 5 5 6 7 5 5 2 3 6 5 5 5 6 3 6 8 8 2 3 4 3 3 3 3 3 7 5 7 6 Twenty-one F6 climbing bean lines introduced from Rwanda, 14 commercial varieties of European origin and 35 F5 climbing bean lines from CIAT were sown in an observation nursery at Kawanda. A survey of economic pests in farmers’ fields in Uganda showed that bean aphids, pod borers and bean fly were the most important insect pests in the surveyed locations, while bean leaf rust, root rot and angular leaf spot are the most important diseases. All farmers interviewed invariably use pesticides in their fields to control the pests and diseases. Results showed that application of inorganic and organic fertilizers increased pod yield from 6.8 t ha-1 to 7.4 t ha-1. Paulista and Teresa were the best yielding commercial varieties with yields of 8.3 and 8.9 t ha-1, respectively. In Rwanda, five F6 lines were selected from Vunikingi x Amy populations. Yield of 58 lines introduced from CIAT and local collections varied from 5 to 12 t ha-1. As expected climbers were better yielding compared to bush varieties. The most promising lines were Boon, Cabbra, G685, Ncekarkonnigia, Saxa, Khaki and Loiret. Survey results showed that major constraints to snap bean production in Rwanda were diseases, pests and weeds. Major diseases included root rots, angular leaf spot, rust and bean common mosaic virus. The main pests are aphids, bean stem maggot, spider mites and crickets. Major weed species were Commelina spp, Digitaria spp, Bidens spp, and Oxalis specie. Diseases were controlled with a range of fungicides. Evaluation of commercial varieties and introductions for reaction to diseases showed that Tarrot was susceptible to rust, Saxa and Cabbra to anthracnose and Ncekarkonnigia to BCMV. In Tanzania, 35 climbing bean lines from CIAT and 5 local bush varieties were sown in observation nursery and to increase seed at Madiira. In Kenya, three snap bean accessions were collected from farmers’ fields in Murang’a district, five from KARI-Thika and three from Madagascar. These were added to the working collection at the University of Nairobi. Thirty-three bush and seven climbing bean lines were sown in observation nurseries and seed increase plots at Mwea and Kabete. One pre-release variety and 12 rust resistant advanced lines were selected at Thika. Thirty four new crosses were made to transfer resistance to root rots, angular leaf spot and common bacterial blight to susceptible commercial varieties (Paulista, Amy, Julia, Teresa, Vernadon, Morgan, Alexandria, Kutuless and Morelli). Sources of resistance included Beldakmi and Beltglade lines 210 with ur genes and L227 which has multiple resistance to root rots, angular leaf spot, rust and common bacterial blight. F1’s were advanced to F2. The characteristics of the 12 lines selected at Moi University in the national performance trials at six locations are presented in Table 111. Flowering was earliest at Thika (37 days) and latest at Njoro (50 days). Duration to first picking was shortest at Thika (56 days) and longest at Njoro (62 days). Picking period varied from 22.6 days at Marigat to 36 days at Njoro and 40 days at Thika. On an average, Lanet and Njoro showed better pod quality scores than Marigat and Eldoret. The pod quality at latter location was poorest, probably due to the poor plant growth in general. The interaction between locations x trial entry was highly significant, suggesting that different entries respond differently to environmental conditions in respect of pod quality. The variety differences for reaction to rust were significant. The entry with the severest symptoms was No. 1, showing a mean score of 7.6. It differed significantly from those that had a score of 6.6 or less. Entry No. 7 had the lowest rust score of 1.2, differing significantly from those having a score of 2.2 or more. On an average, Marigat showed significantly more severe rust symptoms than other locations. The interaction between location x entry was significant, suggesting that possible differences in rust races occur. Variety differences for fresh pod yield were significant. The entry with the lowest yield was No. 3 with an average of 9.6 t ha-1, followed by No.1 with 9.7 t ha-1. These differed only significantly from those that had 12 t ha-1 or more. Entry No. 11 had the highest yield of 13.1 t ha-1, differing significantly from those having a yield of 11 t ha-1 or less. The locations differed significantly pair-wise: Marigat and Njoro had the highest yields; Lanet and Kakamega were intermediate, and Eldoret and Thika lowest. The mean yields per location ranged from 3.1 t per ha-1 at Thika to 19.7 t ha-1 at Marigat. No significant interaction between location and trial entry was observed which indicates that varieties had no differential response to environment. It suggests that the yield adaptation of the entries to different environments was similar. Table 111. Days to flowering, first and last day of pod picking, pod quality, rust score and fresh pod yield of snap bean lines selected at Moi University, Eldoret, Kenya *Line Days to 50% flowering Days to first pod picking Days to last pod picking **Pod quality ***Rust score Fresh Pod yield t ha-1 1 45.1 59.0 89.7 3.8 7.6 9.7 2 43.5 57.1 89.7 3.3 4.6 11.4 3 43.6 57.2 88.9 3.6 2.4 9.6 4 44.8 57.3 89.2 2.3 2.1 12.9 5 43.0 58.1 89.2 3.1 2.1 11.7 6 42.6 56.6 89.4 3.9 6.4 10.6 7 46.1 58.9 89.8 4.0 1.2 12.2 8 43.6 56.7 89.8 3.5 2.3 13.0 9 43.0 56.9 89.3 3.7 4.2 10.3 10 44.4 57.3 89.4 3.9 2.3 11.2 11 45.5 56.8 89.4 2.7 2.0 13.1 12 43.8 59.1 89.4 3.8 4.3 10.3 Mean 44.1 57.6 89.5 3.5 3.5 11.3 CV (%) 6.7 3.57 1.52 12.7 28.7 28.9 LSD .05 1.94 1.35 NS 0.36 0.94 2.2 * Lines 1, 4, 7 and 11 were checks * Pod quality on a scale of 1= best and 5=worst. ** Rust on CIAT scale, 1-3 =resistant, 4-6= intermediate and 7-9=susceptible at three locations- Marigat, Lanet and Njoro. 211 At Kabete, five F3 populations were developed from crosses between a long-day commercial runner snap variety and five short day dry grain type, short-day varieties. The F3 progenies were advanced to F4 and F5 generations at Ol Jorok and Laikipia. Pod set and pod characteristics were the main selection criteria under short day conditions. The population showed considerable segregation for pod traits. Progenies were grouped into six categories (Table 112). Long, straight pods are preferred by exporters. Curved short pods are associated with local short day parental genotypes. F5 lines showed heavy podding at Ol Jorok during the 2008 cropping season. Two to three harvests were made from most of the lines. Runner bean lines showed high levels of resistance to angular leaf spot, root rot, common bacterial blight, anthracnose and frost despite heavy disease pressure which caused severe damage to climbing and bush bean lines in adjacent plots. However, some runner bean lines showed intermediate reactions to rust infection. Table 112. Classification of F4 and F5 runner bean lines based on pod characteristics at Ol Jorok and Laikipia. Pod category Long straight Medium straight Short straight Long curved Medium curved Short curved Number of lines 19 18 14 30 28 26 Conclusion: Although breeding snap beans in eastern Africa is still in the early stages, considerable progress has been made in developing new lines of snap with resistance to major biotic stresses and shortday runner beans in eastern Africa. More than 20 promising bush and climbing snap beans have been selected in preliminary trials from advanced lines and segregating populations. For most of the countries, breeding snap beans is a new but promising innovation. This program was supported by CIAT/PABRA and ASARECA. However, it was temporarily suspended for the last two years due to re-organisation of ASARECA. The program is now expected to re-start late in 2009. Nineteen F5 lines of runner beans with preferred long pods and good pod set under short day conditions were selected. Contributors: Paul Kimani, Steve Beebe (CIAT), Michael Ugen (Uganda), Augustine Musoni (Rwanda), Festo Ngulu (Tanzania), van Rheenen, George Chemining’wa, John Nderitu and Agnes Ndegwa (Kenya). Collaborators: Bean programs at Namulonge (Uganda), SARI, Arusha (Tanzania), University of Nairobi, Moi University and KARI-Thika (Kenya) and ISAR, Rubona (Rwanda), CIAT Bean Program, Cali (Colombia). References CIAT. 2004. Annual IP-1 Report, Cali, Colombia. Kimani, P.M. 2006. Snap beans for income generation by small farmers in East Africa [On line]. Internacional de Agricultura Tropical (CIAT), Kampala, Uganda. 2p. Highlights: CIAT in Africa No.31. Van Rheenen, H.A., Odindo, A.O. and Iruria, D.M. 2003. Anticipated promises and problems for snap bean seed production in Kenya. Proceedings of the workshop on “The seed industry in Kenya at crossroads” held at Nakuru, Kenya from 1st – 3rd July, 2002. 212 Product 4: Strengthened institutions that enhance bean product development and delivery Activity 4.1 Strengthened capacity of NARS: increasing the knowledge and skills of scientists and staff from NARIs, NGOs and Rural Service Providers Highlights: • • • • • • • A total of forty-nine students conducted research activities related to their thesis work, of which twenty eight were at CIAT HQ, and twenty one in Africa. Of these, three PhD and one MSc students were as visiting researchers at HQ. In Latin America, two M.Sc. candidates, and two pre-graduate students completed their research theses. In Africa three PhD, four M.Sc. and one Bs. candidates completed their research theses. A total of thirty-three students continue their studies, as follows: six Ph.D. candidates in Africa and five in Latin America, eight M.Sc. candidates in Africa and four in Latin America, and ten pre-graduate in Latin America. Twelve visiting researchers coming from Colombia, Cuba, Denmark, Guatemala, Hungary, India, Panama and Zambia received training in different disciplines at headquarters. Several courses and workshops were held in Latin America and Africa During this reporting period there was a joint stakeholders meeting for PABRA partners, as well as a joint steering committee meeting for SABRN and ECABREN where they reflected on the progress over the past 4 years, and planned activities to achieve the milestones contributing towards achieving the goals in the final year of the project, as well as to plan for the next phase. A number of students have either registered or started their course work at various universities to sharpen their knowledge and skills in bean research for development. Two new students doing MSc in plant breeding enrolled at the University of Zambia and Penn State University, both from Malawi. Two other students had been accepted for Ph.D. programs at the University of Free State and Massey University – New Zealand. These scientists will add to the existing capacity for bean research in the region. 4.1.1 Degree and non-degree training in Latin America Students: Name Degree Status University Title Ph.D. Candidates Asrat Asfaw Amele Ph.D. Visiting student Wageningen University, Netherlands (M. Blair and I.M. Rao) Genetic investigation of drought tolerance in common beans for Ethiopia and the ECABREN region Louis Butare, ISARRubona Ph.D. Continuing Gembloux Agricultural University, Belgium. (S. Beebe, I.M. Rao and M. Blair) Root development and root health of interspecific progeny of crosses between P. vulgaris and P. coccineus Luz Nayibe Garzón Ph.D. Continuing Anthracnose resistance Homar Gill Ph.D. Visiting student Universidad Nacional de Colombia, Bogota (M.Blair and G. Ligarreto) Universidad de Tamauliplas, Mexico (M. Blair) 213 Research on bean diversity Name Degree Status University Title Lara Ramaekers Ph.D. Continuing University of Leuven, Belgium (M. Blair and I.M. Rao) Biological nitrogen fixation Gloria Santana, CORPOICA Ph.D. Continuing Universidad Nacional de Colombia, Palmira. (M. Blair, F. Morales and G. Ligarreto) Resistance to bean common mosaic virus Assefa Teshale Mamo Ph.D. Visiting student University of Padua, Italy (S. Beebe, M. Blair and J.M. Bueno) Conducting researching in bruchid resistance, drought tolerance and canning quality León Darío Vélez Ph.D. Continuing Universidad Nacional de Colombia, Bogotá. (M. Blair and G. Ligarreto) Inheritance of intercropping ability between common bean and maize M.Sc. Candidates Juana Marcela Cordoba Leonardo Duque M.Sc. M.Sc. Continuing Continuing Universidad Nacional, Bogotá (M. Blair) Universidad del Valle, Cali, Colombia (J.M. Bueno and S. Beebe) Universidad. Nacional, Palmira University of Saskatchewan (M. Blair) Universidad del Valle, Cali, Colombia (J.M. Bueno) SSR development and mapping Evaluation of the use of the oil of limoncillo to repel Bemicia tabaci in bean Victor Mayor Durán M.Sc. Continuing Drought tolerance Hanny El Sadr M.Sc. Orlando Grijalba M. Sc. Visiting student Continuing Hugo Arley Jaimes M.Sc. Completed Universidad Nacional de Colombia, Palmira (J.M. Bueno) Evaluation of bean interspecific hybrids for resistance to Acanthoscelides obtectus Lizzie Kalalokesya M.Sc. Visiting student University of Zambia Studying CBB Resistance breeding in Andean Bean Crosses Paola Sotelo M.Sc. Completed Universidad Nacional de Colombia, Palmira (J.M. Bueno) Inheritance of resistance to bean leaf crumple virus in snap beans Alejandro Chaves B.Sc. Completed Diversity analysis of snap beans Aura Marina Díaz Bravo B.Sc. Completed Universidad Javeriana Bogotá (J.D. Palacio and M. Blair Universidad del Valle (M. Blair) Andrea Carolina Fernandez B.Sc. Continuing Universidad Javeriana, Bogotá (M. Blair) Drought candidate genes for common bean Tannin accumulation in seed coats Development of action thresholds for rational control of Bemisia tabaci biotype B as a pest of sweet pepper Pregraduate students: 214 HPLC analysis of phytate concentration in common bean Name Degree Status University Title Claudia Marcela Franco B.Sc. Continuing Universidad del Valle (M. Blair) SSR mapping Andrea Lorena Herrera B.Sc. Continuing Universidad de Antioquia (M. Blair) Tannin evaluation in common bean Natalia Hurtado B.Sc. Continuing Universidad Javeriana (M. Blair) SSR mapping Paulo Izquierdo B.Sc. Continuing María Alejandra Lozano B.Sc. Continuing Universidad de Tolima (M. Blair) Universidad del Valle (M. Blair) NIRs analysis of common bean nutritional quality HPLC analysis of tannin concentration in bean seed coats Fredy Monserrate B.Sc. Completed Universidad Nacional, Bogotá (G. Hyman and M. Blair) Stability analysis common bean lines Juan Carlos Perez B.Sc. Continuing Universidad Nacional, Palmira (M. Blair and I.M. Rao) Drought tolerance in common bean Alvaro Soler Garzón B.Sc. Continuing Universidad del Tolima Marina Villacís B.Sc. Continuing Universidad del Valle (M. Blair) Genetic diversity of wild core collection of common bean analyzed through fluorescent microsatellites SNP mapping of biofortified Non-degree training of visiting researchers: Name Orlando Chaveco Staff S. Beebe Dates AugustSeptember Institution Unidad de Extensión, Investigación y Capacitación Agropecuaria de Holguín, Cuba Topic Testing materials with high content of iron and zinc and materials tolerant to drought Cristina Cvitanich M. Blair May Institute of Molecular Biology, University of Aarhus Nutrition project Carmen Iliescu M. Blair June Agricultural Biotechnology Center, Godollo, Hungary Evaluation of common bean diversity in Eastern Europe and collaboration on SNP detection Lizzie Kalalokesya M. Blair SeptemberNovember University of Zambia Marker Assisted Selection of VAXderived CBB Resistance in Andean bean crosses Juliana Ines Medina M. Blair June-August Colciencias Positional cloning of the genomic region involved in multiple virus resistance based on the map of Phaseolus vulgaris Audino Melgar M. Blair, S. Beebe November IDIAP, Panama Training on nutritional analysis of common bean 215 Name Staff Dates Institution Topic Natalia Moreno M. Blair JanuaryDecember Colciencias Nutritional quality and diversity of Eastern and Southern African common bean varieties Peter Papp M. Blair April Agricultural Biotechnology Center, Godollo, Hungary Evaluation of common bean diversity in Eastern Europe Elena Perez Vega D. Debouck Pathology November Servicio Regional de Investigación y Desarrollo Agroalimentario, SERIDA, Asturias, Spain Germplasm management; Xanthomonas sp inoculation Karla Ponciano M. Blair July ICTA, Guatemala Training on drought Emigdio Rodriguez M. Blair, S. Beebe November IDIAP, Panama Training on nutritional analysis of common bean Prem Nath Sharma M. Blair OctoberMarch 2009 Hillside Agricultural Research Center Evaluation of common bean diversity in India Courses and Workshops: Date Title Duration (days) Total No. participants No. of female participants No. of CIAT instructors 3-28 March Training course on phenotyping of the Tropical Legumes Project, ICRISAT, Hyderbad, India (TLI) 25 30 25 April Bruchids 1 15 12 1 5 2 0 3 1 15 12 1 8 May 16 May Sampling and identification of whiteflies Insecticide resistance in whiteflies 2 23 May Varietal resistance to insects 1 15 12 1 23 May Sampling methods in bean pests 1 15 12 1 12 June Training in mass rearing methods 1 1 0 2 14 July Pest of beans their control 1 20 5 1 16 July Whitefly mass rearing methods 2 1 1 3 20 Aug. Whitefly insecticide resistance 3 1 1 3 2 Sept. Control of bean insect pests 1 15 5 1 6-10 Sept. CYTED funded project workshop on progress in developing transgenic maize and common bean tolerant to drought 3 12 5 1 216 4.1.2 Degree and non-degree training in Africa Name Degree Status University Title Isaac Fandika Ph.D. Accepted To be decided Virginia Gichuru Ph.D. Completed Massey University, New Zealand Makerere University, Kampala, Uganda Godwill Makunde Ph.D. Registered University of Free State, South Africa Firew Mekbib Ph.D. To defend April 2009 Norwegian University of Life Sciences Clare Mukankusi Ph.D. Completed University of KwaZulu-Natal,RSA John Muthamia Ph.D. Continuing University of Leuven John Nzungize Ph.D. Continuing Gembloux Agricultural University Evaluation of the genetic diversity of the isolates of Pythium spp. in Rwanda for a selection of the varieties of common bean for resistance to the root rot caused by Pythium spp Bernard Okonda Ph.D. Continuing University of Nairobi Genetic variation for nitrogen fixation, micronutrient density and influence of plant growth promoting rhizobacteria in common Musoni Augustine M.Sc. Completed University of Nairobi Steven Buah M.Sc. Continuing Makerere University, Kampala, Uganda Inheritance of fusarium wilt (F. oxysporum f.sp. phaseoli) and selection for multiple disease resistant and marketable climbing bean varieties Phenological and pathogenic characterization of bean landraces from South West Uganda Ph.D. Candidates Symptomatology and characterisation of Pythium spp. of major crops in a bean based cropping system in south-western Uganda Association mapping to quantify genetic diversity in drought adaptation of common bean Genetic enhancement of sorghum diversity through an integrated approach. Breeding beans (Phaseolus vulgaris L) for resistance to Fusarium root rot (Fusarium solani f.sp. phaseoli) and large seed size in Uganda Quantification of the agronomic contribution of the various Plant Growth Promoting Rhizobacteria (PGPR) and VAM singly and in combination on selected bush bean and climbing bean varieties M.Sc. Candidates Virginia Chisale Lizzie Kalolokesya M.Sc. Started Penn State University USA M.Sc. Started University of Zambia 217 To be decided Use of SCAR marker SU91 in marker assisted selection (MAS) for common bacterial blight (CBB) resistance in common bean Name Degree Status University Title Kanyenga Lubobo M.Sc. Continuing Lubumbashi University, DRC Breeding for bean root rots resistance in southern midland of D. R. Congo Francoise Murorunkwere M.Sc. Submitted Makerere University, Kampala, Uganda Jasper Mwesigwa M.Sc. Completed Makerere Pheonah Nabukalu M.Sc. Completed Makerere University, Kampala, Uganda Joseph Orede M.Sc. Continuing University of Nairobi Bello Shano M.Sc. Continuing University of Malawi Geoffrey Wachira M.Sc. Continuing University of Nairobi Felix Waweru M.Sc. Continuing University of Nairobi Improving resistance to bean common mosaic virus and bean common mosaic necrotic virus in common bean using marker assisted selection Characterization of races of C. lindemuthianum in Uganda. Improving resistance to anthracnose in commercial verities in Uganda using marker assisted selection Breeding for common bean for disease resistance through gene pyramiding and phenotyping Evaluation of common bean genotypes for Al and drought resistance Screening common bean cultivars for resistance to bean fly Drought phenotyping in RILs, reference collection and regional bean varieties Pregraduate students: Mackford Maseko Dip. Agric. Completed Natural Resources College; Malawi Non-degree training of visiting researchers: Name Staff Institution Esther Arunga, Uganda Moi University, Kenya Tarcisis Mutuoki Enid Katungi KARI-Katumani Yiteye Abebe Enid Katungi Melkassa 218 Topic Bean pathology research methods and biotechnology Socio-economist, integrated into activities for mentoring, leading the market studies in eastern Kenya Socio-economist, integrated into activities for mentoring, completion of data cleaning for Ethiopia Training Courses and Workshops: Date Title Duration (days) Total No. participants No. Women participants No. of CIAT instructors No. of NARS instructors 9 Jan PABRA quarterly meeting Kampala, Uganda TL II Drought Project Bean and Chickpea Planning Meeting. Nazret Ethiopia Planning workshop for the Bean Drought Project for partners from Central and South Rift Valley, Nyanza and Western Kenya Machakos Kenya TL II Training workshop on drought phenotyping at KARIKatumani, Machakos, Kenya PABRA Stakeholders workshop Kampala, Kenya Participatory Internal Control System and Participatory Monitoring and Evaluation Training PABRA-CIAT-LAGROTECH Nutrition and Health Training workshop for Trainers in Kisumu, western Kenya CEDO Nutrition Training of Trainers with Support from CIAT, Masaka, Uganda Evaluation of PABRA Capacity Building Programme Arusha, Tanzania PABRA Monitoring and Evaluation Arusha, Tanzania Tropical Legumes II - Nangina Social Work Project 2 - Training of Trainers (to build capacity of trainers on basic principles of bean seed production & maintenance of sustainable bean seed systems) Mumias, Kenya 1 8 3 - - 3 14 3 2 3 4 15 3 3 14-15 Jan 17–18 Jan 16 - 19 Jan 20 – 24 Jan 14 - 15 Feb 18 Feb 26-29 Feb 26 Febr 27 Febr 28 Feb Feb - March 4 - 8 March 5 43 12 3 43 17 1 30 13 0 1 26 8 2 1 21 7 2 15 Training of Trainers (Extension staff from partner organizations in Kenya ) on seed production, post harvest management and seed management Training Course on the use of ECOSAUT: A Model for Economic, Social and Environmental Evaluation of Land Use Alternatives, Harare, Zimbabwe 219 5 1 147 52 0 19 0 1 3 Date 7 - 8 March March March–Oct 17-20 March 25 March 27 March 29 March 14 April 17 April 18 April 19 April 30 April1 May 5-16 May 14 May 22-23 May 26-28 May Title Duration (days) Total No. participants No. Women participants No. of CIAT instructors Capacity Building Evaluation of the Participatory Plant BreedingParticipatory Variety Selection (PPBPVS), Mbeya, Tanzania Bean seed system planning workshops in Central Rift Valley of Ethiopia, Southern Ethiopia, North west Ethiopia, East Ethiopia Bean seed production/supply, field data collection (seed production and supply/use) PABRA Steering Committee meeting, Lusaka, Zambia Bean Drought Project Partners’ Meeting Tropical Legume II for Central Rift Valley, Nazereth, Ethiopia Bean Drought Project Partners’ Meeting Tropical Legume II Southern Regions, Awassa, Ethiopia Bean Drought Project Partners’ Meeting Tropical Legume II NorthEast Regions, Debre Brehan, Ethiopia Bean Drought Project Partners’ Meeting Tropical Legume II for West Haraghee, Ethiopia Bean Drought Project Partners’ Meeting Tropical Legume II Southern Regions, Dire Dawa, Ethiopia PABRA Capacity Building Evaluation – Participatory and Monitoring Evaluation, Kakamega, Kenya Evaluation of PABRA Capacity Building PPB-PVS, Kakamega, Kenya Developing Participatory Monitoring and Evaluation Framework, Uganda Training course on breeding and physiology of drought resistance in common bean (TLII project) Seed Aid for Seed Security: Outreach Meeting in Norway TLII Cross-Crop Seed Systems Meeting Nairobi, Kenya Participatory Variety Selection (associated with McKnight and BMZ Projects) 2 24 7 2 7 132 10 1 5 1670 791 0 1 4 20 4 - 1 33 0 1 1 43 0 1 1 16 0 1 1 16 1 1 17 1 1 27 1 1 1 9 1 1 2 29 12 24 5 No. of NARS instructors 5 1 2 3 220 28 8 3 0 Date 28-30 May 29 – 31 May 8-12 June July August 4-5 Sept 11–12 Sept 18-19 Sept 21-25 Sept 22-23 Sept 22-26 Sept 29 Sep4 Oct 28 – 30 Oct 5 Nov 2 – 3 Dec 2 - 3 Dec 5 – 6 Dec 10-12 Dec 13 Dec. 14-15 Dec Title Duration (days) Total No. participants No. Women participants No. of CIAT instructors 3 29 5 3 5 7 3 1 2 1 7 3 1 3 3 14 3 1 2 2 29 3 1 3 2 17 3 1 2 33 6 2 5 24 5 2 2 28 7 1 4 5 23 5 3 0 Participatory Variety selection (Malawi) McKnight Beans Seed Systems: yearly Planning Meeting: Lilongwe, Malawi Organizing and conducting focus group Household questionnaire, the art of interviewing and establishing rapport Organizing and conducting focus group revisited Household questionnaire, the art of interviewing and establishing rapport Review workshops for Bean Drought Project TL II Seed System for partners from Central South rift valley and Nyanza and western Nakuru, Kenya TL II Bean Seed Systems Review Meeting, Kisumu, Kenya Bean seed systems inception and planning meeting, Machakos, Kenya Participatory Plant Breeding and Participatory Variety selection (Training of Trainers), Ethiopia Planning Workshop for Bean Drought Project for Central and eastern Kenya, Machakos, Kenya Participatory Plant Breeding: Training of Trainers (ToT) Course TLII Drought Project : Annual Meeting, full project, Addis Ababa, Ethiopia Participatory Variety selection, Mozambique Seed Security: linking formal and informal sectors, Accra, Ghana TL II Bean Seed System Year II Planning Workshop. Adama, Ethiopia TL II Bean Drought Project Partners Meeting Nazret, Ethiopia TL II Bean Drought Project Partners Meeting- SARI, Awassa, Ethiopia Household questionnaire, the art of interviewing and establishing rapport Market surveys Market surveys No. of NARS instructors 3 4 7 3 19 2 1 1 1 2 42 2 2 2 3 9 0 1 2 1 2 2 7 0 1 1 1 0 0 221 4.1.3 Trips and attendance of Headquarters staff at meetings The Mesoamerican bean breeder and project manager visited the following countries: Date 11-13 April 07-11 May 21-25 My 29 June-05 July 06-08 July 11-15 Aug 27 Sept – 5 Oct 5-8 Oct 9-11 Oct 11-13 Oct 02-05 Dec Destination Costa Rica Malawi Celaya, Guanajuato, Mexico Dakar, Senegal Nairobi Austria, Vienna Ethiopia Bukavu, Dem. Rep. Congo Butare, Rwanda Tanzania Nicaragua Event or purpose LIV PCCMCA meeting and Agro-Salud Workshop Breeding for Drought Workshop Internacional Bean Congress TL1 Annual Meeting TL2 visit to field sites HarvestPlus Workshop TL2 Project meeting Harvest Plus planning meeting Harvest Plus planning meeting Review of progress in selection of drought tolerant beans Review and evaluation of drought trials The Andean breeder/germplasm specialist visited the following countries: Date 11-17 Jan 21-26 Jan Destination San Diego, California Chillán, Chile 21-25 May Celaya, Mexico 8-12 Sept 15-20 Sept 29 Sept-3 Oct 5-8 Oct 9-13 Oct 20-25 Oct Hyderabad, India Bangkok, Thailand Addis Ababa, Ethiopia Bukavu, Dem. Rep. Congo Butare, Rwanda Campinas, Brazil 7-12 Dec Vallarta, Mexico Event or purpose Plant and Animal Genome – presentation of GCP results. collaboration with INIA on diversity assessment of Chilean landraces. Congreso Nacional de Frijol – presentation on marker assisted selection for breeding/producer audience coordination with ADOC sequencing project GCP annual meeting – presentation of results of TL1 project TL2 coordination meeting Harvest Plus planning meeting Harvest Plus planning meeting Congresso Nacional de Pesquisa de Feijão – presentation on bean genomics International Congress on Legume Genetics and Genomics – presentation on diversity of common bean The plant nutritionist: Date 17-29 March 2-10 May Destination ICRISATPatancheru, India Pennsylvania State University, USA Lilongwe, Malawi 6-10 Sept Granada, Spain 11-13 March Event or purpose Training course on phenotyping of the tropical legumes Project (TLI)-Participated as an instructor WUN workshop on abiotic stress factors Training course on breeding and physiology of drought resistance in common bean (TLII project) CYTED funded project workshop on progress in developing transgenic maize and common bean tolerant to drought 222 Awards: • The 2008 CIAT Scientific Poster Award-Second Place Physiological evaluation of drought resistance in elite lines of common bean (Phaseolus vulgaris L.) under field conditions. J. A. Polanía, M. Grajales, C. Cajiao, R. García, J. Ricaurte, S. Beebe and I. M. Rao • Annual Conference of Plant Nutrition Society of Germany-Scientific Poster Award-Second Place The interaction between aluminum toxicity and drought stress in common bean (Phaseolus vulgaris L.). Z. B. Yang, D. Eticha, I. M. Rao and W. Horst 4.1.4 Trips and attendance of African staff at meetings The Plant Pathologist/PABRA Coordinator: Date 21-25 Jan 28 Jan 10-14 Febr 21-22 Febr 24 Febr 5-6 March 18-20 March 16-20 March 4-12 April 2 May 5-6 May 21- 23 May 26-31 May 11-12 June 17-19 June 30 June – 4 July 8 July – 15 Aug 26-30 July 2-4 Sept 27 Sept - 4 Oct 8-10 Oct Nov 7-22 Dec Destination Kampala, Uganda Nairobi, Kenya Gisenyi, Rwanda Aleppo, Syria Arusha, Tanzania Nairobi, Kenya Kampala, Uganda Lusaka, Zambia Cali, Colombia Nairobi, Kenya Rwanda Nairobi, Kenya Japan Arusha, Tanzania Kakamega, Kenya Lausanne, Switzerland Kenya Minneapolis Accra, Ghana Ethiopia Rwanda Burundi Cali, Colombia Event or purpose PABRA Stakeholders Meeting Coordination TSBF Design and PM&E Meeting of LK-PLS (SSACP) System Wide IPM Meeting Visit with Albin Visit With Albin Kirkhouse meeting PABRA SC meeting Knowledge sharing week Coordination (TSBF) Coordination (MINAGRI, ISAR, USAID, CIAT) TLII Meeting Visit JIRCAS, JICA and TICADIV meeting Coordination CRSP Project Planning Meeting Fourth CGIAR Senior Leadership Program Home Leave APS Meeting CGIAR-FARA Partnership Meeting Tropical Legumes II LKPLS Planning Meeting ISABU Planning BoT meeting The SABRN Coordinator/Breeder: Date 15-20 Jan 29 Feb – 04 Mar 06 Mar 5-13 Apr 16-18 June 29 Jun – 5 July 5-10 Oct 25-27 Oct 27 Oct – 4 Nov 15-20 Nov 20-25 Nov Destination Kampala Barcelona Kampala Cali Nairobi Dakar Maputo Harare Johannesburg Luanda Harare Event or purpose PABRA Stakeholders Meeting Legumes CRSP inception meeting Kirk House Trust proposal development on use MAS in beans CIAT Annual review meetings AGRA Soil Health program inception meeting TL-1 annual review meeting in Senegal McKnight CoP annual meeting in Mozambique Follow up on TL-1 and TL-2 agreements in Zimbabwe Follow up on H+ activities in South Africa Follow up on bean research activities in Angola Join the CIAT team during a visit by Dr Pachico to southern Africa 223 The ECABREN Breeder: Date 8-10 Jan 13-16 Jan 17-19 Jan 20-26 Jan 23-24 Febr 5-7 March 11-13 March 4-14 April 5-11 May 30 June- 6 July 7-8 July 25 July-5 Aug 29 Sept -3 Oct 4 -5 Oct Destination Kampala, Uganda Melkassa, Ethiopia Katumani, Kenya Kampala, Uganda Kabete, Thika and Kiboko Kampala, UgandaKampala, Uganda Cali, Colombia in Lilongwe, Malawi Dakar, Senegal Katumani, Kenya Katumani, Embu, Meru, Njoro, Narok, Kisumu, Eldoret, Kakamega and Kitale (Kenya) Thika, Kenya Kampala, Uganda Melkassa and Addis Ababa, Ethiopia Addis Ababa, Ethiopia Awassa, Ethiopia 6-9 Oct 9-11 Oct 11-13 Oct Bukavu, DR Congo Butare, Rwanda Arusha, Tanzania 21-24 Oct Lilongwe, Malawi 24-27 Aug 7-11 Sept 21-27 Sept Nairobi, Kenya 7-11 Nov 14-22 Nov 11-14 Dec Yaounde, Bafossum and Muea, Cameroon Wageningen University, Netherlands Event or purpose PABRA Quarterly Planning meeting TLII planning meeting Drought phenotyping training workshop PABRA stakeholders workshop TL II field sites characterisation with CIAT consultant KT project planning and proposal review workshop PABRA quarterly planning workshop Annual knowledge sharing week TL I annual review and planning workshop Field visits to TL 2 regional nursery National performance trials technical committee monitoring tour in Coast, Central, Eastern, Rift Valley, Nyanza and Western provinces Nutribean Review and Planning workshop PABRA proposal finalization and planning workshop PVS training workshop TL II first annual review and planning workshop Field visit to Southern Agricultural Research Institute bean program in Awassa. HarvestPlus DR Congo planning workshop HarvestPlus Rwanda planning workshop, TL II Tanzania planning meeting and field visits at Selian Agricultural Research Institute SABRN/ECABREN Joint Planning and review workshop Second national workshop and exhibition on the strategy for revitalizing agriculture (SRA) and Vision 2030 Field visits to IRAD/ WECABREN bean research trial sites in Bafossum and Ekona regions, Cameroon Ethical issues in Biofortification workshop, The Monitoring and Evaluation Specialist: Date 8-10 Jan 20-26 Jan 16-20 March 5-11 April 7-9 Oct 23-25 Oct Destination Kampala Kampala Lusaka, Cali Colombia Rome Lilongwe Event or purpose Uganda-PABRA Quarterly Planning meeting Uganda- PABRA stakeholders workshop PABRA Steering Commitee Meeting CIAT Annual Meeting F2F knowledge sharing workshop Joint PABRA Network meeting The Agricultural Economist: Date 22-27 Sept. 29 Sept-3 Oct. 9-10 Feb. Destination Nazareth, Ethiopia Addis Ababa Malawi Event or purpose workshop on participatory variety selection TL2 annual review and planning meeting TL2 breeders meeting 224 Activity 4.2 Strengthen international collaboration through networks (Intra- and internetwork collaboration), bi-lateral relations, and/or joint special projects Highlights: • The Pan African Bean Research Alliance (PABRA) continued to provide funding support to research for development sub-projects within the SABRN 4.2.1 Projects developed in Africa 4.2.1.1 List of ongoing special projects Total amount Amount to Partners (US $) Available in 2008 (US$) 115,000 601,250 502,866 Title Donor Funding period TL1: Improving tropical legume productivity for marginal environments in sub-Saharan Africa (African component) BGMF 2007-2010 115,000 TL2: Enhancing grain legumes’ productivity, production and the incomes of poor farmers in drought-prone areas of subSaharan Africa and South Asia: Seed Systems (African component) BGMF 2007-2010 2,866.084 Getting back to basics: creating impact-oriented bean seed delivery systems for the poor in Malawi, Mozambique and Tanzania McKnight Foundation 2007-2010 US$ 400,000 300,000 100,000 Improved Smallholder food Security, Nutrition and Income through Increased Production and Marketing of Climbing Beans. McKnight Foundation 2007-2010 US$ 400,000 300,000 100,000 Fighting Drought and Aluminium Toxicity: Integrating Genomics, Phenotypic Screening and Participatory Research with Women and Small-Scale Farmers to Development Stress-Resistant Common Bean and Brachiaria for the Tropics BMZ 2006-2009 Title Donor Funding period Increasing Food Security and Rural Incomes in Eastern, Central and Southern Africa through Genetic Improvement of Bush and Climbing Beans (African component) RF 1, 368,000 million seed systems 2005-2008 225 US 63,185 Total amount US 254,000 Amount to Partners (US $) - Available in 2008 (US$) 76,739 Supporting improved nutrition, food security and community empowerment for poverty alleviation – PABRA SDC 2007-2008 US 944,616 944,616 Supporting improved nutrition, food security and community empowerment for poverty alleviation – PABRA III CIDA 2003-2008 US5,298.787 2,231,057 4.2.1.2 Regional research subprojects under SABRN The bean research for development activities within SABRN are largely financed through PABRA, with funding from CIDA-Canada, and SDC-Switzerland. Within PABRA there is strong emphasis on international collaboration through networking both within (SABRN) and between networks (SABRN and ECABREN). Additional activities are funded through special projects - like the McKnight Foundation supported: bean seed systems and climbing bean projects. Through network collaboration different countries implemented research for development activities that contributed to the outputs and outcomes in the PABRA log frame in 2008. Activity 1.1.1 Complete germplasm collection, characterization and mineral analysis for all accessions 1.1.2 Conduct multi-location evaluations and national performance trials Value 1000 3000 650 500 1500 600 800 1500 1000 Country DRC Zambia Angola DRC Malawi Mozambique Swaziland Zambia Zimbabwe 1.1.3 Analyze candidate varieties for minerals and protein in some countries in SABRN 1000 1000 400 500 300 500 500 500 1000 800 500 Angola Malawi Mozambique Malawi Mozambique Zambia Zimbabwe DRC Malawi Mozambique Zimbabwe 1.1.4 Develop descriptors for candidate varieties 1.1.5 Conduct DUS in applicable and present for release: 3 countries in SABRN 226 Activity 1.1.6 produce breeder seed in countries that have released varieties 1.2.1 On-farm evaluations using PVS 1.2.2 Develop descriptors for the new bean varieties 1.2.3 Produce breeders' seed for the new and old bean varieties 1.2.4 Rejuvenate BILFA, Drought and disease nurseries 1.2.8 Combine resistance and select for pyramid (ALS, CBB) in ZA and BSM (MW and ZW) 1.2.10 Selection and testing of climbing beans adapted to mid-altitude (1200 -500 masl) 1.3.1 Identify export market potential including enhancing competitiveness of beans in SABRN 1.3.2 Conduct a bean cross-border trade study across South TZ, South DRC and Zambia 1.4.2 Continue with backcrossing program to improve commercial cultivars - Southern Africa 1.4.3 Strengthen capacity for application of MAS - Bunda 1.4.6 Produce adequate seed for all breeding materials 1.4.7 Production of foundation seed with partners 2.1.1 Validate effectiveness and farmers' acceptance and gender perceptions of promising ISFM and IPDM options with farmers 227 Value 900 1000 400 1000 800 800 2250 2800 1500 2600 3500 1500 3000 1000 700 700 3000 500 1700 3000 1000 1700 1500 1500 7500 1500 850 2000 2000 500 500 3000 1500 Country Angola Malawi Mozambique Swaziland Zambia Zimbabwe Angola DRC Malawi Mozambique South Africa Swaziland Zambia Zimbabwe Zambia Zimbabwe DRC Zimbabwe Angola DRC Malawi Mozambique Zimbabwe Malawi Zambia Angola Mozambique Zambia Zimbabwe Malawi Zimbabwe DRC Zambia 1000 4000 4000 1500 1500 1000 2000 3700 2000 1000 1850 1000 500 1000 1000 Malawi South Africa Malawi Malawi Zambia Zimbabwe DRC Mozambique Zambia Zimbabwe Angola Malawi Mozambique Zambia Zimbabwe Activity Value 2.1.2 Disseminate and promote accepted options with partners for technologies in 10 all countries 2.1.3 Perform cost-benefit tradeoffs analyses and adoption potential of these technologies 3.1.1 Organize, train and technically backstop community seed producers to bulk seeds 3.1.2 Update number, type location and activities of service providers 3.2.3 Facilitate production of promotional and information publications (including publications for SABRN website), transilations in each network 5.5.2 Conduct participatory formulation and evaluation of a basket of diets for improved nutrition - using biofort products 6.1.4. Conduct training workshops on nutrition assessment and linking nutrition support with agricultural extension 8.1.1 Inventory by year products (varietal and non-varietal), promotional materials TOTAL 1500 500 1500 2000 1000 1000 2000 2500 1000 250 1000 1000 1000 950 1000 1000 1000 1000 1000 1000 2000 2000 1500 1000 1000 800 1000 1750 3500 140050 Country Malawi Mozambique Swaziland Zimbabwe Malawi Zimbabwe DRC Mozambique Zimbabwe Angola DRC Mozambique Zimbabwe Angola DRC Malawi Mozambique Swaziland Zambia Zimbabwe DRC Malawi Swaziland Zambia Zimbabwe Swaziland Zimbabwe Angola Mozambique Harvest Plus funded activities Activity Value Country 1. Germplasm collection 4000 4000 3000 Malawi Tanzania Zimbabwe 2. Evaluation of fast trucks lines in various countries 2000 2000 2000 2000 2000 2000 2000 Angola Lesotho Malawi DRC Tanzania Zambia Zimbabwe 228 Activity Value Country 3. Breeding for high Fe combining with other biotic and abiotic stresses 2000 2000 2000 2000 Malawi South Africa Tanzania Zimbabwe 4. Supplies and small equipment: reagents, computer, printer 5000 SABRN TOTAL 40000 Contributor: R. Chirwa Collaborators: R. Buruchara, S. Beebe 4.2.1.3 Projects submitted, Proposals and Concept notes prepared Title Impact and development of Conservation Agriculture techniques in developing countries Donor European commission Supporting Nutrition and health, Food security, Environmental Stresses and Market Challenges that contribute to improve livelihood and create income resource poor small holder families in Sub –Saharan Africa.. CIDA Enhancing productivity, nutrition and incomes through improved marketable climbing bean and biofortified bean varieties Government of Kenya Improving Food and Nutrition Security, and Incomes of Smallholder Farmers in East and Central Africa through increased access to Markets and Technology Innovation Belgium Development Cooperation (BADC) 229 Comments Collaborators are: University of Applied Sciences Eberswalde, Germany; International Food Policy Research Institute (IFPRI), USA International, University of Ghana and Makerere University Participating CIAT technical team include: Enid Katungi and Roger Kirby Funding period 3 years Total amount US 220,000 (CIAT’s budget only) 2009-2013 7.8 million In review 2009-2011 $110,000 Unsuccessful 2008-2011 $3,148,632 Title Climbing out from poverty: Realizing the benefits from high yield potential of Climbing beans for smallholder farmers in Africa Donor JICA Comments Presented to donor in Jan 2008 Funding period Use of marker Assisted Selection in Developing Multiple Disease Resistant Bean Varieties in Malawi - Kirk House Trust Under review by donor (second round) (Research into Use): ‘Partnership Power’: Reaching women and the rural poor with new bean varieties in stress zones. DFID In review £ 282,000 (Food Crisis Proposal) Addressing the Food Crisis with Sustainable Solutions: Getting HighYielding and Adapted Bean Varieties into the Hands and Fields of Stressed African Farmers Swiss Development Cooperation In review $1,920,000 Making seed security more effective in emergency, chronic stress and food crisis contexts OFDA/USAID In review US 727,629 4.2.1.4 2009-12 Total amount US 150,000 New proposals approved Title Donor Supporting Nutrition and health, Food security, Environmental Stresses and Market Challenges that contribute to improve livelihood and create income resource poor small holder families in Sub –Saharan Africa SDC Funding period 2009-2011 230 Total Amount US 3.2 million Amount to partners US$ 2,221,384 Available in 2008 US$ 978,616 4.2.2 Projects developed in Latin America 4.2.2.1 List of ongoing special projects at Headquarters Title Donor 224.000 Amount to Partners (US $) 64.276 Available in 2008 (US$) 125.152 Reducing pesticide use and pesticide resistance in rice and beans in the Andean zone FONTAGRO 2006-2009 Fighting Drought and Aluminium Toxicity: Integrating Genomics, Phenotypic Screening and Participatory Research with Women and Small-Scale Farmers to Development Stress-Resistant Common Bean and Brachiaria for the Tropics BMZ 2006-2009 € 1,100,000 US153,907 US303,233 Biofortified Crops for Improved Human Nutrition – Harvest Plus Challenge Program (Yearly contracts) Gates Foundation World Bank DANIDA, Denmark CIDA 2003-2008 305,000 50,000 255,000 2004-2010 20,000,000 123,855 254,894 Integrated management of whiteflies in the tropics DFID 2005 - 2008 259.788 7.849 22.864 Increasing Food Security and Rural Incomes in Eastern, Central and Southern Africa through Genetic Improvement of Bush and Climbing Beans (Headquarters component) RF 2005-2008 US 254,000 - 10,750 Nutritional Improvement of the important pulse legume, the common bean, through the reduction of seed tannin content, for the benefits of people' diet in Africa and Latin America CIDA/Univ. of Saskatchewan 2007-2010 CAD 225,000 US 32,102 US 34,503 TL1: Improving tropical legume productivity for marginal environments in sub-Saharan Africa (Headquarters component) BMGF grant to GCP 2007-2010 115,000 473,944 TL2: Enhancing grain legumes productivity, production and income of poor farmers in droughtprone areas of sub-Saharan Africa and South Asia (HQ component) BMGF grant to CGIAR 1,104.056 197,701 Combating hidden hunger in Latin America: Biofortified crops with improved vitamin A, essential minerals and quality protein (AgroSalud) Funding period 2007-2010 231 Total amount 1,867,328 3,454.802 Title Donor Variedades de fríjol tolerantes al estrés abiótico de la baja fertilidad y la sequía, y a la sostenibilidad productiva y alimentaria de Centroamérica Red-SICTA, SDC Funding period Total amount 2007- 2008 246,100 Amount to Partners (US $) - Available in 2008 (US$) 45,450 4.2.2.2 Projects submitted, Proposals, and Concept notes prepared Funding period Total amount US 2008-2011 $889,350 Title Donor Comments Extracting the best from a desert species: Mining tepary bean for drought tolerance GCP Concept note not selected for full proposal development Basal root architecture and drought tolerance in common bean GCP Concept note and full proposal approved Integrated experimental and modeling approach to optimize soil water use under limited water GCP Concept note not selected for full proposal development A cross-legume phenotyping effort to identify common traits for superior adaptation to drought GCP Concept note under review 2009-2011 $459,020 Improving tolerance to drought stress in crops WUN Seed grant under review 2009 $48,000 2008-2011 2008-2011 $ 345,000 $905,060 4.2.2.3 New proposals approved Title Donor Biofortificación del Frijol Común (Phaseolus vulgaris L.) en Panamá con Micronutrientes” SENACYT – Panama Improved beans for Africa and Latin America DFID, UK Characterization of bean diversity in Central Europe GCP 232 Funding period Total amount Amount to Partners (US $) Available in 2008 (US$) 2008-2011 12,000 2008 120,690 - 120,690 2008-2009 9,000 - 9,000 7,000 Title Donor Funding period Total amount Dry bean improvement and marker assisted selection for diseases and abiotic stresses in Central America and the Caribbean” GCP 2008-2009 40,120 Capacity Building Needs regarding the Tropical Legume I (TLI) Project BMGF grant to GCP 2008-2009 Obtención y evaluación de Phaseolus vulgaris y Zea mays tolerantes a la sequía CYTED, Spain 2008-2009 $1,000,000 Development of a management system of Bemisia tabaci in paprika and pepper in the Cauca Valley Gracias MADR 2008-2011 58,288 Improvement of Chitti bean in Iran. SPII, Iran Iranian government 2008 18,423 233 Amount to Partners (US $) - Available in 2008 (US$) 40,120 5,904 5,904 - 29,906 16.560 - 18,423 Activity 4.3 Supporting breeding programs in NARS, regional networks, farmers’ associations, and CIALs with germplasm and technical knowledge Highlights: • • • • 4.3.1 More fixed lines with high yield potential and resistance to diseases and cultivars of commercial value for export market were distributed in a regional yield trial to various NARIs partners in different countries. Early generation selections of interspecific crosses for high iron have been realized in ICTA, Guatemala and are pending shipment to CIAT for analysis. Selections from EAP-Honduras have been analyzed and returned to the breeder there. High iron lines are in validation trials in Nicaragua and a variety could be released in the course of 2009. Nutritional analysis of Bolivian germplasm as support for the national genebank and breeding programs Rationale: As part of the Fontagro and Agrosalud funded projects on nutritional breeding, we have developed a strong collaboration with the Bolivian bean-breeding program and the corresponding genebank for this crop. Bolivia is a primary center of diversity for Andean genepool germplasm within the Andean region of South America. The country has the most significant problems of malnutrition on the continent (51% anemia in children under 5 year old and 33.1% in women 15 to 49 years old) and there is a need for higher micronutrients in the diet of most consumers. Common bean, especially biofortified varieties, could provide significant levels of iron and zinc to diets of consumers who eat 10 Kg or more per person per year. Common bean, however, is a relatively new grain crop for the country with most production in the eastern lowlands or sub-Andean valleys of Santa Cruz, some of it for local consumption and some for export to Colombia and other countries in the region. In this region, common bean is growing in importance compared to other pulses such as faba bean, lupines and lentils. Per capita consumption of dry beans has increased particularly among lowland producers and for people living in Santa Cruz and neighboring cities despite the fact that common bean was traditionally consumed previously as fresh green pods. The objective of this research was to screen some of the genotypes held in the national genebank collection at the “Centro de Investigacion Fito-ecogénetico” (which is part of the Pairumani Foundation) for baseline mineral content, with the aim of moving the best genotypes into the breeding program at the Universidad Autónoma Gabriel René Moreno. Materials and Methods: A total of 183 genotypes from 19 collection sites were harvested on the Pairumani research station in Cochabamba, Bolivia in March 2008 and sent to CIAT for mineral analysis. Seed mineral content was evaluated by grinding 3 g of previously crushed, oven-dried grain (two days at 40°C) into a fine powder using a modified Retsch mill with teflon chamber and zirconium grinding balls. Powder was transferred to 30 mL plastic vials and analyzed for both iron and zinc concentration measured in parts per million (ppm) with Atomic Absorption (AAS) spectrophotometry in the Analytical Services laboratory of CIAT. Data was analyzed with the software package Statistix v. 8.0 to determine descriptive statistics and correlation values between minerals. Results and Discussion: Average seed iron and zinc content for the genotypes from Bolivia were 56 ppm and 26 ppm, respectively. The range in iron concentration 34 to 88 ppm, while for zinc the concentrations ranges from 18 to 38 ppm (Table 113, Figure 60). The lines with highest iron are shown in Table 114 and included 19 genotypes with over 65 ppm iron. Zinc concentration was acceptably high in some of the genotypes from this group (for example A48 with 34 ppm, K17 with 35 ppm, J11 with 35 234 ppm and F23 with 38 ppm (Table 114). Correlation between iron and zinc concentrations were highly significant (r=0.52, P<0.0001). Table 113. Descriptive statistics for iron and zinc content in a 183 genotypes from the germplasm collection of Bolivia. Fe ppm 183 56 8.11 65.77 0.60 14.49 34 56 88 0.62 1.41 N Mean SD Variance SE Mean C.V. Minimum Median Maximum Skew Kurtosis Zn ppm 183 26 3.56 12.66 0.26 13.45 18 27 38 0.26 -0.04 Figure 60. Population distribution for iron and zinc content among 183 genotypes from the germplasm collection of Bolivia. 235 Table 114. Best iron levels found in each of 19 collections of genotypes from the germplasm bank of Bolivia. Germplasm collection G8 A19 G7 A11 A48 G1 A16 F23 J11 J23 B28 K2(B) A44 A49 J9 B15 J14 J49 Fe ppm 88 80 78 77 75 75 75 74 72 71 70 68 68 67 66 66 66 66 Zn ppm 33 31 33 30 34 33 28 38 35 33 31 24 27 30 30 28 30 30 J55 65 22 Conclusions and Future Plans: In general the results reflected the higher iron status, but lower zinc status of Andean grain types. While some high iron genotypes were found none were as high as for values seen for improved breeding lines from the biofortification program. Therefore the Bolivian genotypes should be considered as intermediate sources of high-mineral traits that can be used in breeding programs with improved varieties such as the NUA lines. Collaborators: 4.3.2 M.W. Blair, C. Astudillo (SBA-1, CIAT), T. Avila, G. Avila, R. Rios (Fund. Pairumani, Cochabamba); J. Ortubé, T. Anzoátegui, J. Padilla (UAGRM, Santa Cruz) Selection by NARS of segregating populations for nutritional quality 4.3.2.1 Progress in selection of cycle 2 populations in Central America In 2007 elite populations combining high mineral parents with sources of agronomic traits were distributed to collaborating partners in El Salvador, Guatemala, Honduras, and Nicaragua. These populations were created simultaneously with those reported under product 1. These have been selected for local adaptation and for resistance to diseases, especially BGYMV, and have reached the F5 generation in most cases. In Nicaragua the populations have been managed as bulks to date and advanced lines are not yet available for mineral evaluation. Grain is being shipped to Colombia for mineral analysis on the selected families made in El Salvador and Guatemala (Table 115). In the case of the populations selected in a highland environment in Guatemala, it is noteworthy the number of families that are derived 236 from P. polyanthus (G35575) and P. coccineus (G35999). These species are native to this region, they tend to express resistance to most diseases that are endemic to highland areas, and it was hoped that the populations would perform relatively better here. As reported above under Product 1, in Colombia G35575 has given the highest level of iron among all parents used in crosses. Therefore it will be especially interesting to see what levels of iron are obtained from these selections. Table 115. Populations segregating for mineral content, created in CIAT and selected locally by national program breeders in Central America. Country Population Number of selections El Salvador (SER 155 x RCB 234) x (MIB 451 x MIB 487) 37 Guatemala SBCF 16170 SBCF 16175 SBCF 16176 SDFZ 16180 SDFZ 16181 SDFZ 16191 SDFZ 16200 SBCF 16173 (SXB 414 x (ICTA Hunapu x G 35575-2P) (ICTA Hunapu x (ICTA Hunapu x G 35575-2P) (SXB 414 X (ICTA Altense x G 35999-8P) (ICTA Altense X (ICTA Altense x G 35999-8P) Hunapu x (G 23823E x ASC 81) Altense x (G 23818B x ASC 82) (ASC 86 x (G 23823E x G 35575-2P) (ASC 89 x (G 23823E x G 35575-2P) (ASC 91 x (G 23823E x G 35575-2P) (ASC 92 x (G 23823E x G 35575-2P) (ASC 88 x (G 23818B x G 35575-5P) (ASC 91 x (G 23818B x G 35575-5P) (ASC 92 x (G 23818B x G 35575-5P) (ASC 84 x (G 23818B x G 35575-5P) (ASC 87 x (G 23834E x G 35999-8P) (ASC 91 x (G 23834E x G 35999-8P) (G 23834E x G 35999-8P) x ASC 91 (ASC 85 x (G 23834E x G 35999-8P) 2 2 8 1 3 10 2 4 2 1 3 2 5 3 2 2 1 2 1 2 2 5 2 3 6 4 In the Panamerican School at Zamorano, 26 populations to produce red and black seeded cultivars were created on site in 2005 employing parental sources for high iron from CIAT. Selection on these continued during 2007 and 2008 (Table 116). More than 350 lines have been developed (Table 117). Parental materials that were combined with the high iron parents included sources for abiotic stress (drought and low fertility), and important diseases (BGYMV, angular leaf spot and web blight in particular). Samples of 28 advanced lines from the BIOFOR and FEZ populations were sent to CIAT and have been analyzed for minerals, as well as lines in regional trials. Interpretation of the data is pending. 237 Table 116. Chronology of the development and selection of populations and lines for high levels of minerals combined with agronomic characteristics. Zamorano, 2006-08. Planting season z Population No. FEZ 0521-0532 12 712 F3 FES 0551-0554 4 BIOF 1-6 6 BIOF11-14 4 z 06B 07X 07A/B 08A 08B 293 F4 52 F5 97 F6 47 F7 215 F3 73 F4 15 F5 27 F6 16 F7 291 F4 101 F5 20 F6 48 F7 17 F8 4000 F2 523 F3 248 F4 122 F5 X= Dry season (January-April); A= First season (May-August); B= Second season (September-December). Table 117. Numbers of lines developed from crosses between sources of high minerals and elite lines for agronomic and commercial traits. Zamorano 2008. Pedigrees Characters Families Simples: BIOF 1 FE14584-1/PRF9653-16B-3 Fe, Zn, MD, VA, VC 1 F7 BIOF 2 NH21566-7/PRF9653-16B-3 Fe, Zn, MD, VA, VC 6 F7 BIOF3 FE14584-2/EAP9503-32B Fe, Zn, MD, VA, VC 0 F7 BIOF 4 G23818B/EAP9503-32B Fe, Zn, MD, VA, VC 12 F7 BIOF 5 G23823E/EAP9712-13 Fe, Zn, MD, VA, VC 18 F7 BIOF 6 G23834E/ALS9952-27R Fe, Zn, MD, VA, VC 11 F7 FEZ 0521 Amadeus 77//BIOF 1 Fe, Zn, MD, VA, VC 5 F6 FEZ 0522 Cardenal//BIOF 3 Fe, Zn, MD, VA, VC 12 F6 FEZ 0523 DEOHRO//BIOF 4 Fe, Zn, MD, VA, VC 9 F6 FEZ 0524 SRS 6-6//BIOF 1 Fe, Zn, MD, VA, VC, SQ 4 F6 FEZ 0525 MR14292-2//BIOF 3 Fe, Zn, MD, VA,VC, SQ 8 F6 FEZ 0526 MH 2-2//BIOF 1 Fe, Zn, MD, VA, VC, MH 3 F6 Fe, Zn, MD, VA, VC 4 F6 Fe, Zn, MD, VA, VC, MH 10 F6 FEZ 0529 PRF9924-50N//BIOF 5 Fe, Zn, MD, VA, VC 12 F6 FEZ 0530 BCN20-02-94//BIOF 3 Fe, Zn, MD, VA, VC 16 F6 FEZ 0531 Amadeus 77//BIOF 2 Fe, Zn, MD, VA, VC 10 F6 FEZ 0532 Cardenal//BIOF 6 Fe, Zn, MD, VA, VC 4 F6 Triples: FEZ 0527 ALS9952-27R//BIOF 5 FEZ 0528 MH 43-3//BIOF 1 238 Pedigrees Doubles: Characters Families FES 0551 SRS2-4//BIOF 1 Fe, Zn, MD, VA, VC, SQ 3 F6 FES 0552 SRS2-5//BIOF 3 Fe, Zn, MD, VA, VC, SQ 7 F6 FES 0553 SRS2-18//BIOF 1 Fe, Zn, MD, VA, VC, SQ 6 F6 FES 0554 SRS2-20//BIOF 3 Fe, Zn, MD, VA, VC, SQ 11 F6 BIOF 11 FEZ 0521//ALS 0531 Fe, Zn, MD, VA, VC, MA 44 F4 BIOF 12 FEZ 0522//ALS 0532 Fe, Zn, MD, VA, VC, MA 34 F4 BIOF 13 FEZ 0528//ALS 0545 Fe, Zn, MD, VA, VC, MA 75 F4 BIOF 14 FEZ 0529//ALS 0546 Fe, Zn, MD, VA, VC, MA 95 F4 4.3.2.2 Selection of populations in Brazil The bean program of EMBRAPA in the CNPAF station in Goiania, Goias has continued to pursue the development of drought tolerant lines with high mineral content, and has a very effective selection site for drought evaluation in Porangatu. Selection has progressed within populations and families for drought and minerals, both those created in house and those sent from CIAT-Colombia. Numbers and type of selections are summarized below: • 28 F3.5 families from CNPAF populations among 16 elite parents • 113 F1.4 families from 19 populations from CIAT and 6 from CNPAF • 48 F1.5 families from 19 CIAT populations • 90 F8.9 lines from 35 lines sent from CIAT • 13 advanced lines sent from CIAT • 3 interspecific populations from CIAT for drought tolerance • 68 F4 families from CIAT for high minerals The EMBRAPA laboratory for mineral analysis has also reported atomic absorption data on two universal checks sent from CIAT that approach the values with ICP quite satisfactorily (61 with atomic absorption versus 65 with ICP for low iron; 94 versus 102 for high iron). 4.3.3 Evaluation of lines from the 1st cycle of selection in advanced yield trials and validation in Central America Although plans were made during the annual meeting of the PCCMCA in April, 2008 to execute a broad evaluation of advanced lines as selected in previous years, very few trials were salvaged given the very excessive rainfall throughout the growing seasons. Nearly all trials were lost in El Salvador. 4.3.3.1 Nicaragua Nicaragua is the country where the testing of advanced lines in regional trials has advanced the most. In a regional trial the selections from populations for high iron were compared with local check INTA Rojo (Table 118). Across all five environments only two lines compared favorably with INTA Rojo. But in the less favorable environments (Boaca and Las Segovias) with yields that are more typical of those obtained by farmers, more lines yielded comparably or with a modest advantage. However, the widest advantage in relation to INTA Rojo was in the trial in San Isidro, a high yield environment. In Nicaragua it is common for genotypes to present local or regional adaptation, and the varietal release procedure there permits the registration of cultivars for regional use. Grain has been recovered from these trials, and is awaiting chemical analysis in CIAT. 239 Table 118. Yield of lines (kg ha-1) selected from populations with high iron parents in five regions of Nicaragua. 194-15488-30 Carazo (La Compania) 2171 199-15488-32 Carazo (Aguacate) 2110 Boaca (Sta Lucia) 1646 Las Segovias 1274 Sn Isidro Mean 2426 1925 2192 1528 1090 2626 1859 INTA Rojo (check) 2224 2653 1308 1174 1928 1857 181-15488-52 2072 2329 1360 1128 2311 1840 196-15488-30 2142 2355 1278 1054 2150 1796 126-15357-30 1971 2355 1676 962 1887 1770 15357-30 2131 1883 1390 1404 2001 1762 189-15488-30 2119 1976 1386 1178 2152 1762 187-15488-39 1872 2314 1534 1154 1897 1754 MIB 397 1768 1693 1368 1156 2618 1721 428-15094-39-4 2060 2113 1422 1224 1576 1679 202-15488-39 1635 1842 1480 1046 2359 1672 MIB 438 2193 2125 1178 1296 1541 1667 1909 1386 1184 2134 1653 519-15089-22-55 190-15488-30 1687 1693 1240 1320 2108 1610 MIB 396 1579 2097 1056 1038 1970 1548 197-15488-30 2088 1762 1332 834 1428 1489 MIB 395 1728 1732 850 936 1478 1345 In the region of Las Segovias two selections from the AgroSalud project are being evaluated in validation plots on farm (approximately 100 m2 per line). DFSZ 15094-43-5 has been tentatively named INTA Nutritivo, and DFSZ 15094-39-4 has been designated INTA Ferroso. The latter line has performed better in these trials. These names do not imply that these materials have been released, but rather they have been applied to make the lines “user-friendly” in collaboration with farmers. INTA has the intention of releasing a variety in the course of 2009. 4.3.3.2 Honduras In Honduras 20 trials of the 4 most promising lines were distributed to DICTA, FAO, Rural Reconstruction, and FIPAH (an NGO working with farmer research committees). Data are still pending at the time of writing. Grain was recovered from 4 trials for chemical analysis in CIAT (results pending). The yield trial reported from Zamorano station was planted in very favorable conditions (Table 119), and even here, variability in the field was great. Selected lines did not yield as much as elite cultivar Amadeus, but presented a wide margin over local cultivar “Rojo de Seda”. 240 Table 119. Yield, agronomic and commercial value of lines and checks in the regional high mineral nursery. Zamorano 2008. Line Amadeus 77 703-SM 15216-11-4 523-DFBS 15092-04-4 519-DFBS 15089-22-5 428- DFSZ 15094-39-4 Seda LSD 0.05 Yield (kg ha-1) Agronomic value (1-9) Commercial value (1-9) 4,940 4,019 3,646 3,358 3,320 2,474 1,312 3.6 5.3 4.7 3.3 4.7 8.0 0.7 4.0 3.0 4.0 3.5 4.0 2.0 1.0 The 4 lines were subjected to a systematic evaluation of acceptability traits (grain color, textura and viscosity) in Food Science Laboratory of Zamorano (Table 120). Color was determined by the Colorflex Hunter L*a*b. Texture was estimated by the Instron Series IX version 8.12.00. Grain is considered cooked if it requires less than 500 Newtons to be compressed in the texturometer. By this criterion, the 4 lines showed an optimal cooking time of 60 minutes. Minimum broth viscosity should be 13.2 cP at 60 minutes of cooking; by this criterion the lines 523- DFBS 15092-04-4, 429-DFSZ 15094-39-4 and the check Seda produce the best broth. Table 120. Results of the analysis of color, texture, and broth viscosity on four lines and two checks. Zamorano, 2008. Line Color (1-9) 703-SM 15216-11-4 523-DFBS 15092-04-4 519-DFBS 15089-22-5 428- DFSZ 15094-39-4 Amadeus 77 Seda 1 1 1 1 2 1 Texture (N) Viscosity (cP) 300.67 335.33 316.67 345.67 298.67 249.33 11.43 14.17 12.33 13.33 12.70 13.67 4.3.3.3 Brazil Evaluations in Brazil of materials sent from CIAT are normally 1-2 years behind those in other countries, due to the need to pass seed through laboratory and greenhouse quarantine, and then to increase seed coming out of the greenhouse. In 2008 it was possible to report data on the High Mineral Nursery, and to correlate data with those from Central America. Collaborators: Juan Carlos Rosas (EAP), Julio César Villatoro (ICTA), Julio Molina (INTA), Aurelio Llano (INTA), Maria José Peloso (EMBRAPA), S. Beebe, R. Urbina 241 4.3.4 Evaluation of SU91 as a molecular marker for CBB resistance in common bean for Southern Africa Rationale: Common bacterial blight (CBB) is an important foliar and seed-borne disease of Andean beans grown in tropical lowland and mid-elevation areas of Africa, Central America and the Caribbean. The disease is also important in the subtropic and temperate regions of the Americas and Africa during hot, humid summer weather. The disease is caused by the pathogen Xanthomonas axonopodis pv. phaseoli (Xap), which is widespread and part of a complex of Xanthomonad bacterial pathogens attacking many broadleaf and vegetable crops. Races of the Xap pathogen are not recognized. The current level of CBB resistance in Andean genotypes is insufficient and most current Andean varieties are highly susceptible especially in Southern Africa. With this in mind we have implemented marker assisted selection for the major QTL for common bacterial blight resistance tagged by a SCAR marker developed at CIAT called SU91 as part of the breeding program and within an M.Sc. program for SABREN/Univ. of Zambia. As sources, we are using mainly VAX and XAN lines developed in the 1990s, but also are testing RMX lines developed by us in 2004 from crosses of Caribbean type red mottled beans with the sources VAX3 and VAX6. Materials and Methods: Genotypes: A total of 28 genotypes were used in initial testing of the SU91 marker. The control genotypes were the resistance sources RMX2, RMX19, RMX20 (Andean) and VAX3, VAX6 and XAN159 (Mesoamerican), while the other genotypes were the red, cream mottled or red mottled advanced lines CAL143, CMB106, DRK149, RAA21, RMA68, RMA70, RMA71 and RMC57. In addition the drought adapted genotypes SAB575, SAB581, SEQ11, SEQ1003, SEQ1004, SEQ1006 and SEQ1027 were tested as well as the BCMNV sources used in crosses with these previous parents BRB264, BRB265, BRB266 and BRB267. DNA extractions: Two types of DNA extraction were used, namely alkaline extraction (microprep) and proteinase K (miniprep) derived DNA. Once these parental genotypes were evaluated the SU91 marker was tested on a set of gamete selection F1 plants from semester 2008b derived from simple, double and multiple crosses between the parents and using alkaline extraction. Initial marker assisted selection targeted those crosses with VAX3 and VAX6 where the SU91 marker seemed to be more reliable. Results and Discussion: SU91 was found to amplify multiple background bands with alkaline extraction DNA as template compared to miniprep DNA as template when using 2.5 mM MgCl2 (Figure 61). The strongest band was found for the genotypes VAX3, VAX6 and XAN159 at the expected size of 700 bp, but the SU91 marker was also found to be sensitive to template DNA concentration with a weak band of the expected size appearing when concentrations were high in susceptible genotypes, especially with alkaline extraction DNA. Given these initial results, we tried two different dilutions of miniprep and alkaline extraction DNA for VAX3, VAX6 as resistant controls and MAR1 as a susceptible control. The results showed that a dilution of 1:9 produced a strong band in the resistant genotypes and no band in the susceptible genotypes; while a dilution of 1:14 produced a weak band only for VAX6 not for VAX3 or the susceptible check (Figure 62). We then attempted marker assisted selection using alkaline extraction DNAs from a set of 331 gamete selection from the Palmira 2008B season that were used for leaf tissue harvesting 18 days after planting. Amplification of the template DNA was based on a 1:9 dilution and the same PCR conditions as used for the control genotypes. Since the marker is dominant and in cis orientation with the resistance QTL only those plants containing the 700 bp SU91 band and no other background bands were selected. Given this stringent evaluation, a much lower number of positive genotypes (2.1%) were identified than expected based on the pedigrees of the triple, double or multiple crosses evaluated. 242 ladder 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 SU91 700 bp band Figure 61. Sample of SU91 testing on drought tolerance and CBB resistance control genotypes: MAR1 (lanes 1 and 2); XAN159 (3 and 4), SEQ1004 (5 and 6), SAB568 (7 and 8), VAX3 (9 and 10), VAX6 (11 and 12), SAB514 (13 and 14), SEQ1027 (15 and 16). All odd-numbered lanes are the results of PCR amplifications from miniprep DNA while all even-numbered lanes are from alkaline extraction DNA (1:4 dilutions). 1 2 3 4 5 6 7 8 Figure 62. Dilution tests for the SU19 markers: dilution of 1:9 tested on alkaline extraction DNA of MAR1 (lane 1), VAX3 (lane 2), and VAX6 (lane 3), and on miniprep DNA of the latter VAX genotypes (lanes 4 and 5, respectively); followed by dilution of 1:14 tested on alkaline extraction of MAR1 (lane 7), VAX3 (lane 7), and VAX6 (lane 8). 243 Conclusions and Future Plans Initial testing of the SU91 marker shows that it is a difficult marker to use on the widely divergent DNA types we routinely analyze for other markers, therefore further trouble shooting will be necessary, or a new marker will need to be identified. The difficulty in using this marker may derive from the fact that the VAX lines as well as some of the XAN lines (such as XAN159 and XAN309) which are the sources of SU91 tagged resistance were produced by inter-specific hybridization between common bean (Phaseolus vulgaris L.) and tepary bean (P. acutifolius Gray) and evaluated in Mesoamerican genepool material, therefore the PCR amplification product may be difficult to detect in some genetic backgrounds especially from the Andean genepool. Collaborators: M.W. Blair, H.F. Buendia (SBA-1, CIAT) L. Kalalokesya (Univ. of Zambia), R. Chirwa (CIAT-Malawi) 4.3.5 Distribution of seed from CIAT Headquarters Tables 121, 122 and 123 show a summary of Bean Breeding, Andean Breeding and other nurseries distributed from CIAT headquarters to partners and collaborators. Table 121. Nurseries distributed by the Mesoamerican bean breeding section. Purpose Institution/ Collaborators Country 65 Local Evaluation Ministry of Agricultura Roberth Shank Belice Bean lines tolerant to low fertility 2 Local Evaluation Root Biology Center, South China Agricultural Univ. Xialong Yan China Black and red lines, high minerals Black line, high minerals Black lines tolerant to drought Bean lines tolerant to drought and high minerals 8 Local Evaluation Corpoica Cesar Adriana Tofiño Colombia 1 18 7 Local Evaluation Local Evaluation Local Evaluation Black line 1 Local Evaluation Semillas Camerún Gilberto Bastidas Colombia MIB 488 1 Local Evaluation UMATA, Palmira Sandra Salazar Colombia Andean bean lines tolerant to drought 123 Local Evaluation INIAP Esteban Falconi Ecuador Bean lines tolerant to drought and low fertility 24 Local Evaluation National Research Centre Botany Department Magdi Abdelhamid Egypt Description Black and red lines tolerant to drought No. of lines 244 Table 121. cont´d. Purpose Institution/ Collaborators Country 312 Local Evaluation Melkassa Agricultural Research Center Teshale Assefa Ethiopia RILs, MesoAmerican and Andean bean lines tolerant to drought, low fertility, bc-3, ascochyta, and CBB, ALS and anthracnose differentials 192 Local Evaluation Awassa Agricultural Research Center Asrat Asfaw Amele Ethiopia Bean lines tolerant to drought, Fe, low fertility and bc-3 F2 populations drought 85 Local Evaluation Haiti 8 Local Evaluation ORE Eliassaint Magloire University of Nairobi Paul Kimani RILs of BAT 881x G21212 and G 40159 101 F2 populations drought Andean bean lines tolerant to drought RILs of DOR 364 x BAT 477 15 114 Local Evaluation Chitedze Agric. Res. Stat. Rowland Chirwa Malawi Black, white and red bean lines tolerant to drought 21 Local Evaluation INIFAP Fermín Martinez, Alfredo Vargas Mexico Black and red bean lines tolerant to drought, with bc-3 108 Local Evaluation INIFAP Jorge Acosta Mexico VAM (High minerals nursery) 43 Yield evaluation and Nutritional studies IDIAP Emigdio Rodríguez Panama RILs of DOR 364 x BAT 477 100 Local Evaluation Mayagüez University Timoty Porch Puerto Rico Bean lines tolerant to aluminum, drought, low fertility; or with bc3, anthracnose, rust and ALS differentials 394 Local Evaluation ISAR Felicite Nsanzabera Rwanda High mineral bean lines 15 Local Evaluation Institute of Food Science and Nutrition Nicolai Petry, Monika Egli Sweden ALS bean lines 2 Research USDA-ARS Marcial Pastor-Corrales USA Description F4.5 RILs, Mesoamerican and Andean bean lines tolerant to drought, low fertility; and bc-3, ALS and Anthracnose differentials; Zabrotes sources No. of lines Kenya 100 245 Table 122. Nurseries distributed by the Andean bean breeding and Germplasm Characterization section. Description No. of lines BAT 93 , G19833 No. of seeds 1 kg Purpose Institution/ Collaborator Country 2 Mutagenesis Agriculture and Biotechnology Lab. IAEA/Rownak AfzaChikelu Mba Austria RIL s of G2333 x G19839 Accessions and bean lines G2333 and G19839 100 100 1 kg 76 19 2 BNF Evaluation Centre of Microbial And Plant Genetics/ Jos Vanderleyden, Lara Ramaekers Belgium Bush bean lines Climbing bean lines 1 kg 500 g 6 4 Nutritional quality Agronomic testing CORPOICARionegro/ Alejandro Navas Colombia Accessions and bean lines 250 g 30 Agronomic testing CORPOICAValledupar/ Adriana Tofiño Bush and climbing bean lines 250 g 9 Adaptation FIDAR/ José Restrepo 30 9 Disease screening Univ. Nacional de Colombia Luz Nayibe Garzón, Gustavo Ligarreto Climbing bean lines 250 g 12 Adaptation testing at low altitude Institut National Pour L Etude Et La Recherche/ Jean Paul Lodi Lama, Bernard Vanlauwe 4 lines (5 accessions) Acutifolius, coccineus, and lunatus accessions 120 g 20 9 8 Nutritional evaluation 100 30 30 30 100 12 12 124 Drought evaluation Crossing block 100 204 Adaptation trials 40 138 Adaptation trials 40 234 Adaptation trials Bean accessions RILs of DOR364 x BAT477 Anthracnose differentials ALS differentials Bush and climbing bean accessions (reference collection) Kenya and Ethiopia accessions DOR390 x (DOR390 x (G 24423 x DOR390)) lines Accessions (reference collection), 3 reps. 2 trials Adaptation trials 246 Agonomique Univ. of Aarhus, Cristina Cvitanich Awassa Agricultural Research Center/ Asrat Asfaw Amele DRC Denmark Ethiopia Table 122. cont´d. Description RMA lines MBC lines F2 Individual Selections Crosses for Zabrotes with Awash Melka No. of seeds 40 40 30 No. of lines 73 138 332 F2 and F3 populations of crosses for CBB 350 Bush and climbing bean accessions (reference collection) G19833 x BRB lines RIL s of G2333 x G19839 30 219 30 30 181 86 NUA lines NUA lines, 2 reps. 4 trials 50 40 Institution/ Collaborator Adaptation trials Melkassa Agricultural Research Center/ Teshale Assefa Field selection Based on MAS evaluation Country Field selection Thesis trials CIAT-SABRN/ Rowland Chirwa Malawi Root architecture screening IIAM/ Magalhaes Miguel, Celestina Nhagupana Jochua Mozambique 50 64 Adaptation trials Nutritional and Quality evaluation IDIAP/Emigdio Rodríguez Panama 500 g 5 Nutrition analysis CIP/ Wolfang Gruneberg Peru RMA, BRB, and DRK lines, 2 reps. MBC, BRB, and DRK lines, 2 reps. 40 51 Disease screening Uganda 40 96 Agricultural Research Institute Robin Buruchara RILs of G40022 x G40186 RILs of G40186 x G40022 RAD-CERINZA x (RADCERINZA x (G10022 x RAD-CERINZA)) 50 50 79 83 Disease screening University of Nebraska/ James R Steadman USA 50 145 P. acutifolius accessions 30 42 Genetic analysis Virginia State University/ Harbans L Bhardwaj Andean and Mesoamericans accessions RMC bgm-1 + lines DOR 390 x (DOR 390 x (G 24423 x DOR 390)) 20 20 Thesis evaluation 30 20 26 100 INIA-CENIAP/ Miguel Alexis Adrian Perez NUA lines 36 Purpose 247 Venezuela Table 123. Other nurseries distributed Description Macrophomina and fusarium oxisporum Isolates 4.3.6 No. of lines 1 Purpose Institution/ Collaborator Carlos Huertas, Univ. Nacional de Palmira Academic studies Country Colombia Distribution of germplasm within the ECABREN bean network No. of nurseries sent No. of entries Bioforts and Bilfa V lines 2 31 Pythium Root rot 1 Root rot Nursery of 68 Pythium Root rot BC- S5 progenies Root Rot resistant lines, Advanced small, medium and large white lines (16 April 2008) TL II materials Description Purpose Recipients Mr. Gabriel Diasso Burkina Fasso 22 Adaptation and selection trials Research L. Nounamo Cameroon 1 7 Research L. Nounamo Cameroon 1 12 Research L. Nounamo Cameroon 1 17 Adaptation trials in WECABREN Lodi Lama, INERA-Mvuasi, DRC 25 346 1 24 Dr Setegn Gebeyehu R. Otsyula Ethiopia Pythium Root rot RILs Drought phenotyping and selection Research F8 lines having both ALS and Pythium root rot resistance BCS5 F5 (GLP 2 x RWR 719) progenies ALS differentials 1 3 Research R. Otsyula Kenya 1 9 Research R. Otsyula Kenya 1 12 Research Dr Isabella Wagara Kenya Released bush and climbing bean varieties TL II regional nursery (March 2008) 1 6 Mr. Jarvis Njoroge Kenya 25 340 On-farm testing and seed multiplication Drought phenotyping and selection trials John Msacky, SARI Arusha, Tanzania TL II drought nursery 27 620 1 5 RILS, Core Collections & Regional Varieties Anthracnose differentials 3 201 1 12 TL 1 drought phenotying and selection trials M.Sc. research F2 populations from Anthracnose program 1 12 M.Sc. research Dr Roland Chirwa CIAT-SABRN Dr Kiarie Njoroge Univ. of Nairobi Festo Ngulu, SARI Kelvin Kamfwa, Makerere Univ. Dr Kelvin Kamfwa Makerere Univ. Malawi Released varieties Drought phenotyping and selection trials Seed multiplication 248 Country Kenya Kenya Arusha, Tanzania Uganda Uganda 4.3.7 Exchange of germplasm in Southern Africa Bean Research Network (SABRN) Rationale: Some national programs within the SADC region (Angola, south D. R. Congo, Lesotho, Mozambique and Swaziland) still do not have adequate personnel to support breeding programs of their own. The SABRN co-coordinates regional germplasm nurseries and trials, which contain improved lines and released cultivars with the aim of sharing germplasm within the network so that each national program or private sector can benefit from the research that is carried out by others in the region. Materials and Methods: Various countries within SABRN grouping requested for specific nurseries. These nurseries were organized either by market class, or production constraint or plant growth habit. Such nurseries serve as sources of germplasm with good attributes that might be useful to NARS partners. Table 124 below shows the distribution list of the nurseries in the 2008. Table 124. List of nurseries and trials that were distributed in the SABRN, 2008 season Description No. of nurseries sent No. of entries Purpose Recipient Country Southern Africa Regional Bean Yield Trial (SARBYT) 13 20 yield evaluation across countries Southern Africa Regional Bean Evaluation Nursery (SARBEN) 13 100 Adaptation of lines to different environments Bean Improvement for low soil fertility adaptation (BILFA) Drought nursery –small seeded bean lines 1 75 Adaptation to low soil fertility 3 135 Evaluation under drought stress Angola, Swaziland, Zambia Drought nursery – large seeded bean lines 9 21 Evaluation under drought stress Bean stem maggot lines 8 5 Screening for resistance to BSM Sugar bean nursery 3 73 Screening for adaptation Calima bean lines 2 213 Screening for adaptation Angola, Congo Democratic, Lesotho, Malawi, Mozambique, Swaziland, Tanzania, Zimbabwe Angola, Malawi, Mozambique, Swaziland Angola, Lesotho, Mozambique Angola, Mozambique 249 Angola, Congo Democratic, Lesotho, Malawi, Mozambique Soth Africa, Swaziland, Tanzania, Zambia, Zimbabwe Angola, Congo Democratic, Lesotho, Malawi, Mozambique, South Africa, Swaziland, Tanzania, Zambia, Zimbabwe Angola Table 124. cont´d. Description Navy bean lines No. of nurseries sent 6 No. of entries Purpose 45 Screening for adaptation Screening for adaptation Recipient Country Angola, Congo Democratic, Malawi, Mozambique, Tanzania, Zambia Angola, Congo Democratic Angola, Malawi, Mozambique, Swaziland Angola Yellow bean lines 2 3 Bio-fortified bean lines fast/track evaluation 4 43 Yield evaluation Bio-fortified small seeded bean lines Bio-fortified large seeded bean lines Cream/khaki bean lines 1 72 Yield evaluation 3 60 Yield evaluation 3 24 Yield evaluation and adaptation Angular leaf spot nursery 3 45 Observe reaction to ALS Medium climbing bean lines Heavy climbing bean lines 4 13 4 30 Yield evaluation and adaptation Yield evaluation and adaptation to lower attitude warmer environment TL-I regional bean lines evaluation nursery 2 121 Yield evaluation and adaptation Malawi, Zimbabwe, TL-I reference collection evaluation nursery 2 100 Yield evaluation and adaptation Malawi, Zimbabwe, TL-II- drought evaluation nursery 2 772 Screening for drought Zimbabwe, Malawi 4.3.7.1 Angola, Malawi, Mozambique Angola, Congo Democratic, Tanzania Angola, Congo Democratic Angola, Mozambique Angola, Mozambique Southern Africa Regional Bean Yield Trial (SARBYT) Material and Methods: Each set of SARBYT contained 19 test varieties selected for different attributes, including good yield potential, acceptable grain market types and potential for adaptation to various bean production environments across countries in the SADC region. Each country added a local control to make a total of 20 entries. The trial was laid out in a randomized complete block design with 4 replications and data were collected on rainfall, weather, soil characteristics, diseases and grain yield. Results and Discussion: During this reporting period many countries in the SADC region experienced bad weather conditions (drought), which adversely affected productivity of the bean crop. As a result many sites were abandoned, and data is reported from a selected few sites. Disease data were collected on angular leaf spot (ALS), common bacterial blight (CBB), ascochyta blight (ASC), floury leaf spot 250 (FLS), and rust. The ALS disease pressure was highest at Bembeke, Malawi with score rating as high as 8 in some varieties, but there were a few varieties which showed reasonable resistance to ALS with such diseases score ratings of 2-3) on the following varieties: MC12832-129-11, GCI-CAL-172-AR, MN12686-15, CAL143, GCI-LR-171, VTTT923/10-3, MR13557-16-7 and MC13832-129-1. The pressure for CBB was highest at Uyole, Tanzania, where the most susceptible variety had a score of 8. Floury leaf spot was the other disease where some varieties recorded high scores of 8 to 9, at Bembeke, Malawi. In general, a good number of varieties had good resistance to ALS, CBB, ASC and rust, but were susceptible to FLS. The only two varieties which were resistant to all diseases including FLS were: 12832-129-11 and 13832-129-1 (Table 125). Grain yield data showed that Lubumbashi (D. R. Congo), Greytown (South Africa) and Harare (Zimbabwe) were badly affected by drought, and the average site means were less than 1000 kg ha-1, whereas Chitedze and Bembeke (Malawi), Misamfu (Zambia) and Uyole (southern highlands of Tanzania) were less hit by drought and they had better site means, above 1000 kg ha-1. There were 2 carioca varieties (MC13832-129-1 and MC12832-129-1), both with medium seed size, which were among the top yielding varieties across locations. One large red seeded variety (VTTT 924/10-4) also showed good yield potential across sites, with an average grain yield above 1400 kg ha-1, which was similar to the yield of the carioca varieties, and the standard check variety, CAL143. Among the least yielding varieties across locations were those bean varieties which were introduced by ACOS, a commercial firm which exports beans across continents, with the intention of identifying some varieties which could be promoted in SABRN for them to export to Europe. These are the varieties with commercial grain types, which were mostly in cranberry (sugar) and white market classes, but they seem not to be adapted to the environments in most countries, and they are also susceptible to ALS, CBB, FLS and rust in most sites – calling for future breeding activities to incorporate genes for adaptation and resistance to prevalent diseases in the region, into the varieties of commercial value for export to European markets. Contributor: R. Chirwa Collaborators: NARIs Bean Research Teams within SABRN; S. Beebe, R.Buruchara, P. Kimani, M.Blair 251 Table 125. Performance of bean varieties across countries in Southern Africa Bean Yield Trials, 2008. Identity ALS MS 3 3 3 3 3 3 4 2 3 3 4 3 4 3 3 3 5 4 3 5 3 Disease Scores CBB MS LB GT 1 1 3 1 1 3 1 3 4 1 1 2 1 1 4 3 1 3 2 2 3 1 1 3 4 2 3 2 1 3 1 2 3 3 2 3 1 2 3 4 1 3 1 1 2 6 3 3 1 2 3 3 2 4 1 3 3 8 2 3 2 2 3 Grain yield kg ha-1 ASC BB 3 3 3 4 3 3 3 3 4 4 3 3 2 3 4 4 4 3 4 4 3 FLS BB 1 1 7 5 7 7 6 6 7 6 6 7 7 7 5 9 8 6 9 8 6 Rust GT UY 1 1 1 1 1 1 2 1 2 3 1 2 3 2 1 1 4 2 2 2 1 1 2 1 2 2 1 1 1 1 2 5 4 8 6 2 5 5 6 7 2 2 Size (g) 29 27 45 25 41 41 Grain Color BB UY BB CZ BB MS LB GT HR UY Mean MC 12832-129-1 2 1 1 2375 1852 1990 766 822 572 2237 1516 Carioca MC 12832-129-11 2 1 1 2339 1418 1831 629 661 685 2042 1464 Carioca VTTT 924/10-4 4 4 3 2319 1437 1796 763 709 995 2083 1450 Red MR 13557-16-7 3 1 1 2086 1473 1855 635 1200 900 1788 1415 Red CAL 143 2 4 1 2233 1517 1510 661 750 684 2117 1363 Calima GCI-CAL-172-AR 3 5 1 2114 1457 1603 706 628 827 1972 1333 Calima CONTROL 4 4 3 1910 1173 1616 566 1244 561 2004 1316 LOCAL MN 12686-15 2 1 1 2062 1088 1776 802 834 523 1926 1287 30 Red BOUNTY 7 6 6 1911 700 1521 508 589 884 2135 1238 41 Sugar VTTT 924/10-7-1-1 4 5 1 2462 1236 1543 463 616 693 1596 1230 47 Sugar MR 14215-9 4 1 1 2124 1363 1462 542 784 671 1601 1217 24 Red RMA 18 4 4 1 2056 1226 1568 548 583 593 1785 1209 51 Calima VTTT 923/10-3 3 3 1 2260 1120 1256 843 550 713 1585 1180 49 Sugar RMA 20 4 4 3 2015 949 1190 714 889 551 1800 1158 44 Calima GCI-LR-171 3 3 1 1975 997 1692 516 639 429 1365 1088 42 Red CRAN BUSH-ARG. 8 7 7 1969 852 1054 620 689 822 1093 1015 39 Sugar CANNELIN ARG 8 5 5 1138 840 1252 643 816 570 1274 969 42 White CRAN VINE (USA) 4 7 3 1501 1173 1045 546 522 593 1383 934 51 Sugar INNER MONG 7 4 3 1200 766 1098 542 676 703 1108 908 32 Sugar CANNELIN EGY 8 5 7 1339 826 972 573 514 736 1129 843 42 White Mean 4 4 3 1969 1173 1481 629 736 685 1701 1207 SE +/236 90 264 107 85.9 92 96.1 196 CV% 23.9 15.3 17.8 32 23.4 27 11.3 23.2 LSD (5%) 670 258 374 302 244 261 272 Sites: BB-Bembeke-Malawi, CZ-Chitedze-Malawi,MS-Misafu-Zambia, LB-Lubumbashi-Congo, GT-Greytown-South Africa, UY-Uyole-Tanzania, ZM-Zambia, HR-HarareZimbabwe 252 4.3.8 Varietal releases in Latin America and Africa (2006-2008) Latin America: Country Name Origin Year of release Costa Rica Chánguena Nicaragua UCR 55 INTA precoz INTA Seda (Pre-release) MR 13652-39 (Bribri x (VAX 1 x RAB 655) Línea NJBC-20601-1-CM(71) SRC 2-18 DOR 364 x Rojo Seda 2006 2007 2006 2006 Africa: Country Varietal name Line code DRC (west) G 59/1-2* VCB 81013* LIB 1* Kiangara* G59/1-2 VCB 81013 LIB 1 MLV 59/97A 2008 2008 2008 2008 DRC (east) CODMLV 052* CODMLV 056* MLV 224/97A* MLV 198/97A* MLV 59/97A* E8 Lyamungu 85 AFR 708 M22 Local cross Tanzania, CIAT line CIAT line Local cross 2007 2007 2007 2007 2007 2008 2008 2008 2008 Local cross 2008 Local cross Local cross Local cross Local cross Local cross 2008 2008 2008 2008 2008 MAC 13 Kenya Mavuno Kenya Safi Kenya Tamu CODMLV 052 CODMLV 056 MLV 224/97A MLV 198/97A MLV 59/97A New Rosecoco Chelalang Kenya Umoja Super Rosecoco Kenya Red Kidney Kabete Super Kenya Wonder Miezi Mbili Kenya Early Kenya Sugar bean Kenya Safi MAC 64-1 MAC 13-3 MAC 34-5 CIAT cross 2008 2008 2008 2008 DRK 64 DRK 64 CIAT line UBR (91)45-1 UBR (91)45-1 CIAT line ODR ODR Unknown RJ1 RJ1 Local cross,CIAT parents 2008 (pre-release) 2008 (pre-release) 2008 (pre-release) 2008 Kenya M18 L36 L41 E2 E4 E7 Madagascar 253 Source Year of release Country Madagascar Varietal name RI 5-1 Line code RI 5-1 RI 5-5 RI 5-5 RI 5-3 RI 5-3 IL 5-53 IL 5-53 M 211* Kayana* VNB 81010* RWV 1365* MLV 198/97A* M211 Kayana VNB 81010 RWV 1365 MLV 198/97A 2008 2008 2008 2008 2008 Rwanda RWV 1892 RWV 2070 RWV 1892 RWV 1892 RWV 2070 RWV 1892 2007 2007 2007 Tanzania Flor de Mayo CAB 19 Cheupe Selian 06 Cheupe CAB 19 CIAT line CIAT line 2008 2008 2008 Uganda RWR2075 RWR1946 NABE13 NABE14 CIAT line CIAT line 2007 2007 Zambia KID31 C20P30 Kabulangeti Kabale Kapisha Kabulangeti CIAT ZARI Local landrace 2007 2007 2007 Zimbabwe SUG131 CIAT 2007 254 Source Local cross, CIAT parents Local cross, CIAT parents Local cross, CIAT parents Local cross, CIAT parents Year of release 2008 2008 2008 2008 Activity 4.4 Development of sustainable seed systems to support wide dissemination Highlights: • • • 4.4.1 A strategy of marketing small seed packets as a profitable enterprise is being developed with a private seed company in Kenya and shows great promise for reaching thousands of bean growers. An analysis of the effectiveness of training in seed production suggests that participants have significantly improved both technical and communication skills. A seed security assessment methodology has found acceptance at the institutional level among important players such as FAO, USAID and important international NGO’s. Increasing Access to New and Existing Technologies National Bean Research Programmes (NBRPs) and their partners continued to produce and disseminate seed of improved varieties to farmers using different models of seed supply. These models include support to both local seed supply (farmer to farmer-based production, exchange and sale to local market) as well as to integrated local, formal and commercial sector collaborations and enterprises. For instance, in 2008/9 NBRPs in 9 selected countries availed about 181 tones of foundation bean seeds to farmers and commercial seed producers (see Table 126). Table 126. Amounts of foundation seed supplied by NARS in 9 selected PABRA countries. March 2008-Feb. 2009. Country Burundi DRC –Katanga Ethiopia Kenya Madagascar Malawi Mozambique Tanzania Zambia Total No of varieties 7 10 14 4 12 13 8 7 7 82 Amount of foundation seed (kg) 2,560 6,230 53,560 30,292 3,150 5,000 2,925 75,000 2,240 180,957 Sources: NBRPs annual reports- Presented during ECABREN-SABRN Steering committee meetings Oct. 21-24th, 2009. Lilongwe, Malawi. 4.4.2 Linking Participatory Variety Selection and Impact-oriented seed production and supply systems Farmers’ involvement in a seamless set of activities, from PVS, to subsequent on-farm experimentation and information exchange to seed production and delivery, is creating a strong- impact oriented breedingto-seed outreach chain. In 36 sites in Malawi where participatory variety selection activities are taking place, farmers followed the PVS with seed bulking of the most preferred genotypes. Generally farmers started on small-scale, for example, with 80 seeds per variety. In Kaluluma Agricultural Extension Programme area in Kasungu, a farmer group (see Figure 63) under the supervision of a government extension agent, has been increasing the selected varieties since 2007. They started with 20 varieties, each with 80 seeds but by 2008 they had produced 120 kg of assorted varieties, and started to share seed with 4 other communities in the same quantities (80 seeds of each variety), following the manner in which they themselves had received initial stocks. 255 Figure 63. Members of Farm group and their children who received an initial 80 seeds per variety in Dec. 2006 -Malawi-Kasungu (photo taken on 18.10.2008) In Mozambique, foundation seed is being produced in Mutequelesse and Nintulo, in Gurue, under irrigation. Farmers’ Associations and individual farmers of Gurue, Angonia and Tsangano are involved in the production of certified and quality declared seed. A total of four associations with 93 members and 13 individual farmers received the following amounts of foundation seeds of promising and newly released varieties from the NBRP in Mozambique for further increase and supply in their groups and neighborhoods : • Alto Molocue: 20 kg of SUG 131; • Gurue: 55 kg of SUG 131 (Nintulo – 30 kg and Ewarelo – 25 kg); • Malema: 10 kg of PAN 148; • Gurue (Nintulo): 26 kg of PAN 148; • Gurue (Nintulo): 40 kg of VTTT 925/9-1-2; • Angonia (Kanhanja): 60 kg of SUG 131; • Milange (Sede): 40 kg of PC 1459 BC2-RR9 In south highlands of Tanzania, every site (village) where PVS is taking place each farmer group was given 2 kg, and in addition some selected farmer groups and individuals had planted larger plots: a) Ilembo Peasant Group in Mbozi district planted Uyole 04 and Njano on 4.5 ha, b) J Mwampashi’s Ivwanga Group, in Mbozi district planted Uyole 96 on 2.5 ha; and c) a farmer in Kilolo district produced seeds on 2 ha. This PABRA work is linked to a McKnight-funded project. 4.4.3 Marketing of small seed packs Some NARS have also embarked on innovative seed supply models especially facilitating the marketing of small packs (80, 400 and 2000 g) of certified seeds which are more affordable. Linking through the Tropical Legume-II project, this approach is being tried by CIAT and Kenya Agricultural Research Institute (KARI) in Kenya with Leldet Seed Company in partnership with Farm Input Promotions Africa Ltd (FIPS). The price was Ksh 10, 50 and 200 for 80, 400 and 2000 g respectively (1 USD= Ksh 75) of each the four varieties promoted (Kat B1, Kat B9, Kat x56, and Kat x69). Table 127 illustrates results of sales at one of the promotional marketing sites in Kenya. 256 Table 127. Size of marketed bean seed packs and the gender of the buyers in Central Kenya (season starting Oct. 2008 ). Gender of farmers (buyers) Female Male Total 80 469 349 818 (81.3%) Seed pack size (g) bought 400 2000 89 7 71 21 160 (15.9%) 28 (2.78) Total 565 (56.1%) 441 (43.8%) 1,006 (100%) The majority of buyers were women (56.1%) who mainly bought the 80 g pack size (see Figure 64). Figure 64. Female farmers are the majority of buyers of small seed packs supplied by Leldet Seed Company (Kenya). (Season starting Oct 2008). The small pack of 80 g was the most preferred by farmers because it was affordable, and with as little money as Ksh 40 =USD 0.50, a farmer can buy certified seeds of each of the four different varieties. Using lessons learned from PABRA dissemination approaches and efforts in Kenya, the Ministry of Agriculture supplied 120 tons of the four Katumani bean varieties (Kat B1, Kat B9, Kat x 56 and Kat x 69) to 50 districts in Kenya. The supply is through farmers’ groups as loan to the group and the members take the responsibilities to ensure subsequent farmer-to-farmer exchanges as well as repayment of the seeds loan to other farmers. 4.4.4 Skills and knowledge enhancement NARS and CIAT scientists continued key training for partner organizations’ staff. The training covers aspects related to seed systems namely pre- and post-harvest seed management, seed business including seed supply and dissemination. This seed-related training was supplemented by specific Participatory Variety Selection (PVS) training. For instance, in September 2008, CIAT scientists conducted a Trainingof-Trainers (‘ToT’) course on PVS/seed systems for 23 NARS scientists (breeders and social scientists) from 11countries (see Figure 65). 257 Figure 65. 3 NARS scientists and technicians involved in a “Trainer of trainers” (ToT) course on PVS/seed systems held in Ethiopia, Sept. 2008. Some of the scientists have already started using the acquired skills and knowledge. NARS scientists have conducted training for interested staff from partner organizations. In Kenya and Ethiopia, training of trainers was carried out for 147 (52 females and 95 males) extension staff from NGOs, GOs and CBOs. The training covered topics related seed systems (production management, post harvest management, variety evaluation, and seed supply/dissemination at local level). The trained extension staff also imparted the acquired skills to 1670 farmers including 791 women (See Figure 66). Figure 66. Mr. Nicholas Soikan of Maa Aids Awareness Program (MAAP) in Kajiado, a partner organization to Concern Universal training Masai farmers on bean seed bulking and grain production. Maa is the local Masai language. 258 As result of this partnership and local capacity building (exposure to new varieties, imparting knowledge and skills in areas of pre and post harvest crop management), beans are being introduced for the first time and are slowly being adopted in communities where the bean crop was unknown, e.g. among some members of Masai communities in Kenya (see Figure 67) who gradually are become sedentary. Figure 67. Masai Family interested in a new crop (beans) and new varieties with support from MAAP (local NGO) and Concern Universal –Kajiado Kenya (May 2008) 4.4.5 Backstopping to NARS and their partners Backstopping activities include field visits, support to national planning workshops, and writing seed systems proposals. All these activities are aimed at strengthening PABRA members in seed systems and wider impacts. For instance, a field visit in Cameroon helped scientists from NARS and their partners to understand the existing supply and demand for bean seeds (existing varieties, seed policy, seed sector actors, potential partners in variety promotion, and farmers’ and traders’ variety preferences –see Figure 68). This will help in laying down strategies to disseminate seeds of the promising and pre–released varieties in Cameroon. With regard to support to NARS in writing proposals related to seed systems, PABRA scientists assisted NARS in Uganda, Rwanda, D. R. Congo and Zambia. PABRA seed systems team also backstopped several on going PABRA projects on seed systems namely McKnight supported project in Malawi, Mozambique and southern Tanzania, Tropical Legume-II in Kenya and Ethiopia and Alliance for Green Revolution in Africa (AGRA) supported seed systems project in Rwanda (ISAR), Western Kenya (KARI–Kakamega) and Uganda (NARO). Collaborators: J.C. Rubyogo and L. Sperling 259 Figure 68. Female farmers on slope of Mt Cameroon showing their interest in climbing bean varieties introduced by IRAD under PABRA support (November 20, 2008) 4.4.6 Some lessons from keys approaches of PABRA to institutional strengthening with NARS and other partner organizations 4.4.6.1 Partnership development PABRA has had a long and productive history of partnership with diverse stakeholders. The majority of partners are based in-country. The majority of them have common vision, leverage resources to add value to PABRA contribution and allow synergy. This has been documented through the wider impact of bean based technologies across Africa as manifested in improved farmers’ livelihoods (Food security, Incomes, Nutrition and health, Community empowerment) and increased capacity for production, marketing, etc A study on assessing the status of partnership within the PABRA countries was conducted in four countries representing the rest of PABRA members. These countries were Uganda and Ethiopia in ECABREN and Malawi and South Tanzania in SABRN. The study has the following specific objectives: 1. To understand relations between partners - what is working, what is not working and what needs to be improved 2. To map existing network patterns in order to identify which linkages need to be improved 3. To relate partnership process to outcomes 4. To identify gaps and suggest improvement in the future The partnership was evaluated based on the following elements: 1. Agreement on purpose and shared vision of partnership 2. Engagement and commitment of partners 3. Trust and reciprocity 4. Accountability and decision making 5. Partnership arrangement including legal and institutional arrangement 6. Leadership and effective communication Key findings and suggestions for improvement include: • The partnership is very strong in terms of having a shared vision that is agreed on by a majority of the partners in the surveyed countries. Subsequently the roles and responsibilities of the different partners are clearly articulated and understood by most partners. However, the partners 260 • • • • • • • • • • do not have agreed norms, approaches and processes for achieving the objectives of the partnerships. The partnership has high levels of trust and reciprocity at national level with members indicating high levels of honesty, openness, and willingness to help each other The partnership is not strong in terms of partnership arrangements on issues of IPR and resource access In terms of accountability and decision making, while this may be clear between the key partners namely CIAT/PABRA/networks, National Bean Programs, it is not as clear within national and local partners. Communication across the countries needs improvement, especially the extent to which partnership meetings at national level are held with a frequency that ensures full communication and information sharing as well as providing effective feedback mechanisms between partners. Participation of the private sector (apart from seed companies) in the partnership is low in most of the countries. There is a multitude of secondary partners (partners of our partners!). There is potential for widening the scope of partnerships through building better links with secondary partners. A coherent framework for partnership is needed with clear engagement strategies for different types of partners especially private sector and the secondary partners. However this needs to be strategic in order to balance the need for reach and quality of work. A national level forum of partners should be created during which national level works-plans, resource sharing, monitoring and feedback are actualized. This should be included in the coordination mechanisms of the partnerships. There should be mechanisms to promote transparency in resource allocation at this level. Transparent mechanisms for sharing the success of the partnerships and for individual and collective intellectual property rights including authorship should be developed in a participatory process. Collaborators: 4.4.6.2 J.Njuki, P. Sanginga and M. Mapira Analysis of the effectiveness of capacity building in PABRA Rationale: In an effort to strengthen institutional and organizational capacity, CIAT through its Pan African Bean Research Alliance (PABRA) programme, has been running diverse short course trainings in participatory approaches for its partners. During the period between 2003-08, PABRA (in collaboration with its partners) undertook a significant number of training activities aimed at empowering stakeholders with increased knowledge and skills to overcome constraints in technology development, dissemination and use. About 10,556 stakeholders (more than 20% were women) and including the training of trainers, received training in such topics as micronutrient nutrition with bean as the entry point, Participatory variety selection, bean variety development, marker assisted selection (MAS), participatory plant breeding and variety selection, seed systems, bean production, marketing and post-harvest quality assurance, participatory diagnosis, enabling rural innovation (ERI) and gender considerations in research and development, integrated pest and disease management (IPDM), integrated soil and nutrient management, biological nitrogen fixation in legumes, development and design of promotional materials, novel methods in bean research, nutrition, and participatory monitoring and evaluation (PM&E). Fortyeight (48) bean scientists from NARS in the PABRA region enrolled in formal training at M.Sc. and Ph.D. program levels in the areas of cross-border trade, HIV/AIDS and agriculture, agro-enterprise development, community development, bean improvement for disease resistance, molecular and virulence characterization of disease pathogens, participatory research, seed systems and technology transfer. Partners trained included the National Agricultural Research Institutions (NARIs), organizations in the private and public sector, partners from the non - governmental organizations and farmer research groups. 261 This study was initiated following 5 years of consistent capacity building. The study assessed training provided to national partners in selected areas. The findings are reported for decentralized seed system and participatory monitoring and evaluation. The study objectives were to assess changes in individuals trained in terms of changes in their knowledge, attitudes, and practice (KAP); to analyze the outcomes of capacity developments at different levels (individuals, institutions and communities); to determine extent of use and adaptation of the approaches by partners; to assess extent of institutionalization and to draw generic lessons and principles for broad application. Materials and Methods: A semi-structured questionnaire was administered to collect data on: changes in knowledge, attitudes, and practice (KAP) of individuals trained; the outcomes of capacity developments at individual, organization and community levels; target communities; as well as to evaluate the relevance of capacity building activities. The study applied a modified ‘Ripple’ Model as the conceptual framework for assessing change. The Ripple model provides a useful framework to link capacity building activities to outcomes and impacts. Change was assessed at several levels; individual, organization, and target community levels. Data was collected using qualitative and quantitative methods. A modified “Most Significant Change (MSC)” approach was used. MSC is a story based technique that is used to identify and give value to changes that were unintended or unexpected but were nevertheless significant impacts for those involved. Training was defined as an activity that involved theoretical sharing of information and practical interactions intended to develop or impart skills. This kind of learning environment should have taken at least three days for it to be useful to the trainees. Results and Discussion: The number of people assessed was 39. The respondents had different professional backgrounds that included agriculturalists, social scientists, extension, animal health, environmental management and laboratory technician. Additionally respondents also cut across different positions in the organizations (Table 128) Table 128. Positions of people interviewed Management Officers CDF* Technician PM&E 4 9 12 1 Seed Sys 8 5 - Total 12 14 12 1 CDF = community development facilitators Against a scale, all trainings were rated average, reasons for this rating were that methods used by facilitators utilized a variety of practical and theoretical methods and tools, many stakeholders were involved and training materials given were relevant and adequate. Changes were assessed in two main ways. Respondents were required to rate their knowledge and skills before and after the training using pre-determined variables. Results from capacity building in PM&E Results from PM&E indicated that there were significant levels of change in the knowledge and skills on the subject matter covered during trainings (Figure 69). Significant changes were noted in areas of sharing and making use of information, capturing stakeholders’ indicators, periodic reports from stakeholders, presence of functional PM&E plans and frequency of monitoring. Forty two percent (42%) of respondents reported that they had integrated PM&E in other projects. Most commonly integrated aspects in other 262 projects included developing indicators (57%), developing goals and outcomes (27%), and participation by different stakeholders (14%). Only 23% of the individuals trained had implemented PM&E with other organizations they work with. Aspects implemented included determining program outputs and indicators (50%), setting goals and objectives in the form of Monitoring & Evaluation framework for projects (10%) and gender issues (10%). Organizational benefits related to having PM&E systems were assessed. The results show the following as major benefits; existence of monthly/quarterly project review meetings (31%), regular reporting (12%), clearly defined work plans incorporating PM&E (8%), data collection and analysis (8%) and development of measurable indicators (4%). There were other benefits that accrued from implementing PM&E. They also constituted the various changes happening at organizational level. They include: • Increased commitment and performance (73%) • Regular contacts and sharing (53%) • Improved interaction and sharing (50%) • Improved technical skills (29%) • Easy supervision and better understanding of roles (23%) • Increased transparency (20%) • Changes in project budgets (7%) Changes in practice as a result of PM&E training 3 2 Mean scores 1 0 Before After Frequency monitoring & reviewing of project Figure 69. Before After Presence of functional PM&E plans Before After Before After Sharing & Data collection making use of & analysis plan PM&E at diff. levels of information implementation Before After Extent to which stakeholders' indicators are included Before Periodic progress reports from all stakeholders Changes on practice as a result of PM&E training 263 After It can be seen that capacity building has played a major role in facilitating implementation. This is not only in availing capacity but was as well a source of skills for managing stakeholders. Continuous follow up is also a major factor enabling implementation of PM&E. Results from training in decentralized Seed Systems Results from decentralized seed system indicate that there were significant changes among trainees in the use of seed manuals, in involving farmers in testing new varieties, involving stakeholders in dissemination and sharing of roles (Figure 70). Respondents gave examples of the changes observed on individuals trained. The list included: • Increased interaction with communities (92%) • Use of manuals (39%) • Improved interaction with NGOs and CBOs (54%) • Improved interaction with research centers (31%) • Improved interaction with project managers (77%) Individuals that had been trained in Seed Systems felt that there were significant changes at community level, which they attributed to the training received. Outstanding changes were noted in the extent to which communities were supporting other communities, there were new partnerships formed, more farmers were getting involved in technology development and dissemination and there was increased sharing of roles and responsibilities (See Figure 70). Additional benefits within communities included the increased number of farmers involved in seed multiplication (33%), the fact that beans were considered a cash crop (20%), and there was increased exchange of seed among farmers (13%). The farmers interviewed listed additional benefits as follows: • Acquired good agronomic skills (50%) • Access to fertilizers (17%) • Access to improved seed (67%) Farmers attributed these benefits to availability of inputs - seed and fertilizer (50%), trainings (100%) and group marketing (25%). Clearly, building the capacity of the individuals in organizations was translating into benefits for farmers. Changes at organizational level as a result of training in decentralized seed systems were assessed on the basis of indicators for the integration of the approach at the level of organizations, results were recorded in Table 129. Preliminary Conclusions 1. Capacity building in participatory approaches has accrued benefits mostly at individual and community levels 2. A gap was defined between individual empowerment and institutionalization 3. The study reveals scarcity in the diversity of methods for training 4. Limitations on follow up on capacity building activities and limited resources Contributors: E. Lubega, J. Njuki, R. Muthoni, J.C.Rubyogo, P.Mukishi 264 Changes at end-user/beneficiary level in seed systems M ean scores 3 2 1 0 Before Now Before Now Before Now Before Now Before Now Before Now Before Now Increased access Farmer to new involvement in germplasm techn.dev't & dissermination Improved agronomic practices Supported Improved post- Sharing roles & Formation of comm'ts new harvest mgt responsibilities for seed prodn. partnerships influencing other communities Figure 70. Changes at end user –beneficiary level resulting from training in seed systems Table 129. Changes at organizational level resulting from training is seed systems Change in organization Sharing knowledge with other staff within the organization Use of seed manuals and other extension materials Increased number of staff involved in decentralized seed production systems % response 100 100 92 Management appreciating the importance of engaging many partners in seed production / dissemination Other collaborators/partners outside our organization are adopting decentralized seed production system Increased number of partners/organizations involved in the seed production systems 92 Increase in number of promotion materials produced in partnership with other collaborators 75 New proposals and projects have integrated decentralized seed production systems 75 265 85 75 4.4.6.3 Seed systems 1. In 2003 PABRA initiated a ‘Wider Impact’ Approach (WIP) which facilitates both decentralized (informal and formal) and centralized (commercial) seed actors, to produce and supply bean seeds to farmers. Before the WIP was initiated, seed dissemination of improved bean varieties was limited to farmer research groups, or to areas with a presence of development partners e.g. Non-Government (NGOs) or Government organizations (GOs), schools or through parastatals/seed companies, especially for relief seed supplies. A study which assessed the effectiveness of existing seed systems was carried out in Ethiopia, Southern Tanzania and Uganda was carried out between September and December 2007. The information was collected at five levels along the seed supply chain; NARIs, seed companies and parastatals, partner organizations and seed producers (both individual and farmer groups) and grain producers who were the beneficiaries of the seed suppliers. The study has the following objectives To characterize the existing bean seed systems actors and their roles/responsibilities 2. To assess the bean seed production and supply capacities of decentralized seed producers (farm based systems) 3. To assess the potential and drivers of wider dissemination in decentralized seed systems Findings about the WIP 1. As result increased awareness, there is a very high demand of improved bean varieties in all the three countries and more particularly where the farmers have been exposed to varieties. When a variety was still new in certain area, farmer seed producers sold its seeds between 50-100 % above the average grain market price. For instance, NABE 12-C in Kisoro (S-W Uganda) was being sold at UgSh 1200/kg and NABE 13 was being sold at UgSh 1500/ kg in Kabale while the average grain bean price was at UgSH was 1500 /kg (1USD= Ugsh 1760). However as these varieties gained popularity and were available in the local market, the price fell to a level of the grain market in about three to four years and their seeds were no longer attracting premium price. 2. The time lag between variety release/official approval and the wider use was reduced from five years to less than one year especially if the varieties have been released or tested in other bean network countries, e.g. as with Roba in southern Tanzania and several varieties in Ethiopia. The situation was different before 2003 (launch of wider impact). For instance Awasha Melka which is very popular variety in Ethiopia was released in 1999; it stayed on the shelf till 2004 when wider impact approach was launched in Ethiopia. However, new varieties released after wider impact was launched were immediately used by farmers. This was due to institutionalizations of wider impact by NARS and the bean industry. Similar results were found in southern Tanzania where pre-released varieties (Uyole Njano, Calima Uyole, Bilfa 4 and Nyeupe Mpya) are being used by farmers. In Uganda regionally adapted varieties e.g. NABE 13 and 14 (root rot tolerant) and NABE12-C (climbing) which are not supplied by any commercial seed companies are widely disseminated in Kabale and Kisoro Districts (south west of Uganda) where NGOs, CBOs and individual farmers played a role in the dissemination. 3. NARS in Ethiopia and Tanzania have developed the capacity to produce foundation seed to supply to strategic partners. For instance, NARS in Ethiopia doubled their breeder seed supply to partners between 2003 and 2007 period from 4 to 8 tones in that period. During this period, the supply of basic seeds also increased from 40 to 110 tones. Also in southern highlands of Tanzania, since 2004, the bean program at Uyole Research Institute has been supplying 10 tones of foundation of new and preferred bean varieties to strategic partners while the Research Farm sells more than 60 tones of certified seeds of released varieties every year on cash and carry basis. 266 4. The dissemination of improved varieties was more efficient where NARS was engaged in partnership with local service providers e.g. seed company, farmers’ organization, and if there was good social and human capital in farmers groups. For instance in Kisoro (south west Uganda) NARO teamed with the Africa 2000 network (a local NGO) and district extension service to disseminate NABE 12C (MAC 31), a variety released in 2003. In 2007 (4 years after the release), the variety was planted on about ¼ of the district areas where climbing beans are grown. The variety represented about 30% of bean grain market in Kisoro District. 5. Commercial seed companies focused on already very popular bean varieties e.g. K132 and NABE 4 in Uganda which were released 1994 and 1999 respectively. However farmer seed producers supplied both locally adapted or site specific (e.g. root rot tolerant) varieties, and widely popular varieties. According to seed company owners, the production and supply of beans seeds is not profitable, unless there is an opportunistic market such as organised massive seed relief distribution by NGOs/GOs. This market represented about 80% of bean seed sold by seed companies in Uganda. More often, farmers did not participate in the bidding processes and their varieties choices were not considered. Some amounts of seeds were likely to be shipped to neighbouring countries e.g. South Sudan and East DRC. 6. The research farm in Agricultural Research Institute (ARI) Uyole which operates commercially though subsidized avails about 60 tones per year of assorted varieties released by the Uyole Bean Programme. This may due to good linkage with the bean programme and the determination to see the varieties released by their institute in the hands of farmers rather than being driven by simple commercial interests as it would be in the case of commercial seed companies. 7. The multiplier effect of training is relatively high among farmers. In addition to seeds, the trained farmer seed producers also are resource people. They trained other farmers in their groups and beyond; passing on information and knowledge on improved varieties while selling seeds and also about improved management practices. Annually, a trainer-farmer was able to train 35, 14 and 72 in Ethiopia, southern Highlands of Tanzania and Uganda respectively. 8. Farm based seed production and or supply increased business opportunities and incomes at either farmer seed producers or local seed traders e.g. in 2007 in Kabale’ south west Uganda, there were four agro–input suppliers selling seeds of NABE13 and 14 sourced from the Nyamabale Farmer Field School (a farmer group seed producer). The lowest seed sale by the four agro-input was 2 tones while the highest was 3 tones. Their major clients were farmers who were buying between 0.5 to 2 kg. 9. Farmer based (decentralized) seed production/supply respond to farmers variety needs more than commercial seed sector whose clients are mainly NGOs or GOs relief operations. 10. For improving on their seed business opportunities, farmer seed producers promote their seeds through diverse means –local market, farmer to farmer, and/or local development/church organizations. 11. The majority of farmers of seed producers would like to continue in seed production and supply because they find it to be a source of income, and a very profitable enterprise and that the varieties used were higher yielding than their local ones. Contributor: R. Muthoni Collaborators: J.C. Rubyogo and F. Tembo 267 4.4.7 Development of Seed Security Assessment Methodology Background Emergency agricultural assistance seeks to accelerate farmers’ recovery from crises such as drought or short-term conflict, aiming to help them continue with crop production, and reduce vulnerability to future stress. Seed aid is the most common example of this type of assistance, and has been extensively implemented; for instance, the FAO alone managed 400 such projects between 2003 and 2005 (FAO, 2005) and, in response to the current food crisis, has seed aid plans for 48 countries. (http://www.un.org/apps/news/story.asp?NewsID=27313&Cr=Global&Cr1=Food) This type of activity is expanding and involves important sums of money: e.g. Ethiopia has received at least $500 million in emergency seed aid since 1974 (Sperling et al., 2007). Further, ‘emergency’ seed aid is commonly used to help vulnerable households in chronic stress, i.e. the types of populations which public sector agricultural research particularly aims to serve. Research shows that seed system interventions (seed aid) present serious challenges, and even more so more so during crisis periods. As seed is often replanted, even short-term seed-related interventions can have effects over many seasons. Intervention monitoring and evaluation (M+E) also shows that the dominant design of interventions, from the supply-side, as well as their repetitive nature, (for example in Burundi, 26 seasons in a row) can result in a range of negative effects, skewing crop profiles, undermining local and commercial seed markets, and creating farmer dependencies (McGuire and Sperling, 2008; Sperling et al., 2008). For all these reasons, it seems illogical (and unwise) that emergency seed-related assistance has received relatively little attention within research circles. CIAT has aimed to help correct this key research gap, by coordinating an active “Seed Systems in Stress” research program since 2003. The latest research product to emerge from this program is the first-ever ‘Seed System Security Assessment Guide’ (SSSA). The rationale for the guide, its form, and initial use are summarized below. 4.4.7.1 Distinguishing between Seed security and food security Farm families are ‘seed secure’ when they have access to seed of adequate quantity, of acceptable quality, and in time for planting. Helping farmers obtain seed enables them to produce for their own consumption and sale. So fostering seed security contributes to food and livelihood security more generally. While seed security and food security have some elements in common, they are nevertheless quite different. One can have enough seed to sow a plot, but lack sufficient food to eat – for example, during the ‘hungry season’ prior to harvest. Conversely, a household can have adequate food but lack access to seed (or the right seed) for planting. This happens more rarely, but can occur if seed stocks kept in the house become infested with insect pests or are otherwise contaminated, or if a disease outbreak requires a switch to a resistant crop variety. Despite these key differences between food security and seed security, determinations of seed security have nearly always been based, implicitly or explicitly, on food security assessments. Evaluators assess food needs and then just extrapolate seed requirements as part of the aid package. Similarly, they may estimate existing food stocks by measuring harvests or crop losses. If there is a sharp drop in the harvest, they know there will also be a steep decline in food availability. However, this direct link is not necessarily true of seed systems; that is, a production shortfall doesn’t necessarily lead to a seed shortfall. For most cereal crops harvests can drop as much as 80%, and seed would potentially still be available. 268 Ways of calculating seed system needs versus food security needs also differ. We stress the concept of a seed ‘system’ here since assessments of seed security go well beyond tallying up seed needs on a calculator, although that may be part of the work. Attaining seed security means finding ways to support the systems that give farmers ongoing access to seed of the crops and varieties they require. In many cases, this has little to do with delivering seed directly to farmers and a lot to do with supporting and strengthening the channels through which farmers obtain planting materials on their own. 4.4.7.2 Seed System Security Assessment Guide The development of a Seed System Security Assessment Guide emerged from the need for agencies to better understand what happens to farming systems in periods of acute stress, but also in periods of chronic, longer term stress. The guide has also been designed to help organizations (NARS, UN organizations, Humanitarian agencies) explore how to take advantage of development opportunities. So the optic of the assessment guide has been to link relief and recovery operations to developmental strategies, from the start. The challenge of guide development has been to assess the multi-dimensions of seed security per se, including issues of seed availability, seed access and seed quality, in the short and long-term. After years of research, CIAT (with partners, particularly Catholic Relief Services) has developed a method for assessing seed security which blends focus on informal and formal seed channels, and which encourages clear thinking about strategic goals. Seed aid entails making a large number of choices around implementation approaches, which have significant implications, but which rarely have been considered explicitly. For instance, should aid aim to restore the system to the status quo ante, or to strengthen elements of it (e.g. by introducing new crops, or supporting local markets)? Should interventions focus on the most affected crops, those that generate income, or those that can produce food quickly for recovery? Different goals entail distinct strategies (for instance, women may grow different crops from men, HIV-affected households often have serious labor constraints). Seed systems are complex and dynamic. Interventions need to engage with this complexity if they hope to have lasting impact. The ‘Seed System Security Assessment (SSSA) was designed for the ‘lay person’ but reflects considerable specialized agricultural and seed system insight. To develop the method, researchers conceptualized the links between harvests and future seed needs and provided tools for practitioners to calculate these links (See Boxes 1 and 2). Further, research had to pioneer methods for analyzing the functioning of local seed markets, versus food markets, as these often serve primary sources of seed in crisis periods (Sperling, 2008). 269 Box 1. Seed needs from harvests For a given crop (and variety) and area to be planted, it’s easy to calculate the amount of seed a farmer will need for sowing, as well as the size of harvest to be expected. Let PA be the area to be planted by a farmer, in hectares. Let SR be the seeding rate, that is the amount of seed, in kilograms, that needs to be sown for each hectare of the crop and variety in question. Let MR be the multiplication rate of that crop or variety, namely the ratio of harvestable grain to seed sown. Using these three variables, we can determine sowing needs (SN), in kilograms, for the area to be planted, and the expected harvest (H), in kilograms (some of which may be used in the next cropping season as seed), using a few simple formulas: SN = PA x SR H = PA x SR x MR Thus, H = SN x MR A note of caution: The formula for SN assumes a crop is sown only once. However, under certain conditions seeds of an initial sowing may fail to germinate. So farmers may end up planting a crop two or even three times, thus doubling or tripling their sowing needs. Box 2 gives a few examples of seed needs in light of potential harvests of specific crops. A simple calculator, in Microsoft ® Office Excel format, can be downloaded from [www.ciat.cgiar.org/ Africa/seed_manual.htm]. Here’s an example of the inputs and outputs for a hypothetical case. 270 Box 2. A production shortfall does not necessary equal a seed shortfall Drawing on basic agronomic knowledge, and refining it with in-the-field reality, we have examined seed needs as they relate to possible harvests. Basically, the per cent of a normal harvest required to meet the sowing needs in the next season is the inverse of the multiplication rate. As examples, Table 130a shows the basic relationship between harvests and seed need in two crops in Mali, factoring in farmers’ seed sorting and re-sowing rates for this semi-arid context. Table 130b, moves towards greater precision, drawing on actual field data: for a higher and lower potential area in Ethiopia, and contrasting a good versus bad harvest year. The message from both these tables is consistent that a production shortfall is not necessarily equal to a seed shortfall. For many crops analyzed in African contexts (for example, common bean, faba bean, maize, sorghum, groundnut, wheat, tef) harvests can drop as much as 80-90%, and enough seed is potentially available. We add the qualifier ‘potentially’ as the quality of seed harvested has to be adequate and farmers have to be able to save sufficient stocks until sowing time. This may be particularly challenging in regions with just one agricultural season per year. Table 130. Sowing needs in relations to harvests , by household a. Example from Northern Mali Crop Sowing needs (kg/ farmer area; sorting and resowing factored in) Harvest (on normal farmer area) Per cent of harvest needed for seed Pearl Millet Groundnut 10-20 15 kg (1/4 ha) 430 125 kg (1/4 ha) 3.4 12.0 b. Example from Eastern Ethiopia Crop Sorghum Chiro (highland) Sorghum Miesso (Lowland) Surface Area per Household 1/2 ha. 3/4 ha Sowing needs (kg– for area) 7-8 11-12 1250 kg 1600 kg 0.56 to 0.64 0.75 400 kg 260 kg 2.0 4.6 Harvest/yield (good year) Per cent of harvest needed for seed : good year Harvest/yield (bad year) Per cent of harvest needed for seed :bad year 271 As a brief overview, the guide presents a seven step method for understanding seed systems during a crisis and its aftermath, and for identifying what seed-related assistance is needed. It walks the decision maker, relief worker, research through a series of discrete steps. These include analysis of the effects of the disaster on seed systems, identification of possible problems to be addressed, and choice of actions to alleviate the constraints identified. More specifically, the guide is structured to: 1. Identify zones for assessment and possible intervention. 2. Describe the normal status of the crop and seed systems. 3. Describe the broad effects of the disaster on these farming systems. 4. Set goals for agricultural relief and recovery operations based on farmers’ needs. 5. Assess the post-crisis functioning of seed channels to determine whether short-term assistance is needed. 6. Identify any chronic stresses that require longer-term solutions and identify emerging development opportunities. 7. Determine appropriate short- and longer-term responses based on the analysis of priority constraints, opportunities, and farmer needs. Steps 5 and 7 merit special mention Step 5, assessing the functioning of seed channels during a period of stress, is at the heart of the guide. Step 5 leads the assessor through different loops to understand how home production and social networks are functioning during a crisis and in the stressful aftermath; how the local seed and grain markets are holding up or have changed under stress; and possibilities for tapping into the formal seed sector and commercial supplies. These different types of seed channels need to be assessed and then their joint potential for meeting farmers’ needs evaluated. Step 7 matches responses to the situation. It provides decision trees for examining possible interventions and discusses when they may or may not be appropriate. Box 3 gives an example of one decision-making tree. In the accompany text, we focus only on variety quality. (See Sperling, 2008, for full narrative). 4.4.7.3 SSSA Uptake - International Public Good, Responding to Demand The Seed System Security Assessment (SSSA) Guide was published August 2008. It is readily available on line as well: http://www.ciat.cgiar.org/africa/seed_manual.htm It has proved useful for a range of users (the UN, governments, humanitarian agencies, research organizations), and represents a broadly demanded International Public Good (IPG). In terms of reaching varied constituencies, the follow actions have taken place: • USAID (US Government) has posted it on its website. This organization is among the biggest seed aid donors in the world. • The guide has been recommended as the seed security assessment guide in the UN Food and Agriculture Organization (FAO) Seed Unit’s study for the upcoming ‘State of the World on Plant Genetic Resources.’ The FAO Emergency Unit (TCE) is among the leading global advisors on emergency response. • The guide has been reported on several wide-reaching NGO and humanitarian practitioner websites: e.g. ReliefWeb, Catholic Relief Services, and Drylands Coordination Group (Norway). • The guide has also been picked up by several prominent knowledge management groups: Eldis, CTA, SPORE, and UNESCO’s OpenTraining Programs. Hard copies are also being distributed via CIAT, USAID/Government, the FAO and CRS. 272 Box 3: Decision tree for options to respond to problems of poor seed quality Seed of poor quality PROBLEM Lack of appropriate varieties Formal seed system SHORT-TERM RESPONSE Informal seed system Formal seed system Informal seed system DSD or SVF with strong emphasis on the kind of crop/variety to be on offer. Must be able to counter emerging stress. Both modern and/or farmer varieties may be appropriate. Import and distribute healthy or treated seed (via DSD or SVF). Emergency treatment of farmers’ or market seed, depending on problem. Participatory varietal section/breeding to identify crops tolerant to emerging stress. May use modern and farmer varieties as base. LONGER-TERM RESPONSE Poor seed health Reduction of post-harvest losses or deterioration of stored seed by means of granaries and other forms of improved storage. Promotion and awareness raising of existing modern or farmer varieties which are stresstolerant. Routine use of low-cost seed dressings. Small packet distribution or sale of modern tolerant varieties. (Small size increases accessibility.) Training of grain/seed traders and farmers on production, storage, and handling of seed; in some cases, training of commercial suppliers. Variety quality: Farmers do not often experience short-term problems of variety quality, that is, shortages of varieties adapted to their overall conditions. Of course there are cases where crops or specific crop varieties suddenly seem ‘unadapted’ because of marked disease or pest build-up – as with cassava mosaic virus, or root rots in beans, or infestations of the parasitic weed striga in maize and other cereals. More often, short-term concerns over variety quality arise when implementers sense that a potentially useful modern variety, not yet available to farmers, could be made available, and quickly, via emergency aid. Curiously, then, this concern comes not from a ‘problem’, but from the identification of a potential opportunity. In the face of a significant environmental stress, and the need for a short-term response to it, implementers must be careful that what they offer is indeed adapted to the emerging situation. Whether the materials on offer are farmer varieties or improved varieties, they should have been previously tested or grown under the specific conditions now at hand. A cautious but useful approach is to promote a basket of varieties. In the face of adversity, diversity can be the key to encouraging production stability. Over the longer term, farmers may need novel materials, either modern varieties or ones from other local farming systems, to allow them to respond to shifts in their cropping system. These may have been made necessary by environmental changes (like atmospheric warming), rising disease and pest incidence, or inappropriate promotion of unadapted modern varieties. In some cases, farmers may not be able to maintain the levels of purity they desire in their own saved seed or that from the market. So the introduction of local (i.e., nearby) varieties may also contribute to reinvigorating the gene pool farmers’ rely on. In the case of popular or well-known varieties already in use, implementers may wish to concentrate on promotion: making farmers more aware of the varieties, packaging or selling them in user-friendly quantities, and putting them on offer at agricultural and other events. In some instances varietal development (plant breeding, selection, and field trials) may be necessary. Farmer participatory models should be considered for this R&D, especially where growing conditions are stressful. Collaboration between farmers and formal breeders ensures that the varieties eventually selected will actually grow under real production conditions, including farmer management practices, and that they meet local cultural, social, and economic preferences. 273 References FAO. 2005. FAO’s initiatives for capacity building to support the utilization of plant genetic resources for food and agriculture through seed systems and plant breeding and genetic enhancement. Working Group on Plant Genetic Resources for Food and Agriculture, Third Session, 26-28 October 2005. Rome: No. CGRFA/WG-PGR-3/05/4. 12 pp. http://www.fao.org/waicent/FaoInfo/Agricult/AGP/AGPS/pgr/ITWG3rd/pdf/p3w4E.pdf McGuire, S. and Sperling L. Leveraging farmers’ strategies for coping with stress: seed aid in Ethiopia. Global Environmental Change, Vol 18 (4): 679-688. October 2008 Sperling, L., Deressa, A., Assefa, S., Assefa, T., McGuire, S.J., Amsalu, B., Negusse, G., Asfaw, A., Mulugeta, W., Dagne, B., Hailemariam, G., Tenaye, A., Teferra, B., Anchala, C., Admassu, H., Tsehaye, H., Geta, E., Dauro, D., and Molla, Y. 2007. Long-Term Seed Aid in Ethiopia: Past, present and future perspectives. Addis Ababa and Rome: EIAR, CIAT and ODG. Final Project Report prepared for IDRC and USAID-OFDA. 141 pp. . Sperling, L. 2008. When disaster strikes: a http://www.ciat.cgiar.org/africa/seed_manual.htm guide for assessing seed security. Cali: CIAT. Sperling, L., H.D. Cooper and T. Remington. 2008 Moving toward more effective seed aid Journal of Development Studies, Vol 44(4):586-612 April 2008. 274 Activity 4.5 Socio-economic activities Highlights: • 4.5.1 A baseline study to determine the role of beans in drought prone areas of eastern Kenya has been completed. Beans are the second most important food after maize and are critical for food security. Targeting crop breeding and seed delivery efforts to enhance the impact on the livelihoods of the poor in drought-prone regions of sub-Saharan Africa Rationale: The Tropical Legumes-II (TL-2) Project coordinates efforts across six legumes (common bean, graoundnut, cowpea, chickpea, pigeonpea and soybean) to improve the genetic adaptation of these to drought prone areas. With each crop there is a component of social science to establish a baseline description of the target area, and to characterize the role of legumes and the producer and consumer varietal preferences in these areas. The bean component focuses on eastern Kenya and two regions of Ethiopia, and will be closely coordinated with both the seed component and the Participatory Varietal Selection within the same project. Materials and Methods: The major activities of the socio-economic studies inn this component of the project include: 1) regional situational and outlook analysis and baseline studies and 2) the assessment of end-user preferred traits in support of bean breeding and delivery efforts through gap analysis and participation in PVS activities and early adoption studies (in few sites where variety uptake is significant). 3) Targeting for up-scaling: reaching vulnerable groups and mapping of broader impact target domains and 4) capacity building for NARS partners. 4.5.1.1 Regional situational and outlook analysis During the period of reporting, a regional situation and outlook was drafted and shared with scientists in CIAT and NARS, both for peer review and use. A copy of the draft was also sent to the TL-2 Global project manager. The report has also been submitted to CIAT communication and publication unit at Cali for publication on the CIAT website. The regional situational and outlook analysis was conducted in 2008 for four countries from the Eastern and southern Africa targeted for TL-2 project (i.e. Ethiopia, Kenya, Malawi and Tanzania). The report covered the aspects of variety distribution, production trends, utilization, domestic use and trade; market preferred traits, available common bean technologies and their adoption and institutional constraints and outlook for common bean. Data used for this analysis was from reports of the existing surveys conducted in these countries; annual national reports and archived FAO statistics on production and trade statistics (1970-2004). This report is currently undergoing review process and will be published on the CIAT website soon. 4.5.1.2 Socio-economic baseline surveys A socio-economic baseline survey was conducted between June and December 2008 in Eastern province of Kenya, and in Oromia and Southern region of Ethiopia. Eastern Kenya and Oromia region in Ethiopia represent semi-arid, mid-altitude (1000-1500 masl) while SNNP is semi-arid high altitude bean production environment (Wortmann et al., 1998). 275 These socio-economic surveys adopted an approach that accounts for conditions with and without as well as before and after the project and is part of an overall monitoring and evaluation framework aimed at measuring and attributing the short- and long-term impacts of the TL-2 project. The survey involved focus group discussions in the selected communities, individual households interviews and interviews of key informants in the grain markets. The interviews were complemented with transact walks through the selected villages to make direct observations on the farming systems and constraints. The household survey questionnaires were developed to gather data on: (1) basic farm and household characteristics; (2) cropping patterns, input use, production, and yields; (3) gaps between research, extension and farmers; (4) vulnerability (drought, pests, diseases, and prices) and coping strategies; (5) gender roles in input supply, food production and marketing and women’s access to productive assets and financial resources; (6) adoption of common bean varieties and dis-adoption of bean varieties as well as variety trait preferences; and (7) bean seed systems. In addition to the questionnaires, other supporting tools included improved and local seed samples and GPS equipment. A total of 360 farming households selected from 18 villages in the two countries and 120 traders along the value chain (i.e. small collectors, big collectors, retailers, wholesalers, exporter/processors and consumers) in Ethiopia and Kenya were interviewed. Ten markets including Shashamene, the biggest market, were surveyed in Southern region and Oromia in Ethiopia; while five markets from eastern Kenya and one big market, Nairobi, were studied in Kenya (Table 131). In each market common bean varieties were recorded and key informant interviewed. Table 131. Geographical spread of the surveys Description/Country Total No. of districts Number of village/markets Number of households/traders Ethiopia Kenya Market surveys 4 2 12 6 240 120 Ethiopia Kenya 2 2 10 5 60 60 Survey of farming households Data entry has been completed for all 360 sampled households and data entry for the market studies questionnaires is in progress. Of the 360 entered data, 120 taken from Kenya have been fully cleaned and the analysis of this data is in progress. The results from the analysis so far, indicate that farmers in Eastern Kenya engage in a diversity of crops, cropping systems and diverse distinct farming management practices that are integrated with local ecosystems and livelihoods to cope with drought. They dry plant their crops, make terraces to harvest water, intercrop intensively, keep livestock, invest in social capital, work outside their farms for food or wage and undertake petty trade and handcraft but still cannot meet their food requirements the whole year round. On average, each household experiences 5 months of inadequate food supply per year. Drought is ranked the most important constraint to livelihood improvement, causing about 70% yield loss in common beans when it occurs. Nevertheless, common bean is ranked as the second most important food crop after maize, with about 70 percent of the household surveyed growing it primarily for home consumption. Consistent with the existing literature, data from eastern Kenya reveals a great diversity of common bean varieties at community level with each farmer growing about 3 varieties simultaneously on the farm but with as many as 10 varieties on some farms. Popular varieties include GLP2 (allocated 21% of bean area) Mwitemania (allocated 30 % of the 276 area) and KatB1. Household characteristics, as well as variety consumption and production attributes are the driving factors that underlie variety choice and extent of planting. Market imperfections continue to induce farmers to select varieties they would prefer to eat even when such varieties are rated low for drought tolerance. Current trade-offs between consumption and production attributes are very small demonstrating that farming households value both attributes highly and they will be less likely to accept big trade-offs. The implication for the breeding effort is to target to improve both categories 4.5.2 Capacity building of enumerators for baseline study Non-degree training of enumerators was through the surveys. However, special sessions were always held at the beginning of each survey to train enumerators on the questionnaire, the art of interviewing covering establishing rapport and probing, as well as GPS reading. In total five such training sessions were organized at different times and venues (namely in Nazereth, Siraro woreda, and Dale woreda in Ethiopia, and Kari-Katumani in Kenya) during the reporting period. A participatory approach was used to facilitate sharing of experience, to stimulate discussions and to enhance learning. Nineteen trainees attended from: the Melkassa agricultural Research Center; IPMS (an NGO partnering with Awassa Agricultural Research Center and CIAT in seed systems and PVS activities); and government extension offices in the study areas. 277 Publications Book Chapters Arora-Jonsson, Seema, Ballard, Heidi L., Buruchara, Robin, Casolo, Jennifer, Classen, Lauren, DeHose, Judy; Emretsson, Margareta; Fortmann, Louise; Halvarsson, Anne Lundgren; Halvarsson, Ewa; Humphries, Sally; Long, Jonathan; Murphree, Marshall W; Nemarundwe, Nontokozo; Olssen, Anne; Rhee, Steve; Ryen, Anna; Wilmsen, Carl; Wollenberg, Eva. 2008. Conclusions In Louise Fortmann (ed). Participatory Research in Conservation and Rural Livelihoods: Doing Science Together. Blackwell Publishing Ltd. Beebe, S.E., I.M. Rao, M.W. Blair and J.A. Acosta-Gallegos. 2008. Drought resistance phenotyping of common bean. Generation Challenge Program Special Issue on Phenotyping (in press). Buruchara., R.A. 2008. How Participatory Research Convinced a Skeptic. In Louise Fortmann (ed). Participatory Research in Conservation and Rural Livelihoods: Doing Science Together. Blackwell Publishing Ltd. Fortmann, L., H. Ballard, and L. Sperling. 2008. Change around the Edges: Gender Analysis, Feminist Methods and Sciences of Terrestrial Environments. In L. Schiebinger (ed). Gendered Innovations, Stanford University Press. Gepts, P., Aragao, F., Barros, E., Blair, M.W., Brondani, R, Broughton, W., Hernández, G., Kami, J., Lariguet, P., McClean, P., Melotto, M., Miklas, P., Pedrosa-Harand, A., Porch, T., Sánchez, F. 2008. Genomics of Phaseolus beans, a major source of dietary protein and micronutrients in the tropics. In P.H. Moore and R. Ming (eds) Genomics of Tropical Crops, Springer Publ., Chp 5. pp. 113-143. Jansa, J., A.Bationo, E. Frossard, I.M. Rao. 2008. Options for improving plant nutrition to increase common bean productivity in Africa. In: A. Bationo (ed) Fighting Poverty in Sub-Saharan Africa: The Multiple Roles of Legumes in Integrated Soil Fertility Management, Springer-Verlag, New York (in press). Kimani, P.M., Lunze Lubanga, Gideon Rachier and Vicky Ruganzu. 2009. Breeding common bean for tolerance to low fertility acid soils in East and Central Africa. In: Bationo, A. et al (eds). Innovations for the Green Revolution in Africa. Springer Verlag, Dordrecht, The Netherlands (accepted and in press). Mauyo, L. W. J. R. Okalebo, R. A. Kirkby, R. Buruchara, M. Ugen, and H. K. Maritim, 2007. Spatial pricing efficiency and regional market integration of cross-border beans (phaseolus vulgaris) marketing in East Africa: The case of Western Kenya and Eastern Uganda. In p 1027-1033. Bationo et.al. (eds) Advances in Integrated Soil Fertility Management in Sub-Saharan Africa Nandwa, S.M., A. Bationo, S.N. Obanyi, I.M. Rao, N. Sanginga and B. Vanlauwe. 2008. Inter and intraspecific variation of legumes and mechanisms to access and adapt to less available soil phosphorus and rock phosphate. In: A. Bationo (ed) Fighting Poverty in Sub-Saharan Africa: The Multiple Roles of Legumes in Integrated Soil Fertility Management, Springer-Verlag, New York (in press). Teshale Assefa, H. Assefa and P.M. Kimani. 2007. Development of improved haricot bean germplasm for mid- and low altitude sub-humid ecologies of Ethiopia, pages 87-94. In: Food and Forage Legume of Ethiopia: Progress and Prospects. ICARDA, Allepo, Syria. 278 Refereed Journals Akhter, A., M.S.H. Khan, E. Hiroaki, K. Tawaraya, I.M. Rao, P. Wenzl, S. Ishikawa and T. Wagatsuma. 2008. The greater contribution of low-nutrient tolerance to the combined tolerance under highaluminum and low-nutrient stresses for sorghum and maize in a solution culture simulating the nutrient status of tropical acid soils. Soil Science and Plant Nutrition (in press). Astudillo C., Blair, M.W. 2008. Evaluación del contenido de hierro y zinc en semilla y su respuesta al nivel de fósforo en variedades de fríjol colombianas. Agronomía Colombiana 26: 471-476. Beebe, S., I. M. Rao, C. Cajiao, and M. Grajales. 2008. Selection for drought resistance in common bean also improves yield in phosphorus limited and favorable environments. Crop Science 48: 582-592. Blair, M.W., Morales, F.J. 2008. Geminivirus resistance breeding in common bean. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources 3: 1-14. Blair, M.W., Buendía, H.F., Giraldo, M.C., Metais, .I, Peltier, D. 2008. Characterization of AT-rich microsatellites in common bean (Phaseolus vulgaris L.) Theor Appl Genet 118: 91-103. Blair, M.W., Porch, T., Cichy, K., Galeano, C.H., Lariguet, P., Pankurst, C., Broughton, W. 2008. Induced mutants in common bean (Phaseolus vulgaris), and their potential use in nutrition quality breeding and gene discovery. Israel Journal of Plant Sciences 55: 191 – 200. Checa, O.E., Blair, M.W. 2008. Mapping QTL for climbing ability and component traits in common bean (Phaseolus vulgaris L.) Molecular Breeding 22: 201-215. Dwivedi, S.L., Upadhyaya, H.D., Stalker, H.T., Blair, M.W., Bertioli, D., Nielen, S., Ortiz, R. 2008. Enhancing crop gene pools of cereals and legumes with beneficial traits using wild relatives. Plant Breeding Reviews 30: 179-230. Garzón, L.N., Ligaretto, G., Blair, M.W. 2008. Molecular marker assisted backcrossing of anthracnose resistance into Andean climbing beans (Phaseolus vulgaris L.) Crop Science 48:562-570. López-Marín, H.D., I.M. Rao and M.W. Blair. 2008. Quantitative trait loci for aluminum toxicity resistance in common bean (Phaseolus vulgais L.). Theoretical and Applied Genetics (in review). Mauyo,L. W., J. R. Okalebo, R. A. Kirkby, R. Buruchara, M. Ugen and R.O. Musebe. 2007. Legal and institutional constraints to Kenya-Uganda cross-border bean marketing. African Journal of Agricultural Research Vol. 2 (11), pp. 578-582 Mauyo, L. W., J. R. Okalebo, R. A. Kirkby, R. Buruchara, M. Ugen, C.T. Mengist, V.E. Anjichi and R.O. Musebe. 2007. Technical efficiency and regional market integration of cross-border bean marketing in western Kenya and eastern Uganda. African Journal of Business Management pp. 077084 McGuire, S. and Sperling, L. 2008. Leveraging farmers’ strategies for coping with stress: seed aid in Ethiopia. Global Environmental Change, Vol 18 (4): 679-688. Montoya, C.A., Leterme, P., Beebe, S., Souffrant, W.B., Mollé, D., and Lalle`s, J.P. 2008. Phaseolin type and heat treatment influence the biochemistry of protein digestion in the rat intestine. British Journal of Nutrition, 99, 531–539. 279 Montoya, C.A., Leterme, P., Victoria, N.F., Toro, O., Souffrant, W.B., Beebe, S., and Lallès, J.P. 2008. Susceptibility of Phaseolin to in Vitro Proteolysis Is Highly Variable across Common Bean Varieties (Phaseolus vulgaris). J. Agric. Food Chem., 56, 2183–2191. Mwang'ombe, A.W., Wagara, N. Kimenju, J.W., Buruchara, R.A. 2007. Occurrence and Severity of Angular Leaf Spot of Common Bean in Kenya as Influenced by Geographical Location, Altitude and Agroecological Zones. Plant Pathology Journal. 6: 235-241 Odeny, D.A., S. M. Githiri and P.M. Kimani. 2009. Inheritance of resistance to fusarium wilt in pigeonpea, cajanus cajan (L.) Millsp. J. Animal and Plant Sciences 2: 89-95. Polanía, J., I.M. Rao, S. Beebe, and R. García. 2008. Desarrollo y distribución de raices bajo estrés por sequía en frijol común usando tubos con suelo en condiciones de invernadero. Agronomía Colombiana (in review). Rangel, A.F., I M. Rao and W.J. Horst. 2008. Cellular distribution and binding state of aluminum in root apices of common bean (Phaseolus vulgaris L.) genotypes differing in aluminum resistance. Physiologia Plantarum (published online on 5 November 2008). Rao, I.M., P. Wenzl, A. Arango, J. Miles, T. Watanabe, T. Shinano, M. Osaki, T. Wagatsuma, G. Manrique, S. Beebe, J. Tohme, M. Ishitani, A. Rangel and W. Horst. 2008. Advances in developing screening methods and improving aluminum resistance in common bean and Brachiaria. Braz. J. Agric. Res. (in review). Remans, R., S. Beebe, M.W. Blair, G. Manrique, I.M. Rao, A. Croonenborghs, R.T. Gutierrez, M. ElHoweity, J. Michiels and J. Vanderleyden. 2008. Detection of quantitative trait loci for root responsiveness to auxin producing plant growth promoting bacteria in common bean (Phaseolus vulgaris L.). Plant and Soil 302:149-161. Rivera, M., E. Amézquita, I. Rao and J. C. Menjivar. 2008. Análisis de la variabilidad especial y temporal del contenido de humedad en el suelo de diferentes sistemas de uso de suelo. Acta Agronómica (in review). Rubyogo, J.C., L. Sperling , R. Muthoni and R. Buruchara. Bean seed delivery in sub-Saharan Africa: the power of partnerships. peer reviewed journal: Society and Natural Resources (accepted July 2008). Forthcoming 2009. Schlueter, J.A., Goicoechea, J.L., Collura, K., Gill, N., Lin, J-Y., Yu, Y., Vallejos, E., Muñoz, M., Blair, M.W., Tohme, J, Tomkins, J., McClean, P., Wing, R., Jackson, S.A. 2008. BAC-end sequence analysis and a draft physical map of the common bean (Phaseolus vulgaris L.) genome. Tropical Plant Biology 1: 40-48. Sperling, L., H.D. Cooper, and T. Remington. 2008. Moving toward more effective seed aid. Journal of Development Studies, Vol 44(4):586-612. Wagara, I.N. A. W. Mwangombe, J. W. Kimenju, and R. A. Buruchara, 2007. Variation in aggressiveness of Phaeoisariopsis griseola and angular leaf spot development in common bean J. Trop. Microbiol. Biotechnol. 3:3-13 280 Zhang, X., Blair M.W., Wang, S. 2008. Genetic diversity of Chinese Common bean (Phaseolus vulgaris L.) landraces assessed with simple sequence repeat (SSR) markers. Theor Appl Genet 117:629–640. Non -Refereed Journals Blair, M.W., Buendía, H.F., Díaz, L.M., Díaz, J.M., Giraldo, M.C., Tovar, E., Duque, M.C., Beebe, S.E., Debouck, D.G. 2008. Utilization of microsatellite markers in diversity assessments for common bean. Annual Report of the Bean Improvement Cooperative 51: 12-13. Blair, M.W., Caldas, G.V., Muñoz, C., Bett, K.E. 2008. Evaluation of condensed tannins in tepary bean genotypes. Annual Report of the Bean Improvement Cooperative 51: 130-131. Blair, M.W., Iriarte, G., Beebe, S.E. 2008. Utilization of wild accessions to improve common bean (Phaseolus vulgaris) varieties for yield and other agronomic characteristics. Grain Legumes 50: 8-9. Blair, M.W., Namayanja, A., Kimani, P., Checa, O., Cajiao, C., Kornegay, K. 2008. Development and testing of mid-elevation, commercial-type, Andean climbing beans. Annual Report of the Bean Improvement Cooperative 51: 124-125. Porch, T.G., Blair, M.W., Lariguet, P., Broughton, W. 2008. Mutagenesis of common bean genotype BAT 93 for the generation of a mutant population for TILLING. Annual Report of the Bean Improvement Cooperative 51: 16-17. Workshops and Conferences Blair, M.W. 2008. Advances in Common Bean Genomics. Presented at IV International Congress on Legume Genetics and Genomics, in Vallarta, Mexico, 7-12 Dec. Blair, M.W., A. Asfaw, G. Makunde. 2008. Advances for the common bean TL1 project. Presented at Tropical Legumes I meeting, 2 July. Blair, M.W. Bean Genomics/ Genetics at CIAT. 2008. Presented at INIA- Quilamapu, Chile, 23 Jan. Blair, M.W. 2008. Breeding medium – large seeded Andean beans for high minerals. Presented at Harvest Plus bean meetings in Bukavu, DR Congo, 8 Oct., and Butare, Rwanda, 12 Oct. Blair, M.W. 2008. Genômica do Feijoeiro no CIAT. IX Congresso Nacional de Pesquisa de Feijão, in Campinas, Brazil, 21 Oct. Blair, M.W. 2008. Improving common bean productivity for drought prone environments in sub-Saharan Africa. GCP Annual Research Meeting in Bangkok, Thailand, 15-20 Sept. Blair, M.W., and Beebe, S. 2008. Marcadores Moleculares para el Mejoramiento de Frijol Común. Primer Congreso Internacional y Feria de Frijol in Celaya, Guanajuato, México, 22 May. Blair, M.W. 2008. Microsatellite diversity of cultivated common bean (Phaseolus vulgaris L.). - CIAT internal seminar, 23 April. Blair, M.W. 2008. Population structure in cultivated common bean (Phaseolus vulgaris L.). IV International Conference on Legume Genomics and Genetics in Vallarta, Mex., 6 Dec. 281 Blair, M.W. 2008. Potential of the Common Bean reference collection (diversity structure and drought tolerance performance assessment). ADOC meeting – ICRISAT, Hyedrabad, AP, India, 10-12 Sept. Blair, M.W. 2008. Race structure and relationships among “ecotypes” in cultivated common bean (Phaseolus vulgaris L.). Plant and Animal Genome, San Diego, California, 11-16 Jan. Buruchara, R. A. 2008. Contributing towards reducing hunger and poverty in Africa: CIAT’s approach, experience and opportunities. Presentation at JIRCAs, Tokyo, Japan, May 2008 Buruchara, R. A. 2008. ISFM-based crop production systems for major impact zones in sub-Saharan Africa. Presentation at the Round Table |Meeting on Agricultural Research for African Development May, 2008, University of Tokyo. Kimani, P.M., S. Beebe, M. Blair, R. Chirwa and I. Rao. 2008. Improving productivity of common bean and incomes for the poor in marginal environments of sub-Saharan Africa: Overview of TL I and II projects. Drought phenotyping workshop, 4-17 May 2008 Lilongwe, Malawi. Kimani, P. M., G. Mbugua and P. Okwiri. 2008. Breeding beans for drought resistance in East and Central Africa region. Drought phenotyping workshop, 4-17 May 2008 Lilongwe, Malawi. Kimani, P.M. 2008. Characterisation in drought testing sites in East and Central Africa. Drought phenotyping workshop, 4-17 May 2008, Lilongwe, Malawi. Kimani, P.M. 2008. Future breeding for drought resistance in eastern Africa. Drought phenotyping workshop, 4-17 May 2008, Lilongwe, Malawi. Kimani, P.M., R. Chirwa, A. Namayanja, C. Ruradama, S. Gebeyehu, N. Mbikayi and Lodi Lama. 2008. Breeding better bean varieties for African farmers: Achievements and Future directions. PABRA Stakeholders Workshop, 21-25 January 2008, Kampala, Uganda Kimani, P.M., S. Beebe and M. Blair. 2008. Breeding Micronutrient Dense Bean Varieties in East and Central Africa. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M., S. Beebe, Nkonko Mbikayi and M. Blair. 2008. Screening bean germplasm for micronutrients. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. 2008. Genotype x environment interactions for micronutrient density and variety release. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. 2008. Breeding micronutrient dense beans in ECABREN: Objectives, activities and Milestones. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M., Ben Okonda, S. Beebe and J.P. Keter. 2008. Influence of fertilization with inorganic macroelements on micronutrient density and agronomic traits in common bean genotypes. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. 282 Kimani, P.M. and R. Chirwa. 2008. Future Breeding for Drought Resistance, Better Nutrition and Health in PABRA. Pan African Bean Research Alliance Annual workshop, 20-24 October 2008, Lilongwe, Malawi Kimani, P.M. 2008. New Research Directions in PABRA: Implications for WECABREN. IRADWECABREN Collaborative Bean Research Program Workshop, 16-21 November 2008, Bafoussam, Cameroon. Kimani, P.M. 2008. Agronomic management for maximising micronutrient density in beans. HarvestPlus Regional Review and Planning Workshop, 6-9 October 2008, Bukavu, DR Congo. Kimani, P.M. 2008. Improving food security and quality for low input farmers in the East African Highlands: Lessons Learnt. Nutribean Review and Planning Workshop, 24-27 August 2008, Nyeri, Kenya. Muthoni R, Barungi M, 2008. Achievements in PABRA in terms of Effects and Impacts for the Period 2003 and 2007.Internal Technical manuscript. PABRA M&E Kampala. Uganda Muthoni R, 2008 PABRA Past to Current 2003 – 2008, Presentation to CIAT DG, Dr G. Hawtin. November 26, 2008 Muthoni R, 2008 Highlights from the CIAT Bean Program in Africa, Presentation to the JIRCAS strategic visits to CGIAR centers in Africa led by Dr. Kensuke Okada. June 20, 2008. Muthoni R, 2008 Highlights from the PABRA Program, Presentation to Swiss Development Cooperation Representatives led by Dr. Willi Graf Deza. July 25, 2008 Rubyogo, J.C., and L. Sperling, 2008. Developing seed systems in Africa . In Robert Chambers, Ian Scoones and John Thompson eds. Farmer First Revisited : Farmer Participatory Research and Development Twenty Years on. Workshop held Institute of Development Studies, University of Sussex, Brighton, UK. 12-14 December, Sussex: IDS Sperling , L , S. Nagoda and A. Tveteraas. 2008. Moving from emergency seed aid to seed security linking relief with development. Workshop organized by the Drylands Coordination Group Norway and Caritas Norway, in collaboration with Norad and The Norwegian Ministry of Foreign Affairs. Oslo, Norway, DCG Proceedings No. 24, 14 May. Proceedings, Posters, and Others Proceedings Chirwa, R. M., M. Pyndji and R. Buruchara. 2008. CIAT-PABRA Management and Organization – An assessment of strengths, weaknesses, opportunities and threats. A paper presented at a PABRA Stakeholders Workshop, Kampala, Uganda , 15-20 January Chirwa, R. M, R. Buruchara. 2008. CIAT’s Pan Africa Bean Research Alliance (PABRA) – An Overview. A paper presented at a Grain Legumes CRSP inception Workshop, Barcelona, Spain, 29 Feb. - 4 March 283 Chirwa, R. M., J. M. Bokosi and E. Mazuma. 2008. Use of Marker Assisted Selection in Developing Bean Varieties for multiple disease resistance in Malawi. A paper presented at a Meeting organized by Kirkhouse Trust in Kampala, Uganda 6-7 March Chirwa, R. M. 2008. The Status of Southern Africa Bean Research Network – Progress Towards Achieving Targets in the Current Phase. A paper presented at the PABRA Steering Committee Meeting, Lusaka, Zambia, 17-19 March . Chirwa, R. M., D. Fourie and G. Makunde. 2008. Bean breeding for drought resistance in SABRN. A paper presented at the TL-II training workshop held at MIM, Lilongwe, Malawi, 5-16 May Chirwa, R. M. 2008. Future bean breeding for drought resistance in SABRN. A paper presented at the TL-II training workshop held at MIM, Lilongwe, Malawi, 5-16 May Chirwa, R. M., E. Mazuma and J. C. Rubyogo. 2008. Getting back to basics: creating impact -oriented bean seed delivery systems for the poor (and others) in Malawi. A paper presented at the PVS training Workshop for NARS partners, Mponela, Malawi, 26-27 May Chirwa, R. M., H. Tefera and M. Siambi. 2008. Current Status of the Legume Industry: Bean, Soybean, Groundnut & Goal of the Legume Platform. Presented at the 1st RIU-Legume Platform Meeting Held at NASFAM Conference Room, Lilongwe, Malawi, 5th June Gomonda, R.W.J, I.M.G Phiri, R. Chirwa and C. Mwale. 2008. Improving Soil Fertility: Key Programmes, Strategies and Challenges in Malawi. Presented at the Soil Health Program launch workshop, held at Windsor Golf Hotel, Nairobi Kenya 16-18 June Chirwa, R. M., J. C. Rubyogo, L. Sperling, E. Mazuma, M. Amane and C. Madata. 2008. Getting back to basics: creating impact -oriented bean seed delivery systems for the poor (and others) in Malawi, Mozambique and Tanzania - A progress report. A paper presented at the McKnight’s Legumes CCRP community of practice workshop held at Hotel VIP, Maputo, Mozambique, 6-9 Oct. Chirwa, R. M. 2008. The status of bean research activities in the SABRN. A paper presented at the SABRN/ECABREN joint SC meeting held at Lilongwe Hotel, Lilongwe, 22-24 Oct. Chirwa, R.M, C. Mwale, A. R. Saka, and Ian Kumwenda. 2008. Alliance for a Green Revolution in Africa - Soil Health Program Business Planning Process. A Country Report for Malawi. October Horst, W.J., A.F. Rangel, D. Eticha, M. Ishitani and I.M. Rao. 2008. Aluminum toxicity and resistance in Phaseolus vulgaris – physiology drives molecular biology. Proceedings of the 7th International Symposium on Plant-Soil Interactions at Low pH, Guangzhou, China, 17-21 May. Posters Asfaw, A., M.W. Blair. 2008. Population Genetic Structure of Common Bean (Phaseolus vulgaris L.) Landraces from Ethiopia and Kenya. Plant Animal Genome, San Diego, California, 11-17 Jan. Becerra, V., M. Paredes, C. Rojo, M.W. Blair, J. Tay. 2008. Morphological, agronomical and genetic characterization of a core collection of common bean (Phaseolus vulgaris L.): Race Chile. IV International Conference on Legume Genomics and Genetics, Chillán, Chile, 21-26 Jan. 284 Blair, M.W., H.F. Buendía, L. Díaz, J.M. Díaz, M.C. Giraldo, E. Tovar, M.C. Duque, S.E. Beebe, D. Debouck. 2008. Microsatellite marker diversity in common bean (Phaseolus vulgaris L.). Plant Animal Genome, San Diego, California, 11-17 Jan. Checa, O.E., M.W. Blair. 2008. Mapping QTL for climbing ability and component traits in common bean (phaseolus vulgaris L.) – CIAT posters. Diaz, A., G.V. Caldas, M.W. Blair, 2008, Cuantificación de taninos condensados e identificación de QTLs asociados a su contenido en una población de frijol comun (P. vulgaris). Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Kimani, P.M., John Nderitu and Levi Akundabweni. 2008. Towards Vision 2030: New Bean Varieties for improved productivity, food and nutrition security and wealth creation. 9-12 November 2008, Strategy for Revitalising Agriculture, Second National Workshop, Safari Park Hotel, Nairobi (Award winning poster presentation). Presented to H.E the President, H.E. Vice-President and Hon Minister for Agriculture. Kimani, P.M., A. Mwang’ombe and J. W. Kimenju. 2008. New Varieties from University of Nairobi Bean Program. 9-12 November 2008, Strategy for Revitalising Agriculture, Second National Workshop, Safari Park Hotel, Nairobi (Award Winning poster presentation). Presented to H.E the President, H.E. Vice-President and Hon Minister for Agriculture. Lozano, M.A., G.V. Caldas, M.W. Blair. 2008. Cuantificación de fitatos por espectroscopía visible en 16 genotipos de una población de fríjol común (P. vulgaris l.) sembrada en suelos con alto y bajo fósforo. Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Makumba, W., R. Chirwa, J.C. Rubyogo, R. S. Weldesemayat and M. Jonasse. 2008. Improving smallholders food security, nutrition and income through increased production and marketing of climbing beans in Malawi and Mozambique – presented in Mozambique Njuki. J and Muthoni R, 2008.Participatory Monitoring & Evaluation for Institutional Learning and Community Empowerment. Knowledge Sharing week April 7-11, 2008. CALI Ortiz, D., H. Pachón, M.W. Blair, D. Gutiérrez, C. Araujo, J. Restrepo. 2008. Evaluación del valor nutricional de micronutrientes en una receta típica (fríjol sancochado) preparada con fríjoles nutricionalmente mejorados. Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Ortiz, D., H. Pachón, M.W. Blair, D. Gutiérrez , C. Araujo, J. Restrepo. 2008. Evaluación de la calidad proteica de recetas preparadas con cultivos de maíz mejorado nutricionalmente. Congreso Panamericano de Semillas, Cartagena, Colombia, 14-18 Oct. Papp, P., T. Gollenar, L. Holly, M.L. Warburton, M.W. Blair, G.B. Kiss. 2008. Evaluation of allelic diversity in maize and common bean germplasm, GCP annual meeting, Thailand, 15-20 Sept. Rubyogo J.C., F. Tembo., R. Chirwa. E. Mazuma, M. Amane. and C. Madata. 2008. Collaborative research program for creating impact oriented bean seed delivery systems for the poor in Malawi, Mozambique and Tanzania – presented in Mozambique Yang, Z.B., D. Eticha, I.M. Rao and W. Horst. 2008. The interaction between aluminum toxicity and drought stress in common bean (Phaseolus vulgaris L.). Poster paper presented at the Annual Meeting of the German Society of Plant Nutrition in Limbergerhof/Speyer, Germany, 23-24 Sept. 285 Others International Newsletters Sperling, L. and S. McGuire, 2008 Seed aid in Ethiopia. Anthropology News 49(7):52 Guides and Handbooks Buruchara, R. A., C. Mukankusi and K. Ampofo. Pests and Diseases of Common Bean and their Management in Africa. Handbook for Small Scale Seed Producers (in Press) Sperling, Louise, 2008. When Disaster Strikes: A Guide to Assessing Seed System Security. Cali, Colombia: International Center for Tropical Agriculture Brochures PABRA Outlook: Issue 3. Media Campaign May/June 2008: Seed Aid, with, CIAT Communications unit, CG Communication Unit and Burness Communications. Based on Seed AID work of L. Sperling, Tom Remington and other partners Wires Asian News International (India) Reuters (Nature….) (which linked to Science) Broadcast BBC Network Africa South African Broadcasting Corporation (SABC) Channel Africa Print Hindustan Times (India) New Vision (Uganda) Bistandaktuelt (Norway) Online Africa Science News Service Agricultural Biodiversity Blog Andhranews.net (India) DailyIndia.com KTIC Rural Radio Online Malaysia Sun Online Nature News NewKerala.com (India) Star Online (Malaysia) Thaindian.com (India) TopNews.in (India) Webindia123.com Editorial contributions I.M. Rao served on the scientific committee of the editorial board of the journal, Agronomia Colombiana, and a reviewer to the journals: Crop Science, Agroforestry Systems and Acta Agronomica. 286 Donors Belgium: Belgian Administration for Development Cooperation (BADC) European Commission (EC) K.U. Leuven Canada: Canadian International Development Agency (CIDA) International Development Research Center (IDRC) Chile: Universidad de Chile Colombia: CORPOICA Ministerio de Agricultura y Desarrollo Rural (MADR) CORPOICA/COLCIENCIAS Universidad Nacional de Colombia ECOFONDO Denmark: Danish International Development Agency (DANIDA) Germany: Bundes Ministerium Für Wirtschaftliche Zusammenarbeit und Entwicklung German Federal Ministry for Economic Cooperation and Development (BMZ) German Agency for Technical Cooperation (GTZ) Israel: The Volcani Center Peru: Government of Peru (Ministry of Agriculture) Instituto Peruano de Leguminosas de Grano, IPL Switzerland: Eidgenössische Technische Hochschule-Zentrum – Zentrum für Internationale Landwirtshaft (ETHZ-ZIL) Swiss Development Cooperation (SDC) United Kingdom: Department for International Development (DFID) USA Banco Interamericano de Desarrollo (BID) Bill and Melinda Gates Foundation Fondo Regional de Tecnología Agropecuaria (FONTAGRO) Generation Challenge Program (GCP) Instituto Interamericano de Cooperación para la Agricultura (IICA) Rockefeller Foundation The McKnight Foundation United States Agency for International Development (USAID) USAID/Office of Foreign Disaster Assistance World Bank Contracts BID/IICA Project approved by FONTAGRO CORPOICA, C.I. La Selva, Rionegro, Antioquia, Colombia COSUDE-PROMPEX CIAT Bean Project, Peru COSUDE-PRONALAG Technical Assistance, Bolivia FIDAR INIAP - CORPOINIAP, Ecuador Ministerio de Agricultura - Instituto Nacional de Investigación Agraria (INIA), Peru UDENAR - Universidad de Nariño, Facultad de Ciencias Agrícolas, Pasto, Colombia Universidad Nacional de Colombia 287 Partners Collaborating with Headquarters Ing. Oscar Vizgarra, EEA “Obispo Colombres”, Tucumán, Argentina Dr. Teresa Fowles, University of Adelaide, Australia Dr. Robin Graham, University of Adelaide, Australia Dr. Alain Goossens, University of Ghent, Belgium Dr. Jean-Pierre Busogoro, Agricultural University of Gembloux, Belgium Dr. Jozef Vanderleyden, Catholic University of Leuven, Belgium Dr. Roseline Remans, Centrum voor Microbiele, Belgium Ing. Hernán Campos, CIF “La Violeta”, Cochabamba, Bolivia Ing. Ruddy Meneses Arce, CIF “La Violeta”, Cochabamba, Bolivia Ing. Juan Ortubé, Instituto de Investigaciones Agrícolas “El Vallecito”, Santa Cruz, Bolivia Ing. Carlos Rivadeneira, Universidad Autónoma Gabriel Rene Moreno, Santa Cruz, Bolivia Ing. Tito Anzoátegui, Instituto de Investigaciones Agrícolas “El Vallecito”, Santa Cruz, Bolivia Ing. Teresa Avila, Centro Fitoecogenético Pairumani, Bolivia Dr. Gonzalo Avila, Centro Fitoecogenético Pairumani, Bolivia Ing. Ximena Reyes, Centro Fitoecogenético Pairumani, Bolivia Dr. Maria Jose. Peloso, EMBRAPA-Centro Nacional de Pesquisa Arroz e Feijao, Brazil Dr. Rosana Brondani, EMBRAPA-Centro Nacional de Pequisa Arroz e Feijao, Brazil Dra. Priscila Zaczuk Bassinello, EMBRAPA-Centro Nacional de Pequisa Arroz e Feijao, Brazil Dr. Murillo Lobo Junior, EMBRAPA Arroz e Feijão, Brazil Dr. Patrice Dion, Laval University, Quebec, Canada Dr. Andre Levesque, Agriculture and Agri-Food, Ottawa, Canada Dr. Art Schaafsma, University of Guelph, Canada Dr. K. Bett, University of Saskatchewan, Canada Prof. Manuel Pinto, University of Chile, Santiago, Chile Dr. Shumin Wang, Chinese Academy of Agricultural Sciences, China Dr. Zhide Geng, Yunnan Academy of Agricultural Sciences (YAAS), China Dr. Ana Julia Colmenares Dulcey, Universidad del Valle, Cali, Colombia Dr. Beatriz Gracia, Universidad del Valle, Cali, Colombia Dr. Mildrey Mosquera, Universidad del Valle, Cali, Colombia Dr. Cecilia de la Plata, Universidad del Valle, Cali, Colombia Dr. Alberto Pradilla, Universidad del Valle, Cali, Colombia Dr. Oscar Checa, University of Nariño, Pasto, Colombi Universidad Nacional de Colombia, Palmira, Colombia Dr. Gustavo Ligarreto, Universidad Nacional de Colombia, Bogotá, Colombia Dr. O. Oliveros, Universidad Nacional de Colombia, Bogotá, Colombia Dr. Aristóbulo López, CORPOICA, Tibaitatá, Colombia Dr. Mario Lobo, CORPOICA, Rionegro, Antioquia, Colombia Dr. Alejandro Navas, CORPOICA, Rionegro, Antioquia, Colombia Blga. Gloria Esperanza Santana, CORPOICA, Rionegro, Antioquia, Colombia Ing. José Restrepo, FIDAR - Cali, Colombia Dr. Carlos Manuel Araya, National University, Heredia, Costa Rica Ing. Juan Carlos Hernández, MAG, Costa Rica Ing. Rodolfo Araya, University of Costa Rica, San Jose, Costa Rica Ing. Juan German Hernandez, EE “La Renee”, Instituto de Suelos, MINAG, Quivicán, Cuba Ing. Orlando Chaveco, ETIAH, Holguín, Cuba Ing. Sandra Miranda, INCA, Cuba Ing. Julio César Nin, IDIAF, San Juan de la Maguana, Dominican Republic Ing. Graciela Godoy, IDIAF, San Juan de la Maguana, Dominican Republic Ing. Eduardo Peralta, INIAP, Ecuador 288 Ing. Carlos Monar, INIAP, Ecuador Ing. Carlos Atilio Perez (deceased), CENTA, El Salvador Mr. Asrat Asfaw Amele, SARI, Ethiopia Mr. Teshale Assefa, EIAR, Ethiopia Dr. Jean Jacques Drevon, INRA, Montpellier, France Prof. Walter Horst, University of Hannover, Hannover, Germany Ing. Julio César Villatoro, ICTA, Guatemala, Guatemala Ing. Mousson Finnegan, ORE, Camp Perrin, Haiti Ing. Eliassaint Magloire, ORE, Camp Perrin, Haiti Ing. Levael Eugene, USAID-PADF Project, Haiti Ing. Emmanuel H. Prophète, CRDA, Ministry of Agriculture, Port-au-Prince, Haiti Ing. Danilo Escoto, DICTA, Tegucigalpa, Honduras Dr. Juan Carlos Rosas, EAP (Zamorano), Honduras Dr. Jorge Acosta, INIFAP, Texcoco, Edo. Mexico, México Ing. Javier Cumpian Gutierrez, Veracruz, México Dr. Raul Diaz Plazas, INIFAP, Mérida, Yucatán, México Dr. Ramón Garza, INIFAP, Texcoco, México Ing. Ernesto Lopez, INIFAP, Veracruz, México Dr. Gina Hernández, Universidad Autónoma de México, México Mrs. Celestina Nhagupana Jochua, IIAM, Maputo, Mozambique Mr. Magalhaes Amade Miguel, IIAM, Maputo, Mozambique Ing. Aurelio Llano, INTA/CNIA, Managua, Nicaragua Ing. Mauricio Guzmán, INTA, Nicaragua Dr. Octavio Menocal, INTA, Nicaragua Ing. MSc. Julio Molina, INTA, Estelí, Nicaragua Ing. Norman Alfaro Castellón, CIPRES, Nicaragua Dr. Oscar Gómez, UNA, Nicaragua Ing. Emigdio Rodríguez Quiel, IDIAP, David, Chiriquí, Panama Dr. Gruneberg, Wolfgang, CIP, Peru Dr. Thomas ZumFelde, CIP, Peru Ing. Angel Valladolid, INIA, Lima, Peru Ing. Miriham Gamarra, INIA, Lima, Peru Dr. James Beaver, University of Puerto Rico, Mayaguez, Puerto Rico Dr. Deidre Fourie, ARC-Grain Crops Research Institute, South Africa Dr. Merion Liebenberg, ARC-Grain Crops Research Institute, South Africa Dr. Bill Broughton, University of Geneva, Switzerland Dr. Emmanuel Frossard, ETH University Zurich, Switzerland Eng. Joseph Ndunguru, Plant Protection Division, Tanzania. Dr. Bonnie McClafferty, HarvestPlus, IFPRI, Washington D.C., USA Dr. Howarth Bouis, HarvestPlus, IFPRI, Washington D.C., USA Dr Ross Welch, USDA, Cornell University campus, USA Dr Ray Glahn, USDA, Cornell University campus, USA Dr. M. Grusak, USDA, USA Dr. T. Porch, USDA, USA Dr. Mark Brick, Colorado State University, USA Dr. Henry Thompson, Colorado State University, USA Dr. Theresa Fulton, Cornell University, USA Dr. Steve Kresovich, Cornell University, USA Dr. James D. Kelly, Michigan State University, USA Dr. Jonathan Lynch, Pennsylvania State University, USA Ing. María Elena Morros, INIA, Estado de Lara, Venezuela 289 Ing. Miguel Pérez, INIA, Maracay, Venezuela Ing. Ramiro de la Cruz, INIA, Estado de Lara, Venezuela. Dr. Godwill Makunde, Crop Breeding Institute, Zimbabwe Mr. Walter Manyangarirwa, Africa University, Zimbabwe Institutional Partners in Africa National Research and Extension Programs: Democratic Republic of Congo, Ethiopia, Burundi, Rwanda, Uganda, Kenya, Tanzania, Sudan, Madagascar, Malawi, Zimbabwe, Lesotho, Mozambique, Republic of South Africa, Swaziland, Angola, Zambia. Universities: Country D R Congo Kenya Malawi Mozambique Norway South Africa Swaziland Tanzania The Netherlands Uganda United Kingdom Zambia Zimbabwe NGOs: • University of Lubumbashi • • • • • • • • • • • • • • • • • University of Nairobi Moi University University of Malawi – Bunda College of Agriculture University of Mondrane Norway University of Life Sciences University of Free State University of Pretoria University of KwaZulu Natal University of Swaziland Sokoine University of Agriculture Wageningen University University of Alemaya Overseas Development Group/University of East Anglia University of Zambia University of Zimbabwe Africa University Midland University Country Angola D R Congo Ethiopia Kenya Lesotho • • • • • • • • • • • • • • • • World Vision International -WVI World Vision International -WVI National Seed Company 13 Community Based Organizations Association des Producteurs de Semences de Katanga (APSK) PRODEL (local NGO) Church based organizations Catholic Relief Services Self Help Development International CARE Rural Farm Alternative Organization Catholic Relief Services Nargina Social Work Group Concern Universal Participatory Ecological Land Use and Management (PELUM) US Canada 290 • • • • • • • • • • Mozambique • • • Rwanda • • Swaziland • Tanzania • • • • • • • • Uganda • • • • Zambia • • • Zimbabwe • Glo World Vision International-WVI Concern Universal –CU Plan International Harvest Help and Find your Feet CARE International Concern World Wide Canadian Physician for Relief and Development (CPAR) Small Holding Coffee Trust Funds Action Aid World Vision International-WVI CARE International Concern Worldwide CARE World Vision International-WVI World Vision International-WVI Action Aid Adventist Development and Relief Agency –ADRA Agric. Development Trusts in Mbeya Catholic Relief Services Lay Volunteers International Agency and Christian Council Mbozi, Ileje and Isangati Consortium/foundation (MIICO) Save the Children VECO AfriCare CARE World Vision International-WVI CARE International Plan International-Zambia Lutheran World Foundation Harvest Help (UK) Zimbabwe Farmers’ Union Malawi CBOs: in more than 14 countries. Regional Institutions: ASARECA, Food, Agriculture and Natural Resources (FANR), CORAF. Private Sector: Country Kenya Lesotho Madagascar Malawi South Africa Swaziland Tanzania Company • Lagrotech Seed Co • PANNAR • CTHA • Demeter Farm • SeedCo-Malawi • PANNAR • Umlimi Lokhomile Seed Co • PANNAR • Dodoma Transport Agency Ltd • Masware Farm Seed Co 291 Zambia Zimbabwe • • • • • • Kamano Seed Co Sunnyside farm Pristine Seeds Progeny Seeds PANNAR Seed Co Europe Institutions: Horticultural Research International (HRI)(UK), NRI (UK), Central Science Laboratory (UK), Agri-Food and Food Canada (Canada). Other CGIAR centers and programs: AHI (ICRAF), ICRISAT, IPGRI, WARDA, IRRI, CIMMYT. Others: Tanzania Italy and Ethiopia The MEDIAE Company FAIDA-MALI Himo Environment Management Trust (HEM) Food and Agriculture Organizations (FAO), UN agency 292 List of NARS and Collaborating Partners in Africa Country Angola Name Salvador Miguel Antonio Chicapa Dovala NARs Instituto de Investigacao Agronomica Burundi Dismas Nsengiyumva ISABU, Bujumbura D.R. Congo Lubobo Kanyenga Mbikayi, Nkoko Lunze Lubanga INERA Kipopo INERA, PNL, Mulungu INERA, PNL, Mulungu Ethiopia Teshale Assefa Asrat Asfaw Amele EARO-NARC, Melkassa Ethiopia Institute of Agricultural Research (EIAR) Southern Agricultural Research Institute (SARI) Amhara Agricultural Research Institute (ARARI) Tigray Agricultural Research Insitute (TARI) Kenya David Karanja KARI, Machakos Lesotho Simon Bereng Agricultural Research Department Madagascar Herimihamina Andriamazaoro DRA, FOFIFA Malawi Charles Kapapa Elisa Mazuma I. Fandika A. Mwakikunga Dr Sleshe James M. Bokosi Chitedze Research Station Department of Agricultural Research Services Kasinthula Research Station Chenachena Sub Research Station ICRAF Bunda College of Agriculture, University of Malawi Northern Corridor Ministry of Agriculture Ekwendeni Hospital Action Aid Care International Demeter Farm Edward R. Shela B.J.V. Msukwa Lizzie Shumba Edson Musopole Alfred Kambwiri Jimmy Demitri Mozambique Manuel Amane Maria Jonase Patricio F. Benito Nuria Chavez Francisco C. Edwardo Instituto de Investigacao Agraria, IIAM, Maputo Lutheran World Federation APLA District Department of Agriculture DDA Tsangano Rwanda Augustin Musoni, ISAR-Rubona, Butare South Africa Andries Liebenberg Deidre Fourie Antony Jarvie Agricultural Research Institute Council – Grain Crops Institute (ARC-GCI) PANNAR Research Services Pty (LTD) 293 Country Sudan Name Elsadig S. Mohamed NARs Wad Medani Swaziland Zodwa Mamba Agricultural Research Division Tanzania Catherine Madata Agricultural Reserarch Institute Festus Ngulu SARI, Arusha Zainabu S. Majubwa ADP-Mbozi Benson J. Minja LAELA Agriculture Centre Shila Mwashala Caritas Mollo Prison Farm Rhodes Family Farm K.K.K.T. Church Isangati Agriculture Development Organization Uganda Michael Ugen NAARI, NARO Zambia Kennedy Muimui Horemans Desire Frans M Kanenga Kennedy Geoffery Kayama Aubrey Chulu Zambia Agricultural Research Institute KAMANO Seed Company Ltd Food Legume Research Program Self- Help Africa Plan –Zambia Zimbabwe Godwill Makunde Cain Manzira Crop Breeding Institute Crop Breeding Institute 294 Project Staff List Senior staff Beebe, Stephen, PhD, Breeder, Geneticist, Bean Project Manager Blair, Matthew, PhD, Germplasm Characterization Specialist/Bean Breeder Buruchara, Robin, PhD, Plant Pathologist, CIAT-Africa Coordinator (stationed in Kampala, Uganda) Chirwa, Rowland, PhD, Plant Breeder/SABRN Coordinator (stationed in Lilongwe, Malawi) Kimani, Paul, PhD, Plant Breeder for ECABREN (University of Nairobi/CIAT) (stationed in Nairobi, Kenya) Morales, Francisco, PhD, Virologist (now has left CIAT) Muthoni, Rachel, BSc, MPA, Monitoring and Evaluation Specialist, (stationed in Kampala, Uganda) Njuki, Jemimah, PhD, ERI Specialist, (previously stationed in Zimbabwe; now has left CIAT) Nyang’aya, Martha, MSc, Nutrition (stationed in Kampala, Uganda) Pyndji, Mukishi, PhD, Plant Pathologist, ECABREN Coordinator (previously stationed in Arusha, Tanzania; now has left CIAT) Rao, Idupulapati, PhD, Plant Nutritionist/Physiologist Rubyogo, Jean Claude, BSc., Seed System Specialist (stationed in Malawi) Sperling, Louise, PhD, Social Scientist, (stationed in Rome, Italy) Postdoctoral Fellow Katungi, Enid, PhD, Agricultural Economics (stationed in Kampala, Uganda) *Rangel, Andres F., Plant Nutrition and Physiology Administrative staff Administrative staff gives support to all CIAT Africa staff at each location regardless of the project they belong to Bakulama, Samuel (stationed in Kampala, Uganda) Basemera, Jolly, Administrative Assistant, B.A.M.A. (stationed in Kampala, Uganda) Dossi, Susan, Administrative Assistant (stationed in Lilongwe, Malawi) Gambo, Abdalla, Security Guard (stationed in Arusha, Tanzania) Giraldo, Isabel Cristina, Economist, Administrative Assistant, CIAT-Bean Project Manager *Gondwe, Zifa, Driver (stationed in Lilongwe, Malawi) Kauwa, Anthony, Driver/Mechanic (stationed in Lilongwe, Malawi) *Mawanda, Limani, Office Assistant (stationed in Lilongwe, Malawi) Magombo, Sinosi, Office Assistant (stationed in Lilongwe, Malawi) Mkagula, Peter, Driver (stationed in Lilongwe, Malawi) Nassozi, Sarah, A.C.I.S., Regional Finance and Administration Officer (stationed in Kampala, Uganda) *Ngondo, Ella Administrative Assistant (staioned in Lilongwe, Malawi) Ngwira, Evelyn, Accounts Assistant (staioned in Lilongwe, Malawi) Shirima, Julita, Office Assistant (staioned in Arusha, Tanzania) Travas, Betty, Administrative Assistant (staioned in Arusha, Tanzania) Yoasmu Mugarura (stationed in Kampala, Uganda) Research associates and assistants Acam Catherine, Lab Technician (stationed in Kampala, Uganda) Astudillo, Carolina, Biol. Germplasm Characterization Lab. Buah, Stephen, MSc, (stationed in Kampala, Uganda) Buendía, Héctor Fabio, Ing. Agr., Germplasm Characterization Lab. Bueno, Juan Miguel, Ing. Agr., M.Sc., Entomology Cajiao, César, Ing. Agr., Mesoamerican Bean Genetics Caldas, Gina Viviana, Chem., Germplasm Characterization Lab. Castaño, Mauricio, Ing. Agr., Virology Research Unit Cortés, María Luisa, Ing. Agr., M.Sc., Mesoamerican Bean Genetics Galeano, Carlos Hernando, Ing. Agr., Germplasm Characterization Lab. Grajales, Miguel Angel, Agric., Tech., Mesoamerican Bean Genetics Jara, Carlos Eduardo, Ing. Agr., M.Sc., Phytopathology 295 Kalolokesya, Lizzie, Bean Breeding (staioned in Lilongwe, Malawi), on study leave *López, Hernán, Biol., Germplasm Characterization Lab. Lubyobera John, Lab attendant, (stationed in Kampala, Uganda) Magaleta, Ruth (stationed in Lilongwe, Malawi), on study leave Maina, David (stationed in Nairobi, Kenya) Male Alan (stationed in Kampala, Uganda) *Medina Zapata, Juliana Ines, Biol., COLCIENCIAS Researcher Monserrate Fredy Alexander, Ing. Agr., Andean Bean Genetics Mukankusi Clare Mugisha (stationed in Kampala, Uganda) Mutungu, Esther, (stationed in Nairobi, Kenya) *Navia, Mónica, Biol., Phytopathology Polanía, José Arnulfo, Ing. Agr. Plant Nutrition Ricaurte, José Jaumer, Ing. Agr., MSc., Plant Nutrition Rivera, Mariela, Ing. Agr., MSc., Plant Nutrition Rodríguez, Isaura, Ing. Agr., Entomology *Sangole, Noel, ERI (staioned in Lilongwe, Malawi) Tembo, Frank (staioned in Lilongwe, Malawi) *Wachira, Geoffrey, MSc, (stationed in Nairobi, Kenya) Wambugu, John (stationed in Nairobi, Kenya) Secretaries Gómez, Julia, Plant Nutrition (25%) Tibalikwana, Mabel, Executive Secretary (stationed in Kampala, Uganda) Technicians Castellanos, Guillermo, Phytopathology Cerón, Carlos Alberto, Mesoamerican Bean Genetics Cuasquer, Juan Bosco, Phytopathology Díaz, Orlando, Entomology Guerrero, Alberto Fabio, Mesoamerican and Andean Bean Genetics * Gómez, Ivan, Germplasm Characterization Hoyos, Agobardo, Germplasm Characterization Joaqui, Orlando, Mesoamerican Bean Genetics Maseko Mafuta, MacFord, (stationed in Lilongwe, Malawi) Melo, Edifonso, Plant Nutrition Molina Jarden, Plant Nutrition Morales, Héctor, Entomology Musoke, Steven, Field Technician (stationed in Kampala, Uganda) Mwase, Archangel (stationed in Lilongwe, Malawi) Ortíz, Guillermo, Germplasm Characterization Otero, Martin, Plant Nutrition Sebuliba, Sulaiman (stationed in Kampala, Uganda) *Solarte, Olmedo, Phytopathology Valor, Jose Flower, Entomology Vargas, Luis Alberto, Mesoamerican Bean Genetics *Zuleta, Jesús María, Mesoamerican Bean Genetics *Left in 2008 296 Acronyms and Abbreviations used ADRA AFLP AHI ALS ANOVA APS ARC/GCRI ARI ARS ASA ASARECA ASCOLFI AU AUDPC BBK BCMV BCMNV BEAN BGMV BGYMV BILFA BIWADA BLCrV BMGF BMZ-GTZ BOT BSM CAAS CARE CBB CBOs CCF CD CENARGEN CENIAP CENTA CG CGIAR CGS CIAL CIALCA CIAS CIAT CIDA CIFP Adventist Development and Relief Agency Amplified Fragment Length Polymorphism African Highlands Ecoregional Programme (led by ICRAF) Angular Leaf Spot Analysis of Variance American Phitopathology Society Agricultural Research Council, Grain Crops Research Institute, South Africa Agricultural Research Institute Agricultural Research Station American Society of Agronomy Association for Strengthening Agricultural Research in East and Central Africa Asociación Colombiana de Fitopatología y Ciencias Afines Alemaya University, Ethiopia Area Under Disease Progress Curve Bembeke Bean Common Mosaic Virus Bean Common Moasic Necrosis Virus Bridging Expectations, Accomplishments and further Needs Bean Golden Mosaic Virus Bean Golden Yellow Mosaic Virus Bean Improvement for Low soil Fertility in Africa Bean Improvement for Water Deficits in Africa Bean Leaf Crumple Virus Bill and Melinda Gates Foundation German Federal Ministry for Economic Cooperation and Development (BMZ) German Agency for Technical Cooperation (GTZ) Board of Trustees Bean Steam Magot Chinese Academy of Agricultural Sciences Cooperative for American Remittances Everywhere Common Bacterial Blight Community Based Organizations Consolidated Conceptual Framework Congo Democratic Centro Nacional de Investigaciones en Recursos Genéticos y Biotecnología Centro Nacional de Investigaciones Agropecuarias, Venezuela Centro Nacional de Tecnología Agropecuaria, El Salvador Consultative Group Consultative Group on International Agricultural Research Competitive Grant System Comité de Investigación Agrícola Local Consortium for Improved Agriculture-based Livelihoods in Central Africa Centro de Investigaciones Agrícolas del Sureste, SEA, Dominican Republic Center for International Tropical Agriculture Canadian International Development Agency Centro de Investigaciones Fitoecogenéticas Pairumani, Bolivia 297 CIM CIMMYT CIP CIPRES CMAD CN COLCIENCIAS CORAF/ WECARD CORFOCIAL CORPOICA COSUDE CPAR CRDA CRIA CRSP CRS CSSA CTHA CTZ CU CV CYTED DANIDA DAO DARTS DEL DFID DGDC DICTA DNA DR DRC DRD DSI DTF DTM EAP EARO ECA ECAPAPA ECABREN EEA EIAR EMBRAPA ERI ESA ETIAH CIAT Malawi International Maize and Wheat Improvement Centre Centro Internacional de la Papa, Peru Centro de Investigación y Promoción de Desarrollo Rural y Social Community Mobilization Against Desertification Care Norway Instituto Colombiano para el Desarrollo de la Ciencia y la Tecnología “Francisco José de Caldas” Conférence des Responsables de Recherche Agricole en Afrique de l’Ouest et du Centre/West and Central African Council for Agricultural Research and Development Corporación para el Fomento de los Comités de Investigación Agrícola Local Corporación Colombiana de Investigación Agropecuaria Cooperación Suiza para el Desarrollo Canadian Physician for Relief and Development Centre Recherche de Agriculture, Haiti Centro Regional de Investigaciones Agrícolas Collaborative Research Support Project Catholic Relief Services Crop Science Society of America Centre Technique Horticole d´Antananarivo Chitedze Concern Universal Coefficient of Variance Latin American Program for the Development of Science and Technology The Danish Agency for Development Assistance District Agricultural Office Department of Agricultural Research and Technical Services Delmas Department for International Development General Directorate for Development Cooperation, Belgium Dirección de Investigación de Ciencias y Tecnología Agrícola, Honduras DeoxyriboNucleic Acid Democratic Republic Democratic Republic of Congo Directorate of Research and Development Drought Susceptibility Index Days to Flowering Days to Maturity Escuela Agrícola Panamericana, Honduras Ethiopian Agricultural Research Organization East and Central Africa Eastern and Central Africa Programme for Agricultural Policy Analysis Eastern and Central Africa Bean Research Network Estación Experimental Agrícola Ethiopia Institute of Agricultural Research Empresa Brasilera de Pesquisa Agropecuaria Enabling Rural Innovation East and Southern Africa Estación Territorial de Investigaciones Agropecuarias de Holguín 298 FA FAIDA FANR FAO FARM FC FENALCE FIDAR FIPAH FIPS FLS FONTAGRO FOFIFA GCP GIS HAAS HEM HIV/AIDS HPLC HRE HRI IACR IAEA IBFA ICA ICDPM ICIPE ICRAF ICRISAT ICTA ICP IDIAF IDIAP IDRC IIA IIAM IICA IITA-SARRNET INCA INERA INHA INIA INIA INIAP INIFAP INM INPRHU INRA INTA Farm Africa (just a name in Swahili) Food, Agriculture and Natural Resources Food and Agriculture Organization an NGO, Ethiopia Field capacity Federación Nacional de cultivadores de Cereales Fundación para la Investigación y Desarrollo Agrícola Fundación para la Investigación Participativa con Agricultores de Honduras Farm Inputs Promotion Africa Floury leaf spot Fondo Regional de Tecnología Agropecuaria Centre National de la Recherche Appliqué au Développement Rural, Madagascar Generation Challenge Program Geographical Information System Harbin Agricultural Academy of Sciences Himo Environmental Management Trust Human Immuno Deficiency Virus/Acquired Immune Deficiency Syndrome High Performance Liquid Chromatography Harare Horticultural Research Institute (UK) Rothamsted (UK) International Atomic Energy Agency Ikulwe Bean Farmers Association (Uganda) Instituto Colombiano Agropecuario Integrated Crop Disease and Pest Management Centre for Research in Agro-Forestry International Center for Research in Agroforestry, Kenya International Crops Research Institute for Semi-Arid Tropics Instituto de Ciencia y Tecnología Agrícolas, Guatemala Inductive Coupling Plasma Instituto Dominicano de Investigaciones Agropecuarias y Forestales Instituto de Investigación Agropecuaria, Panama International Development Research Center Instituto de Investigaciones Agrícolas Instituto de Investigacao Agraria, Maputo, Mozambique Instituto Interamericano de Cooperación para la Agricultura International Institute for Tropical Agriculture - Southern Africa Regional Root Crops Research Network Instituto Nacional de Ciencias Agrícolas, Cuba Institut National des Etudes sur la Recherche Agronomique, D.R. Congo Instituto de Nutrición La Habana, Cuba Instituto National de Investigacao Agronomica (Mozambique) Instituto Nacional de Investigación Agrícola Instituto Nacional Autónomo de Investigaciones Agropecuarias, Ecuador Instituto Nacional de Investigac. Forestales y Agropecuarias, Mexico Integrated Nutrient Management Instituto de Promoción Humana Institut National de Recherche Agronomique Instituto Nicaragüense de Tecnología Agropecuaria 299 IPDM IPGRI IPL IPM IPRA IRRI ISABU ISAR ISEM ISISCAN ITA JEEP KARI KEPHIS KHAK KIS LAI LSD MAC MAFP MAG MARDI MAS MASL MATF MDG MEIA MIICO MIP MoA MT MTP MU MW NAARI NARC NARI NARO NARS NAV NBPR NBPGR NEPAD NGOs NIRS NN NPP NRI Integrated Pest and Disease Management International Plant Genetic Resources Institute Instituto Peruano de Leguminosas de Grano Integrated Pest Management Investigación Participativa en Agricultura/ Participatory Research in Agriculture of CIAT International Rice Research Institute, Philippines Institut des Sciences Agronomiques du Burundi Institut des Sciences Agronomiques du Rwanda Integrated Soil Ecosystem Management a complete Routine Analysis Software designed for operation of FOSS NIR insturments in lab Instituto Técnico Agrícola Joint External Evaluation of PABRA Kenya Agricultural Research Institute Kenya Plant Health Inspectorate Khaki Kisanga Leaf area index Least significant difference Medium Altitude Climbers Ministério de Agricultura, Florestas e Pescas (República Democrática de Timor-Leste) Ministerio de Agricultura y Ganadería Maruku Agricultural Reseanch and Development Institute Marker Assisted Selection Meters above sea level Maendeleo Agricultural Trust Funds Millennium Development Goals Microparticles Enzymatic Immuno Assays Mbozi, Ileje and Isangati Consortium/foundation Manejo Integrado de Plagas/Integrated Pest Management Ministry of Agriculture Metric ton Medium Term Plan Makerere University, Uganda Malawi Namulonge Agricultural and Animal Production Research Institute Nazareth Agricultural Research Center National Agricultural Research Institute National Agricultural Research Organization, Uganda National Agricultural Research Systems Navy National Bean Research Programmes National Bureau of Plant Genetic Resources The New Partnership for Africa’s Development Non-Governmental Organizations Near InfraRed Spectrophotometry Non nodulating Networks, Programs and Project, Natural Resources Institute (UK) 300 NRM NUA OFDA OPV ORE ppm PABRA PADF PANNAR PBS PCCMCA PCR PHI PM&E PNL PPB PRGA PRIAM PROFRIZA PROMPEX PRONALAG PVS QPM QTL RAZ RBM REDSO/ESA REU RF RIL RM RD RL RR RT RSP RSSP SABRN SACCAR SADC SARBEN SARBYT SARI SAS SC SCAR SDC Natural Resource Management Andean Nutrition Office of Foreign Disaster Assistance Open Pollinated Variety Organization for the Rehabilitation of the Envirnoment, Haiti parts per million Pan-Africa Bean Research Alliance Pan American Development Foundation Seed Company Potato Bean Sweet Programa Cooperativo Centroamericano para el Mejoramiento de Cultivos Alimenticios Polymerase chain reaction Pod harvest index Participatory Monitoring & Evaluation Programme Nacional Legumineuse Participatory Plant Breeding Participatory Research and Gender Analysis Participatory Research for Improved Agroecosystem Managemente project, Africa Proyecto Regional de Frijol para la Zona Andina Comisión para la Promoción de Exportaciones Programa Nacional de Leguminosas Alimenticias Participatory Variety Selection Quality Protein Maize project Quantitative trait loci Resistance to Acanthoscelides and Zabrotres Result based monitoring Regional Economic Development Services Office for East and Southern Africa Reach-End Users The Rockefeller Foundation Recombinant inbred line Red Mottled Root diameter Root length Root rot Root tips Regional Support Programme Rural Sector Support Program Southern Africa Bean Research Network Southern African Centre for Cooperation in Agricultural and Natural Resources Research and Training Southern Africa Development Council Southern Africa Regional Bean Evaluation Nursery Southern Africa Regional Bean Yield Trial Selian Agricultural Research Institute, Ethiopia Statistical Analysis System Steering Committee Sequence-characterized amplified regions Swiss Development Cooperation 301 SEA SENA SENASEM SERIDA SGRP SICTA SPAD SRL SSACP SSSA SSSN SUG SWMnet t TARS TILLING TNC TPRI TSBF TSG TSP TWFP TZ UAGRM UK UMATA UN UNA USAID USDA Uyo VICARIBE VIVA VLIR-UOS WARDA WECABREN WINISI WVI ZW Secretaría de Estado de Agricultura, Dominican Republic Servicio Nacional de Aprendizaje Service National des Semences Servicio Regional de Investigación y Desarrollo Agroalimentario, Spain Systemwide Genetic Resources Programme Sistema de Integración Centroamericano de Tecnología Agropecuaria Chlorophyll meter to determine leaf chlorophyll content Specific root length Sub-Saharan Africa Challenge Program Seed System Security Assessment SADC Seed Security Network Sugar Soil water management network ton Tropical Agricultural Research Station Target Induced Local Lessions in Genomics Total nonstructure carbohydrates Tropical Pesticides Research Institute, Tanzania Tropical Soils Biology and Fertility Program, Kenya Technical Support Group Triple Super Phosphate Tropical Whitefly Project Tanzania Universidad Autónoma Gabriel René Moreno United Kingdom Unidad Municipal de Asistencia Técnica Agropecuaria United Nations Universidad Nacional Agraria, Nicaragua United States Agency for International Development United States Department of Agriculture Uyole Vivero Caribeño de grano Andino Vivero Internacional de Volubles Andinos Vlaamse Interunivesitaire Raad-University Development Co-operation The Africa Rice Center West, Eastern and Central Africa Bean Research Network A calibration software package from ISI World Vision International Zimbabwe 302