Transverse cotyledonal cracking (TVC) was characterised at physiological, biochemical and molecular levels in three common bean (Phaseolus vulgaris L.) cultivars: Imbali, Tongaat and Tokai. The incidence of TVC was determined visually on ten-day old seedlings and was expressed as the number of cracks per plant. The effect of Ca++ on TVC incidence was examined by enhancing the calcium content of seed cotyledons using calcium salts in seed priming and coating. EDAX was used to quantify the cotyledonal calcium content. Activities of the pectinolytic enzymes polygalacturonase (PG) and pectin methylesterase (PME) were assayed at dry seed, VC, R4 and R6 phenophases of the bean plant, and zymogram electrophoresis was used to identify the two enzymes. The results showed that cultivars Imbali, Tongaat, and Tokai were susceptible resistant and intermediate, respectively, with respect to TVC incidence. Calcium uptake was higher in Tongaat cultivar (P= 0.05) compared with Imbali and Tokai, irrespective of calcium salt enhancement. Vigour was significantly reduced (P< 0.01) by both coating and priming. TVC significantly lowered yield (P= 0.05) in the susceptible Imbali cultivar. Zymogram analysis identified lytic bands at ~45kDa (PG gel) and ~30kDa (PME gel). The activity profile of PME was similar for all cultivars. However, PG activity of susceptible Imbali was high in dry seeds and at the R4 and R6 stages of plant development, whereas the more resistant cultivars displayed high activity at the VC stage only. It is concluded that high PG activity at R4 stage is a more reliable determinant of green bean propensity to cotyledonal cracking. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

Common bean (Phaseolus vulgaris L.) is an important legume crop in Kenya and is a cheap source of proteins. The small scale farmers in Kenya produce common bean under low agricultural input systems and this predisposes the crop to pests and diseases. Among the diseases, angular leaf spot (ALS) is a major constraint to common bean production and contributes to yield losses as high as 80%. The causative pathogen Pseudocercospora griseola (Sacc.) Crous & Braun is highly variable and several races have been reported. There are few common bean genotypes with resistance to this disease. Therefore breeding for resistance to ALS is important for the country. This study was carried out to; i) evaluate the common bean production systems, constraints and farmer varietal preferences in Kenya, ii) evaluate local landraces and selected introductions of common bean for yield performance and reaction to ALS, iii) study the genetics of resistance to ALS in common bean and iv) develop a breeding method for durable resistance to ALS in common bean. To determine the common bean production systems, farmers’ preferred traits and their knowledge on common bean constraints including ALS, a survey was conducted in Kiambu county using a semi-structured questionnaire, interviews, and focus group discussions. The study revealed that farmers cultivate common beans during the short and long rain seasons. However, they experience better yields in the short rains due to reduced disease incidence. The majority of the farmers (71%) intercrop common bean and this ensures maximum utilisation of space. A high percentage (70%) of the farmers utilise their retained seed for production. The farmers identified ALS as one of the most important constraints to production. The only preventative measure they undertake to control the disease is weeding. The farmers reported that they would prefer improved varieties that were resistant to ALS. Farmers have a preference for particular common bean traits that include high yield (80%), resistance to insect-pests and diseases (72%), type I growth habit (52%), early maturity (68%), seed size and colour (21%) and cooking time (20%). These should be incorporated in breeding programmes. Two hundred common bean landraces and market class varieties were evaluated for ALS resistance in a nethouse at University of Nairobi, Kabete Field Station and for ALS resistance and yield in the field in KARI-Tigoni. The results showed that disease severity scores for the genotypes were similar in the two locations, with the top three resistant genotypes being Minoire, GBK 028123 and Murangazi with disease severity scores of 2.9, 2.9 and 3.2 in Kabete and 2.6, 2.8, and 2.9 in Thika respectively. These resistant genotypes can be used as sources of resistance in a breeding programme or they can be used as resistant varieties. All the market class varieties were susceptible to ALS (disease severity score 6.7-8.0). There was a non-significant correlation between disease and yield most likely because most of the resistant genotypes were exotic and hence not adapted to the local conditions. There was also a non-significant correlation between disease and seed size. The two hundred common bean genotypes were evaluated for yield at University of Nairobi, Kabete Field Station and KARI-Thika. The results indicated that the 2011 and 2012 seasons had similar mean yields and that yields at Kabete were higher than at KARI-Thika. The highest yielding genotypes across the two locations were; GLP 2 (766 kg ha-1), Nyirakanyobure (660 kg ha-1), GBK 028110 (654 kg ha-1), GLP 585 (630 kg ha-1) and Mukwararaye (630 kg ha-1). There was a significant genotype x environment interaction and hence it is important for breeders to carry out stability analysis, so as to recommend varieties for a wide range of environments. To study the genetics of ALS resistance in common bean, three inter-gene pool crosses: Super-rosecoco x Mexico 54, Wairimu x G10909 and Wairimu x Mexico 54 were made. The resistant genotypes were Mexico 54 and G10909, while Super-rosecoco and Wairimu were susceptible. The generations F1, F2, BC1P1 and BC1P2 for each of the crosses were developed. The parents P1, P2 and the five generations of each cross were evaluated for resistance to ALS in Kabete Field Station. Results showed that both dominance and additive gene action were important in the expression of resistance to ALS. However, additive gene action was predominant over dominance gene action. There was a moderately high narrow sense heritability estimate (52.9-71.7%). The minimum number of genes controlling resistance to ALS was between 2 and 3. The predominance of additive gene effects and the moderately high narrow sense heritability estimates recorded imply that progress in resistance to ALS could be made through selection in the early segregating generations. A double cross followed by selection against resistant genotypes was used to develop a method to breed for durable resistance to ALS in common bean. The method was used to accumulate minor genes of ALS resistance into single genotypes. Four intermediate resistant landraces were used to develop a double cross population that was screened using a mixture of ALS races. Selection in F1 and F2 population was done on the basis of intermediate resistance (disease severity score 4.0-6.0), while selection from F3 population was based on resistance (disease severity score 1.0-3.0). Ten advanced F4 lines along with their parents were evaluated for ALS resistance. The F4 advanced lines had a significantly improved resistance to ALS compared to their parents. Hence the method was successful in accumulating minor genes for resistance thus showing significant breeding progress in breeding for durable resistance. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.

The common bean (Phaseolus vulgaris L.) is an important crop grown widely in Uganda. It is also an important source of income for smallholder farmers particularly women. Despite its importance, production in the cool highland regions is constrained by anthracnose disease which causes losses in both the quantity and the quality of beans produced. The principal aim of this research was to elucidate on the status of dry bean anthracnose and the genetics governing its resistance. A participatory rural appraisal study was conducted to explore farmers' knowledge, experience, problems and cultivar preferences in association with managing dry bean anthracnose disease. This study revealed that anthracnose is an important constraint to production which is not controlled in any way. Although farmers have varying cultivar preferences, they use mostly home saved seed and only 1% could access improved seed. The study suggested the need for practical approaches in the provision of quality anthracnose resistant seed in consideration of farmers' preferences and the dynamics of their rural livelihoods. A study was conducted to determine the variability of the anthracnose (Colletotrichum lindemuthianum) pathogen in some of the major bean growing regions of Uganda. Use was made of a set of 12 internationally accepted anthracnose differential cultivars to identify the physiological races present. The results obtained indicated the presence of eight races with one race (767) being dominant and most aggressive. Differential cultivars AB 136 and G2333 were resistant to all the eight races, and can be utilised as potential sources of resistant genes. A germplasm collection of mostly Ugandan accessions was screened for anthracnose resistance. Using the area under disease progression curve as the tool for assessing disease severity, eleven accessions were identified that posses good levels of anthracnose resistance. The yield loss attributed to the anthracnose disease was determined on three susceptible Ugandan market-class dry bean cultivars and two resistant cultivars. The results showed that the yield of susceptible cultivars was reduced by about 40% and an almost equivalent yield was lost due to poor quality seed. In comparison, the yield lost by the resistant cultivars was not significant. The study suggested the use of resistant cultivars as the best solution in combating anthracnose resistance. Three susceptible Ugandan market class dry bean cultivars and six resistant cultivars were used for the study of the inheritance of resistance to the anthracnose pathotype 767 in a complete 9x9 diallel design. The results clearly indicated that the resistance was predominately conditioned by additive gene action. It was also established that epistatic gene action was important. More than one pair of genes displaying partial dominance were responsible for determining resistance and the maternal effect did not have an influence on resistance. Additionally, the result showed that some of susceptible cultivars combined very well with the resistant cultivars and that anthracnose resistance heritability estimates in both the narrow and broad sense were high. These results suggested that the use of simple pedigree breeding procedures such as backcross selection could be useful in improving anthracnose resistance levels in the Ugandan market class varieties. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

Bean fly (Ophiomyia spp.) is a major pest of common bean (Phaseolus vulgaris L.) throughout eastern and southern Africa. In the semi-arid areas, apart from drought, the insect pest is reported to cause high crop losses up to 100%, particularly when drought occurs and under low soil fertility. Host-plant resistance is part of the integrated pest management strategies that have been widely employed against major insect pests of tropical legumes. However, information regarding its use in control of bean fly in common bean is limited. Therefore, the objectives of this study were to: (1) validate farmers’ perceptions of major constraints responsible for yield losses, particularly the major insect pests of beans; (2) asses the level of adoption of improved bean varieties and determine factors that influence farmers’ preferences of the varieties and criteria for selection; (3) identify sources of resistance to bean fly available in landraces; (4) determine the nature of gene action controlling bean fly resistance and seed yield in common bean; (5) describe a procedure for generating optimal bean fly populations for artificial cage screening for study of the mechanisms of resistance available in common bean against bean fly. Farmers considered drought and insect pest problems as main causes for low yields. The adoption rate for improved varieties was high but self-sufficiency in beans stood at 23% in the dry transitional (DT) agro-ecology and at 18% in the dry mid-altitude (DM) agroecology, respectively. Drought, earliness, yield stability, and insect pest resistance were the factors determining the choice of varieties by farmers. Bean fly (Ophiomyia spp.), African bollworm (Helicoverpa armigera) and bean aphid (Aphis fabae) were identified as key crop pests of beans limiting yield. The study to identify new sources of resistance included 64 genotypes consisting of landraces, bean fly resistant lines and local checks. The experiment was done under drought stressed (DS) and non-stressed (NS) environments and two bean fly treatments (insecticide sprayed and natural infestation) for three cropping seasons between 2008 and 2009. Genotypes differed in their reaction to natural bean fly attack under drought stressed (DS) and non-stressed environments (NS) over different cropping seasons. However, the effect of bean fly appeared to vary between the long rains (LR) and short rains (SR). It was observed that an increase in the number of pupae per stem resulted in a higher plant mortality. The range of seed yield was from 345 to1704 kg ha-1 under natural infestation and from 591 to 2659 kg ha-1 under insecticide protection. Seed yield loss ranged from 3 to 69 %. The resistance of most of the bean fly resistant lines seemed to be ineffective in presence of DS. To determine the nature of gene action controlling the inheritance of resistance to bean fly, four parents with known reaction to bean fly were crossed with four locally adapted genotypes in an 8 x 8 half-diallel mating design. Similarly, two resistant and two susceptible parents were selected and crossed to produce populations for generations means and variance components analysis. Results revealed that both general combining ability (GCA) and specific combining ability (SCA) mean squares were significant (p A 0.05) for all four traits studied, except SCA for stem damage during one cropping season. Among the parents, GBK 047858 was the best general combiner for all the traits studied across seasons except for stem damage during LR 2009. Genotypes GBK 047821 and Kat x 69 (a locally adapted variety) were generally good general combiners for resistance traits as well as seed yield. General predictability ratio values ranging from 0.63 to 0.90 were obtained for plant mortality, stem damage, pupae in stem and seed yield across cropping seasons. These results established the predominance of additive gene effects (fixable variation) over the non-additive effects in controlling the traits. Low to moderate narrow sense heritability values ranging from 0.22 to 0.45 were obtained for pupae in stem. Such heritability estimates indicate that although additive gene components were critical in the inheritance of resistance for the trait, non-additive gene action was also important in addition to the environmental effects. A major disadvantage in screening for resistance to bean fly in common bean by controlled means in net cages has been the lack of a method to use for raising adequate fly populations for screening. Due to this problem, a simple procedure for raising sufficient numbers of adult bean flies required for screening was described. Through this method, up to 62 % emergence of the adult flies was achieved. Moreover, the flies retained their ability to infest bean plants. To determine the presence of antibiosis and antixenosis mechanisms of resistance in common bean, five genotypes [CC 888 (G15430), GBK 047821, GBK 047858, Ikinimba and Macho (G22501)] and two local check varieties (Kat B1 and Kat B9) were screened under free-choice in outdoor net cages and no-choice conditions in net cages placed in a shadehouse. All the five resistant genotypes tested had relatively long internodes. It was established that long internode was a morphological trait associated with reduced pupation rate in bean stems, hence an antixenosis component of resistance. Both ovipositional non-preference and antibiosis mechanisms were found to exist in three genotypes namely CC 888 (G45430), GBK 047858 and Macho (G22501). These genotypes were resistant when they were subjected to bean fly under both free-choice and no-choice conditions. They had fewer feeding/oviposition punctures, low number of pupae in the stem, reduced damage to the stems and low percent plant mortality. The remaining genotypes, Ikinimba and GBK 047821 only expressed antixenosis. To maximize the effectiveness of host-plant resistance against bean fly, multiple insect resistances should be incorporated into a single bean genotype in order to ensure durability. However, this should be within the background of integrated pest management strategy. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

Dry bean (Phaseolus vulgaris L.) is economically and nutritionally an important legume, not only in Malawi, but in many parts of Africa and Latin America. Unfortunately, two bruchid species (Acanthoscelides obtectus Say, and Zabrotes subfasciatus Boheman) are known to cause extensive damage in storage, reducing the economic importance, food value and planting value of the crop. The aim of this study was to: i) ascertain farmers’ perceptions of the importance of bruchids as storage pests, and to identify their preferred varietal traits in dry beans; ii) screen Malawian dry bean landraces for effective and adaptable sources of resistance to the two bruchid species; iii) determine the gene action and inheritance of bruchid resistance. Farmers’ perceptions on the importance of the two bruchid species to beans both in the field and in storage were established using a participatory rural appraisal (PRA) in three extension planning areas (EPAs) in Lilongwe agricultural development division (ADD). Results confirmed that the two bruchid species are important storage pests, causing serious storage losses among smallholder farmers. In the absence of any control measures, farmers indicated that more than 50% of their stored beans could be lost to bruchids. Indigenous bruchid control measures are not very effective, making it necessary to search for other control methods. It was also clear from the PRA results that breeders need to consider both agronomic and culinary traits in bean cultivar development. This would enhance uptake of newly developed varieties. To address the problem of bruchid damage experienced by smallholder farmers, a total of 135 dry bean genotypes, comprising 77 landraces and 58 improved varieties (obtained from collaborating partners) were tested under laboratory infestation (nochoice test methods) and field infestation (free-choice test methods). The objective of this study was to identify effective sources of resistance to the two bruchid species. Results of the study showed that there was a wide variation among the genotypes for resistance to the two bruchid species. Overall results showed that 88% of the genotypes ranged from susceptible to highly susceptible to Z. subfasciatus and only 12% of the genotypes were moderately resistant to resistant. Genotype screening for resistance to A. obtectus showed that only 12.5% were resistant, whereas 87.5% were moderately to highly susceptible. All of the improved genotypes were 100% susceptible to A. obtectus in storage. One landrace, KK35, consistently showed a high level of resistance to both bruchids under laboratory infestation, with results similar to the resistant checks (SMARC 2 and SMARC 4), while another landrace, KK90, displayed stable resistance under both laboratory and field infestation. However, performance of most genotypes was not consistent with field and laboratory screenings, suggesting that mechanisms of bruchid resistance in the field are different from that in the laboratory and field screening should always be used to validate laboratory screening. Resistance in the field was not influenced by morphological traits. The seed coat played a significant role in conferring resistance to both bruchid species in the laboratory, whereas arcelin did not play any significant role in conferring resistance in the landraces. The inheritance of resistance to A. obtectus was studied in a 6 x 6 complete diallel mating design, involving crosses of selected Malawian dry bean landraces. The F1 crosses, their reciprocals, and six parents were infested with seven F1 generation (1 to 3 d old) insects of A. obtectus in a laboratory, no-choice test. There were significant differences among genotypes for general combining ability (GCA) and specific combining ability (SCA). However, SCA accounted for 81% of the sum of squares for the crosses, indicating predominance of the non-additive gene action contributing to bruchid resistance. A chi-square test for a single gene model showed that 5 of the 13 F2 populations fitted the 1:2:1 segregation ratio of resistant, intermediate and susceptible classes, respectively indicating partial dominance. The eight F2 populations did not conform to the two gene model of 1:4:6:4:1 segregation ratio of resistant, moderately resistant, moderately susceptible, susceptible and highly susceptible classes, respectively. Average degree of dominance was in the partial dominance range in five F3 populations, but in general resistance was controlled by over-dominance gene action in the F2 populations. The additive-dominance model was adequate to explain the variation among genotypes indicating that epistatic effects were not important in controlling the bruchid resistance. The frequency distribution of the 13 F3 populations for resistance to A. obtectus provided evidence for transgressive segregation, suggesting that resistance is conditioned by more than one gene. Reciprocal differences were not significant in the F2 generation seed; but were significant in four crosses in the F3 generation seed for adult bruchid emergence, suggesting that maternal effects or cytoplasmic gene effects also played a role in the inheritance of resistance to the common bean weevil. Through this study, important sources of bruchid resistance in dry bean have been identified in Malawian landraces (KK35, KK90 and KK73). These resistant sources will be used in a breeding programme to develop bruchid resistant bean cultivars, as well as improve resistance in susceptible commercial bean cultivars currently grown by farmers in Malawi. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2007.