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Resistances to bean common mosaic virus, halo, common and Ascochyta blight, angular leaf spot, anthracnose and rust pathogens of beans in South Africa were combined by reverse dichotomous crossing. Full resistance to Uromyces appendiculatus from Carioca 80 was conditioned by a single dominant gene. Partially dominant resistance to Phaeoisariopsis griseola was conditioned by a single gene in Carioca 80 and two genes in PAl 127. Differences in aggressiveness of isolates of Phoma exigua var. exigua were found. Different levels of Ascochyta blight resistance were found in the glasshouse, but field testing showed little difference after flowering. Inoculations of differential cultivars indicated the presence of at least eight races of U. appendiculatus and the a-Brazil race of Colletotrichum lindemuthianum . Inoculations of the old set of halo blight differential cultivars identified races 1 and 2. Forty-five lines with partial resistance to rust were obtained by recurrent selection. Very highly significant differences were noted between ratings of percentage leaf area affected by rust and yield of 23 cultivars planted in field trials. Significant genotype x environment interaction was noted for rust ratings. Ratings at different dates within a trial were correlated with one another, showing few ratings are required per trial, and a correlation of -0.678 between yield and rust rating was found. Inheritance of partial resistance and improved yield of eight cultivars crossed in a full diallel was mostly due to additive effects but non-additive effects were also very highly significant. Reciprocal effects were not significant for yield and rust ratings. Genotype x environment interactions were significant for rust ratings and yield. High estimates of narrow-sense heritability for rust resistance were obtained. No relationship between resistance and time to flowering, pustule size, leaf hairs and stomata was found. Latent periods in unifoliate leaves did not correlate with resistance but a closer match was found in the fourth trifoliate leaves. Inoculations with three additional single-pustule isolates of the 23 parent cultivars indicated the cultivars had similar levels of resistance. Ring necrosis was found in nine cultivars or crosses with them. The ring reaction was conditioned by a single dominant gene and possibly by the epistatic interaction of two dominant genes in Carioca 80. Differences in symptom severity in plants derived from Epicure indicated the possibility of additional gene interaction. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 1994.

LD2668 .T4 1954 T64 / Master of Science

In this study, we have demonstrated that inoculation of bean seeds with non-pathogenic binucleate Rhizoctonia (np-BNR) at sowing protected bean seedlings from infection of R. solani. Using quantitative real-time RT-PCR (QRT-PCR), transcript levels of defense genes encoding 1,3-beta-glucanase (GLUC), phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) in one-week old bean seedlings was monitored during np-BNR and R. solani interaction. The results revealed that protection effect of np-BNR correspond to a systemic suppression of these three defense genes' expression from significant higher level elicited by R. solani to the level of non-infected plants. This indicates that bio-protection by np-BNR isolates is not correlated to activation of these three defense genes' expression. Similar suppression was achieved for pre-colonization of bean seedlings with arbuscular mycorrhizal (AM) Glomus introradices on GLUC gene expression, although the AM fungus did not significantly reduce rot symptoms. Possible mechanisms implicated in down-regulation during plant-pathogen and np-BNR or AM interaction are discussed.

A time course study was conducted to monitor disease development and expression of the defense-related genes phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), and hydroxyproline-rich glycoprotein (HRGP) in bean (Phaseolus vulgaris L.) plants colonized by the arbuscular mycorrhizal (AM) fungus Glomus intraradices , and post-infected with the soil-borne pathogen Rhizoctonia solani. Pre-colonization of bean plants by the AM fungus did not significantly reduce the severity of rot symptoms. RNA blot analysis revealed a systemic increase in transcript levels of the four defense-related genes in response to R. solani infection. On the other hand, pre-colonization of bean plants with G. intraradices elicited no change in PAL, CHS and CHI transcripts, but an increase of HRGP transcripts in leaves was detected. A differential and systemic alteration in the expression of all four defense genes was observed in AM beans post-infected with R. solani. Depending on the time after infection with R. solani and the tissue examined, varying responses from stimulation, suppression, to no change in transcript levels were detected.

The effect of the arbuscular mycorrhizal symbiosis at reducing the incidence of root diseases has received considerable attention. However, information on the role of mycorrhizae in reducing disease incidence of Fusarium root rot of beans (Phaseolus vulgaris), caused by the root pathogen Fusarium solani f. sp. phaseoli, is scarce. A study was undertaken to investigate how the arbuscular mycorrhizal fungus (AMF) Glomus intraradices affects disease development and population number of F. solani f. sp. phaseoli in the mycorhizosphere of bean plants growing in an experimental microcosm unit. This newly designed unit facilitated the spatial monitoring and quantification of both the symbiont and pathogen in different ecological soil regions of the mycorrhizosphere using compartmentation based on a physical segregation of roots, colonized or not by AMF (rhizosphere), AMF mycelium alone (mycosphere), or none (bulk soil). To study the interaction between both organisms, the experimental set-up consisted of a randomized complete block design using bean seedlings pre-colonized or not for 28 days by G. intraradices and infected or not for 6 days with F. solani f. sp. phaseoli. Monitoring of population number of the symbiont and the pathogen in bean plants and in the different mycorrhizosphere soil compartments was achieved with quantitative real-time PCR using specific molecular probes for each fungus, and with cultivation-dependant or morphological based methods. The results of this study indicated that non-mycorrhizal bean plants infected with the pathogen had typical root rot symptoms while infected plants that were pre-colonized by G. intraradices remained free of disease. The population number of F. solani f. sp. phaseoli was significantly reduced in the root system and in each of the mycorrhizosphere soil compartments of mycorrhizal infected plants. The mycorrhizosphere population of G. intraradices was not significantly modified, although the p

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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.