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Small-sieve snap beans (Phaseolus vulgaris L.) are an important source of income for smallholder farmers in East Africa. In this region as well as in other tropical and subtropical environments, common bean rust, caused by Uromyces appendiculatus (Pers.:Pers.), and heat stress reduce the yield and quality of snap beans. Small-sieve rust-resistant snap beans and that are heat-tolerant were developed using heat-tolerant snap bean breeding lines that had broad-spectrum rust resistance conditioned by the combination of the Andean Ur-4 and Mesoamerican Ur-11 genes. The Ur-4 and Ur-11 rust gene combination confers resistance to 90 races of the hypervariable pathogen from different parts of the world, including East Africa, that are maintained at Beltsville, MD. Four breeding lines each having the combination of the two rust genes were crossed in a 4 × 5 diallel mating design with five susceptible small-sieve cultivars to give 20 F1 hybrids. The hybrid combinations were advanced through the F2, F3, and F4 generations with selection for heat tolerance, rust resistance, and pod quality to develop lines combining these traits. Twenty F5 breeding lines that had the combination of these traits were selected and evaluated in East Africa at four field sites selected on the basis of differences in altitude, climate, and virulence diversity of the bean rust pathogen and in Puerto Rico at a field site characterized by high temperatures. There was a significant positive correlation between ranks of heat stress influenced yield components (seeds per pod and total yield) at the hot field site and the controlled high-temperature (32/27 °C) greenhouse. Four of the breeding lines developed, L5, L9, L13, and L17, combined heat tolerance and rust resistance in the desired plant type with high yield and good pod quality. These lines are the first known small-sieve snap beans with the combination of traits for heat tolerance and broad-spectrum rust resistance conferred by the Ur-4 and Ur-11 genes. These results demonstrate ability to combine heat tolerance and rust resistance as important traits for adaptation of specific market classes of common bean to tropical and subtropical environments through targeted selection of multiple traits in controlled environments.
The color of vegetables is an important factor in consumer food choices and in cultivar choice by growers and processors for production. In absorbing a broad spectrum of light, leaves support plant development by influencing factors such as biomass accumulation, chlorophyll content, and reproductive growth. The edible organ of the snap bean (Phaseolus vulgaris L.) is the pod, and its color is not only one of the most important traits for commercial consideration, but also influences phytonutrient content. Although chlorophyll provides the base color, other compounds such as carotenoids and flavonoids may affect leaf and pod color. Darker yellow- or blue-green pods are preferred for processing, but there is more leeway for fresh market, with lighter-colored pods being acceptable. This research characterized leaf and pod color variation in the 378-member Snap Bean Association Panel. Leaf and pod colors were measured with a colorimeter using the L*a*b* scale, which was then transformed to L* (lightness), C* (chroma), and H° (hue angle) for analysis. Both green and wax bean accessions had predominantly green leaves, even though both exterior and interior colors of pods varied by accession. The leaves at the upper level in the canopy were lighter than lower and middle-level leaves. C* of leaves was similar across environments but leaves from the field were greener than leaves of greenhouse-grown plants when converted to Royal Horticultural Society (RHS) values, even though they had similar H°. L* did not differ for corresponding leaf positions of both field and greenhouse leaves. Purple pods were darker (lowest L*) and yellow pods were lighter (highest L*). Although wax beans had similar exterior and interior colors, accessions with purple exterior of pods had green interiors. Green pods were generally two times higher for L* and lower in C* compared with leaves. Pod interior L* was darker than exterior in both years. Pod exterior L* was not significantly different among accessions, whereas pod interior L* differed significantly between years. Broad sense heritabilities ranged from 0.69 to 0.88 for L*, 0.12 to 0. 87 for C*, and 0.81 to 0.89 for H°. Although greater variation was observed in pods than leaves, lower heritability was determined. Moderate correlations between leaf L* and the interior and exterior pod L* implies that it would be possible to select for pod color on the basis of leaf color, with verification using standard cultivars.
Circumlineatus (cl) in common bean (Phaseolus vulgaris L.) is identified by a precipitation line in the seedcoat at the boundary of the white and colored zones. Cl is recessive and is expressed in partly colored seedcoats (t) with restricted patterns such as virgarcus. In this study, amplified fragment length polymorphism (AFLP) and single nucleotide polymorphism (SNP) markers, and the common bean genome sequence were used in combination with bulk segregant analysis and bidirectional selective genotyping to identify the genetic location of Cl. Markers were identified that cosegregated with Cl using Cl/Cl and cl/cl F3 and F5 progeny bulks from the cross t z cl G b v virgarcus BC3 5-593 × t z sel Cl G b v sellatus BC3 5-593. Two bands from an AFLP primer combination, which yielded unambiguous polymorphisms between the bulks, were cloned and sequenced. The two sequences were used to interrogate the common bean whole genome sequence identifying a region also found through cosegregation analysis using bidirectional selective genotyping with SNPs. Thus, the Cl gene was localized on Pv09 using cosegregating AFLP and SNP markers, and the physical location was confirmed with the whole genome sequence.
Common bean rust disease (caused by Uromyces appendiculatus) and high temperatures (heat stress) limit snap bean (Phaseolus vulgaris) production in many tropical and temperate regions. We have developed snap bean lines combining broad-spectrum rust resistance with heat tolerance for tropical agroecosystems. Eight breeding populations were developed by hybridizing BelJersey-RR-15 and BelFla-RR-1 (each possessing the Ur-4 and Ur-11 rust resistance genes) and the heat-tolerant snap bean breeding lines HT601, HT603, HT608, and HT611. F2–F4 generations of the populations were evaluated under greenhouse conditions and selected for heat tolerance while simultaneously selecting for the rust resistance genes Ur-4 and Ur-11. Three heat-tolerant F5 lines, which were homozygous for Ur-4 and Ur-11 genes, were selected together with a rust-resistant but heat-sensitive control. These and 12 cultivars adapted to different geographical regions were evaluated for their reaction to rust and yield at six contrasting field sites in eastern Africa and their response to high temperature verified in Puerto Rico. Rust incidence and severity was high at three of the trial sites in eastern Africa. Two of the 12 cultivars were resistant to rust at most of these sites, and three of the four breeding lines were resistant at all sites. The Ur-11 gene effectively conferred rust resistance at all sites. Yield in Puerto Rico was strongly correlated (R 2 = 0.71, P < 0.001) with that of the hottest site in eastern Africa, highlighting the similarity in genotypic response to high temperatures at the two distinct sites. The newly developed rust-resistant and heat-tolerant breeding lines showed stable yield at the eastern Africa sites with contrasting mean temperatures compared with the cultivars presently grown in the region. Two of these lines, HT1 and HT2, were confirmed to be homozygous for Ur-4 and Ur-11 and with high heat tolerance under both greenhouse and field environments. This research validates the effectiveness of targeted rust resistance gene combinations for tropical environments and the effective selection of high temperature tolerance traits correlating across multiple environments. The breeding lines HT1 and HT2 developed in this research could be used to improve snap beans for the tropics and other environments with similar constraints.
Common bean (Phaseolus vulgaris) is the major food legume worldwide, making it an important target for novel approaches of genetic analysis. This study evaluated the use of ethyl methane sulfonate (EMS) for the generation of a mutant population for targeted induced local lesions in genomes (TILLING) in common bean. TILLING is a powerful reverse genetics approach that uses a large mutant population for identification of mutants in loci of interest. Based on overall survival, development, and yield of treated seed, 40 mm EMS was found to be an appropriate concentration for the generation of a mutant population in common bean genotype BAT 93. Higher concentrations of EMS resulted in survival rates of less than 10% and lower concentrations resulted in the generation of fewer mutants. Based on TILLING results from other species, a population of 5000 lines is estimated to be sufficient for saturation of the common bean genome. Phenotypic mutation frequencies and the isolation of targeted mutations in the BAT 93 mutant population indicate that mutagenesis was effective.