Race Structure and Molecular Diversity of Colletotrichum lindemuthianum of Common Bean in Zambia.

Anthracnose, caused by the fungus Colletotrichum lindemuthianum, is a major disease of common bean (Phaseolus vulgaris L.) worldwide. C. lindemuthianum is genetically highly variable, and understanding the pathogen's diversity and distribution is a key step in developing common bean varieties with durable anthracnose resistance. The objectives of this study were to (i) characterize the race structure of C. lindemuthianum in Zambia and (ii) assess the molecular diversity of C. lindemuthianum in Zambia. A field survey was conducted in 20 bean-growing districts in Zambia to collect anthracnose symptomatic bean plants. A total of 103 C. lindemuthianum isolates were collected and characterized based on their reactions on 12 common bean race differential cultivars. RAM and ERIC-BOX DNA markers were used to assess molecular diversity of 60 isolates. A total of 58 races were characterized from the 103 isolates. Race 5 was the least virulent, and race 1631 was the most virulent based on their reaction on the 12 race differential cultivars. Race 19 had the highest recovery frequency (11%) and was the most extensively dispersed among the 22 bean-growing districts from where the isolates were collected. Only six races had previously been reported in Zambia, and 52 races were identified as new races reported for the first time in Zambia. Two races were virulent only on Andean cultivars, 11 races were virulent only on Middle American cultivars, and 45 races were virulent on both Andean and Middle American cultivars. No individual isolate showed pathogenicity on all the differential cultivars, and no isolate overcame the Co-4, Co-5, and Co-7 resistance gene pyramid that naturally exists in G2333. Phylogenetic analysis categorized the 60 isolates in six major clusters and six subclusters. The 60 isolates showed high genetic heterogeneity among and within a race of the same virulence. The study has revealed the existence of both Andean and Middle American races and extensive molecular diversity of C. lindemuthianum in Zambia. The knowledge on the race structure of C. lindemuthianum that this study has provided will be valuable for making breeding decisions on the host plant resistance genes required for developing common bean varieties with durable resistance to anthracnose in Zambia.

Genome-wide association analysis of anthracnose resistance in the Yellow Bean Collection of Common Bean.

Anthracnose caused by Colletotrichum lindemuthianum is a major disease of common bean (Phaseolus vulgaris) worldwide. Yellow beans are a major market class of common bean especially in eastern and southern Africa. The Yellow Bean Collection (YBC), which is comprised of 255 genotypes, and has not been used previously in genetic studies on anthracnose, is an excellent genetic resource for understanding the extent of anthracnose resistance and its genetic architecture in the yellow bean market class. The objectives of this study were i) evaluate the YBC for resistance to races 5, 19, 39, 51, 81, 183, 1050 and 1105 of C. lindemuthianum. and ii) conduct genome-wide association analysis to identify genomic regions and candidate genes associated with resistance to C. lindemuthianum. The YBC was genotyped with 72,866 SNPs, and genome-wide association analysis was conducted using Mixed Linear Model in TASSEL. Andean and Middle American genotypes with superior levels of resistance to the eight races were identified. YBC278 was the only one among 255 genotypes that was highly resistant to all eight races. Resistance to anthracnose in the YBC was controlled by major-effect loci on chromosomes Pv01, Pv03, Pv04, Pv05 and Pv07. The genomic region on Pv01, which overlapped with the Andean locus Co-1 provided resistance to races 81, 1050 and 1105. Significant SNPs for resistance to race 39 were identified on Pv02. The genomic region on Pv04, which overlapped with known major-effect loci Co-3, Co-15, Co-16, Co-y and Co-z, provided resistance to races 5, 19, 51 and 183. Novel genomic regions for resistance to race 39 were identified on Pv05 and Pv07. Plant resistance genes (R genes) with NB-ARC and LRR domains, which occurred in clusters, were identified as positional candidate genes for genomic regions on Pv02 and Pv04.

Identification of quantitative trait loci for drought tolerance in Bukoba/Kijivu Andean mapping population of common bean.

KEY MESSAGE: Quantitative Trait Loci "hotspots" for drought tolerance were identified on chromosomes Pv06, Pv07 and Pv10 of common bean. Drought is a major production constraint of common bean (Phaseolus vulgaris L.) worldwide. The objective of this study was to identify the Quantitative Trait Loci (QTL) for drought tolerance in an Andean population of Recombinant Inbred Lines (RILs). A total of 155 F5:7 RILs derived from a cross between Kijivu (drought tolerant) and Bukoba (drought susceptible) were evaluated for drought tolerance in field and pot experiments. Four field experiments were conducted at three locations in Zambia in 2020 and 2021. All field trials were conducted in the dry season under irrigation. The 155 RILs were genotyped with 11,292 SNPs, and composite interval mapping was conducted to identify QTL for drought tolerance. Seed yield for Kijivu under drought stress was consistently higher than for Bukoba across all four field trials. A total of 60 QTL were identified for morphological, agronomic, and physiological traits under drought stress and non-stress conditions. However, the majority of these QTL were specific to drought stress. QTL "hotspots" for drought tolerance were identified on chromosomes Pv06, Pv07, and Pv10. Extensive co-localizations for agronomic and morpho-physiological traits under drought stress were observed at the three drought-tolerance QTL hotspots. Additionally, these three QTL hotspots overlapped with previously identified QTL for drought tolerance, while several others identified QTL are novel. The three identified QTL hotspots could be used in future marker-assisted selection for drought tolerance in common bean.

Genome-Wide Identification of Powdery Mildew Resistance in Common Bean (Phaseolus vulgaris L.).

Genome-wide association studies (GWAS) have been utilized to detect genetic variations related to several agronomic traits and disease resistance in common bean. However, its application in the powdery mildew (PM) disease to identify candidate genes and their location in the common bean genome has not been fully addressed. Single-nucleotide polymorphism (SNP) genotyping with a BeadChip containing 5398 SNPs was used to detect genetic variations related to PM disease resistance in a panel of 211 genotypes grown under two field conditions for two consecutive years. Significant SNPs identified on chromosomes Pv04 and Pv10 were repeatable, ensuring the phenotypic data's reliability and the causal relationship. A cluster of resistance genes was revealed on the Pv04 of the common bean genome, coiled-coil-nucleotide-binding site-leucine-rich repeat (CC-NBS-LRR, CNL), and Toll/interleukin-1 receptor-nucleotide-binding site-leucine-rich repeat type (TIR-NBS-LRR, TNL)-like resistance genes were identified. Furthermore, two resistance genes, Phavu_010G1320001g and Phavu_010G136800g, were also identified on Pv10. Further sequence analysis showed that these genes were homologs to the disease-resistance protein (RLM1A-like) and the putative disease-resistance protein (At4g11170.1) in Arabidopsis. Significant SNPs related to two LRR receptor-like kinases (RLK) were only identified on Pv11 in 2018. Many genes encoding the auxin-responsive protein, TIFY10A protein, growth-regulating factor five-like, ubiquitin-like protein, and cell wall RBR3-like protein related to PM disease resistance were identified nearby significant SNPs. These results suggested that the resistance to PM pathogen involves a network of many genes constitutively co-expressed.

Characterization of Colletotrichum lindemuthianum Races in Zambia and Evaluation of the CIAT Phaseolus Core Collection for Resistance to Anthracnose.

Colletotrichum lindemuthianum, the causal pathogen of common bean (Phaseolus vulgaris) anthracnose, is highly variable. Therefore, understanding its race structure and identification of new sources of resistance is necessary for the development of varieties with durable resistance. The objectives of this study were (i) to characterize isolates of C. lindemuthianum collected from three major bean-growing regions in Zambia and (ii) evaluate the International Center for Tropical Agriculture (CIAT) Phaseolus core collection for resistance to C. lindemuthianum races 37, 73, and 566 and a blend of 20 races. Isolates collected from three major bean-growing districts in Zambia, namely Mporokoso, Mpika, and Mbala, were characterized as races 37, 73, and 566, respectively. A subset of the CIAT core collection comprising 885 accessions of common bean, 13 accessions of scarlet runner bean (Phaseolus coccineus), and 11 accessions of year bean (Phaseolus dumosus) was evaluated for resistance to races 37, 73, and 566 and a blend of 20 races in a greenhouse at the University of Zambia, Lusaka, Zambia. Totals of 72, 66, 48, and 9% of P. vulgaris accessions evaluated were highly resistant to races 37, 73, and 566 and a blend of 20 races, respectively. Also, accessions of P. coccineus and P. dumosus highly resistant to races 37, 73, and 566 were identified. Only eight of the 331 P. vulgaris accessions were highly resistant to all three individual races (37, 73, and 566) and to a blend of 20 races. These eight accessions constitute a valuable breeding resource for developing varieties with durable resistance to C. lindemuthianum.

Genetic variability and genome-wide association analysis of flavor and texture in cooked beans (Phaseolus vulgaris L.).

KEY MESSAGE: Cooked bean flavor and texture vary within and across 20 Andean seed types; SNPs are significantly associated with total flavor, beany, earthy, starchy, bitter, seed-coat perception, and cotyledon texture. Common dry beans are a nutritious food recognized as a staple globally, but their consumption is low in the USA. Improving bean flavor and texture through breeding has the potential to improve consumer acceptance and suitability for new end-use products. Little is known about genetic variability and inheritance of bean sensory characteristics. A total of 430 genotypes of the Andean Diversity Panel representing twenty seed types were grown in three locations, and cooked seeds were evaluated by a trained sensory panel for flavor and texture attribute intensities, including total flavor, beany, vegetative, earthy, starchy, sweet, bitter, seed-coat perception, and cotyledon texture. Extensive variation in sensory attributes was found across and within seed types. A set of genotypes was identified that exhibit extreme attribute intensities generally stable across all three environments. seed-coat perception and total flavor intensity had the highest broad-sense heritability (0.39 and 0.38, respectively), while earthy and vegetative intensities exhibited the lowest (0.14 and 0.15, respectively). Starchy and sweet flavors were positively correlated and highest in white bean genotypes according to principal component analysis. SNPs associated with total flavor intensity (six SNPs across three chromosomes), beany (five SNPs across four chromosomes), earthy (three SNPs across two chromosomes), starchy (one SNP), bitter (one SNP), seed-coat perception (three SNPs across two chromosomes), and cotyledon texture (two SNPs across two chromosomes) were detected. These findings lay a foundation for incorporating flavor and texture in breeding programs for the development of new varieties that entice growers, consumers, and product developers alike.

Identification of quantitative trait loci for symbiotic nitrogen fixation in common bean.

KEY MESSAGE: QTL were identified for symbiotic nitrogen fixation in common bean. These QTL were detected in both greenhouse and field studies, and many overlapped with previously reported QTL in diverse mapping populations. Common bean (Phaseolus vulgaris L.) productivity can be improved through the genetic enhancement of its symbiotic nitrogen fixation (SNF) capacity. This study was aimed at understanding the genetic architecture of SNF through QTL analysis of a recombinant inbred line (RIL) population contrasting for SNF potential. The mapping population consisted of 188 F4:5 RILs derived from a cross of Solwezi and AO-1012-29-3-3A that were evaluated for SNF in the greenhouse and field in Zambia. The population was genotyped with 5398 single-nucleotide polymorphism (SNP) markers. QTL for shoot biomass, nitrogen percentage in shoot biomass, nitrogen percentage in seed, total nitrogen derived from atmosphere (Ndfa) and percentage of nitrogen derived from the atmosphere (%Ndfa) were identified. Three QTL for %Ndfa were identified on chromosomes Pv01, Pv04 and Pv09. Five QTL for Ndfa were identified on Pv04, Pv06, Pv07, Pv09 and Pv11. The QTL Ndfa9.1SA identified in the current study overlapped with a previously reported QTL for SNF. A major QTL Ndfa7.1DB, SA (R2 = 14.9%) was consistently identified in two greenhouse studies and overlapped with previously reported QTL. The QTL Ndfa4.2SA identified from the greenhouse experiment is novel and overlapped with the QTL %NB4.3SA, %NS4.2SA and %Ndfa4.2SA from the field experiment. These QTL identified in both greenhouse and field experiments, which overlap with previously reported QTL, could potentially be deployed by marker-assisted breeding to accelerate development of bean cultivars with enhanced SNF.

Transcriptome analysis of two recombinant inbred lines of common bean contrasting for symbiotic nitrogen fixation.

Common bean (Phaseolus vulgaris L.) fixes atmospheric nitrogen (N2) through symbiotic nitrogen fixation (SNF) at levels lower than other grain legume crops. An understanding of the genes and molecular mechanisms underlying SNF will enable more effective strategies for the genetic improvement of SNF traits in common bean. In this study, transcriptome profiling was used to identify genes and molecular mechanisms underlying SNF differences between two common bean recombinant inbred lines that differed in their N-fixing abilities. Differential gene expression and functional enrichment analyses were performed on leaves, nodules and roots of the two lines when grown under N-fixing and non-fixing conditions. Receptor kinases, transmembrane transporters, and transcription factors were among the differentially expressed genes identified under N-fixing conditions, but not under non-fixing conditions. Genes up-regulated in the stronger nitrogen fixer, SA36, included those involved in molecular functions such as purine nucleoside binding, oxidoreductase and transmembrane receptor activities in nodules, and transport activity in roots. Transcription factors identified in this study are candidates for future work aimed at understanding the functional role of these genes in SNF. Information generated in this study will support the development of gene-based markers to accelerate genetic improvement of SNF in common bean.

Transcriptome Profiling of the Phaseolus vulgaris - Colletotrichum lindemuthianum Pathosystem.

Bean (Phaseolus vulgaris) anthracnose caused by the hemi-biotrophic pathogen Colletotrichum lindemuthianum is a major factor limiting production worldwide. Although sources of resistance have been identified and characterized, the early molecular events in the host-pathogen interface have not been investigated. In the current study, we conducted a comprehensive transcriptome analysis using Illumina sequencing of two near isogenic lines (NILs) differing for the presence of the Co-1 gene on chromosome Pv01 during a time course following infection with race 73 of C. lindemuthianum. From this, we identified 3,250 significantly differentially expressed genes (DEGs) within and between the NILs over the time course of infection. During the biotrophic phase the majority of DEGs were up regulated in the susceptible NIL, whereas more DEGs were up-regulated in the resistant NIL during the necrotrophic phase. Various defense related genes, such as those encoding PR proteins, peroxidases, lipoxygenases were up regulated in the resistant NIL. Conversely, genes encoding sugar transporters were up-regulated in the susceptible NIL during the later stages of infection. Additionally, numerous transcription factors (TFs) and candidate genes within the vicinity of the Co-1 locus were differentially expressed, suggesting a global reprogramming of gene expression in and around the Co-1 locus. Through this analysis, we reduced the previous number of candidate genes reported at the Co-1 locus from eight to three. These results suggest the dynamic nature of P. vulgaris-C. lindemuthianum interaction at the transcriptomic level and reflect the role of both pathogen and effector triggered immunity on changes in plant gene expression.

Genome-Wide Association Study of Anthracnose Resistance in Andean Beans (Phaseolus vulgaris).

Anthracnose is a seed-borne disease of common bean (Phaseolus vulgaris L.) caused by the fungus Colletotrichum lindemuthianum, and the pathogen is cosmopolitan in distribution. The objectives of this study were to identify new sources of anthracnose resistance in a diverse panel of 230 Andean beans comprised of multiple seed types and market classes from the Americas, Africa, and Europe, and explore the genetic basis of this resistance using genome-wide association mapping analysis (GWAS). Twenty-eight of the 230 lines tested were resistant to six out of the eight races screened, but only one cultivar Uyole98 was resistant to all eight races (7, 39, 55, 65, 73, 109, 2047, and 3481) included in the study. Outputs from the GWAS indicated major quantitative trait loci (QTL) for resistance on chromosomes, Pv01, Pv02, and Pv04 and two minor QTL on Pv10 and Pv11. Candidate genes associated with the significant SNPs were detected on all five chromosomes. An independent QTL study was conducted to confirm the physical location of the Co-1 locus identified on Pv01 in an F4:6 recombinant inbred line (RIL) population. Resistance was determined to be conditioned by the single dominant gene Co-1 that mapped between 50.16 and 50.30 Mb on Pv01, and an InDel marker (NDSU_IND_1_50.2219) tightly linked to the gene was developed. The information reported will provide breeders with new and diverse sources of resistance and genomic regions to target in the development of anthracnose resistance in Andean beans.

Genome-wide association analysis of symbiotic nitrogen fixation in common bean.

KEY MESSAGE: Significant SNPs and candidate genes for symbiotic nitrogen fixation (SNF) and related traits were identified on Pv03, Pv07 and Pv09 chromosomes of common bean. A genome-wide association study (GWAS) was conducted to explore the genetic basis of variation for symbiotic nitrogen fixation (SNF) and related traits in the Andean Diversity Panel (ADP) comprising 259 common bean (Phaseolus vulgaris) genotypes. The ADP was evaluated for SNF and related traits in both greenhouse and field experiments. After accounting for population structure and cryptic relatedness, significant SNPs were identified on chromosomes Pv03, Pv07 and Pv09 for nitrogen derived from atmosphere (Ndfa) in the shoot at flowering, and for Ndfa in seed. The SNPs for Ndfa in shoot and Ndfa in seed co-localized on Pv03 and Pv09. Two genes Phvul.007G050500 and Phvul.009G136200 that code for leucine-rich repeat receptor-like protein kinases (LRR-RLK) were identified as candidate genes for Ndfa. LRR-RLK genes play a key role in signal transduction required for nodule formation. Significant SNPs identified in this study could potentially be used in marker-assisted breeding to accelerate genetic improvement of common bean for SNF.

Genome-Wide Association Study of Agronomic Traits in Common Bean.

A genome-wide association study (GWAS) using a global Andean diversity panel (ADP) of 237 genotypes of common bean (Phaseolus vulgaris L.) was conducted to gain insight into the genetic architecture of phenology, biomass, yield components, and seed yield traits. The panel was evaluated for 2 yr in field trials in Michigan and genotyped with 5398 single nucleotide polymorphism (SNP) markers. After correcting for population structure and cryptic relatedness, significant SNP markers associated with several agronomic traits were identified. Positional candidate genes, including Phvul.001G221100 on P. vulgaris (Pv) chromosome 01, associated with days to flowering and maturity were identified. Significant SNPs for seed yield were identified on Pv03 and Pv09 and colocalized with quantitative trait loci (QTL) for yield from previous studies conducted in several environments and contrasting genetic backgrounds. The majority of germplasm carrying the alleles with positive effects on seed yield was of African origin and largely underutilized in US breeding programs. The study provided insights into the genetic architecture of agronomic traits in Andean beans.