Bean Chromosomes Research Articles

Biochemical Defense Arsenal, Genes/QTLs and Transcripts for Imparting Anthracnose Resistance in Common bean (Phaseolus vulgaris L.)

Anthracnose (ANT), caused by Colletotrichum lindemuthianum, is the most devastating disease affecting common bean (Phaseolus vulgaris L.), leading to significant yield losses in the Western Himalayas. The study provides a comprehensive understanding of ANT resistance via trait phenotyping, biochemical profiling, genome-wide association studies (GWASs), and RNA sequencing. The assessment of bean association mapping panel in different environments revealed a diverse spectrum of resistance levels. Biochemical analysis revealed distinctive defense responses against ANT infection among different genotypes. GWAS approach identified 24 significant marker‒trait associations (MTAs) distributed across all 11 bean chromosomes. Notably, 03 MTAs (BMr205, BMr269 and BMr244) present on chromosome Pv07 were validated for ANT, and the remaining MTAs were novel MTAs for ANT. Transcriptome sequencing of resistant (PBG-3) and susceptible (PBG-26) genotypes under mock and 120-hour post inoculation conditions revealed key differentially expressed genes, such as leucine-rich repeat domain-containing protein (PHAVU_007G087700g), NB-ARC domain-containing protein (PHAVU_003G002500g) and transcription factors pivotal for disease resistance. The expression patterns of four genes (PHAVU_007G087700g, PHAVU_003G002500g, PHAVU_007G056100g and PHAVU_003G003000g) were validated through quantitative reverse transcription polymerase chain reaction (qRT‒PCR). Furthermore, the integration of GWAS-identified candidate genes with transcriptomics and cross-referencing with previous studies validated overlapping regions and common candidate genes, enriching our understanding of the genetic basis of ANT resistance. Therefore, the results offer a holistic perspective on ANT resistance in common bean, providing a foundation for targeted breeding efforts. The identified potential candidate genes and associated pathways will contribute valuable insights into the development of ANT resistant common bean varieties.

Alterations in Gene Expression during Incompatible Interaction between Amendoim Cavalo Common Bean and Colletotrichum lindemuthianum.

Anthracnose, caused by the fungus Colletotrichum lindemuthianum, poses a significant and widespread threat to the common bean crop. The use of plant genetic resistance has proven to be the most effective strategy for managing anthracnose disease. The Amendoim Cavalo (AC) Andean cultivar has resistance against multiple races of C. lindemuthianum, which is conferred by the Co-AC gene. Fine mapping of this resistance gene to common bean chromosome Pv01 enabled the identification of Phvul.001G244300, Phvul.001G244400, and Phvul.001G244500 candidate genes for further validation. In this study, the relative expression of Co-AC candidate genes was assessed, as well as other putative genes in the vicinity of this locus and known resistance genes, in the AC cultivar following inoculation with the race 73 of C. lindemuthianum. Gene expression analysis revealed significantly higher expression levels of Phvul.001G244500. Notably, Phvul.001G244500 encodes a putative Basic Helix-Loop-Helix transcription factor, suggesting its involvement in the regulation of defense responses. Furthermore, a significant modulation of the expression of defense-related genes PR1a, PR1b, and PR2 was observed in a time-course experiment. These findings contribute to the development of improved strategies for breeding anthracnose-resistant common bean cultivars, thereby mitigating the impact of this pathogen on crop yields and ensuring sustainable bean production.

Inheritance and genetic mapping of the first CPMMV tolerance locus in common bean

AbstractAlthough cowpea mild mottle virus (CPMMV) has been reported in Brazil since 1983, it has only become a significant concern for researchers and farmers in recent years. The objective of this work was to investigate the genetic basis of CPMMV tolerance, mapping and reporting the first loci associated with this trait in common bean (cv. BRS Sublime). Phenotypic assays were carried out on 180 individual plants (F2 generation) and 180 F2:3 progenies comprising 12 plants per family and their parents (BRS Sublime, tolerant parent × CNFCT 16207, susceptible parent). CPMMV was mechanically inoculated and symptoms were evaluated at 35 days after inoculation, using a 1–5 scoring scale. A linkage map was constructed using 1695 single‐nucleotide polymorphism (SNP) and SilicoDArT markers that segregated in the F2 and F2:3 generations as expected. Markers were distributed across 11 common bean chromosomes, resulting in a total length of 2864 cM, with an average distance between markers of 1.8 cM. Phenotypic observations revealed that tolerance in cv. BRS Sublime is controlled by a single dominant gene. The main effect quantitative trait locus (QTL; CPMMV.Pv08) associated with CPMMV tolerance was identified in the terminal region on chromosome 8 (Pv08). This QTL explained approximately 77% of phenotypic variation, indicating that the inheritance of tolerance to CPMMV is monogenic, controlled by a major locus. As far as we know, this study represents the first investigation into the inheritance and genetic mapping of CPMMV tolerance in common bean, with potential for the development of elite lines with multiple virus resistance/tolerance.

Genome-wide association study for yield-related traits in faba bean (Vicia faba L.).

Yield is the most complex trait to improve crop production, and identifying the genetic determinants for high yield is a major issue in breeding new varieties. In faba bean (Vicia faba L.), quantitative trait loci (QTLs) have previously been detected in studies of biparental mapping populations, but the genes controlling the main trait components remain largely unknown. In this study, we investigated for the first time the genetic control of six faba bean yield-related traits: shattering (SH), pods per plant (PP), seeds per pod (SP), seeds per plant (SPL), 100-seed weight (HSW), and plot yield (PY), using a genome-wide association study (GWAS) on a worldwide collection of 352 homozygous faba bean accessions with the aim of identifying markers associated with them. Phenotyping was carried out in field trials at three locations (Spain, United Kingdom, and Serbia) over 2 years. The faba bean panel was genotyped with the Affymetrix faba bean SNP-chip yielding 22,867 SNP markers. The GWAS analysis identified 112 marker-trait associations (MTAs) in 97 candidate genes, distributed over the six faba bean chromosomes. Eight MTAs were detected in at least two environments, and five were associated with multiple traits. The next step will be to validate these candidates in different genetic backgrounds to provide resources for marker-assisted breeding of faba bean yield.

Combining GWAS and comparative genomics to fine map candidate genes for days to flowering in mung bean

BackgroundMung bean (Vigna radiata (L.) Wilczek), is an important pulse crop in the global south. Early flowering and maturation are advantageous traits for adaptation to northern and southern latitudes. This study investigates the genetic basis of the Days-to-Flowering trait (DTF) in mung bean, combining genome-wide association studies (GWAS) in mung bean and comparisons with orthologous genes involved with control of DTF responses in soybean (Glycine max (L) Merr) and Arabidopsis (Arabidopsis thaliana).ResultsThe most significant associations for DTF were on mung bean chromosomes 1, 2, and 4. Only the SNPs on chromosomes 1 and 4 were heavily investigated using downstream analysis. The chromosome 1 DTF association is tightly linked with a cluster of locally duplicated FERONIA (FER) receptor-like protein kinase genes, and the SNP occurs within one of the FERONIA genes. In Arabidopsis, an orthologous FERONIA gene (AT3G51550), has been reported to regulate the expression of the FLOWERING LOCUS C (FLC). For the chromosome 4 DTF locus, the strongest candidates are Vradi04g00002773 and Vradi04g00002778, orthologous to the Arabidopsis PhyA and PIF3 genes, encoding phytochrome A (a photoreceptor protein sensitive to red to far-red light) and phytochrome-interacting factor 3, respectively. The soybean PhyA orthologs include the classical loci E3 and E4 (genes GmPhyA3, Glyma.19G224200, and GmPhyA2, Glyma.20G090000). The mung bean PhyA ortholog has been previously reported as a candidate for DTF in studies conducted in South Korea.ConclusionThe top two identified SNPs accounted for a significant proportion (~ 65%) of the phenotypic variability in mung bean DTF by the six significant SNPs (39.61%), with a broad-sense heritability of 0.93. The strong associations of DTF with genes that have orthologs with analogous functions in soybean and Arabidopsis provide strong circumstantial evidence that these genes are causal for this trait. The three reported loci and candidate genes provide useful targets for marker-assisted breeding in mung beans.

Mapping yield and yield-related traits using diverse common bean germplasm.

Common bean (bean) is one of the most important legume crops, and mapping genes for yield and yield-related traits is essential for its improvement. However, yield is a complex trait that is typically controlled by many loci in crop genomes. The objective of this research was to identify regions in the bean genome associated with yield and a number of yield-related traits using a collection of 121 diverse bean genotypes with different yields. The beans were evaluated in replicated trials at two locations, over two years. Significant variation among genotypes was identified for all traits analyzed in the four environments. The collection was genotyped with the BARCBean6K_3 chip (5,398 SNPs), two yield/antiyield gene-based markers, and seven markers previously associated with resistance to common bacterial blight (CBB), including a Niemann-Pick polymorphism (NPP) gene-based marker. Over 90% of the single-nucleotide polymorphisms (SNPs) were polymorphic and separated the panel into two main groups of small-seeded and large-seeded beans, reflecting their Mesoamerican and Andean origins. Thirty-nine significant marker-trait associations (MTAs) were identified between 31 SNPs and 15 analyzed traits on all 11 bean chromosomes. Some of these MTAs confirmed genome regions previously associated with the yield and yield-related traits in bean, but a number of associations were not reported previously, especially those with derived traits. Over 600 candidate genes with different functional annotations were identified for the analyzed traits in the 200-Kb region centered on significant SNPs. Fourteen SNPs were identified within the gene model sequences, and five additional SNPs significantly associated with five different traits were located at less than 0.6Kb from the candidate genes. The work confirmed associations between two yield/antiyield gene-based markers (AYD1m and AYD2m) on chromosome Pv09 with yield and identified their association with a number of yield-related traits, including seed weight. The results also confirmed the usefulness of the NPP marker in screening for CBB resistance. Since disease resistance and yield measurements are environmentally dependent and labor-intensive, the three gene-based markers (CBB- and two yield-related) and quantitative trait loci (QTL) that were validated in this work may be useful tools for simplifying and accelerating the selection of high-yielding and CBB-resistant bean cultivars.

Genetic control of pod morphological traits and pod edibility in a common bean RIL population

Key messageQTL mapping, association analysis, and colocation study with previously reported QTL revealed three main regions controlling pod morphological traits and two loci for edible pod characteristics on the common bean chromosomes Pv01 and Pv06.Bean pod phenotype is a complex characteristic defined by the combination of different traits that determine the potential use of a genotype as a snap bean. In this study, the TUM RIL population derived from a cross between ‘TU’ (dry) and ‘Musica’ (snap) was used to investigate the genetic control of pod phenotype. The character was dissected into pod morphological traits (PMTs) and edible pod characteristics (EPC). The results revealed 35 QTL for PMTs located on seven chromosomes, suggesting a strong QTL colocation on chromosomes Pv01 and Pv06. Some QTL were colocated with previously reported QTL, leading to the mapping of 15 consensus regions associated with bean PMTs. Analysis of EPC of cooked beans revealed that two major loci with epistatic effect, located on chromosomes Pv01 and Pv06, are involved in the genetic control of this trait. An association study using a subset of the Spanish Diversity Panel (snap vs. non-snap) detected 23 genomic regions, with three regions being mapped at a position similar to those of two loci identified in the TUM population. The results demonstrated the relevant roles of Pv01 and Pv06 in the modulation of bean pod phenotype. Gene ontology enrichment analysis revealed a significant overrepresentation of genes regulating the phenylpropanoid metabolic process and auxin response in regions associated with PMTs and EPC, respectively. Both biological functions converged in the lignin biosynthetic pathway, suggesting the key role of the pathway in the genetic control of bean pod phenotype.

Genome-wide analysis and characterization of the LRR-RLK gene family provides insights into anthracnose resistance in common bean

Anthracnose, caused by the hemibiotrophic fungus Colletotrichum lindemuthianum, is a damaging disease of common beans that can drastically reduce crop yield. The most effective strategy to manage anthracnose is the use of resistant cultivars. There are many resistance loci that have been identified, mapped and associated with markers in common bean chromosomes. The Leucine-rich repeat kinase receptor protein (LRR-RLK) family is a diverse group of transmembrane receptors, which potentially recognizes pathogen-associated molecular patterns and activates an immune response. In this study, we performed in silico analyses to identify, classify, and characterize common bean LRR-RLKs, also evaluating their expression profile in response to the infection by C. lindemuthianum. By analyzing the entire genome of Phaseolus vulgaris, we could identify and classify 230 LRR-RLKs into 15 different subfamilies. The analyses of gene structures, conserved domains and motifs suggest that LRR-RLKs from the same subfamily are consistent in their exon/intron organization and composition. LRR-RLK genes were found along the 11 chromosomes of the species, including regions of proximity with anthracnose resistance markers. By investigating the duplication events within the LRR-RLK family, we associated the importance of such a family with an expansion resulting from a strong stabilizing selection. Promoter analysis was also performed, highlighting cis-elements associated with the plant response to biotic stress. With regard to the expression pattern of LRR-RLKs in response to the infection by C. lindemuthianum, we could point out several differentially expressed genes in this subfamily, which were associated to specific molecular patterns of LRR-RLKs. Our work provides a broad analysis of the LRR-RLK family in P. vulgaris, allowing an in-depth structural and functional characterization of genes and proteins of this family. From specific expression patterns related to anthracnose response, we could infer a direct participation of RLK-LRR genes in the mechanisms of resistance to anthracnose, highlighting important subfamilies for further investigations.

Whole-genome resequencing of common bean elite breeding lines

The expansion of bean genome technologies has prompted new perspectives on generating resources and knowledge essential to research and implementing biotechnological tools for the practical operations of plant breeding programs. This study aimed to resequence the entire genome (whole genome sequencing—WGS) of 40 bean genotypes selected based on their significance in breeding programs worldwide, with the objective of generating an extensive database for the identification of single nucleotide polymorphisms (SNPs). Over 6 million SNPs were identified, distributed across the 11 bean chromosomes. After quality variant filtering, 420,509 high-quality SNPs were established, with an average of 38,228 SNPs per chromosome. These variants were categorized based on their predicted effects, revealing that the majority exerted a modifier impact on non-coding genome regions (94.68%). Notably, a significant proportion of SNPs occurred in intergenic regions (62.89%) and at least one SNP was identified in 58.63% of the genes annotated in the bean genome. Of particular interest, 7841 SNPs were identified in 85% of the putative plant disease defense-related genes, presenting a valuable resource for crop breeding efforts. These findings provide a foundation for the development of innovative and broadly applicable technologies for the routine selection of superior genotypes in global bean improvement and germplasm characterization programs.

The ricebean genome provides insight into Vigna genome evolution and facilitates genetic enhancement.

The ricebean genome provides insight into Vigna genome evolution and facilitates genetic enhancement.

Construction of a high-density genetic map for faba bean (Vicia faba L.) and quantitative trait loci mapping of seed-related traits.

Faba bean (Vicia faba L.) is a valuable legume crop and data on its seed-related traits is required for yield and quality improvements. However, basic research on faba bean is lagging compared to that of other major crops. In this study, an F2 faba bean population, including 121 plants derived from the cross WY7×TCX7, was genotyped using the Faba_bean_130 K targeted next-generation sequencing genotyping platform. The data were used to construct the first ultra-dense faba bean genetic map consisting of 12,023 single nucleotide polymorphisms markers covering 1,182.65 cM with an average distance of 0.098 cM. The map consisted of 6 linkage groups, which is consistent with the 6 faba bean chromosome pairs. A total of 65 quantitative trait loci (QTL) for seed-related traits were identified (3 for 100-seed weight, 28 for seed shape, 12 for seed coat color, and 22 for nutritional quality). Furthermore, 333 candidate genes that are likely to participate in the regulation of seed-related traits were also identified. Our research findings can provide a basis for future faba bean marker-assisted breeding and be helpful to further modify and improve the reference genome.

Multi-Environment Genome-Wide Association Studies of Yield Traits in Common Bean (Phaseolus vulgaris L.) × Tepary Bean (P. acutifolius A. Gray) Interspecific Advanced Lines in Humid and Dry Colombian Caribbean Subregions

Assessing interspecific adaptive genetic variation across environmental gradients offers insight into the scale of habitat-dependent heritable heterotic effects, which may ultimately enable pre-breeding for abiotic stress tolerance and novel climates. However, environmentally dependent allelic effects are often bypassed by intra-specific single-locality genome-wide associations studies (GWAS). Therefore, in order to bridge this gap, this study aimed at coupling an advanced panel of drought/heat susceptible common bean (Phaseolus vulgaris L.) × tolerant tepary bean (P. acutifolius A. Gray) interspecific lines with last-generation multi-environment GWAS algorithms to identify novel sources of heat and drought tolerance to the humid and dry subregions of the Caribbean coast of Colombia, where the common bean typically exhibits maladaptation to extreme weather. A total of 87 advanced lines with interspecific ancestries were genotyped by sequencing (GBS), leading to the discovery of 15,645 single-nucleotide polymorphism (SNP) markers. Five yield traits were recorded for each genotype and inputted in modern GWAS algorithms (i.e., FarmCPU and BLINK) to identify the putative associated loci across four localities in coastal Colombia. Best-fit models revealed 47 significant quantitative trait nucleotides (QTNs) distributed in all 11 common bean chromosomes. A total of 90 flanking candidate genes were identified using 1-kb genomic windows centered in each associated SNP marker. Pathway-enriched analyses were done using the mapped output of the GWAS for each yield trait. Some genes were directly linked to the drought tolerance response; morphological, physiological, and metabolic regulation; signal transduction; and fatty acid and phospholipid metabolism. We conclude that habitat-dependent interspecific polygenic effects are likely sufficient to boost common bean adaptation to the severe climate in coastal Colombia via introgression breeding. Environmental-dependent polygenic adaptation may be due to contrasting levels of selection and the deleterious load across localities. This work offers putative associated loci for marker-assisted and genomic selection targeting the common bean’s neo-tropical lowland adaptation to drought and heat.

Genetic fine‐mapping of anthracnose disease‐resistance allele Co‐14 present in the Andean common bean cultivar AND 277

AbstractAnthracnose (ANT) is among the common bean fungal diseases responsible for significant yield and grain quality losses. Durable genetic resistance is the primary ANT control method due to the pathogen's high variability. Genetic studies showed that the common bean chromosome Pv01 contains multiple disease resistance genes, including the ANT resistance loci Co‐1 (Co‐12, Co‐13, Co‐14, Co‐15, Co‐1HY, and Co‐1x), Co‐AC, Co‐14, Co‐Pa, Co‐Perla, Co‐w, Co‐x, and CoPv01CDRK. This work aimed to: (i) perform the genetic fine‐mapping of the Co‐14 allele present in the cultivar AND 277, using recombinant inbred lines derived from the cross between Rudá × AND 277; and (ii) identify candidate resistance genes in the Co‐14 allele based on the common bean reference genome. Initially, the Co‐14 allele was mapped between the single‐nucleotide polymorphism markers ss715645251 and ss715645250 at a distance of 2.0 and 19.6 cM, respectively. Fine‐mapping localized Co‐14 between the markers ss715645251 and BARCPVSSR01356, spanning a 40.51 kb region at the end of Pv01. Two genes are described within the Co‐14 region in the reference genome, Phvul.001G243800 and Phvul.001G243900. The linkage between the Co‐14 allele and the markers ss715645251 and BARCPVSSR01356 will be essential for plant breeding programs during the resistance gene transfer to elite cultivars via marker‐assisted selection (MAS). Identifying and functionally analyzing candidate resistance genes in this region will allow more efficient MAS by developing accurate markers for ANT resistance.

Delineating meta-quantitative trait loci for anthracnose resistance in common bean (Phaseolus vulgaris L.).

Anthracnose, caused by the fungus Colletotrichum lindemuthianum, is one of the devastating disease affecting common bean production and productivity worldwide. Several quantitative trait loci (QTLs) for anthracnose resistance have been identified. In order to make use of these QTLs in common bean breeding programs, a detailed meta-QTL (MQTL) analysis has been conducted. For the MQTL analysis, 92 QTLs related to anthracnose disease reported in 18 different earlier studies involving 16 mapping populations were compiled and projected on to the consensus map. This meta-analysis led to the identification of 11 MQTLs (each involving QTLs from at least two different studies) on 06 bean chromosomes and 10 QTL hotspots each involving multiple QTLs from an individual study on 07 chromosomes. The confidence interval (CI) of the identified MQTLs was found 3.51 times lower than the CI of initial QTLs. Marker-trait associations (MTAs) reported in published genome-wide association studies (GWAS) were used to validate nine of the 11 identified MQTLs, with MQTL4.1 overlapping with as many as 40 MTAs. Functional annotation of the 11 MQTL regions revealed 1,251 genes including several R genes (such as those encoding for NBS-LRR domain-containing proteins, protein kinases, etc.) and other defense related genes. The MQTLs, QTL hotspots and the potential candidate genes identified during the present study will prove useful in common bean marker-assisted breeding programs and in basic studies involving fine mapping and cloning of genomic regions associated with anthracnose resistance in common beans.

Exploring the GRAS gene family in common bean (Phaseolus vulgaris L.): characterization, evolutionary relationships, and expression analyses in response to abiotic stresses.

Genome-wide identification reveals 55 PvuGRAS genes belonging to 16 subfamilies and their gene structures and evolutionary relationships were characterized. Expression analyses highlight their prominence in plant growth, development and abiotic stress responses. GRAS proteins comprise a plant-specific transcription factor family involved in multiple growth regulatory pathways and environmental cues including abiotic/biotic stresses. Despite its crucial importance, characterization of this gene family is still elusive in common bean. A systematic genome-wide scan identified 55 PvuGRAS genes unevenly anchored to the 11 common bean chromosomes. Segmental duplication appeared to be the key driving force behind expansion of this gene family that underwent purifying selection during evolution. Computational investigation unraveled their intronless organization and identified similar motif composition within the same subfamily. Phylogenetic analyses clustered the PvuGRAS proteins into 16 phylogenetic clades and established extensive orthologous relationships with Arabidopsis and rice. Analysis of the upstream promoter region uncovered cis-elements responsive to growth, development, and abiotic stresses that may account for their differential expression. The identified SSRs could serve as putative molecular markers facilitating future breeding programs. 37 PvuGRAS transcripts were post-transcriptionally regulated by different miRNA families, miR171 being the major player preferentially targeting members of the HAM subfamily. Global expression profile based on RNA-seq data indicates a clade specific expression pattern in various tissues and developmental stages. Additionally, nine PvuGRAS genes were chosen for further qPCR analyses under drought, salt, and cold stress suggesting their involvement in acclimation to environmental stimuli. Combined, the present results significantly contribute to the current understanding of the complexity and biological function of the PvuGRAS gene family. The resources generated will provide a solid foundation in future endeavors for genetic improvement in common bean.

Genome‐wide association study (GWAS) of Phaseolus vulgaris resistance to Xanthomonas citri pv. fuscans

AbstractCommon bean is one of the main legumes used for food worldwide. Among the main limitations for its production is common bacterial blight (CBB) caused by Xanthomonas citri pv. fuscans (Xcf). The most desirable method to control the disease is the use of plant genetic resistance, but resistance against Xcf has not been sufficiently investigated. Genetic studies have shown that resistance to CBB is of quantitative type, but no gene associated with resistance has been identified. Here, we aimed to determine the resistance to CBB of 103 Mesoamerican common bean cultivars by evaluating disease severity and the area under the disease progress curve (AUDPC) after inoculation with a Xcf strain. In addition, 80 of the evaluated cultivars were used to investigate the association of single nucleotide polymorphisms (SNPs) with the resistance phenotype in a genome‐wide association study (GWAS). Twenty‐nine cultivars were classified as highly resistant, 15 of which are shown to be resistant to CBB for the first time in this work. A positive correlation between AUDPC and disease severity was observed. The 10 SNPs with the highest −log10(p) values in the GWAS are located on bean chromosomes Pv01, Pv03, Pv06, Pv07, Pv08, Pv09, Pv10, and Pv11 within or near genes predicted to code for biochemical functions such as serine/threonine‐protein kinase, glutamine synthase, lectin domain protein, MADS‐domain transcription factor, and pentatricopeptide repeat protein, previously associated with immunity. This work identifies common bean germplasm resistant to Xcf, molecular markers for germplasm selection, and genes potentially involved in the resistance response.

First approach to pod dehiscence in faba bean: genetic and histological analyses

Pod dehiscence causes important yield losses in cultivated crops and therefore has been a key trait strongly selected against in crop domestication. In spite of the growing knowledge on the genetic basis of dehiscence in different crops, no information is available so far for faba bean. Here we conduct the first comprehensive study for faba bean pod dehiscence by combining, linkage mapping, comparative genomics, QTL analysis and histological examination of mature pods. Mapping of dehiscence-related genes revealed conservation of syntenic blocks among different legumes. Three QTLs were identified in faba bean chromosomes II, IV and VI, although none of them was stable across years. Histological analysis supports the convergent phenotypic evolution previously reported in cereals and related legume species but revealed a more complex pattern in faba bean. Contrary to common bean and soybean, the faba bean dehiscence zone appears to show functional equivalence to that described in crucifers. The lignified wall fiber layer, which is absent in the paucijuga primitive line Vf27, or less lignified and vacuolated in other dehiscent lines, appears to act as the major force triggering pod dehiscence in this species. While our findings, provide new insight into the mechanisms underlying faba bean dehiscence, full understanding of the molecular bases will require further studies combining precise phenotyping with genomic analysis.

Integration of anthracnose resistance loci and RLK and NBS‐LRR‐encoding genes in the Phaseolus vulgaris L. genome

AbstractThe most effective strategy to manage bean anthracnose (ANT), caused by Colletotrichum lindemuthianum, is the use of resistant cultivars. There are more than 20 ANT resistance genes that have been identified and mapped in common bean (Phaseolus vulgaris L.) chromosomes. Moreover, quantitative resistance loci (QRLs) have been described through genome‐wide association studies (GWAS). Identification of pathogen‐responsive genes and proteins on a molecular level provides a better understanding of metabolic pathways involved in ANT resistance. In this study, we investigated typical resistance proteins located close to ANT resistance loci in the common bean reference genome. Among them, we checked for proteins with nucleotide‐binding and leucine‐rich repeat (NBS‐LRR) domains and kinase domains since most resistance genes encode proteins with these domains. In addition, proteins with kinase domains are known to operate as pattern‐recognition receptors that recognize pathogen‐associated molecular patterns (PAMPs) and activate an immune response. Based on the common bean reference genome (Version 2.1), the regions 500‐kb upstream and downstream of the physical position of each ANT resistance locus were considered for a candidate gene search. Thus, an integrated map of ANT resistance loci and candidate genes (encoding defense response‐related proteins) was constructed. This map contains candidate genes for all ANT resistance genes and QRLs previously described in the literature. A total of 256 NBS‐LRR proteins and 200 protein kinases were detected. The functions of the identified candidate genes in ANT response should be validated and studies should be performed to understand how they interact with metabolic pathways.

Last-Generation Genome-Environment Associations Reveal the Genetic Basis of Heat Tolerance in Common Bean (Phaseolus vulgaris L.).

Genome–environment associations (GEAs) are a powerful strategy for the study of adaptive traits in wild plant populations, yet they still lack behind in the use of modern statistical methods as the ones suggested for genome-wide association studies (GWASs). In order to bridge this gap, we couple GEA with last-generation GWAS algorithms in common bean to identify novel sources of heat tolerance across naturally heterogeneous ecosystems. Common bean (Phaseolus vulgaris L.) is the most important legume for human consumption, and breeding it for resistance to heat stress is key because annual increases in atmospheric temperature are causing decreases in yield of up to 9% for every 1°C. A total of 78 geo-referenced wild accessions, spanning the two gene pools of common bean, were genotyped by sequencing (GBS), leading to the discovery of 23,373 single-nucleotide polymorphism (SNP) markers. Three indices of heat stress were developed for each accession and inputted in last-generation algorithms (i.e. SUPER, FarmCPU, and BLINK) to identify putative associated loci with the environmental heterogeneity in heat stress. Best-fit models revealed 120 significantly associated alleles distributed in all 11 common bean chromosomes. Flanking candidate genes were identified using 1-kb genomic windows centered in each associated SNP marker. Some of these genes were directly linked to heat-responsive pathways, such as the activation of heat shock proteins (MED23, MED25, HSFB1, HSP40, and HSP20). We also found protein domains related to thermostability in plants such as S1 and Zinc finger A20 and AN1. Other genes were related to biological processes that may correlate with plant tolerance to high temperature, such as time to flowering (MED25, MBD9, and PAP), germination and seedling development (Pkinase_Tyr, Ankyrin-B, and Family Glicosil-hydrolase), cell wall stability (GAE6), and signaling pathway of abiotic stress via abscisic acid (histone-like transcription factors NFYB and phospholipase C) and auxin (Auxin response factor and AUX_IAA). This work offers putative associated loci for marker-assisted and genomic selection for heat tolerance in common bean. It also demonstrates that it is feasible to identify genome-wide environmental associations with modest sample sizes by using a combination of various carefully chosen environmental indices and last-generation GWAS algorithms.

Development and validation of a robust KASP marker for zt2 locus in faba bean (Vicia faba)

AbstractFaba bean has the potential to become a key food and feed protein crop in many areas of the world. The presence of tannins in its seed coat has limited the deployment of this crop as feed and food. The expression of either of the two recessive genes, zt1 and zt2, causes a great reduction of tannins from the seed coat and results in a white flower phenotype. Molecular markers linked to these loci are fundamental tools for speeding up the breeding of low‐tannin varieties. The main aim of this study was to develop and validate a robust molecular marker linked to the zt2 locus. We used 176 recombinant inbred lines of the Disco/2 × ILB 938/2 cross at F6 and genotyped those using 257 SNP (single nucleotide polymorphism) markers. An SNP marker associated with zt2 locus was found on faba bean chromosome 3 and was used to develop a high‐throughput low‐cost KASP (kompetitive allele‐specific PCR) marker. The KASP marker can successfully discriminate low‐tannin faba beans carrying zt2 from those carrying zt1 and wild‐type alleles.