YUCCA, belonging to the class B flavin-dependent monooxygenases, catalyzes the rate-limiting step for endogenous auxin synthesis and is implicated in plant-growth regulation and stress response. Systematic analysis of the YUCCA gene family and its stress response benefits the dissection of regulation mechanisms and breeding applications. In this study, 12 YUCCA genes were identified from the mungbean (Vigna radiata L.) genome and were named based on their similarity to AtYUCCAs. Phylogenetic analysis revealed that the 12 VrYUCCAs could be divided into 4 subfamilies. The evidence from enzymatic assays in vitro and transgenetic Arabidopsis in vivo indicated that all the isolated VrYUCCAs had biological activity in response to IAA synthesis. Expression pattern analysis showed that functional redundancy and divergence existed in the VrYUCCA gene family. Four VrYUCCAs were expressed in most tissues, and five VrYUCCAs were specifically highly expressed in the floral organs. The response toward five stresses, namely, auxin (indole-3-acetic acid, IAA), salinity, drought, high temperatures, and cold, was also investigated here. Five VrYUCCAs responded to IAA in the root, while only VrYUCCA8a was induced in the leaf. VrYUCCA2a, VrYUCCA6a, VrYUCCA8a, VrYUCCA8b, and VrYUCCA10 seemed to dominate under abiotic stresses, due to their sensitivity to the other four treatments. However, the response modes of the VrYUCCAs varied, indicating that they may regulate different stresses in distinct ways to finely adjust IAA content. The comprehensive analysis of the VrYUCCAs in this study lays a solid foundation for further investigation of VrYUCCA genes’ mechanisms and applications in breeding.

The azuki bean weevil (Callosobruchus chinensis L.) is an insect pest responsible for serious postharvest seed loss in leguminous crops. In this study, we performed quantitative trait locus (QTL) mapping of seed resistance to C. chinensis in moth bean (Vigna aconitifolia [Jaqc.] Maréchal). An F₂ population of 188 plants developed by crossing resistant accession ‘TN67’ (wild type from India; male parent) and susceptible accession ‘IPCMO056’ (cultivated type from India; female parent) was used for mapping. Seeds of the F₂ population from 2014 and F_2:3 populations from 2016 and 2017 were bioassayed with C. chinensis, and the percentage of damaged seeds and progress of infestation severity were measured. Segregation analysis suggested that C. chinensis resistance in TN176 is controlled by a single dominant gene, designated as Rcc. QTL analysis revealed one principal and one modifying QTL for the resistance, named qVacBrc2.1 and qVacBrc5.1, respectively. qVacBrc2.1 was located on linkage group 2 between simple sequence repeat markers CEDG261 and DMB-SSR160 and accounted for 50.41% to 64.23% of resistance-related traits, depending on the trait and population. Comparative genomic analysis suggested that qVacBrc2.1 is the same as QTL Brc2.1 conferring C. chinensis resistance in wild azuki bean (V. nepalensis Tateishi and Maxted). Markers CEDG261 and DMB-SSR160 should be useful for marker-assisted selection for C. chinensis resistance in moth bean.

Mung bean is vulnerable to bruchids (Callosobruchus spp.), resulting in low quality and losses worldwide. Developing resistant cultivars is the most effective, economical, and eco-friendly way to protect mung bean seeds from the damage of bruchids. Previously, we identified two gene loci that are tightly linked with bruchid resistance, which makes new cultivar development possible using molecular assisted selection (MAS). In the present study, marker-assisted backcross (MABC) breeding was employed to introgress the bruchid resistance gene VrPGIP2 locus from the donor parent V2802 into a popular cultivar, Kamphaeng Saen 1. Markers VrBR-SSR013 and DMB-SSR158 were used for foreground selection, and phenotypic selection was used for background selection. Three advanced inbred lines (R67-11, R67-22, and R92-15) carrying the resistance gene VrPGIP2 were developed from the BC₃F₆ population. Further analysis showed that the line R67-22 performed best; it was highly resistant to bruchids and showed excellent agronomic traits in the field. Therefore, R67-22 could be further evaluated in large-scale trials for release as a variety.

Faba bean is an important legume crop consumed as a vegetable or snack food, and its green cotyledons could present an attractive color for consumers. A mutation in SGR causes stay-green in plants. In this study, vfsgr was identified from a green-cotyledon-mutant faba bean, SNB7, by homologous blast between the SGR of pea and the transcriptome of faba bean. Sequence analysis revealed that a SNP at position 513 of the CDS of VfSGR caused a pre-stop codon, resulting in a shorter protein in the green-cotyledon faba bean SNB7. A dCaps marker was developed according to the SNP that caused the pre-stop, and this marker was completely associated with the color of the cotyledon of faba bean. SNB7 stayed green during dark treatment, while the expression level of VfSGR increased during dark-induced senescence in the yellow-cotyledon faba bean HST. Transient expression of VfSGR in Nicotiana. benthamiana leaves resulted in chlorophyll degradation. These results indicate that vfsgr is the gene responsible for the stay-green of faba bean, and the dCaps marker developed in this study provides a molecular tool for the breeding of green-cotyledon faba beans.

WRKY-, PHD-, and MYB-like proteins are three important types of transcription factors in mungbeans, and play an important role in development and stress resistance. The genes’ structures and characteristics were clearly reported and were shown to contain the conservative WRKYGQK heptapeptide sequence, Cys4-His-cys3 zinc binding motif, and HTH (helix) tryptophan cluster W structure, respectively. Knowledge on the response of these genes to salt stress is largely unknown. To address this issue, 83 VrWRKYs, 47 VrPHDs, and 149 VrMYBs were identified by using comparative genomics, transcriptomics, and molecular biology methods in mungbeans. An intraspecific synteny analysis revealed that the three gene families had strong co-linearity and an interspecies synteny analysis showed that mungbean and Arabidopsis were relatively close in genetic relationship. Moreover, 20, 10, and 20 genes showed significantly different expression levels after 15 days of salt treatment (p < 0.05; Log₂ FC > 0.5), respectively. Additionally, in the qRT-PCR analysis, VrPHD14 had varying degrees of response to NaCl and PEG treatments after 12 h. VrWRKY49 was upregulated by ABA treatment, especially in the beginning (within 24 h). VrMYB96 was significantly upregulated in the early stages of ABA, NaCl, and PEG stress treatments (during the first 4 h). VrWRKY38 was significantly upregulated by ABA and NaCl treatments, but downregulated by PEG treatment. We also constructed a gene network centered on the seven DEGs under NaCl treatment; the results showed that VrWRKY38 was in the center of the PPI network and most of the homologous Arabidopsis genes of the interacted genes were reported to have response to biological stress. Candidate genes identified in this study provide abundant gene resources for the study of salt tolerance in mungbeans.

Heterosis has been utilized in crops for a long time, and although crop fertility is the basis for the utilization of heterosis, there is limited information concerning the genic male sterility (GMS) of mungbean. Therefore, based on the genic male sterile mutant, M₁, obtained by EMS mutagenesis of the Weilyu cultivar, this project used multi-omics analysis to detect the male sterile site and further identify its candidate gene, and then the mechanism of the candidate gene was discussed. As a result, one locus region (Chr5: 6,835,001–6,935,000 bp) associated with GMS was identified, and nine genes were found within the 100 Kb region. The candidate gene, VrCYCA1, around the above loci had a TATA box deletion approximately 4.7 Kb upstream of the gene, and this was evidenced by comparative genomics, transcriptome analysis, and RT-qPCR analysis. The expression level of VrCYCA1 was significantly downregulated (log₂FC = −2.06, p-value = 0.025) in the ms lines compared with the control group. Moreover, 6653 genes showed differential expression between the Weilyu lines and mutant lines as well as 165 metabolites with significant differences in their concentration levels. Among those differentially expresses genes, 226 were annotated with functional categories involved in flowering and endosperm development, and six genes had protein–protein interactions with VrCYCA1. Seven categories of metabolites and seven genes participated in the relationship between reproductive growth and vegetative growth, which might have caused the sterility of mungbean in the mutant plants. This study used multi-omics data to mine a mungbean GMS-related gene, VrCYCA1, and constructed a GMS genetic network to explore the molecular mechanism of VrCYCA1. The results lay a solid foundation for further molecular biology research and utilization in mungbean male sterility.