Vegetable soybean ((Glycine max (L.) Merr.), commonly referred to as edamame, holds significant agricultural importance in China as a legume vegetable harvested at the pod-filling stage (R6).  The visual appeal of vegetable soybeans is crucial for consumer preference and marketability, and it depends on factors such as pod length, pod width, and pod color.  This study cultivated 264 vegetable soybeans in Nanjing, Huai’an, and Nantong, Jiangsu Province, China to assess pod traits using PlantPhenoM, a system for pod phenotypic identification and analysis.  The results revealed a variability range of 8.64 to 30.00% in appearance quality traits among the vegetable soybeans.  Leveraging phenotypic data and employing a genome-wide association study (GWAS) identified 525 SNPs significantly linked to the appearance quality traits in different regions.  In addition, five candidate genes (Glyma.04G004700, Glyma.15G051600, Glyma.18G225700, Glyma.18G225900, and Glyma.18G272300) associated with target traits were identified, and KASP markers for S04_372771 (pod length), S18_51477324 (pod width), and S18_55553200 (pod color) were developed.  This study offers valuable insights for breeding superior vegetable soybean varieties and lays the groundwork for exploring candidate genes and molecular markers related to appearance and quality traits in vegetable soybeans.

The number of pods per plant (PP) is strongly correlated with seed yield, and identifying genes that regulate PP could enhance the yield of mung bean (Vigna radiata (L.) Wilczek), providing valuable insights for molecular breeding.  In this study, VrKNAT6 was identified through genome-wide association and multiomics analyses.  Chr3-14344673 (P=3.02E-10~8.80E-07) was found to be significantly associated with PP using EMMAX, CMLM, GEMMA, GLM, and 3VmrMLM.  Among the 12 genes located within a 100 kb region near Chr3-14344673 on chromosome 3, EVM0027029 (VrKNAT6) is homologous to known PP development-related genes in Oryza sativa and Arabidopsis thaliana.  Overexpression of VrKNAT6^H1 significantly increased rosette numbers, branch numbers, PP, and the 1,000-seed weight in transgenic Arabidopsis lines.  Furthermore, when overexpressed in mung bean hairy roots and Arabidopsis, VrKNAT6^H1 was found to participate in jasmonic acid (JA) synthesis through physical interaction with VrATH1.  This interaction partly explains the differences in branch numbers between VrKNAT6^H1-overexpressing Arabidopsis lines and the control.  Additionally, the expression of JA synthetase-related genes was significantly elevated in the positive VrKNAT6^H1 lines.  Based on the multiomics analysis results, we propose a molecular regulatory mechanism for VrKNAT6^H1, suggesting that it is a JA synthesis-related gene that could be utilized in mung bean high-yield molecular breeding.

Mungbean (Vigna radiata L. (Wilczek)) is an important food legume crop.  The utilization of heterosis based on male sterile lines can help increase mungbean yields, yet genetic studies on mungbean male sterility are rare.  Therefore, it is of great significance to explore the male sterility genes in mungbean.  In this study, a no-pollen male sterile mutant vrnpms (Vigna radiata no pollen male sterility) was identified in mungbean.  Gene mapping was conducted using F₂ populations derived from the cross between vrnpms and V2709.  The gene controlling the male sterility was mapped to a 426.65 kb region on chromosome 6.  A candidate gene VrMYB80 (EVM0016947), encoding a protein homologous to MYB80 transcription factors, exhibits a 52-kb deletion in vrnpms, resulting in a truncated protein lacking the C’-terminus.  A molecular marker linked to the male sterility phenotype was developed based on the deletion in vrnpms.  Functional complementation in Arabidopsis demonstrated that VrMYB80 could restore fertility in the myb80 mutant.  Subcellular localization showed that VrMYB80 was located in the nucleus. Transcriptional activation assays revealed that the C’-terminus of VrMYB80 was the transcriptional activation domain.  The result of in-situ hybridization indicated that VrMYB80 is expressed in the anther tapetum.  The expression level of downstream VrMS1 was down regulated in vrnpms, indicating that Vrmyb80 with the truncated C’-terminal transcriptional activation domain failed to activate downstream genes, which was the reason of sterility of vrnpms.  The findings of this study contribute to unraveling the molecular genetic mechanism underlying pollen development in legume crops and pave the way for utilizing heterosis in mungbean.

Soil salinization severely impairs mungbean (Vigna radiata (L.) Wilczek) seedling uniformity and productivity. In this study, genome-wide association study (GWAS) was conducted using a natural population of 374 mungbean accessions and 4,875,143 SNPs. By evaluating the population under two independent environments and applying two statistical models, we identified a significant SNP (Chr01_26769549) associated with relative germination traits under salt stress. Based on this locus, a Kompetitive Allele-Specific PCR (KASP) marker was successfully developed for marker assisted selection. Integrated haplotype and expression analyses confirmed polygalacturonase gene VrPG1 as a key candidate gene regulating salt tolerance during seed germination. Two haplotypes of VrPG1 (Hap1/Hap2) were identified, with a mutation in the Hap1 promoter region enhancing its transcriptional activity. Overexpression of VrPG1 in Arabidopsis thaliana significantly increased germination rates under salt stress by promoting endosperm cell wall softening. Salt-tolerant mungbean varieties exhibit higher polygalacturonase activity and earlier loosening of thin-walled cell walls during the germination period, which promotes seed imbibition and radicle emergence. Collectively, these findings demonstrate that VrPG1 enhances salt tolerance during germination through cell wall remodeling. This study provides novel genetic targets and efficient marker-assisted selection tools for breeding salt-tolerant mungbean. This study provides novel genetic targets and efficient marker-assisted selection tools for breeding salt-tolerant mungbean varieties.

The bean bug, Riptortus pedestris, is a major pest of soybeans in East Asian countries. Male-released aggregation pheromones attract both adults and nymphs, offering potential for eco-friendly pest control. However, the molecular mechanisms underlying the detection of the aggregation pheromones remain unclear. In the present study, functional analysis using the Xenopus oocyte expression system demonstrated that two ORs (OR23h and OR109d) were responsible for sensing aggregation pheromones, with the primary component (E)-2-hexenyl (E)-2-hexenoate (E2HE2H) being shared by the two ORs. Further quantitative PCR (qPCR) profiling indicated that OR109d was expressed only in male antennae, while OR23h was expressed in both sexes at similar levels. RNA interference assays demonstrated that dsOR23h-treatment significantly reduced the Electroantennographic (EAG) response of (E)-2-hexenyl (Z)-3-hexenoate (E2HZ3H) in both sexes. Furthermore, simultaneous RNAi knockdown of the two ORs significantly reduced the male EAG response to E2HE2H and abolished male attraction to this compound. These results were consistent with the sex expression profile, demonstrating the sex and functional differentiation between the two ORs. Taken together, this study characterizes the ORs responsible for chemical perception and the associated aggregation behaviors driven by these pheromones. Thus, this study enhances our understanding of olfactory signaling in a hemipteran insect and contributes to the knowledge required for improved pest management.

Enhancing grain weight of wheat is a crucial strategy for improving yields in the Huaibei Plain (HP). However, the impacts and regulatory mechanisms of different irrigation regimes on wheat grain formation in the HP remained poorly understood. Therefore, a two-year field experiment was conducted to explore three treatments on wheat’s source-sink relationship and grain formation: rain-fed (RI, no irrigation, 202.5 kg ha^-1 N applied at sowing), conventional flood irrigation (CI, 60 mm irrigation at jointing stage, 112.5 kg ha^-1 N at sowing+90 kg ha^-1 N with irrigation), and micro-sprinkler irrigation (MI, irrigation based on 0–40 cm soil layer water deficit at jointing, booting and anthesis stages, 112.5 kg ha^-1 N at sowing+30 kg ha^-1 N at each irrigation). The results indicated that, compared with RI and CI, MI significantly increased the chlorophyll content and enhanced the activity of sucrose phosphate synthase (SPS) in flag leaf at 4 days after anthesis (DAA 4), and these parameters in CI were higher than those in RI. The sucrose and soluble sugar content in grain of MI were the highest at DAA 4. Additionally, at DAA 4, compared with RI, both CI and MI significantly elevated the content of indole propionic acid+zeatin nucleoside (IPA+ZR) and gibberellin (GA) in grain, while reducing the content of auxin (IAA) and abscisic acid (ABA). And the highest endosperm cells number was observed in MI. At the grain filling stage, MI exhibited the slowest chlorophyll degradation rate and the highest activities of ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) and SPS in flag leaf, resulting in more sugar accumulation in the leaf and grain. Moreover, MI showed the highest IAA and lowest ABA levels in grain, and maintained the highest starch synthase activity during the filling stage, promoting the starch accumulation. Compared to CI and RI, MI significantly increased 1,000-grain weight by 4.99–5.55% and 7.33–11.51%, and grain yield by 4.99–11.60% and 15.60–39.14% over the two years, respectively. Overall, micro-sprinkler irrigation can optimize the water and nitrogen supply for wheat, effectively enhancing the source capacity in the early stage and the sink capacity in the late stage of grain development, thereby increasing grain weight and achieving high yield in the HP.

Walnut is an important economic woody oil tree species, and anthracnose caused by Colletotrichum gloeosporioides is a devastating disease affecting walnut production in China. The MAPK-WRKY signaling pathway plays an important role in regulating plant disease resistance. However, the MAPK-WRKY pathway in walnut and the mechanism involved in anthracnose resistance remain unclear. Using 'Taile' and 'Xiangling' with significant differences in anthracnose resistance as materials, we identified a potential JrMAPK3-JrWRKY22 pathway related to anthracnose resistance through transcriptomics. Further analysis using yeast two-hybrid, bimolecular fluorescence complementation, pull-down, and in vitro phosphorylation assays revealed that JrWRKY22 interacts with and is phosphorylated by JrMAPK3. Transient injection results in walnut fruit revealed that overexpression of JrWRKY22 can inhibit Colletotrichum gloeosporioides infection, increase fruit anthracnose resistance, and significantly promote the expression of the β-1,3-glucanase gene JrGLU of the PR-2 family and the pathogenesis-related gene JrPR1. In contrast, silencing JrWRKY22 resulted in a significant increase in lesion size caused by Colletotrichum gloeosporioides and a corresponding decrease in gene expression levels. A dual-luciferase assay confirmed that JrWRKY22 can activate the promoter activity of JrPR1 and JrGLU and that phosphorylation by JrMAPK3 increases this activation. Further analysis using yeast one-hybrid assays, ChIP-PCR, and EMSA demonstrated that JrWRKY22 can bind to W-box elements in the promoters of JrPR1 and JrGLU. These findings elucidate the molecular mechanism by which the JrMAPK3-JrWRKY22 module increases walnut anthracnose resistance, broaden the understanding of resistance mechanisms, and provide a scientific basis for molecular breeding in the context of walnut disease resistance.

The yield and quality of soybean (Glycine max [L.] Merr.), a globally important food and cash crop, are affected by a variety of environmental factors, one of which is low-light stress. In previous work, we demonstrated that knockout of the SEIPIN homolog FA9, whose encoded protein participates in lipid-droplet formation, reduced oil content and increased protein content in seeds. In this study, we compared the responses of gmfa9 knockout mutants (fa9-KO) and wild-type (WT) Dongnong 50 (DN50) soybean to low-light stress. Vegetative indices (GNDVI and CIGreen), spectral reflectance, and chlorophyll-fluorescence parameters were higher in fa9-KO than in the WT under low light. fa9-KO exhibited prolonged reproductive development and more pronounced morphological changes than WT soybean under low-light conditions; under all light conditions, it produced larger seeds with higher protein and sugar contents but had a lower total seed weight per plant. Transcriptome and RT-qPCR analyses demonstrated that FA9 mutation was associated with the downregulation of multiple genes involved in fatty acid synthesis and photosynthetic light reactions. These findings suggest that FA9 is an important genetic resource for breeding high-quality soybean varieties adapted to low-light environments.

Bud dormancy is a key adaptive strategy in perennial plants, enabling them to survive in adverse environmental conditions. However, it generates challenges in crop cultivation, especially in fruit crops like apple, where synchronized bud break is crucial for consistent growth and yield. The synthetic cytokinin 6-benzylaminopurine (6-BA) promotes dormancy release, but its molecular and metabolic mechanisms remain poorly understood. This study investigates dormancy release in Xianheng 01 apple rootstock nursery plants through integrated transcriptomic, metabolomics, and hormonal analyses. Dormant buds were treated with 6-BA, followed by morphological, biochemical, and molecular profiling conducted over 30 days. 6-BA treatment increased plant height and leaf emergence by increasing the level of endogenous hormones such as cytokinins (DHZR/IPA) and decrease in level of abscisic acid (ABA). Transcriptomics analysis identified 7,009 differentially expressed genes (DEGs) in response to 6-BA treatment. The cytokinin-responsive gene A-ARR8 exhibited a distinct expression pattern, remained upregulated at 1, 3 and 6 d post-treatment but downregulated at 11 d. In contrast ABA related genes SnRK2a/b, PP2C and ABF3 were consistently downregulated throughout the treatment period. Metabolomics analysis identified 2,053 metabolites, showing early-phase dominance of phenylpropanoids and flavonoids, followed by a shift towards ABC transporter-mediated nutrient mobilization. Conjoint analysis highlighted coordinated activation of secondary metabolite biosynthesis and cytokinin signaling. These results demonstrate that 6-BA induces dormancy release through cytokinin-ABA antagonism and phased metabolic reprogramming from stress protection to growth promotion. Our findings provide a comprehensive framework for optimizing dormancy management in apple cultivation and highlight 6-BA as an effective agrochemical for enhancing temperate fruit production.