Mung bean (Vigna radiata) is a globally important legume crop valued for its short growing cycle, nitrogen-fixing capacity and high nutritional value, particularly in developing countries. Here we report a comprehensive graph-based pan-genome assembled from 11 genetically diverse global accessions. The framework captures 75,268 gene families (50.86% core, 35.19% dispensable and 13.95% private) and 66,862 nonredundant structural variants. Integrating these structural variants and single nucleotide polymorphisms, genome-wide association studies across five environments identified candidate genes for 20 agronomic traits, underscoring the pivotal roles of these variants in driving mung bean domestication and improvement. Mechanistically, we demonstrate that a 68-bp promoter insertion in VrTIFY6B and a 136-bp promoter deletion in VrPGIP1 regulate flavonoid content and confer bruchid resistance, respectively. These genomic resources and actionable functional variants provide a powerful toolkit to accelerate mung bean improvement through marker-assisted breeding, genomic selection and genome editing to address global food security.

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.

Cassava (Manihot esculenta Crantz) exhibits high photosynthetic efficiency and remarkable starch accumulation in its storage roots. The effective loading of photosynthates into the phloem from mesophyll cells in leaves is a critical determinant of yield; however, this process remains poorly understood. In this study, we propose a theoretical model of apoplastic sucrose phloem loading in cassava based on a multitechnique approach. The concentration of primary photoassimilates in leaf veins, analyzed using a [14C]CO2 tracer, and the existence of few plasmodesmata between bundle sheath cells/phloem parenchyma cells and sieve element-companion cell (SE-CC) complexes in minor veins suggest characteristic apoplastic phloem loading in cassava. We identified 5 sucrose transporters (MeSUTs) in the cassava genome, among which MeSUT1a exhibited the highest expression and the strongest sucrose intake activity. Subcellular localization analyses showed that MeSUT1a specifically localizes in the plasma membrane of the SE-CC complexes and that sugar transporter (MeSWEET2a) localizes on parenchyma cells, suggesting potential functional synergy. Interference with MeSUT1a expression led to a reduction in sucrose loading efficiency by more than 50%, resulting in abnormal sucrose and transient starch accumulation. This interference subsequently impaired chloroplast development and leaf photosynthesis, ultimately reducing storage root yield and starch content. RNA-seq analysis of MeSUT1a transgenic lines further revealed remarkable transcriptional changes in genes associated with sugar transport, carbohydrate metabolism, and photosynthesis. These results establish that MeSUT1a is essential for driving sucrose phloem loading and plays a key role in the distribution of photosynthetic assimilates and the coordination of source-sink dynamics in cassava.

Mungbean (Vigna radiata) is an important cash crop, yet the production is significantly compromised by continuous cropping. Beneficial microbial inoculation offers a promising strategy to alleviate the stresses through rhizosphere modulation and host physiological reprogramming. This study evaluated the efficacy of two biological control agents, Bacillus subtilis (B. subtilis) and Trichoderma harzianum (T. harzianum), in promoting mungbean growth under continuous-cropping conditions. Both individual applications of B. subtilis and T. harzianum significantly improved plant biomass, root system architecture, and yield. Combined metagenomic and transcriptomic analyses were conducted to unravel the underlying mechanisms. According to metagenomic analysis, both B. subtilis and T. harzianum were responsible for significant changes in beta diversity without significantly affecting the alpha diversity of the rhizosphere microbial community. T. harzianum recruited Chitinophagaceae unclassified, Abditibacterium, Hydrogenophilaceae unclassified, Methylophilaceae unclassified, and Chimaeribacter, while Bs recruited Candidatus Saccharibacteria unclassified. Transcriptomic analysis indicated that T. harzianum induced more extensive transcriptional reprogramming than B. subtilis. The enrichment analysis revealed both shared and distinct responses triggered by the two treatments. These findings suggest that B. subtilis and T. harzianum alleviate continuous-cropping stress through distinct yet complementary mechanisms involving rhizosphere microbiome modulation and mungbean transcriptional reprogramming. This study provides a sustainable strategy for legume cultivation.

Traditional methods for identifying salt tolerance levels in soybean varieties are often cumbersome, time-consuming, and labor-intensive. These challenges are further exacerbated by the limited utility of chlorophyll fluorescence imaging phenotype data, which are insufficiently diverse and difficult to analyze. Additionally, the corresponding parameter text data have not been fully explored and utilized. In this study, salt stress experiments were conducted on 178 soybean varieties, and a multimodal dataset comprising chlorophyll fluorescence images and corresponding textual data was constructed using a chlorophyll fluorescence imaging instrument. A novel gated mechanism network for learnable image-text interaction (Mm-VitnNet) is proposed, which enables global cross-modal interaction between image and text data. The model introduces a gated mechanism to dynamically regulate the fusion intensity of cross-modal information and incorporates two learnable tokens that focus on feature learning for each individual modality. This approach effectively mitigates interference between modalities while preserving modality-specific features, thereby enhancing model performance. The proposed model demonstrates an accuracy rate of 98.97%, significantly outperforming typical models: it improves by 1.09 and 2.33 percentage points compared to CNN-based models such as EfficientNetV2-s (97.88%) and MobileNetV2 (96.64%), respectively, and by 3.21 and 2.60 percentage points compared to Transformer-based Swin Transformer_tiny (95.76%) and hybrid models like MobileViT_S (96.37%), respectively. The model has 10.22M parameters and a computational cost (FLOPs) of 1.84G, which is significantly lower than models like VGG and ResNet50, and only slightly higher than some lightweight CNNs, achieving an effective balance between accuracy and efficiency. The improved model demonstrates notable performance in identifying samples with varying salt tolerance levels, even under limited computational resources, ensuring reliable classification performance. Moreover, this multimodal non-destructive identification method based on chlorophyll fluorescence technology offers an efficient and feasible approach for assessing the salt tolerance levels of soybeans, while also advancing agricultural phenotyping towards greater precision and intelligence.

Mungbean is an important legume and protein source, but its productivity is severely limited by the mungbean yellow mosaic India virus (MYMIV), and no MYMIV resistance gene has been cloned and functionally validated in mungbean. This study conducted extensive phenotypic variation research on the resistance of a newly developed mungbean multiparent advanced-generation intercross (MAGIC) population. Through an integrated genome-wide association study (GWAS), transcriptome analysis, and induced expression analysis, the candidate gene for MYMIV resistance was identified as VrADH, encoding alcohol dehydrogenase. Haplotype analysis revealed natural variation in VrADH, with VrADH Hap1 being the elite haplotype that has undergone selection in regions severely affected by MYMIV. Functional validation demonstrated that VrADH significantly enhanced resistance by limiting excessive reactive oxygen species accumulation and reducing viral proliferation. Collectively, our results indicate that VrADH can contribute to MYMIV resistance, providing a valuable genetic resource for future molecular breeding and resistance improvement in mungbean.

Cassava (Manihot esculenta Crantz) exhibits exceptional tolerance to infertile soils and contains abundant cyanogenic glucosides (CGs). Previous research has indicated that CGs can serve as a significant reservoir of organic nitrogen in plants. However, the extent to which its high-CG content contributes to efficient nitrogen utilisation and adaptation to low nitrogen (N) in cassava remains to be further elucidated. This study represents the first identification of MeHNL11 as a bifunctional protein. In response to N deficiency, the hydroxynitrile lyase activity of MeHNL11 promotes the generation and accumulation of cyanide and the Cys245 residue of MeHNL11 is critical for its nuclear oligomerization, in which the protein functions as a transcription factor. Following the cyanide transmission into the nucleus, the oligomeric form of MeHNL11 dissociates into monomers, leading to a dramatic upregulation of MeCAS1b transcription. This regulatory mechanism helps sustain intracellular cyanide homeostasis within cassava and facilitates the synthesis of primary N metabolites, thereby alleviating N deficiency. The exogenous application of the cyanide antidote hydroxocobalamin (COB) inhibited cyanide assimilation by MeCAS1b, leading to exacerbated N deficiency symptoms, such as leaf yellowing and a significant reduction in the contents of NH4 + and free amino acids (AA) in cassava seedlings under low-N conditions (LN). Our research demonstrates that the MeHNL11-MeCAS1b module plays a pivotal role in CG recycling, offering new insights into the underlying mechanisms governing cassava's exceptional tolerance to low N stress.

Callosobruchus maculatus (cowpea weevil) is an insect pest that causes significant yield loss in cultivated black gram (Vigna mungo var. mungo) during storage. A previous study showed that seed resistance to C. maculatus in wild black gram (V. mungo var. silvestris) accession ‘TC2210’ is controlled by two linked QTLs, qVmunBr6.1 and qVmunBr6.2. However, none of these QTL have been validated, and the molecular basis of these QTLs is not yet known. The objectives of this study are to validate qVmunBr6.2 and identify candidate gene(s) for bruchid resistance at this locus using wild black gram accession ‘TVNu1076’ as the source of resistance. QTL mapping using an F2 population of the cross ‘Chai Nat 80’ (cultivated black gram) × TVNu1076 showed that qVmunBr6.2 controls the resistance in TVNu1076. Fine mapping using a large F2:3 population of 1,144 plants located the qVmunBr6.2 to the marker interval VmBru-SSR74 and VmBru-SSR98. The markers VmBru-SSR74 and VmBru-SSR98 corresponded to a genome region of 9.27 Kb containing two genes including VmunHev encoding hevamine-A, a known enzyme involved biotic stress resistance. Compared with the wild black gram TVNu1076, sequence alignment of VmunHev revealed two one-base pair substitutions that cause amino acid changes in hevamine-A in the cultivated black grams. Gene expression analysis revealed that VmunHev expressed in the seeds. These results showed that VmunHev is a strong candidate gene conferring seed resistance to C. maculatus in wild black gram. The results present in this study provide insight into the genetic basis of C. maculatus resistance in wild black gram.

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.

Aflatoxin B1 (AFB1) is a carcinogenic mycotoxin synthesized by Aspergillus flavus, a pathogen that commonly contaminates food products. Currently, efficient and safe methods for the removal of AFB1 from food are still lacking. Although various microorganisms have been screened for degrading AFB1, these approaches commonly involve the transformation of AFB1 into other toxic aflatoxins. In this study, the yeast Meyerozyma caribbica NJC41 was found to inhibit A. flavus mycelial growth and AFB1 biosynthesis up to 43.4% and 61.4%, respectively. NJC41 (1 & times; 107 cells/mL) degraded AFB1 by 76.0% within 12 h. The primary degradation product was aflatoxin P1 (AFP1), a metabolite of significantly reduced toxicity, while a smaller portion was converted to aflatoxin M1 (AFM1). The resulting aqueous solution showed no significant toxicity in the zebrafish model, with histopathological analysis confirming the absence of liver damage. Application of NJC41 at 1 & times; 107 cells/mL inhibited A. flavus-caused rot symptoms in kiwifruit by 91.4% (based on lesion area) and reduced AFB1 level by 96.1% via combined inhibition of biosynthesis and enzymatic degradation, demonstrating its potential for practical AFB1 control. This study reveals the first method for the management of AFB1 in fruits and confirms for the first time the ability of M. caribbica to inhibit A. flavus growth and degrade AFB1. NJC41 exhibited unique properties, converting primarily AFB1 into non-toxic AFP1, a degradation product only observed previously when AFB1 decomposition was catalyzed by ozone or larval cytochrome P450 oxidases.

[Objective]Protein and threonine are the core nutrients of mung bean seeds,which have a critical impact on the nutritional quality of mung beans.Among them,mung bean protein is a high-quality protein source for vegetarians due to its easy digestion,absorption,and rich biological activity,which is of great value to human health.This study aims to explore genetic loci and candidate genes significantly associated with the protein content and relative content of threonine in mung bean seeds,providing a theoretical basis for genetic improvement of protein and threonine content in mung bean seeds.[Method]This study used Kjeldahl nitrogen determination method and liquid chromatography-mass spectrometry to determine the protein content and relative content of threonine in 279 mung bean varieties planted in 2022.The general linear model of Tassel5 software was used for genome-wide association study to explore genetic loci related to protein content and relative content of threonine in mung bean seeds.By combining linkage disequilibrium analysis,transcriptome comparative analysis,and comparative genomics,key candidate genes related to nitrogen metabolism were identified.[Result]Six key loci located on chromosome 3 were identified,which are related to the protein content and relative content of threonine in mung bean seeds,and can explain 6.06%to 12.20%of phenotypic variation.These 6 loci are closely linked and can be mainly divided into two haplotypes,with Hap2 being the dominant haplotype.Then,KASP molecular marker was designed for the key candidate locus SLG03_966208(A/G),which can successfully classify 80 mung bean varieties into three genotypes:AA,GG,and AG,with a detection accuracy rate of 87.5%.The key candidate gene EVM0000757 was identified within the locus linkage region,and the homologous gene AT5G65750 in Arabidopsis was significantly associated with nitrogen metabolism.Its gene expression was significantly different between high and low protein varieties(P=1.84E-03,|log2FC|=1.39),making it a reliable candidate gene.[Conclusion]279 mung bean varieties were identified for their grain protein content and relative content of threonine.Six SNP loci were detected within the 939 296-1 039 749 bp region of chromosome 3.A KASP molecular marker for the SLG03_966208(A/G)locus was developed,which can distinguish the high and low levels of protein and threonine content in mung bean grains.A key candidate gene related to nitrogen metabolism,EVM0000757,was identified.

As a major commercial legume crop, soybean ranks among the world's most significant sources of edible oil and plant protein. We previously identified a SEIPIN homologue (FA9) at the fatty acid 9 locus that promotes fatty acid accumulation in soybean. To examine the detailed molecular mechanisms by which FA9 regulates lipid metabolism, we performed single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (stRNA-seq) of wild-type and FA9-knockout soybean seeds at the late maturity stage. scRNA-seq analysis identified 26 transcriptional clusters and revealed the spatial distribution of FA9 in seeds, in which the deletion of FA9 altered lipid and storage-related transcriptional programmes. On the basis of single-cell sequencing and immunoprecipitation-mass spectrometry (IP-MS), the vesicle-associated membrane protein (VAMP)-associated protein (VAP) was identified, and subsequent experiments demonstrated that FA9 interacts specifically with VAP via its N-terminal FFAT motif at the endoplasmic reticulum. Seeds of vap knockout (vap-KO1 and vap-KO2) and fa9 vap double knockout (fa9 vap-KO) lines, created by CRISPR-Cas9 gene editing, had higher protein contents and lower total fatty acid contents than wild-type soybean, whereas overexpression of FA9 and VAP enhanced lipid droplet formation in Nicotiana benthamiana. These findings reveal that FA9 interacts with VAP to promote lipid droplet biogenesis and lipid transport, thereby driving fatty acid accumulation in soybean seeds. This research provides new insight into the molecular mechanisms that regulate seed oil synthesis and identifies potential target genes for improvement of soybean oil quality through molecular breeding.

Plant height is a key agronomic trait in soybean (Glycine max L.), as it directly influences the number of nodes on the main stem, pod number per plant, yield, and lodging resistance. Identifying genes that regulate plant height is therefore essential for optimizing plant architecture and enhancing soybean productivity. Through a genome-wide association study (GWAS), we identified qGPH3 as a region associated with soybean plant height that co-located with qBPH16, a quantitative trait locus (QTL) derived from a bulked segregant analysis by sequencing (BSA-seq) and mapping to chromosome 17. RT-qPCR analysis revealed that Glyma.17 g082100 expression levels are significantly higher in the dwarf parent Wandou 15 than in the tall parent Xudou 18 used for the QTL mapping. We detected four haplotypes for this gene, with soybean accessions carrying Glyma.17g082100Hap3 being the shortest, whereas those with Glyma.17g082100Hap4 were the tallest. Functional validation using gene editing and overexpression in transgenic lines confirmed Glyma.17 g082100 as the causal gene regulating soybean height underlying qGPH3 and qBPH6. This bHLH transcription factor gene, designated GmBIM1, is highly expressed in developing stems and leaf buds and encodes a nucleus-localized protein. Our findings identify useful genetic resources for improving soybean yield potential through targeted breeding of plant architecture.

The yam Dioscorea alata L. is widely cultivated globally. Purple-fleshed varieties of this important crop have enhanced market value due to their high anthocyanin contents, but how anthocyanin biosynthesis in D. alata tubers is regulated remains poorly understood. In this study, we identified and functionally validated key transcription factors that regulate anthocyanin biosynthesis based on a comparative transcriptome and metabolome analysis of three D. alata cultivars with different colored tubers (dark purple, light purple, and white). The anthocyanin glycoside cyanidin-3-O-(2′′-O-glucosyl) glucoside was abundant during early tuber development, and we determined that its accumulation is regulated in opposite manners by two R2R3-MYB transcription factors: DaMYB75 and DaMYB56. Yeast two-hybrid and bimolecular fluorescence complementation assays in Nicotiana benthamiana and co-expression assays in D. alata demonstrated that DaMYB75 promotes anthocyanin biosynthesis by specifically activating the promoter of the late anthocyanin biosynthesis gene DaANS and enhancing its expression through an interaction with DabHLH72. By contrast, DaMYB56 is a negative regulator of anthocyanin biosynthesis that binds to the DaANS promoter together with DabHLH72. Furthermore, the methylation levels of the DaMYB75 promoter were significantly lower in purple tubers than in white tubers. These findings shed light on the regulation of anthocyanin biosynthesis by MYBs and provide the basis for genetically improving anthocyanin content in D. alata.

Bacillus amyloliquefaciens (BA) has a positive effect in alleviating salt stress and promoting plant growth. However, the mechanism by which BA enhances plant stress resistance and improves nutrient removal efficiency in ecological ditch systems under high salt stress remains unclear. The experiment constructed two ecological ditch of [P (substrate + plants) and PM (substrate + plants + BA)], and four different salinity treatments (5 g/L, 10 g/L, 15 g/L and 20 g/L) were set up to investigate the removal efficiency of nitrogen and phosphorus in water, and plant growth and physiological indicators, antioxidant enzyme activities and inorganic salt ion concentrations under different salinity stress by adding BA. It also clarified the relationship between plant growth physiology and the response of water nutrient salts. The results show that the PM treatments for purifying nutrients performed significantly better than the P treatments under different salinity stress, the removal rates of TN, TP and NH4⁺-N increased by 13.61% - 22.50%, and the effect was the best at a salinity of 15 g/L. Bacillus amyloliquefaciens enhanced the salt tolerance by increasing the activities of superoxide dismutase(SOD), peroxidase(POD) and catalase(CAT) of plants, reducing the content of malondialdehyde(MAD), lowering the concentration of Na+ and increasing the concentration of K+, improving the growth and activity of plant roots. Meanwhile, through a comprehensive evaluation, it was determined that BA has the most significant promoting effect on the growth of reed, and stronger plant root system is the key to improving the overall purification efficiency of the system.

Drought stress is a major global challenge in crop production. The occurrence of drought during seed germination directly affects crop establishment and final yield. In this work, we performed whole-genome resequencing of 414 mungbean (Vigna radiata (L.) R. Wilczek) germplasms and identified 4,875,142 high-quality single-nucleotide polymorphisms (SNPs). Two seed germination traits of the mungbean panel under drought stress across two environments were evaluated via a genome-wide association study (GWAS), and the results revealed a significant quantitative trait locus (QTL). Additionally, kompetitive allele-specific PCR (KASP) markers that can identify the germination characteristics of mungbean seeds under drought stress were developed and verified. Moreover, among the nine candidate genes near this site, the expression levels of different haplotypes of VrERF088 differed. Interestingly, the only ABA-responsive element in the VrERF088 promoter was mutated. The results demonstrated that VrABI5 activates the transcriptional activity of the Hap1 promoter by binding to the ABRE motif. The overexpression of VrERF088 in Arabidopsis thaliana significantly reduced the germination rate of plants under drought stress by increasing the expression of the ABA response genes AtPM6 and AtLEA4 and reducing the expression of the germination-related genes AtEXPAs, resulting in hypersensitivity to ABA. Our results indicate that mutation of the ABRE motif in the VrERF088 promoter results in a relatively high germination rate under drought stress.

Mungbean (Vigna radiata L.) is a nutritionally diverse food, associated with human health, such as heat dissipation. Although the genetic basis of yield-related traits in major crops has been thoroughly investigated, there is limited related gene information on mungbean. To address this issue, in this study, we first reported the high-quality genome assembly of cultivar Sulv1 Its size was 473.67 Mb, and the N50 value (11.3 Mb) was larger than those in previous studies. We then performed domestication analyses and genome-wide association studies on days to flowering and 100 seed weight. In domestication analyses, 20 differentially expressed genes were identified from 1022 selected regions and their Arabidopsis homologous genes were confirmed in Arabidopsis, such as LFR, GA20OX, and ACC2. In particular, EVM0030058 significantly increased the synthesis of isovitexin in mungbean hairy roots, and 40 μM isovitexin showed good effects in reducing inflammation in vitro. The domestication loci Chr3-47 757 894 was significantly associated with days to flowering and/or 100-seed weight. Tts candidate genes, VrLFR, a homolog of LFR in Arabidopsis, and verified by real-time PCR and haplotype analysis and found to be negatively correlated and differentially expressed in large- and small-seeded accessions. Finally, VrLFRH1 overexpression lines increased seed-size-related traits and promoted early flowering in transgenic Arabidopsis compared to Col-0. The p35S::VrLFR lines recovered the phenotype of lfr mutants. VrLFR was shown to be available for early maturing and high yielding breeding. Moreover, 100-seed weight showed a subtle positive correlative trend with isovitexin content (r = 0.385, P = 0.092). These findings establish a solid experimental foundation for anti-inflammatory studies on mungbean and provide a theoretical basis for its high-yield and high-quality molecular breeding.

To understand the mechanism of sweetness formation in vegetable soybean seeds, an integrative transcriptomic and metabolomic analysis was conducted using high sweetness (HS) and low sweetness (LS) varieties selected from 287 resources based on electronic tongue evaluation. The HS variety exhibited significantly higher levels of soluble sugars (58.91 mg/g) and free amino acids (54.15 mg/g). Transcriptomic results indicated that DEGs correlated with glycolysis/gluconeogenesis, starch and sucrose metabolism, and biosynthesis of amino acids pathways were significantly up-regulated in the HS variety. Metabolomic analysis showed that DAMs were significantly enriched in the biosynthesis of secondary metabolites, the amino acid biosynthesis, and the pentose phosphate pathway. Co-expression network analysis further demonstrated correlations between DEGs and DAMs related to glycolysis/gluconeogenesis and amino acid biosynthesis. Eight candidate genes related to sweetness formation were identified through transcriptomic data and validated by RT-qPCR. The present findings represent a fundamental advance in understanding the regulatory mechanisms underlying the sweetness of vegetable soybeans.

High soil salinity has become a key factor affecting the yield and quality of alfalfa. Calmodulin 1 (CaM1) gene is involved in salinity stress signal transduction, which plays a positive or negative role in regulating plant salinity tolerance. Nevertheless, the specific function of MsCaM1 in alfalfa remains unknown. This study showed that the MsCaM1 gene contains a complete open reading frame of 450 bp, encoding a protein of 149 amino acids. Subcellular localization analysis revealed that the MsCaM1 protein is located to the nucleus. Meanwhile, the expression of MsCaM1 gene showed increased trend by salinity treatment in leaves of salinity tolerance variety GN5. The heterologous expression of MsCaM1 in Arabidopsis resulted in increased germination energy, germination percentage, plant height, and shoot fresh weight compared with wild-type. Moreover, the shoots of transgenic Arabidopsis plants under NaCl treatment displayed better growth compared to wild-type plants. In transgenic Arabidopsis, the MDA content significantly decreased by NaCl-treated, while the SOD activity remained unchanged, contrasting with the wild-type, where MDA content unchanged and SOD activity decreased. Moreover, the transgenic Arabidopsis plants have lower O2− level under salinity stress compared to wild-type. Furthermore, MsCaM1 expression significantly affected the expression of the AtCaM1 genes. These results indicate that MsCaM1 may act as a positive regulator of growth and salt tolerance in Arabidopsis. These findings could contribute to the understanding of the role of MsCaM1 in alfalfa.