Soybean is the world's foremost oilseed crop, and leaf morphology significantly influences yield potential by affecting light interception, canopy structure, and photosynthetic efficiency. In this study, leaf length, leaf width, maximum leaf width, leaf apex opening angle, and leaf area were measured in 216 soybean accessions, and genome-wide association studies (GWAS) were conducted using genomic resequencing data to identify genetic variants associated with leaf morphological traits. A total of 824 SNP loci were found to be significantly associated with leaf shape, and 130 candidate genes were identified in the genomic regions flanking these significant loci. KEGG enrichment analysis revealed that the above candidate genes were significantly enriched in arginine biosynthesis (ko00220), nitrogen metabolism (ko00910), carbon metabolism (ko01200), pyruvate metabolism (ko00620), glycolysis/glycogenolysis (ko00010), starch and sucrose metabolism (ko00500), plant-pathogen interaction (ko04626), and amino acid biosynthesis (ko01230). By combining KEGG and GO enrichment analysis as well as expression level analysis, four candidate genes related to leaf shape (Glyma.10G141600, Glyma.13G062700, Glyma.16G041200 and Glyma.20G115500) were identified. Further, through candidate gene association analysis, it was found that the Glyma.10G141600 gene was divided into two major haplotypes. The leaf area of haplotype 1 was significantly smaller than that of haplotype 2. Subsequently, the cutting amplification polymorphism sequence (CAPS) molecular marker was developed. The marker Chr.10:37502955 can effectively distinguish the differences in leaf size through enzymatic digestion technology, and has excellent typing ability and application potential. The above results can provide a theoretical basis for molecular-assisted selection (MAS) of soybean leaf morphology.

Carboxylesterases (CXEs) participate in diverse plant metabolic processes, including isoflavone biosynthesis. However, the soybean GmCXE subfamily remains poorly characterized, especially in relation to root isoflavone accumulation and the response to Fusarium oxysporum. Here, fifty-six putative GmCXE genes were identified in the soybean genome and classified into three major phylogenetic clades. Analyses of gene structure, conserved motifs, protein domains, and promoter cis-elements revealed conserved features as well as potential functional divergence among subfamily members. Collinearity and duplication analyses indicated that segmental duplication was the main driver of GmCXE subfamily expansion. Tissue-specific expression profiling and RT-qPCR validation selected five root-expressed genes as candidates associated with isoflavone accumulation. SNP variation analysis and allelic group analysis of 209 soybean accessions further prioritized GmCXE54 as a candidate gene for root isoflavone accumulation. Allelic groups defined by a putative promoter SNP, Chr.20-rs39215413, showed significant differences in root daidzein and total isoflavone contents, with accessions carrying the C allele exhibiting higher levels of both traits than those carrying the T allele. Functional analysis in soybean hairy roots showed that GmCXE54 overexpression increased daidzein and total isoflavone accumulation. At 3 h after F. oxysporum inoculation, GmCXE2, GmCXE39, and GmCXE54 were induced, with GmCXE54 showing the strongest response in the resistant accession ZD27. These findings clarify GmCXE subfamily evolution and identify GmCXE54 as a candidate gene associated with root isoflavone accumulation and early F. oxysporum response, offering new perspectives for improving soybean isoflavone-related traits and investigating root response mechanisms.

Genetic transformation is an essential tool for functional genomics and genetic improvement in soybean (Glycine max L.). However, its application remains limited by strong genotype dependence, low transformation efficiency, and poor regeneration. Progress has recently been made to optimize Agrobacterium-mediated systems, physical DNA delivery methods, and tissue culture–free transformation systems. The introduction of developmental regulators, including WUSCHEL (WUS), BABY BOOM (BBM), WUSCHEL-related homeobox (WOX), and GROWTH-REGULATING FACTOR (GRF)–GRF-Interacting Factor (GIF) complexes, has improved the transformation efficiency and expanded the range of amenable soybean genotypes. This review summarizes the recent advances in soybean transformation technologies and regeneration-promoting factors, analyzes the major factors influencing transformation efficiency, and discusses future prospects for high-efficiency, genotype-flexible, and exogenous DNA-free gene- editing and breeding in soybean.

The content and composition of fatty acids are crucial determinants of soybean nutritional quality. In this study, we conducted an expression genome-wide association analysis (eGWAS) using 298 soybean germplasm accessions. We identified 904,984 high-quality SNP markers (MAF > 0.02, missing data ≤ 10

Phosphofructokinase (PFK), a core metabolic enzyme conserved across all organisms, plays a pivotal role in plant growth and development as well as in the regulation of responses to abiotic stresses. While the PFK gene family has been characterized in a variety of plant species and a partial identification in soybean has been reported recently, the complete repertoire and the functional roles of this gene family in soybean remain to be fully elucidated, with inconsistencies in gene number observed across different studies. In this study, we further performed a systematic genome-wide screening of soybean and identified a total of 27 GmPFK family members. Phylogenetic tree analysis further classified these members into three distinct subfamilies. Conserved structure analysis revealed that all GmPFKs contain the PFK domain (PF00365). Real-time quantitative PCR (qRT-PCR) analysis demonstrated that the GmPFK6 gene exhibited a significant upregulation of expression under salt stress. Furthermore, the GmPFK6 gene was cloned and introduced into soybean hairy roots. The results demonstrated that under salt stress, the root length and fresh weight of hairy roots from the control lines were significantly greater than those from the transgenic lines. In addition, under salt stress, the superoxide dismutase (SOD) activity in the control lines was significantly higher than that in the transgenic lines, while the malondialdehyde (MDA) content in the transgenic lines was significantly lower than that in the control lines.

Fructose-1,6-bisphosphate aldolase (FBA), as a key enzyme in energy metabolism, is widely present in organisms. Although FBA genes have been characterized in numerous plant species, their functional roles in soybean remain poorly understood. In this study, a total of 14 GmFBA genes were identified at the genome-wide level. Additionally, phylogenetic analysis divided all GmFBA family members into two divergent subclades. qRT-PCR analysis revealed that most GmFBA genes were markedly responsive to PEG6000 and salt stresses, among which GmFBA5 exhibited the strongest induction under salinity treatment. The heterologous expression of GmFBA5 in yeast was performed. The results showed that GmFBA5 markedly enhanced salt tolerance of yeast cells. Association analysis identified two distinct haplotypes of GmFBA5, and Haplotype 2 significantly enhances salt tolerance in soybean. These findings provide new insights into the potential role of GmFBA5 in soybean salt stress responses and provide candidate genetic resources for salt tolerance improvement.

Fusarium root rot, primarily caused by Fusarium solani, is a damaging soil-borne disease that restricts soybean growth and reduces yield. In the present study, a panel of 330 soybean germplasm accessions was inoculated with F. solani. Disease responses were evaluated using the disease severity index (DSI). The soybean accessions displayed substantial variation in their susceptibility to F. solani. Based on DSI, 39 accessions were classified as highly resistant, 94 as resistant, 125 as susceptible, and 72 as highly susceptible, accounting for 11.82%, 28.48%, 37.88%, and 21.82% of the panel, respectively. The genome-wide association study (GWAS) was performed using a genotyping dataset of 627,436 high-quality single nucleotide polymorphisms (SNPs) and two models, Fixed and random model Circulating Probability Unification (FarmCPU) and mixed linear model (MLM). Both GWAS models detected putative SNP associations across seven chromosomes. Based on SNP allelic-effect analysis and gene function annotation, eight genes were prioritized and subsequently evaluated by quantitative reverse transcription PCR (qRT-PCR) for their responses to F. solani infection. Two Kompetitive allele-specific PCR (KASP) markers, KASP-S13_37431242 and KASP-S13_37529208, were developed from resistance-associated SNPs on chromosome 13 and evaluated across the diverse soybean association panel used in this study. Accessions carrying the favorable genotypes of these markers were enriched for resistant germplasm, with positive predictive values of 65.52% and 66.67%, respectively, indicating their potential value for preliminary favorable-allele tracking and germplasm prioritization. Collectively, these results improve our understanding of the genomic components underlying soybean responses to F. solani. The identified loci, candidate genes, and KASP assays provide a basis for further functional validation and the development of multi-locus strategies for improving soybean resistance to Fusarium root rot.

Soybean root rot caused by Fusarium graminearum is an important soil-borne disease. It hinders seedling establishment and ultimately reduces soybean yield. Resistant germplasm and reliable molecular markers are therefore needed for resistance breeding. In this study, 336 soybean accessions were evaluated for resistance to F. graminearum root rot using the disease severity index (DSI), which ranged from 5.71 to 100.00 across the association panel. Genome-wide association analysis was performed using resequencing-based single nucleotide polymorphism (SNP) data with mixed linear model (MLM) and Fixed and random model Circulating Probability Unification (FarmCPU) models, which detected 117 and 113 candidate resistance-associated SNPs, respectively. Among these, 105 shared SNPs were used to define candidate genomic intervals containing 247 annotated genes. Based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, functional annotation, and allelic-effect analysis, six candidate genes and their associated exonic SNPs were prioritized. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed infection-responsive expression patterns for all six candidate genes, with Glyma.17g202500 and Glyma.18g266700 showing stronger induction in the resistant accession. Two SNPs in these genes were converted into Kompetitive allele-specific PCR (KASP) assays. KASP-S17_32244510 and KASP-S18_55105706 were successfully developed for genotype screening, with screening efficiencies of 73.08% and 74.29%, respectively. These findings identify useful genetic targets and molecular markers for improving soybean resistance to root rot caused by F. graminearum.

Soybean is crucial to food processing and agricultural output. However, pests and diseases can easily impact soybeans, reducing their production. Soybean cyst nematode (SCN) is a soilborne pathogen that has a large geographic range, a long lifespan, and the potential to inflict substantial harm to the soybean industry. Persistent use of major resistance genes leads to a progressive loss of resistance; therefore, continuous identification of new soybean strains and genes is essential for continued sustainable soybean production. In this research, the SCN-resistant and SCN-sensitive germplasm DN-L10 and Heinong 37 were inoculated with SCN 3. After stress treatment, the stressed roots were collected for RNA-Seq analysis. The sequencing results screened out the differentially expressed gene GmbHLH18. The GmbHLH18 gene was cloned, and the overexpression vector pCAMBIA3300-GmbHLH18 was constructed. Agrobacterium infected soybean hairy roots and genetically modified the roots of DN50 soybeans, and transgenic root seedlings were obtained. The transgenically identified root seedlings were transplanted in soil infested with SCN 3, and resistance to root nematodes was determined by magenta staining. The secondary and tertiary structures of the protein, phosphorylation sites, as well as the hydrophilicity related to the GmbHLH18 gene were analyzed. Subsequently, the recombinant subcellular localization vector pCAMBIA1302-GmbHLH18 was employed. Agrobacterium was injected into tobacco leaves, and organelle-specific expression was observed. Finally, stress resistance-related indexes of the roots of overexpressing plants and WT plants under SCN 3 stress were measured. The results showed that overexpression and subcellular localization vectors were successfully constructed and transformed into Agrobacterium K599 and GV3101, respectively. The encoded protein had 1149 amino acids, a molecular weight of 95.76 kDa, an isoelectric point of 5.04, 60 phosphorylation sites, a tertiary structure of a-helix (36.39%), random coil (53.40%), extended chain (8.64%), and corner (1.57%), and was hydrophilic. The protein that the gene encoded was a nuclear-localized protein, according to the results of subcellular localization analysis. Moreover, the Agrobacterium-induced hairy root test revealed that the number of overexpressed pCAMBIA3300-GmbHLH18 transgenic roots in the unit area of DN50 was substantially lower than in the control group, which at first suggested that the gene had partial resistance to SCN 3. Stress resistance-related indexes suggest that the contents of POD, SOD, and proline in the overexpressing root significantly increase after SCN 3 stress, demonstrating that this gene can enhance the plant’s resistance to the SCN 3 pathogen. Future research could focus on further elucidating the molecular mechanism underlying the gene’s resistance to SCN 3 and exploring its potential application in breeding soybean varieties with enhanced resistance.

Although miR395 is well established as a key regulator of sulfur (S) metabolism, its potential role in integrating light signaling with S homeostasis to coordinate secondary metabolite biosynthesis remains underexplored. Here, we identify a regulatory module comprising mdm-miR395, its target MdAPS1, and the transcription factor MdWRKY26, which integrates light and S signaling to promote anthocyanin and ascorbic acid (AsA) accumulation in Malus. Subcellular localization analysis revealed dual localization of MdAPS1 in the cell membrane and mitochondrial, suggesting its role in sensing extracellular S concentrations and modulating assimilation efficiency. Transient suppression of miR395 in apple fruit peel enhanced light-induced anthocyanin accumulation, whereas miR395 overexpression produced opposite effects, underscoring its regulatory role in S homeostasis. Genetic manipulation demonstrated that MdAPS1 positively regulates reduced glutathione (GSH) levels, with overexpression increasing anthocyanin and AsA accumulation. Notably, MdWRKY26 was identified as a novel positive regulator of both GSH and anthocyanin accumulation. Exogenous GSH application significantly enhanced pigmentation in apple fruits and plantlets. Our findings reveal that light signals modulate S homeostasis through the miR395-MdAPS1 and MdWRKY26, and establish a GSH-mediated regulatory node linking light signals to anthocyanin and AsA accumulation in Malus, providing a basis for optimizing S nutrition to improve apple fruit coloration.

Fatty acid transporters (FAXs) play an important role in fatty acid synthesis by facilitating transport fatty acids from the plastid to the endoplasmic reticulum. This process is essential for providing precursor substances necessary for triglycerides (TAGs). Although FAX genes have been identified in variety of plant species, the identification and molecular functions of the GmFAX gene members in soybean are still unclear. In this study, soybean FAX genes were identified through the utilization of the Phytozome (v13) and NCBI online websites. Subsequently, phylogenetic trees, expression patterns, gene structures, and qRT-PCR were analyzed. A total of eight GmFAX members were identified at the whole genome level, and further phylogenetic analysis revealed that these members can be categorized into four subfamilies. In addition, all members of GmFAX contain a highly conserved domain Tmemb_14. Through qRT-PCR analysis, it was found that the expression level of the GmFAX8 gene is relatively high in leaves and stems. Further investigation revealed that the total fatty acid content in hairy roots overexpressing the GmFAX8 gene was significantly greater than that observed in the control strain. The results presented above suggest that the GmFAX8 gene may play an important role in the accumulation of oil within soybeans.

Soybean (Glycine max) is a major crop for grain and oil production worldwide. However, soybean cyst nematode (SCN) disease is a serious threat to soybean cultivation, causing major challenges for agriculture in China and globally. In this study, 306 soybean germplasms were evaluated for resistance to SCN HG type 0 using the female index (FI), a metric quantifying nematode reproduction on soybean roots. Furthermore, we performed a genome-wide association analysis study using 1,332,548 high-density single nucleotide polymorphism (SNP) markers. Through this analysis, the study identified quantitative trait nucleotides (QTN) and pinpointed candidate genes that are linked to resistance against SCN HG type 0. A total of 77 SNPs and 117 candidate genes associated with soybean resistance to SCN HG type 0 were identified. Subsequently, two kompetitive allele-specific PCR (KASP) markers, S19_rs8522772 and S19_rs8384176, along with four cleaved amplified polymorphic sequence (CAPS) markers (S19_rs838271, S19_rs8522589, S19_rs8466511 and S19_rs8481473) were developed. These markers were found to be closely linked to soybean resistance to SCN based on the results of beneficial allele analysis of candidate genes. These markers not only expanded the soybean germplasm resource, but also provided a solid foundation for molecular breeding of soybean varieties resistant to SCN.

The plant stress response mechanism is activated by biotic and abiotic stresses, but its continuous activation typically affects growth. The role of ferritin in regulating biomass accumulation has been extensively characterized in diverse plant species; however, the underlying mechanisms through which it contributes to salt stress tolerance and Fusarium resistance remain poorly understood. Here, we confirm that overexpression of ferritin leads to iron accumulation and Fe3+ sequestration in both aboveground and roots, activating the iron uptake and transport system. More importantly, GmFER1 enhances salt stress tolerance and Fusarium resistance. First, GmFER1 is localized in chloroplasts and significantly induced by salt stress and Fusarium infection. Overexpression of GmFER1 increases soybean yield per plant by enhancing net photosynthetic rate and Rubisco enzyme activity, without activating the reactive oxygen scavenging mechanism. Under salt stress, GmFER1 enhances resistance by improving the activities of SOD and CAT enzymes, as well as Na+ efflux capacity. Under Fusarium infection, GmFER1 enhances resistance to the pathogen by boosting antioxidant capacity. Moreover, iron-deficiency tests revealed that increased CAT and SOD activities under salt stress are linked to iron ions accumulation. Lastly, we analysed the effects of GmFER1 gene variation on salt tolerance, disease resistance and 23 agronomic traits related to yield and quality. Further analysis of GmFER1 gene variation revealed that the Hap2 haplotypes could potentially enhance salt resistance, disease resistance, pod number and oil content in soybean. Our research offers a new way to reduce growth penalties while boosting plant resistance to salt stress and Fusarium infection.

Rhizoctonia solani root rot (RSRR) is a major disease that significantly reduces soybean yields, causing substantial economic losses to global soybean production. To elucidate the genetic basis of RSRR resistance, 310 soybean germplasm accessions were evaluated using the disease severity index (DSI) following inoculation with R. solani. Among these accessions, 46.13% were susceptible, and only 2.26% exhibited high resistance. Utilizing resequencing data consisting of 738,561 Single Nucleotide Polymorphism (SNP) loci, a genome-wide association study (GWAS) was performed by integrating both general linear model (GLM) and mixed linear model (MLM) approaches, resulting in the identification of 21 SNPs significantly associated with resistance on chromosomes 3, 13, 15, 16, 17, and 18, and six candidate genes. RT-qPCR expression analysis revealed that four genes, including Glyma.03G166300, Glyma.03G168100, Glyma.13G212700, and Glyma.13G212300, were significantly upregulated in resistant genotypes after inoculation. Furthermore, Cleaved Amplified Polymorphic Sequences (CAPS) and Kompetitive Allele Specific PCR (KASP) molecular markers were successfully developed based on the RSRR-associated SNPs S3_38086892, S3_38247290, and S13_32595026, providing effective tools for marker-assisted selection (MAS). The findings strengthen our genetic knowledge concerning RSRR resistance and contribute to the molecular breeding of resistant soybean cultivars.

Using the cotyledonary node method, four traits related to callus induction rate were identified in 185 soybean germplasm resources. Cultivation of callus tissue is crucial for soybean (Glycine max (L.) Merr.) genetic transformation and functional genomics studies. Identifying genes associated with the induction rate of soybean callus tissue is therefore essential for biotechnological breeding and for understanding the molecular genetic mechanisms of soybean regeneration. The efficiency of genetic transformation impacts the breeding rate of soybeans, with its success rate dependent on the soybean regeneration system. Subsequently, whole genome association analysis (GWAS) and multidimensional functional validation were conducted. GWAS identified 66 significantly associated SNP loci corresponding to the four traits. Expression analysis in extreme phenotypes highlighted four candidate genes: Glyma.12G164100 (GmARF1), Glyma.12G164700 (GmPPR), Glyma.02G006200 (GmERF1), and Glyma.19G128800 (GmAECC1), which positively regulate callus formation. Overexpression and gene-editing assays in hairy roots confirmed that these genes significantly enhanced callus formation rate and density, with GmARF1 exerting the most prominent effect. Hormone profiling revealed elevated levels of gibberellin (GA), auxin (IAA), cytokinin (CTK), and other phytohormones in transgenic lines, consistent with enhanced responsiveness to exogenous GA. Overall, the results suggest that these four candidate genes may promote soybean regeneration, with GmARF1 showing the most pronounced effect. These results provide valuable genetic resources for improving soybean regeneration efficiency and accelerating genetic transformation-based breeding.

Fusarium root rot, caused by Fusarium equiseti, poses a significant threat to soybean production. This study aimed to explore the genetic basis of resistance to Fusarium equiseti root rot (FERR) by evaluating the resistance phenotype of 346 soybean germplasms and conducting a genome-wide association study (GWAS) using 698,949 SNP markers obtained from soybean germplasm resequencing data. GWAS analysis identified 101 SNPs significantly associated with FERR resistance, distributed across nine chromosomes, with the highest number of SNPs on chromosomes 13 and 20. Further gene-based association and allele variation analyses identified candidate genes whose mutations are closely related to FERR resistance. To accelerate soybean FERR resistance breeding screening, we developed CAPS markers S13_14464319-CAPS1 and S15_9215524-CAPS2, targeting these SNP sites, and KASP markers based on the S15_9205620-G/A, providing an effective tool for marker-assisted selection (MAS). This study offers a valuable theoretical foundation and molecular marker resources for the functional validation of FERR resistance genes and soybean disease resistance breeding.

Obesity, characterized by excessive fat accumulation, represents a global health crisis closely linked to metabolic disorders such as type 2 diabetes, hypertension, and atherosclerosis. tRNA-derived small RNAs (tsRNAs) have recently emerged as important epigenetic regulators, yet their roles in fat deposition remain poorly characterized. This study aims to identify tsRNAs that influence fat accumulation and to elucidate their molecular mechanisms, with a focus on tRF‑Gly‑GCC‑037 (tRF‑Gly) as a candidate regulator of adipocyte differentiation. Visceral adipose tissue was collected from obese and lean pigs for comprehensive tRF and tiRNA sequencing. Differential expression analysis identified tRF‑Gly as a highly abundant candidate in obese samples. Functional assays in 3T3‑L1 preadipocytes included both overexpression and knockdown of tRF‑Gly, followed by lipid accumulation measurements and assessment of key adipogenic markers (CEBPα and PPARγ) by quantitative real-time PCR (qRT‑PCR) and western blot. Mechanistically, dual‑luciferase reporter assays, RNA immunoprecipitation (RIP), and nuclear–cytoplasmic protein fractionation were performed to examine how tRF‑Gly modulates the RAC1/JNK2/β‑catenin signaling axis. tRF‑Gly was significantly upregulated in visceral adipose tissue from obese pigs and ranked among the most abundant tsRNAs. Overexpression of tRF‑Gly in 3T3‑L1 cells and in C57BL/6 mice promoted lipid accumulation and increased CEBPα and PPARγ expression, whereas tRF‑Gly knockdown reduced lipid deposition. Mechanistically, tRF-Gly was suggested to bind RAC1 mRNA with AGO3 involvement, leading to RAC1 silencing. Consistently, RAC1 knockdown phenocopied the adipogenic effects of tRF-Gly, whereas RAC1 overexpression reversed these effects. Furthermore, RAC1 deficiency disrupted the RAC1/JNK2/β‑catenin complex, impaired β‑catenin nuclear translocation, and suppressed Wnt/β‑catenin signaling. Our findings demonstrate that tRF‑Gly functions as a key regulator of fat accumulation. By silencing RAC1 via AGO3, tRF‑Gly disrupts RAC1/JNK2/β‑catenin complex assembly, prevents β‑catenin nuclear translocation, and downregulates Wnt/β‑catenin signaling, thereby promoting lipid deposition. This study uncovers a novel epigenetic mechanism by which tRF‑Gly controls fat accumulation and suggests that targeting tRF‑Gly may represent a therapeutic strategy for obesity and related metabolic disorders.

The bile acid: sodium symporter (BASS) plays a pivotal role in plant growth, development, and response to abiotic stress. In this study, eight members of the BASSs group were identified through genome-wide screening. Additionally, employing phylogenetic analysis allowed for the categorization of the BASS gene family into five distinct subfamilies. The localization of GmBASSs in young tobacco leaves was primarily observed within chloroplasts using confocal microscopy. The qRT-PCR analysis revealed that a significant response was observed in the majority of GmBASSs genes toward abiotic stress, such as NaCl, NaHCO3, and 20% PEG6000. The GmBASSs gene was also observed to display distinct expression patterns across various tissue. It is noteworthy that the GmBASS2-2 gene exhibits a remarkably high level of upregulation in response to salt stress, suggesting its pivotal role in mediating salt stress responses. The GmBASS2-2 gene was cloned and overexpressed in soybean hair roots and Saccharomyces cerevisiae. We found the GmBASS2-2 overexpressing Saccharomyces cerevisiae grew better than the control (pYES3 empty vector) under salt stress. The transgenic soybean plants overexpressing the GmBASS2-2 gene demonstrated superior growth compared to control plants. Gene-based association analysis showed that a total of 2 haplotypes were identified for the GmBASS2-2 gene, and Hap2 haplotype has emerged as a superior allele for enhancing salt tolerance. The above findings suggest that the GmBASS2-2 gene might be involved in the process of soybean tolerance to salt stress. The findings of this study may provide valuable insights into the regulatory mechanisms of GmBASS2-2 in response to salt stress in soybean.

The ABI3 transcription factor is a key regulator in plant growth and development. Through transcriptome analysis of the resistant soybean cultivar ‘Dongnong L10′ and the susceptible cultivar ‘Heinong 37′ exposed to soybean cyst nematode race 3 (SCN 3) stress, the differentially expressed gene GmABI3VP1 was identified. The GmABI3VP1 gene was then cloned and analyzed through bioinformatics, subcellular localization, and qRT-PCR analysis of resistant and susceptible soybean germplasms, as well as overexpression and gene editing of soybean hairy roots followed by SCN 3 identification analysis. It was found that the protein encoded by GmABI3VP1 is an acidic and hydrophilic protein with transmembrane domains. It has a collinear relationship with Arabidopsis and is widely distributed in plants. Through the analysis of promoter elements, it was shown that this gene contains multiple hormone-responsive promoter elements like ABRE/ABRE3a/ABRE/4a/as-1 and stress-responsive elements such as Myb/MYC/MYc. Transient expression in tobacco indicated that the GmABI3VP1 gene is located in the nucleus. The transcription of GmABI3VP1 responds to the stress of SCN, and its transcriptional level is relatively high in the roots of resistant materials. Genetic transformation mediated by Agrobacterium rhizogenes was used to obtain GmABI3VP1 gene overexpressed and CRISPR-Cas9 gene-edited soybean hairy roots. In comparison to the wild type (WT), the density of nematodes per area was notably lower in hairy roots overexpressing (OX) the gene, whereas the density of SCN per unit area (per cm of lateral root length) significantly increased in gene-edited (KO) soybean hairy roots. Through SCN phenotyping, GmABI3VP1 was identified as a contributor to SCN 3 resistance. This study provides initial insights into the role of the GmABI3VP1 gene in SCN resistance, establishing a robust basis for future research on the mechanisms underlying SCN disease resistance and offering valuable genetic reservoirs for SCN 3 resistance.