Drought is a significant abiotic stress that limits crop production. Soybeans [Glycine max (L.) Merr.] are regarded as drought-sensitive. In the present study, the drought responses of wild soybean accessions were evaluated at different growth stages. Based on the leaf wilting index of 411 accessions at the vegetative stage, seven highly tolerant (HT) and 24 tolerant (T) accessions were identified, although most wild soybeans were classified as moderate (M), sensitive (S), and highly sensitive (HS) genotypes. In addition, with selected wild soybeans, stomatal density decreased in HT accessions but increased in HS accessions under drought conditions at the vegetative stage. However, for cultivated soybeans, the stomatal density of the drought-tolerant and drought-sensitive were not significantly different between the two conditions. The expression levels of drought-related transcriptional factors indicated that the HT genotype showed a higher expression level of drought-related genes than that of the HS genotype at the vegetative stage. At the reproductive stages, 12 wild soybeans randomly selected from HT, T, S, and HS based on assessment at the vegetative stage showed consistent drought responses with seed yield, root development, and water status. However, the extent of the detrimental effect of drought on the germination rates and root length of 165 wild soybeans at the germination stage varied depending on the genotype, indicating that there may not be a robust correlation between phenotypic measurements at the germination stage and drought-related assessments at the two growth stages. The information from this study can provide useful breeding materials for the development of drought-tolerant cultivars from wild soybeans.

Drought stress, which is becoming more prevalent due to climate change, is a significant abiotic factor that adversely impacts crop production and yield stability. Cultivated soybean (Glycine max), a versatile crop for humans and animals, exhibits sensitivity to drought, resulting in reduced growth and development under drought conditions. However, few genetic studies have assessed wild soybean’s (Glycine soja) response to drought stress. In this work, we conducted a genome-wide association study (GWAS) and analysis of wild soybean accessions to identify loci responsible for drought tolerance at the vegetative (n = 187) and the germination stages (n = 135) using the available resequencing data. The GWAS analysis of the leaf wilting score (LWS) identified eight single-nucleotide polymorphisms (SNPs) on chromosomes 10, 11, and 19. Of these, wild soybeans with both SNPs on chromosomes 10 (adenine) and 11 (thymine) produced lower LWS, indicating that these SNPs have an important role in the genetic effect on LWS for drought tolerance at the vegetative stage. At the germination stage, nine SNPs associated with five phenotypic measurements were identified on chromosomes 6, 9, 10, 13, 16, and 17, and the genomic regions identified at the germination stage were different from those identified for the LWS, supporting our previous finding that there may not be a robust correlation between the genes influencing phenotypes at the germination and vegetative stages. This research will benefit marker-assisted breeding programs aimed at enhancing drought tolerance in soybeans.

Cultivated soybean is an important legume crop that is generally sensitive to flooding stress, including submergence and waterlogging treatments. Wild soybeans, the ancestor of cultivated soybeans, have been potential genetic resources for resistance to abiotic or biotic stresses. The present study aimed to evaluate 163 wild soybean accessions for foliar damages at the early vegetative stage and 105 accessions for germination rates, normal seedling rates, and electrical conductivity at the germination stage under submergence stress. In addition, a genome-wide association study (GWAS) was conducted to identify genomic regions associated with phenotypic measurements at these two growth stages by using MLM and FarmCPU models with publicly available genotypic data. The phenotypic evaluation revealed six and three accessions were tolerant to submergence at the early vegetative and germination stages, respectively. Notably, only one wild soybean accession showed a tolerance reaction to submergence at two stages. Through GWAS analysis, 16 and 20 SNPs across different chromosomes were determined for the submergence-related traits at the early vegetative and germination stages, respectively. Based on the linkage disequilibrium block on the detected genomic regions, ten and four putative genes were identified at the early vegetative and germination stages, respectively. Of these genes, certain genes may be related to submergence stress in wild soybeans. Further studies should be performed to validate the function of these putative genes in the responses of wild soybeans to submergence stress.

This method article presents the Nested Mixed Methods Survey (NMMS), a five-step procedure that embeds open-ended qualitative questions, including a final “super-open” question for unsolicited data, within a quantitative survey instrument. Participant identification codes enable optional individual-level triangulation. The design extends data-linked nesting (Schatz, 2012) by collecting qualitative responses from the full sample concurrently rather than through resource-intensive follow-up interviews. Evidence of efficiency is provided from an application with 108 pre-service teacher trainees in Vietnam (100% closed-ended response rate; 63% open-ended engagement). Validation with 108 participants demonstrated that the procedure produces both a dataset suitable for statistical analysis and qualitative insights, with individual-level triangulation revealing corroboration and productive discrepancy across data types. The NMMS is intended for researchers working under time, funding, or training constraints who require both breadth and a meaningful degree of depth in a single administration.