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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. [ABSTRACT FROM AUTHOR]

Summary: 2,3‐Dihydro‐2,5‐dihydroxy‐6‐methyl‐4H‐pyran‐4‐one (DDMP) saponins are one of the major saponin groups that are widely distributed in legumes such as pea, barrel medic, chickpea, and soybean. The steps involved in DDMP saponin biosynthesis remain uncharacterized at the molecular level.We isolated two recessive mutants that lack DDMP saponins from an ethyl methanesulfonate‐induced mutant population of soybean cultivar Pungsannamul.Segregation analysis showed that the production of DDMP saponins is controlled by a single locus, named Sg‐9. The locus was physically mapped to a 130‐kb region on chromosome 16. Nucleotide sequence analysis of candidate genes in the region revealed that each mutant has a single‐nucleotide polymorphism in the Glyma.16G033700 encoding a UDP‐glycosyltransferase UGT73B4. Enzyme assays and mass spectrum‐coupled chromatographic analysis reveal that the Sg‐9 protein has glycosyltransferase activity, converting sapogenins and group B saponins to glycosylated products, and that mutant proteins had only partial activities. The tissue‐specific expression profile of Sg‐9 matches the accumulation pattern of DDMP saponins.This is the first report on a new gene and its function in the biosynthesis of DDMP saponins. Our findings indicate that Sg‐9 encodes a putative DDMP transferase that plays a critical role in the biosynthesis of DDMP saponins. [ABSTRACT FROM AUTHOR]

In soybean, triterpenoid saponin is one of the major secondary metabolites and is further classified into group A and DDMP saponins. Although they have known health benefits for humans and animals, acetylation of group A saponins causes bitterness and gives an astringent taste to soy products. Therefore, several studies are being conducted to eliminate acetylated group A saponins. Previous studies have isolated and characterized the Sg-5 (Glyma.15g243300) gene, which encodes the cytochrome P450 72A69 enzyme and is responsible for soyasapogenol A biosynthesis. In this study, we elucidated the molecular identity of a novel mutant of Glycine soja, ′CWS5095′. Phenotypic analysis using TLC and LC-PDA/MS/MS showed that the mutant ′CWS5095′ did not produce any group A saponins. Segregation analysis showed that the absence of group A saponins is controlled by a single recessive allele. The locus was mapped on chromosome 15 (4.3 Mb) between Affx-89193969 and Affx-89134397 where the previously identified Glyma.15g243300 gene is positioned. Sequence analysis of the coding region for the Glyma.15g243300 gene revealed the presence of four SNPs in ′CWS5095′ compared to the control lines. One of these four SNPs (G1127A) leads to the amino acid change Arg376Lys in the motif, which is invariably conserved among the CYP450 superfamily proteins. Co-segregation analysis showed that the missense mutation (Arg376Lys) was tightly linked with the absence of group A saponins in ′CWS5095′. Even though Arg and Lys have similar chemical features, the 3D modelled protein structure indicates that the replacement of Arg with Lys may cause a loss-of-function of the Sg-5 protein by inhibiting the stable binding of a heme cofactor to the CYP72A69 apoenzyme. [ABSTRACT FROM AUTHOR]

In soybean, flavonoid 3′5′-hydroxylase (F3′5′H) and dihydroflavonol-4-reductase (DFR) play a crucial role in the production of anthocyanin pigments. Loss-of-function of the W1 locus, which encodes the former, or W3 and W4, which encode the latter, always produces white flowers. In this study, we searched for new genetic components responsible for the production of white flowers in soybean and isolated four white-flowered mutant lines, i.e., two Glycine soja accessions (CW12700 and CW13381) and two EMS-induced mutants of Glycine max (PE1837 and PE636). F3′5′H expression in CW12700, PE1837, and PE636 was normal, whereas that in CW13381 was aberrant and missing the third exon. Sequence analysis of F3′5′H of CW13381 revealed the presence of an indel (~90-bp AT-repeat) in the second intron. In addition, the F3′5′H of CW12700, PE1837, and PE636 harbored unique single-nucleotide substitutions. The single nucleotide polymorphisms resulted in substitutions of amino acid residues located in or near the SRS4 domain of F3′5′H, which is essential for substrate recognition. 3D structure modeling of F3′5′H indicated that the substitutions could interfere with an interaction between the substrate and heme group and compromise the conformation of the active site of F3′5′H. Recombination analysis revealed a tight correlation between all of the mutant alleles at the W1 locus and white flower color. On the basis of the characterization of the new mutant alleles, we discussed the biological implications of F3′5′H and DFR in the determination of flower colors in soybean. [ABSTRACT FROM AUTHOR]

Cultivated soybeans [Glycinemax (L.) Merr.] have various flower colours such as dark purple, purple, light purple, pink, magenta, near white and white. About one-third of the soybean accessions in the United States Department of Agriculture – Germplasm Resource Information Network (USDA-GRIN) Soybean Germplasm Collections have white flowers and are the second dominant accessions after the purple-flowered accessions. Earlier studies have shown that the w1 recessive allele of the W1 gene encoding flavonoid 3′,5′-hydroxylase produces white flowers. In the present study, we aimed to understand why the white-flowered accessions have become abundant among the cultivated soybeans and what their genetic and regional origin is. For this purpose, 99 landraces with white flowers and 39 landraces with purple flowers from eight Asian countries and Russia were analysed with regard to the nucleotide sequences of the W1 locus. We not only found that the w1 alleles of the 99 white-flowered landraces were identical to those of the white-flowered Williams 82, but also found that these w1 alleles displayed no polymorphism at all. By carrying out a phylogenetic analysis, we were able to identify a group with W1 alleles from which the w1 allele might have diverged. [ABSTRACT FROM AUTHOR]