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Background <p>Anthocyanins are essential flavonoid pigments that contribute to plant coloration, stress resilience, and nutritional quality. Despite their importance, the mechanisms underlying anthocyanin transport and accumulation in soybean (<i>Glycine max</i>) remain poorly understood. Here, we performed a comprehensive genome-wide characterization of the glutathione S-transferase (GST) gene family in soybean and functionally investigated GmGSTF members potentially associated with anthocyanin stabilization, intracellular trafficking, and vacuolar sequestration.</p> Results <p>A total of 104 <i>GmGST</i> genes were identified and classified into seven subfamilies, including 14 Phi-class GmGSTF members. These genes exhibited diverse exon-intron structures, conserved motif compositions, and uneven chromosomal distributions, with segmental duplication likely contributing substantially to family expansion. Phylogenetic and comparative amino acid sequence analyses showed that GmGSTF8 and GmGSTF11 clustered closely with previously characterized anthocyanin-associated GSTs and contained conserved GST-N-Phi and GST-C-Phi domains. Expression profiling revealed tissue-specific and stress-responsive patterns among <i>GmGSTF</i> genes. Notably, <i>GmGSTF8</i> and <i>GmGSTF11</i> showed coordinated expression with two key anthocyanin biosynthetic genes <i>GmUGT78K2</i> and <i>GmF3′5′H</i>, respectively, and their expression profiles were closely associated with anthocyanin accumulation across soybean tissues. These phylogenetic and co-expression results prompted us to further investigate whether GmGSTF8 and GmGSTF11 have conserved anthocyanin-associated functions. Subcellular localization analysis showed that both GmGSTF8 and GmGSTF11 were mainly distributed in the cytoplasm. Ectopic expression of GmGSTF8 or GmGSTF11 in <i>Arabidopsis thaliana tt19</i> mutants restored stem-base pigmentation and increased relative anthocyanin content in the tt19 background; seed coat pigmentation was also partially restored. These results indicate that GmGSTF8 and GmGSTF11 are conserved anthocyanin-associated GST proteins that may function as carrier- or chaperone-like proteins involved in anthocyanin stabilization and vacuolar sequestration.</p> Conclusion <p>Taken together, this study provides systematic characterization and heterologous functional evidence for soybean GSTF members associated with anthocyanin accumulation and identifies GmGSTF8 and GmGSTF11 as candidate targets for future studies aimed at improving flavonoid accumulation in soybean. These findings provide a valuable framework for future mechanistic and metabolic engineering studies in legumes.</p>

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Genome-wide characterization of the glutathione S-transferase gene family identifies GmGSTF8 and GmGSTF11 as anthocyanin-associated GSTs in soybean

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月11日

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  • Taotao Han,
  • Tianhao Lu,
  • Xinyue Tang,
  • Mingli Sha,
  • Wenwei Chu,
  • Guopeng Miao

摘要

Background

Anthocyanins are essential flavonoid pigments that contribute to plant coloration, stress resilience, and nutritional quality. Despite their importance, the mechanisms underlying anthocyanin transport and accumulation in soybean (Glycine max) remain poorly understood. Here, we performed a comprehensive genome-wide characterization of the glutathione S-transferase (GST) gene family in soybean and functionally investigated GmGSTF members potentially associated with anthocyanin stabilization, intracellular trafficking, and vacuolar sequestration.

Results

A total of 104 GmGST genes were identified and classified into seven subfamilies, including 14 Phi-class GmGSTF members. These genes exhibited diverse exon-intron structures, conserved motif compositions, and uneven chromosomal distributions, with segmental duplication likely contributing substantially to family expansion. Phylogenetic and comparative amino acid sequence analyses showed that GmGSTF8 and GmGSTF11 clustered closely with previously characterized anthocyanin-associated GSTs and contained conserved GST-N-Phi and GST-C-Phi domains. Expression profiling revealed tissue-specific and stress-responsive patterns among GmGSTF genes. Notably, GmGSTF8 and GmGSTF11 showed coordinated expression with two key anthocyanin biosynthetic genes GmUGT78K2 and GmF3′5′H, respectively, and their expression profiles were closely associated with anthocyanin accumulation across soybean tissues. These phylogenetic and co-expression results prompted us to further investigate whether GmGSTF8 and GmGSTF11 have conserved anthocyanin-associated functions. Subcellular localization analysis showed that both GmGSTF8 and GmGSTF11 were mainly distributed in the cytoplasm. Ectopic expression of GmGSTF8 or GmGSTF11 in Arabidopsis thaliana tt19 mutants restored stem-base pigmentation and increased relative anthocyanin content in the tt19 background; seed coat pigmentation was also partially restored. These results indicate that GmGSTF8 and GmGSTF11 are conserved anthocyanin-associated GST proteins that may function as carrier- or chaperone-like proteins involved in anthocyanin stabilization and vacuolar sequestration.

Conclusion

Taken together, this study provides systematic characterization and heterologous functional evidence for soybean GSTF members associated with anthocyanin accumulation and identifies GmGSTF8 and GmGSTF11 as candidate targets for future studies aimed at improving flavonoid accumulation in soybean. These findings provide a valuable framework for future mechanistic and metabolic engineering studies in legumes.