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<p>As the staple crop in plateau agriculture, Tibetan hulless barley (<i>Hordeum vulgare</i> L. var. <i>nudum</i>) experiences dual stresses from challenging environments (abiotic stress) and pathogenic attacks (biotic stress). TGACG-binding transcription factors (TGAs) serve as master regulators orchestrating plant growth, developmental processes, and defense responses against biotic and abiotic stresses, with particularly pivotal functions in regulating plant immunity. Here, 10 <i>HvnTGAs</i> were identified in the Qingke genome, which were phylogenetically classified into five distinct clades. HvnTGA proteins possessed conserved structural domains, characteristic motif compositions, and three-dimensional architectures typical of the <i>TGA</i> family. Subcellular localization assays confirmed that both HvnTGA2 and HvnTGA6 were localized to the nucleus. RNA-seq profiling revealed that eight out of ten <i>HvnTGA</i> genes were differentially expressed following powdery mildew infection. The expression of <i>HvnTGA2</i> and <i>HvnTGA6</i> was specifically upregulated by ethephon (ETH) and methyl jasmonate (MeJA) treatment. Yeast two-hybrid (Y2H) assays demonstrated that both HvnTGA2 and HvnTGA6 could form homodimers. Furthermore, heterodimeric interactions were detected between HvnTGA2 and HvnTGA4, HvnTGA2 and HvnTGA6, as well as HvnTGA4 and HvnTGA6. Interaction assays based on split luciferase complementation assay (SLCA) confirmed the interaction between HvnTGA2 and HvnTGA6, as well as the self-interaction of HvnTGA6. The silencing of <i>HvnTGA2</i> resulted in reduced expression of <i>HvnPR-4</i>, enhanced susceptibility of Qingke to <i>Pyrenophora graminea</i>, and accumulation of ROS. This study comprehensively elucidates the regulatory mechanisms of <i>TGA</i> transcription factors in Qingke, a highland-adapted crop, provides novel molecular insights into plant-pathogen interactions.</p>

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Unveiling the TGA gene family in Qingke (Hordeum vulgare L. var. nudum) and the role of HvnTGA2 in host-pathogen interaction

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年4月10日

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  • Shuai Wang,
  • Hailing Lu,
  • Kai Xu,
  • Yuping Zhao,
  • Li Wang,
  • Jianping Si,
  • Fengning Liu,
  • Tao He,
  • Le Wang

摘要

As the staple crop in plateau agriculture, Tibetan hulless barley (Hordeum vulgare L. var. nudum) experiences dual stresses from challenging environments (abiotic stress) and pathogenic attacks (biotic stress). TGACG-binding transcription factors (TGAs) serve as master regulators orchestrating plant growth, developmental processes, and defense responses against biotic and abiotic stresses, with particularly pivotal functions in regulating plant immunity. Here, 10 HvnTGAs were identified in the Qingke genome, which were phylogenetically classified into five distinct clades. HvnTGA proteins possessed conserved structural domains, characteristic motif compositions, and three-dimensional architectures typical of the TGA family. Subcellular localization assays confirmed that both HvnTGA2 and HvnTGA6 were localized to the nucleus. RNA-seq profiling revealed that eight out of ten HvnTGA genes were differentially expressed following powdery mildew infection. The expression of HvnTGA2 and HvnTGA6 was specifically upregulated by ethephon (ETH) and methyl jasmonate (MeJA) treatment. Yeast two-hybrid (Y2H) assays demonstrated that both HvnTGA2 and HvnTGA6 could form homodimers. Furthermore, heterodimeric interactions were detected between HvnTGA2 and HvnTGA4, HvnTGA2 and HvnTGA6, as well as HvnTGA4 and HvnTGA6. Interaction assays based on split luciferase complementation assay (SLCA) confirmed the interaction between HvnTGA2 and HvnTGA6, as well as the self-interaction of HvnTGA6. The silencing of HvnTGA2 resulted in reduced expression of HvnPR-4, enhanced susceptibility of Qingke to Pyrenophora graminea, and accumulation of ROS. This study comprehensively elucidates the regulatory mechanisms of TGA transcription factors in Qingke, a highland-adapted crop, provides novel molecular insights into plant-pathogen interactions.