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<p>Salt stress critically impairs plant growth and development; therefore, elucidating salt-tolerance genes and their regulatory mechanisms is essential for advancing molecular breeding. In our previous study, a GRAS transcription factor, BpGRAS19, was identified within the salt-responsive gene regulatory network of birch (<i>Betula platyphylla</i>). Phylogenetic analysis and conserved domain prediction validated BpGRAS19 as a member of the GRAS family, which contains a highly conserved GRAS domain. Subcellular localization experiments and quantitative real-time PCR (RT-qPCR) analyses further showed that BpGRAS19 is a nuclear-localized protein with tissue-specific expression patterns, and its expression is significantly induced by salt stress in birch. Transient transformation assays confirmed that BpGRAS19 functions as a positive regulator of salt tolerance in birch. In contrast, CRISPR/Cas9-mediated knockout of BpGRAS19 remarkably reduced salt tolerance, which was associated with impaired reactive oxygen species (ROS) scavenging capacity. Yeast one-hybrid (Y1H) assays indicated that BpGRAS19 specifically binds to characterized (MBS, W-box) and novel cis-acting elements (UN1: ACCGAGC; UN2: CGACATC), thereby expanding the DNA-binding motif of GRAS transcription factors. Taken together, these results demonstrate that BpGRAS19 improves salt tolerance in birch by mitigating cell membrane damage and enhancing ROS scavenging efficiency. This study lays a foundation for understanding the molecular mechanisms underlying salt tolerance in woody plants and provides theoretical support for salt-resistant breeding of birch.</p>

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The GRAS transcription factor BpGRAS19 confers salt stress tolerance in birch by regulating ROS scavenging

  • 期刊论文

  • 发表日期：

    2026年7月17日

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  • Furun Hao,
  • Qiang Wang,
  • Weiqi Li,
  • Wenli Zhu,
  • Lei Li,
  • Xiaoman Cui,
  • Xiaoyu Ji

摘要

Salt stress critically impairs plant growth and development; therefore, elucidating salt-tolerance genes and their regulatory mechanisms is essential for advancing molecular breeding. In our previous study, a GRAS transcription factor, BpGRAS19, was identified within the salt-responsive gene regulatory network of birch (Betula platyphylla). Phylogenetic analysis and conserved domain prediction validated BpGRAS19 as a member of the GRAS family, which contains a highly conserved GRAS domain. Subcellular localization experiments and quantitative real-time PCR (RT-qPCR) analyses further showed that BpGRAS19 is a nuclear-localized protein with tissue-specific expression patterns, and its expression is significantly induced by salt stress in birch. Transient transformation assays confirmed that BpGRAS19 functions as a positive regulator of salt tolerance in birch. In contrast, CRISPR/Cas9-mediated knockout of BpGRAS19 remarkably reduced salt tolerance, which was associated with impaired reactive oxygen species (ROS) scavenging capacity. Yeast one-hybrid (Y1H) assays indicated that BpGRAS19 specifically binds to characterized (MBS, W-box) and novel cis-acting elements (UN1: ACCGAGC; UN2: CGACATC), thereby expanding the DNA-binding motif of GRAS transcription factors. Taken together, these results demonstrate that BpGRAS19 improves salt tolerance in birch by mitigating cell membrane damage and enhancing ROS scavenging efficiency. This study lays a foundation for understanding the molecular mechanisms underlying salt tolerance in woody plants and provides theoretical support for salt-resistant breeding of birch.