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<p>Low-temperature stress severely limits grapevine cultivation. <i>Vitis amurensis</i> Rupr., a cold-hardy wild grape species, serves as a valuable germplasm resource for breeding. In this study, a basic helix-loop-helix (<i>bHLH</i>) transcription factor gene, <i>VabHLH014</i>, was identified from <i>V. amurensis</i> ‘Zuoshan-1’. The expression of <i>VabHLH014</i> was strongly induced by low temperature, methyl jasmonate (MeJA), ethylene (ETH), and abscisic acid (ABA), with its promoter containing multiple stress-responsive cis-elements (ABRE, ARE, MYC). Subcellular localization confirmed VabHLH014 as a nuclear protein. Functional analysis through overexpression of <i>VabHLH014</i> in <i>Arabidopsis thaliana</i>, <i>Vitis vinifera</i> ‘Pinot Noir’ calli, and <i>V. vinifera</i> ‘Chardonnay’ plants revealed a significant reduction in cold tolerance compared to wild-type controls. Transgenic lines exhibited exacerbated freezing injury, increased malondialdehyde (MDA) accumulation, higher electrolyte leakage, decreased antioxidant enzyme activities (SOD, POD, CAT), and reduced proline levels. Moreover, expression of key cold-responsive genes (<i>CBF1/2/3</i>, <i>RD29A</i>, <i>NCED3</i>, <i>KIN1</i> in Arabidopsis; <i>VvCBF1/2/3</i>, <i>VvRD29B</i>, <i>VvNCED1</i>, <i>VvKIN2</i> in grapevine) was significantly downregulated in the transgenic background. Transcriptome profiling of wild-type and transgenic ‘Chardonnay’ before and after low-temperature treatment further revealed extensive changes in gene expression. Collectively, these results demonstrate that VabHLH014 negatively regulates cold tolerance and provide new insights into the molecular mechanisms of cold stress adaptation in grapevines.</p>

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VabHLH014, a stress-responsive gene from Vitis amurensis, suppresses cold tolerance in transgenic Arabidopsis and grapevine

  • 期刊论文

  • 发表日期：

    2026年1月21日

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  • Shan Li,
  • Chengnan Li,
  • Wenling Ma,
  • Ningning Guo,
  • Shouming Shan,
  • Wenkong Yao,
  • Xiao Yin

摘要

Low-temperature stress severely limits grapevine cultivation. Vitis amurensis Rupr., a cold-hardy wild grape species, serves as a valuable germplasm resource for breeding. In this study, a basic helix-loop-helix (bHLH) transcription factor gene, VabHLH014, was identified from V. amurensis ‘Zuoshan-1’. The expression of VabHLH014 was strongly induced by low temperature, methyl jasmonate (MeJA), ethylene (ETH), and abscisic acid (ABA), with its promoter containing multiple stress-responsive cis-elements (ABRE, ARE, MYC). Subcellular localization confirmed VabHLH014 as a nuclear protein. Functional analysis through overexpression of VabHLH014 in Arabidopsis thaliana, Vitis vinifera ‘Pinot Noir’ calli, and V. vinifera ‘Chardonnay’ plants revealed a significant reduction in cold tolerance compared to wild-type controls. Transgenic lines exhibited exacerbated freezing injury, increased malondialdehyde (MDA) accumulation, higher electrolyte leakage, decreased antioxidant enzyme activities (SOD, POD, CAT), and reduced proline levels. Moreover, expression of key cold-responsive genes (CBF1/2/3, RD29A, NCED3, KIN1 in Arabidopsis; VvCBF1/2/3, VvRD29B, VvNCED1, VvKIN2 in grapevine) was significantly downregulated in the transgenic background. Transcriptome profiling of wild-type and transgenic ‘Chardonnay’ before and after low-temperature treatment further revealed extensive changes in gene expression. Collectively, these results demonstrate that VabHLH014 negatively regulates cold tolerance and provide new insights into the molecular mechanisms of cold stress adaptation in grapevines.