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<p>Rising temperatures linked to global climate change pose a major threat to crop yield stability. Sexual reproduction in angiosperms is mediated by the female gametophyte and the male gametophyte (pollen), whose development, viability, and function are highly vulnerable to heat stress. We previously reported that the constitutive overexpression of the <i>C-repeat Binding Factor 1</i> (<i>CBF1</i>) enhanced heat tolerance but significantly delayed reproduction in Arabidopsis. In this study, we further exploited three promoters―<i>pOsRMP1</i>, <i>pOsLPS1</i>, and <i>pTDF1</i>―to drive <i>CBF1</i> overexpression specifically in male reproductive cells. The transgenic lines with pollen-specific <i>CBF1</i> overexpression displayed normal vegetative growth and transition to reproduction. Initial screening under heat stress conditions revealed that the transgenic line harboring the <i>OsRMP1</i> promoter exhibited the smallest reduction in silique length. Therefore, we focused further analyses, including pollen germination assays and gene expression profiling, on the <i>pOsRMP1::CBF1</i> line. RT-qPCR analyses revealed that targeted expression of <i>CBF1</i> significantly upregulated multiple stress-responsive genes, including heat responsive transcription factors, reactive oxygen species-scavenging enzymes, and ABA-responsive signaling components, indicating coordinated activation of cellular protective mechanisms. Taken together, our results demonstrate that targeted expression of <i>CBF1</i> using the <i>OsRMP1</i> promoter enhances heat tolerance in reproductive tissues, providing a promising strategy for developing climate resilient crops.</p>

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Targeted Overexpression of CBF1 Using a Pollen-specific Promoter Enhances Fertility in Heat-stressed Arabidopsis

  • 期刊论文

  • 发表日期：

    2025年8月6日

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  • Sang Dae Yun,
  • Eun Young Kim,
  • Sung-Aeong Oh,
  • Soon Ki Park

摘要

Rising temperatures linked to global climate change pose a major threat to crop yield stability. Sexual reproduction in angiosperms is mediated by the female gametophyte and the male gametophyte (pollen), whose development, viability, and function are highly vulnerable to heat stress. We previously reported that the constitutive overexpression of the C-repeat Binding Factor 1 (CBF1) enhanced heat tolerance but significantly delayed reproduction in Arabidopsis. In this study, we further exploited three promoters―pOsRMP1, pOsLPS1, and pTDF1―to drive CBF1 overexpression specifically in male reproductive cells. The transgenic lines with pollen-specific CBF1 overexpression displayed normal vegetative growth and transition to reproduction. Initial screening under heat stress conditions revealed that the transgenic line harboring the OsRMP1 promoter exhibited the smallest reduction in silique length. Therefore, we focused further analyses, including pollen germination assays and gene expression profiling, on the pOsRMP1::CBF1 line. RT-qPCR analyses revealed that targeted expression of CBF1 significantly upregulated multiple stress-responsive genes, including heat responsive transcription factors, reactive oxygen species-scavenging enzymes, and ABA-responsive signaling components, indicating coordinated activation of cellular protective mechanisms. Taken together, our results demonstrate that targeted expression of CBF1 using the OsRMP1 promoter enhances heat tolerance in reproductive tissues, providing a promising strategy for developing climate resilient crops.