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<p>Adventitious root (AR) is critical for the clonal propagation, yet the molecular mechanisms underlying this process remain poorly understood of woody plants. Auxin Response Factors (ARFs), central transcriptional regulators of auxin signaling, are hypothesized to govern AR development. Here, we systematically identified and characterized the <i>ARF</i> gene family in <i>C. bungei</i> to elucidate its roles in AR ontogeny. Through integrated <i>de novo</i> transcriptome assembly and PacBio sequencing database, 17 <i>CbARF</i> genes were identified, all harboring conserved B3 DNA-binding and Auxin_resp domains. Phylogenetic analysis classified these genes into six clades, with lineage-specific expansion observed in Classes III and V. Structural variations, including the absence of the C-terminal dimerization domain in four members (CbARF3, CbARF10, CbARF16, and CbARF6.3), might imply altered interaction dynamics with Aux/IAA repressors. Spatiotemporal expression profiling via transcriptomics and qRT-PCR revealed stage- and tissue-specific expression patterns during AR development. <i>CbARF5</i> and <i>CbARF6.3/6.4</i> exhibited peak expression during early AR induction (callus formation), while <i>CbARF6.1/6.2</i> and <i>CbARF19</i> showed progressive downregulation. Yeast assays confirmed the transcriptional autoactivation capacity of <i>CbARF5</i>, <i>CbARF6</i> paralogs, and <i>CbARF19</i>, highlighting their roles as autonomous regulators. These findings underscore the potential sequence and functional divergence through auxin-mediated transcriptional networks in AR initiation and elongation between <i>CbARF1</i> and <i>CbARF6</i> paralogs respectively. Our study provides comprehensive characterization of the <i>CbARF</i> family, offering molecular insights to advance the development of high-efficiency propagation technologies for <i>C. bungei</i> and related woody species.</p>

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Gene Identification and Functional Characterization of Catalpa bungei Auxin Response Factor Family in Adventitious Root Development

  • 期刊论文

  • 发表日期：

    2026年2月2日

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  • Jingyi Liu,
  • Weijie Si,
  • Hongying Li,
  • Jian Xu,
  • Fuqiang Yang,
  • Yifan Li,
  • Shiqi Niu,
  • Ziyuan Hao

摘要

Adventitious root (AR) is critical for the clonal propagation, yet the molecular mechanisms underlying this process remain poorly understood of woody plants. Auxin Response Factors (ARFs), central transcriptional regulators of auxin signaling, are hypothesized to govern AR development. Here, we systematically identified and characterized the ARF gene family in C. bungei to elucidate its roles in AR ontogeny. Through integrated de novo transcriptome assembly and PacBio sequencing database, 17 CbARF genes were identified, all harboring conserved B3 DNA-binding and Auxin_resp domains. Phylogenetic analysis classified these genes into six clades, with lineage-specific expansion observed in Classes III and V. Structural variations, including the absence of the C-terminal dimerization domain in four members (CbARF3, CbARF10, CbARF16, and CbARF6.3), might imply altered interaction dynamics with Aux/IAA repressors. Spatiotemporal expression profiling via transcriptomics and qRT-PCR revealed stage- and tissue-specific expression patterns during AR development. CbARF5 and CbARF6.3/6.4 exhibited peak expression during early AR induction (callus formation), while CbARF6.1/6.2 and CbARF19 showed progressive downregulation. Yeast assays confirmed the transcriptional autoactivation capacity of CbARF5, CbARF6 paralogs, and CbARF19, highlighting their roles as autonomous regulators. These findings underscore the potential sequence and functional divergence through auxin-mediated transcriptional networks in AR initiation and elongation between CbARF1 and CbARF6 paralogs respectively. Our study provides comprehensive characterization of the CbARF family, offering molecular insights to advance the development of high-efficiency propagation technologies for C. bungei and related woody species.