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<p>The wound-induced protein (WIP) is a member of the A1d subfamily of C2H2 zinc finger proteins and plays a crucial role in plant growth and development. Foxtail millet (<i>Setaria italica</i>) serves as a model organism for research on C4 crops. To date, the WIP family has been identified in several plant species, including <i>Arabidopsis thaliana</i> and tomato (<i>Solanum lycopersicum</i> L.), but it has not yet been reported in foxtail millet. In this study, we conducted bioinformatics analysis to identify five <i>SiWIP</i> genes in the whole genome of foxtail millet, and further examined their chromosomal distribution, gene structure, <i>cis</i>-elements, and conserved protein motifs. The analysis of tissue expression patterns of <i>SiWIP3</i>, <i>SiWIP4</i> and <i>SiWIP5</i> members, as well as their response to exogenous hormone treatments, indicates that <i>SiWIPs</i> demonstrate tissue specificity and exhibit distinct reactions to exogenous hormones. Notably, <i>SiWIP3</i> displays a certain level of sensitivity to exogenous hormones and shows the highest expression in roots. Further research on SiWIP3 has revealed its nuclear localization, and the overexpression of <i>SiWIP3</i> in <i>Arabidopsis</i> results in a reduction in leaf size, slower plant growth, and suppressed flowering. However, the inhibition was relieved by the exogenous application of gibberellin (GA<sub>3</sub>), leading to leaf expansion and promotion of flowering. RT-qPCR results revealed alterations in the expression levels of GA pathway related genes in <i>Arabidopsis</i> overexpressed lines, both before and after GA<sub>3</sub> treatment. In addition, the dual luciferase assay demonstrated that SiKNOX1 or SiBPC6 significantly suppressed the expression of the <i>SiWIP3</i>. Protein interactions between SiWIP3 and SiTCP19 were confirmed through yeast two-hybrid and luciferase complementation imaging assay experiments. <i>SiWIP3</i> may play a crucial role in GA pathway, thereby regulating the growth and development in foxtail millet. This research provides a fundamental basis for further elucidating the mechanism of WIP protein in plant growth and development.</p>

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Genome-wide identification of the WIP family in foxtail millet (Setaria italica) and functional analysis of SiWIP3 in inhibiting growth in transgenic Arabidopsis thaliana

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年10月22日

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  • Xiling Chang,
  • Tianqi Song ,
  • Jiandong Ren,
  • Xiaoxing Wang,
  • Jianfei Zhou,
  • Yaning Bu,
  • Xiaohong Chen,
  • Yuxin Wang,
  • Siyi Li,
  • Xinpeng Lei,
  • Jishan Xiang,
  • Xiaoke Zhang

摘要

The wound-induced protein (WIP) is a member of the A1d subfamily of C2H2 zinc finger proteins and plays a crucial role in plant growth and development. Foxtail millet (Setaria italica) serves as a model organism for research on C4 crops. To date, the WIP family has been identified in several plant species, including Arabidopsis thaliana and tomato (Solanum lycopersicum L.), but it has not yet been reported in foxtail millet. In this study, we conducted bioinformatics analysis to identify five SiWIP genes in the whole genome of foxtail millet, and further examined their chromosomal distribution, gene structure, cis-elements, and conserved protein motifs. The analysis of tissue expression patterns of SiWIP3, SiWIP4 and SiWIP5 members, as well as their response to exogenous hormone treatments, indicates that SiWIPs demonstrate tissue specificity and exhibit distinct reactions to exogenous hormones. Notably, SiWIP3 displays a certain level of sensitivity to exogenous hormones and shows the highest expression in roots. Further research on SiWIP3 has revealed its nuclear localization, and the overexpression of SiWIP3 in Arabidopsis results in a reduction in leaf size, slower plant growth, and suppressed flowering. However, the inhibition was relieved by the exogenous application of gibberellin (GA3), leading to leaf expansion and promotion of flowering. RT-qPCR results revealed alterations in the expression levels of GA pathway related genes in Arabidopsis overexpressed lines, both before and after GA3 treatment. In addition, the dual luciferase assay demonstrated that SiKNOX1 or SiBPC6 significantly suppressed the expression of the SiWIP3. Protein interactions between SiWIP3 and SiTCP19 were confirmed through yeast two-hybrid and luciferase complementation imaging assay experiments. SiWIP3 may play a crucial role in GA pathway, thereby regulating the growth and development in foxtail millet. This research provides a fundamental basis for further elucidating the mechanism of WIP protein in plant growth and development.