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中国农学通报

中国农学通报 ›› 2025, Vol. 41 ›› Issue (5): 50-55.doi: 10.11924/j.issn.1000-6850.casb2024-0208

• 生物科学 • 上一篇    下一篇

芦竹独脚金内酯受体基因克隆及功能验证

陈虞超1,2(), 郭生虎1,2, 田莉1,2, 钟楠1,2, 杨建国1,2, 甘晓燕1,2()   

  1. 1 宁夏农林科学院农业生物技术研究中心，银川 750002
    2 宁夏农业生物技术重点实验室，银川 750002
  • 收稿日期:2024-03-28 修回日期:2024-06-15 出版日期:2025-02-15 发布日期:2025-02-11
  • 通讯作者:
    甘晓燕，女，1982年出生，宁夏中宁人，副研究员，硕士研究生，研究方向：农业生物技术。通信地址：750002 宁夏银川市金凤区黄河东路590号 宁夏农林科学院农业生物技术研究中心，Tel：0951-6886758，E-mail：。
  • 作者简介:

    陈虞超，男，1982年出生，湖北孝感人，副研究员，博士研究生，研究方向：分子生物学。通信地址：750002 宁夏银川市金凤区黄河东路590号 宁夏农林科学院农业生物技术研究中心，Tel：0951-6886758，E-mail：。

  • 基金资助:
    宁夏重点研发计划项目“新型禾本科牧草芦竹高效繁育及饲料加工技术研究”(2022BBF03033)

Clone and Functional Characterization of Strigolactone Receptor Gene in Arundo donax L.

CHEN Yuchao1,2(), GUO Shenghu1,2, TIAN Li1,2, Zhong Nan1,2, YANG Jianguo1,2, GAN Xiaoyan1,2()   

  1. 1 Agricultural Biotechnology Centre, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002
    2 Key Laboratory of Agricultural Biotechnology of Ningxia, Yinchuan 750002
  • Received:2024-03-28 Revised:2024-06-15 Published:2025-02-15 Online:2025-02-11

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摘要：

本研究旨在探究芦竹(Arundo donax L)分蘖调控的分子机制，通过克隆其独脚金内酯(SL)受体基因AdD14，并进行生物信息学分析、基因功能验证。以芦竹幼苗为材料，同源克隆其SL受体基因AdD14（Genbank登陆号：OR915727）；利用ExPASy、MEGA等软件对基因进行生物信息学分析；利用其原核表达蛋白体外催化的方法验证基因功能。结果发现，AdD14开放阅读框全长816 bp，编码分子量为29.75 kDa的271个氨基酸残基。AdD14蛋白预测属于不稳定的疏水性蛋白，不存在跨膜结构，无信号肽。系统进化树显示，AdD14与其他种类植物D14蛋白具有较高的同源性，保留了SL受体保守结构域和催化三联体，可水解SL受体荧光探针YLG(Yoshimulatone Green)，显示AdD14为芦竹SL受体基因。本研究分离了芦竹AdD14基因，其编码蛋白与其他植物的SL受体D14蛋白结构及功能相一致，初步证实其为芦竹SL受体基因，为深入揭示芦竹分蘖调控分子机制奠定了一定基础。

关键词: 芦竹, 独脚金内酯受体, 基因克隆, AdD14基因, 基因功能验证, 生物信息学, 分蘖调控

Abstract:

In order to explore the molecular mechanism of the regulation of tiller in Arundo donax L., we cloned AdD14, a SL receptor gene in Arundo donax L., and conducted bioinformatics analysis and gene function verification. The SL receptor gene AdD14 (Genbank login number: OR915727) was homologous cloned from Arundo donax L. seedlings. ExPASy, MEGA and other software were used for bioinformatics analysis of the genes. The function of the gene was verified by the method of in vitro catalysis of the prokaryotic expression protein. The results showed that the AdD14 open reading frame was 816bp long and encoded 271 amino acid residues with a molecular weight of 29.75kDa. AdD14 protein was predicted to be an unstable hydrophobic protein with no transmembrane structure and no signal peptide. The phylogenetic tree showed that AdD14 had high homology with D14 protein of other species of plants, retained the conserved domain of SL receptor and catalytic triad, and could hydrolyze the fluorescent probe YLG (Yoshimulatone Green) of SL receptor, indicating that AdD14 was the SL receptor gene of Arundo donax L.. In this study, the AdD14 gene of Arundo donax L. was isolated, and its encoding protein was consistent with the structure and function of SL receptor D14 protein of other plants. It was initially confirmed that it was the SL receptor gene of Arundo donax L., which laid a certain foundation for further revealing the molecular mechanism of tiller regulation in Arundo donax L..

Key words: Arundo donax L., strigolactone receptor, gene clone, AdD14 gene, gene function verification, bioinformatics, tiller regulation