华北农学报 ›› 2025, Vol. 40 ›› Issue (3): 166-172. doi: 10.7668/hbnxb.20195409

所属专题: 植物保护 生物技术

• 资源环境·植物保护 • 上一篇    下一篇

豆伞滑刃线虫类FMRF酰胺多肽基因的克隆与分析

林世锋1,2, 王仁刚1,2, , 王自力1,2, 李莉3, 陈越男3, 吴沙沙3, 刘国琴4,   

  1. 1 贵州省烟草科学研究院,贵州 贵阳 550081
    2 烟草行业烟草分子遗传重点实验室,贵州 贵阳 550081
    3 贵州师范大学,贵州 贵阳 550081
    4 贵州大学,贵州 贵阳 550081
  • 收稿日期:2024-09-04 出版日期:2025-07-04
  • 通讯作者:
    王仁刚(1976—),男,辽宁营口人,副研究员,硕士,主要从事分子植物病理学研究。
    刘国琴(1975—),女,贵州遵义人,教授,博士,主要从事植物生理学研究。
  • 作者简介:

    林世锋(1978—),男,黑龙江牡丹江人,副研究员,博士,主要从事分子植物病理学研究。

  • 基金资助:
    中国烟草总公司贵州省公司科技项目(2023XM04); 国家自然科学基金项目(32060342); 贵州师范大学学术新苗基金项目(黔师新苗[2021]A13号); 贵州省科技厅农业攻关项目(黔科合NY[2013]3041); 贵州省林业科研项目(黔林科合[2014]02号)

Cloning and Analysis of a FMRFamide-like Neuropeptide Gene from Bursaphelenchus doui

LIN Shifeng1,2, WANG Rengang1,2, , WANG Zili1,2, LI Li3, CHEN Yuenan3, WU Shasha3, LIU Guoqin4,   

  1. 1 Guizhou Academy of Tobacco Science,Guiyang 550081,China
    2 Key Laboratory of Molecular Genetics of CNTC,Guiyang 550081,China
    3 Guizhou Normal University,Guiyang 550081,China
    4 Guizhou University,Guiyang 550081,China
  • Received:2024-09-04 Published:2025-07-04

摘要:

为探究豆伞滑刃线虫与寄主植物互作的分子机制,通过构建豆伞滑刃线虫体前端特异cDNA文库与RACE技术相结合的方法克隆获得1个豆伞滑刃线虫类FMRF酰胺多肽基因Bd-FLP-12,并应用生物信息学方法进行序列分析,应用Southern杂交和半定量RT-PCR方法分别检测基因拷贝数和在不同发育阶段的表达水平。序列分析表明,Bd-FLP-12编码蛋白由90个氨基酸残基组成,其中含有1个信号肽序列和1个FLP-12成熟肽序列,无跨膜结构域,属于分泌型蛋白。根据Southern杂交结果推测,Bd-FLP-12基因在豆伞滑刃线虫基因组中为单拷贝。半定量RT-PCR结果显示,Bd-FLP-12在豆伞滑刃线虫各龄期均有表达,但在卵期相对比较微弱。综上,首次在豆伞滑刃线虫中克隆获得Bd-FLP-12基因全长序列,解析了该基因的结构、性质和表达特征,为进一步研究该基因的功能奠定了基础。

关键词: 豆伞滑刃线虫, 类FMRF酰胺多肽, 生物信息学, Southern杂交, 表达分析

Abstract:

In order to explore the molecular mechanism of the interaction between Bursaphelenchus doui and host plants,an FMRFamide-like neuropeptide gene,named Bd-FLP-12,was cloned from B.doui by using RACE-PCR technique in a specific cDNA library of the anterior end of B.doui.Further,the nucleotide sequence and amino acid sequence of Bd-FLP-12 were analyzed using bioinformatics methods,the gene copy number was identified by Southern Blot,and the developmental expression pattern was investigated by semi-quantitative RT-PCR.The Bd-FLP-12 gene encoded a protein of 90 amino acids with a mature peptide sequence of FLP-12 and a signal peptide sequence but no transmembrane structure,indicating that its encoded protein was a secretory protein.Southern Blot indicated that the Bd-FLP-12 gene was a single copy gene in the B.doui genome,and the RT-PCR result showed that the transcription level of Bd-FLP-12 was lower in the egg stage than in the other stages.In summary,the full-length sequence of the Bd-FLP-12 gene was cloned for the first time in B.doui,and the structure,nature and expression of the gene were characterized,which could serve as a foundation for further study of the gene function.

Key words: Bursaphelenchus doui, FMRFamide-like neuropeptide, Bioinformatics, Southern Blot, Expression analysis

中图分类号: 

引用本文

林世锋, 王仁刚, 王自力, 李莉, 陈越男, 吴沙沙, 刘国琴. 豆伞滑刃线虫类FMRF酰胺多肽基因的克隆与分析[J]. 华北农学报, 2025, 40(3): 166-172. doi: 10.7668/hbnxb.20195409.

LIN Shifeng, WANG Rengang, WANG Zili, LI Li, CHEN Yuenan, WU Shasha, LIU Guoqin. Cloning and Analysis of a FMRFamide-like Neuropeptide Gene from Bursaphelenchus doui[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 166-172. doi: 10.7668/hbnxb.20195409.