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中国生物防治学报 ›› 2021, Vol. 37 ›› Issue (6): 1353-1362.DOI: 10.16409/j.cnki.2095-039x.2021.06.022

• 研究简报 • 上一篇    

青枯雷尔氏菌Ⅲ型分泌系统hrcN基因的功能分析

周大祥1,2, 龙泉洲2, 殷幼平1, 熊书3   

  1. 1. 重庆大学生命科学学院/重庆市基因功能与调控重点实验室, 重庆 400300;
    2. 重庆三峡学院生物与食品工程学院, 万州 404120;
    3. 重庆三峡医药高等专科学校基础医学部, 万州 404120
  • 收稿日期:2021-06-11 发布日期:2021-12-15
  • 通讯作者: 熊书,硕士,讲师,E-mail:24594294@qq.com。
  • 作者简介:周大祥,副教授,博士研究生,E-mail:dqzhou79@163.com。
  • 基金资助:
    重庆市自然科学基金(cstc2016jcyjA0198)

Functional Analysis of Type III Secretion System hrcN Gene in Ralstonia solanacearum

ZHOU Daxiang1,2, LONG Quanzhou2, YIN Youping1, XIONG Shu3   

  1. 1. College of Life Science, Chongqing University/Chongqing Key Lab of Genetic Function and Regulation, Chongqing 400030, China;
    2. College of Biology and Food Engineering, Chongqing Three Gorges University, Wanzhou 404120, China;
    3. Department of Basic Medicine, Chongqing Three Gorges Medical College, Wanzhou 404120, China
  • Received:2021-06-11 Published:2021-12-15

摘要: hrcN是植物病原菌Ⅲ型分泌系统基因(T3SS)的结构基因之一,对病原菌的致病力有重要影响。本研究克隆青枯雷尔氏菌CBM613的hrcN基因并作生物信息学分析,结果显示该基因长度为1320 bp,共编码439个氨基酸,预测HrcN蛋白无信号肽,无跨膜区域,整体呈弱亲水性,该蛋白为ATP酶,定位于细胞内膜和细胞质。基因敲除结果显示,菌株CBM613的hrcN突变体与野生型菌株在富营养培养基上的生长速率差异不明显,而在贫营养培养基上前者的生长速率显著快于后者。敲除hrcN基因后,青枯雷尔氏菌的运动能力无明显变化,生物被膜形成能力显著降低。以敲除突变型和野生型青枯雷尔氏菌株接种番茄,与后者相比,前者的致病力下降,病情指数降低,病程延长,但并未完全丧失致病力,表明hrcN基因是青枯雷尔氏菌致病性相关的重要基因。本研究为进一步探索青枯雷尔氏菌致病机制及防治具有重要意义。

关键词: 青枯雷尔氏菌, T3SS, hrcN, 基因敲除, 功能分析

Abstract: The gene hrcN is one of the structural genes of the type III secretion system (T3SS) in plant pathogens, and has an important influence on pathogenicity of pathogens. In this study, the gene hrcN was cloned from Ralstonia solanacearum strain CBM613 to perform bioinformatics analysis. The results showed that the gene was 1320 bp in length, encoding 439 amino acids. The protein HrcN was predicted to have no signal peptide and transmembrane region, and to be weakly hydrophilic. The protein HrcN is an ATPase, which is located in the cell inner membrane and cytoplasm. The growth rate of the hrcN mutant had no significant difference with the wild-type strain in eutrophic media, but was significantly faster than that of the latter in the poor nutrition media After the gene hrcN was removed, motility of the hrcN mutant and the wild-type strain had no significant change, but the biofilm formation ability of the hrcN mutant was significantly reduced. Moreover, pathogenicity of the hrcN mutant was decreased compared with the wild-type strain, and disease index was decreased, meanwhile the course of disease was prolonged, but its pathogenicity was not completely lost. The results indicated that the gene hrcN is an important gene associated with the pathogenicity of the R. solanacearum. Our study is of great significance to further explore the pathogenic mechanism of R. solanacearum and biocontrol of bacterial wilt.

Key words: Ralstonia solanacearum, T3SS, hrcN, gene knock-out, functional analysis

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