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西瓜噬酸菌Ⅲ型分泌系统hrcS基因功能分析

引用本文：白庆荣,范泽慧,关巍,杨琳琳,杨玉文,赵廷昌.西瓜噬酸菌Ⅲ型分泌系统hrcS基因功能分析.植物保护学报,2023,50(2):350-358

DOI：10.13802/j.cnki.zwbhxb.2023.2021053

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作者	单位	E-mail
白庆荣	吉林农业大学植物保护学院, 长春 130118 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193
范泽慧	吉林农业大学植物保护学院, 长春 130118 中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193
关巍	中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193	wguan@ippcaas.cn
杨琳琳	中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193
杨玉文	中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193
赵廷昌	中国农业科学院植物保护研究所, 植物病虫害生物学国家重点实验室, 北京 100193	zhaotgcg@163.com

中文摘要:为明确编码Ⅲ型分泌系统(type Ⅲ secretion system,T3SS)中重要核心蛋白的hrcS基因对西瓜噬酸菌Acidovorax citrulli致病力的影响,以西瓜噬酸菌野生型菌株Aac5为对象,通过基因敲除构建hrcS基因缺失突变菌株ΔhrcS及其互补菌株ΔhrcS-comp,测定其致病力及致病相关基因表达量、烟草过敏性反应能力、运动能力及运动能力相关基因表达量、生物膜形成能力和离体生长能力。结果显示,与野生型菌株Aac5相比,hrcS基因缺失突变菌株丧失了对寄主的致病能力和引起非寄主烟草过敏性反应能力,运动能力显著增强了191.75%,生物膜形成能力显著下降了43.90%,离体生长能力在达到对数期后呈下降趋势。hrcS基因缺失使西瓜噬酸菌T3SS中hrpG、hrpX和hrpE基因以及鞭毛fliC基因表达量显著上调,分别为野生型菌株的2.27倍、2.38倍、1.26倍和1.77倍,hrcQ基因表达量显著下调,仅为野生型菌株的50.75%。表明hrcS基因在维持西瓜噬酸菌致病能力中发挥着重要作用。

中文关键词:西瓜噬酸菌 Ⅲ型分泌系统 hrcS基因基因功能致病力

Functional analysis of hrcS gene in type Ⅲ secretion system of bacterial pathogen Acidovorax citrulli

Author Name	Affiliation	E-mail
Bai Qingrong	College of Plant Protection, Jilin Agricultural University, Changchun 130118, Jilin Province, China State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Fan Zehui	College of Plant Protection, Jilin Agricultural University, Changchun 130118, Jilin Province, China State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Guan Wei	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China	wguan@ippcaas.cn
Yang Linlin	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Yang Yuwen	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
Zhao Tingchang	State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China	zhaotgcg@163.com

Abstract:In order to understand the role of hrcS gene encoding core protein of type III secretion system (T3SS) in the pathogenicity of Acidovorax citrulli, the hrcS deletion strain ΔhrcS, and its complementation strain ΔhrcS-comp were derived from Aac5, a wild type strain of A. citrulli in this study. The pathogenicity and the expression levels of genes involved in pathogenicity, hypersensitive response on tobacco, motility and the expression levels of genes involved in motility, biofilm formation, in vitro growth rate of the A. citrulli strains were determined. The results showed that the hrcS deletion mutant strain lost its virulence to the watermelon seedlings and the ability to induce hypersensitive reaction on non-host Nicotiana tabacum. Compared to the wild type strain Aac5, the swimming motility of the mutant strain significantly increased by 191.75%, while the biofilm formation decreased by 43.90%, and the in vitro growth rate decreased after logarithmic phase. The hrcS gene deletion significantly increased the expression level of T3SS genes hrpG, hrpX, hrpE and flagellin gene fliC by 2.27, 2.38, 1.26 and 1.77 times, respectively. However, the expression level of T3SS gene hrcQ was significantly reduced by 50.75%. These results indicated that the gene hrcS plays an important role in maintaining the pathogenicity of A. citrulli.

keywords:Acidovorax citrulli type Ⅲ secretion system (T3SS) hrcS gene function pathogenicity

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