大豆黄单胞杆菌基因hrpG的突变及其对致病性影响
Mutation of Xanthomonas hrpG Gene and Its Effect on Pathogenicity
刘冬旭 1周红阳 1郭广雨 2孙雨田 1马胜男 1苏安玉 3辛大伟 1王锦辉1
作者信息
- 1. 东北农业大学农学院,哈尔滨,150030
- 2. 哈尔滨市农业科学院粮食与经济作物分院,哈尔滨,150028
- 3. 东北农业大学资源与环境学院,哈尔滨,150030
- 折叠
摘要
大豆细菌性斑疹病是贻害大豆稳产、高产的主要病害之一,其影响遍及全球.该病害的病原之一是革兰氏阴性菌黄单胞杆菌的致病变种 Xanthomonas axonopodis pv.glycines(Xag)和 Xanthomonas campestris pv.glycinea(NaKmo)Dye(Xcg).革兰氏阴性病原菌大多通过hrp基因编码的Ⅲ型分泌系统(type Ⅲ secretion systems,T3SS)将毒力因子注入寄主细胞中,从而导致寄主发病.本研究采用的细菌性斑疹病致病菌Xagneau-001,分离自黑龙江佳木斯大豆主产区,该菌株已经完成全基因组测序.在基因组注释过程中发现其基因组中含有hrp基因家族中的hrpG基因,本课题组针对hrpG基因的致病性开展深入研究.通过三亲杂交的方法成功构建突变体菌株Xag△hrpG,并利用PCR检测结合DNA测序结果证实突变体Xag△hrpG中hrpG基因已成功敲除.将野生型菌株Xagneau001、突变菌株Xag△hrpG共同接种12个遗传背景不同品种的大豆资源,测定突变体Xag△hrpG对大豆的致病力的影响.结果显示大豆接种Xag△hrpG后斑疹病发病时间滞后、进程减缓,发病程度显著降低.统计接菌4 d后单位面积病叶的病原菌数量显示,突变体Xag△hrpG的病原菌数量显著低于野生型Xagneau001.这表明与野生型Xagneau001相比,Xag△hrpG突变体对于大豆的致病力大幅减弱,指示hrpG基因正向调控Xagneau001对大豆致病力.
Abstract
Soybean bacterial spot disease is one of the major diseases that seriously damage the soybean yield.The pathogens of this disease are Xanthomonas axonopodis pv.glycines(Xag)and Xanthomonas campestris pv.glycinea(NaKmo)Dye(Xcg),which are gram-negative bacteria of Xanthomonas.Gram-negative pathogens cause Type Ⅲ secretion systems(T3SS),host-dependent on the hrp gene,to inject virulence factors into host cells.This study is of great significance to advance the breeding progress of soybean bacterial spot disease.This study used Xagneau001,a bacterial leaf spot pathogen,isolated from Jiamusi region.Its genome had been sequenced,and the hrpG gene was found in genome.Further research on pathogenicity of gene hrpG is carried out in our subject group.The mutant Xag△hrpG was constructed by triparental mating,and the hrpG gene was successfully knocked out by PCR detection and DNA sequencing.12 genetic backgrounds of different soybean germplasms were inoculated the wild type strain Xagneau001 and the mutant strain Xag△hrpG,respectively.The results showed that after inoculation of soybean with Xag△hrpG,the onset time of the spotted disease was prolonged,the progress was slowed down,and the pathogenicity was significantly reduced.The number of pathogenic bacteria per unit area of diseased leaves after 4 days was counted.The number of pathogenic Xag△hrpG pathogens was lower than that of wild-type Xagneau001.This result shows that Xag△hrpG is less pathogenic to soybean than wild-type Xagneau001.The hrpG enhances the pathogenicity of soybean bacterial spot disease.
关键词
大豆/细菌性斑疹病/hrpG基因/致病能力Key words
Soybean/Soybean bacterial pustule/hrpG/Pathogenic ability引用本文复制引用
基金项目
中国博士后基金项目(2020M681072)
国家自然科学基金项目(32072014)
国家自然科学基金项目(U20A2027)
出版年
2024
NETL
NSTL
万方数据