刺芹侧耳软腐病病原菌分泌系统组成、蛋白互作分析及效应蛋白预测  

Secretion System Composition,Protein Interaction Analysis and Effector Proteins Prediction of Pleurotus eryngii Soft Rot Pathogen

作　　者：谷彤彤 赵爽[1,2] 宋爽 刘宇[1,2] 荣成博 GU Tong-Tong;ZHAO Shuang;SONG Shuang;LIU Yu;RONG Cheng-Bo(Institute of Plant Protection,Beijing Academy of Agriculture and Forestry Sciences,Beijing 100097,China;Beijing Engineering Research Center for Edible Mushroom,Beijing 100097,China)

机构地区：[1]北京市农林科学院植物保护研究所,北京100097 [2]北京市食用菌工程技术研究中心,北京100097

出　　处：《农业生物技术学报》2022年第7期1368-1384,共17页Journal of Agricultural Biotechnology

基　　金：北京市自然科学基金(6202008);北京市农林科学院创新能力建设专项(KJCX20200105);北京市农林科学农产品加工与营养健康协同创新中心项目(KJCX201915)。

摘　　要：软腐病是刺芹侧耳(Pleurotus eryngii)(商品名:杏鲍菇)的主要病害,北京欧文氏菌(Erwinia beijingensis)是该病的病原菌,目前该病原菌的致病机理尚不明确。本研究采用三代Nanopore与二代Illumina平台结合的方法获得了北京欧文氏菌的全基因组信息(0 gap),并进行了基因功能注释和分泌系统组成分析。在STRING网站(https://www.string-db.org/)预测了分泌系统蛋白的相互作用并绘制了蛋白互作网络;用SecReT6和Effectivet3软件预测了分泌的效应蛋白。北京欧文氏菌基因组大小为4.11 Mb,GC含量为50.17%,具有编码基因3815个。北京欧文氏菌具有Ⅰ型(typeⅠsecretion system,T1SS)、Ⅲ型(T3SS)、Ⅳ型(T4SS)和Ⅵ型分泌系统(T6SS)。T1SS由3个蛋白组成,包含内膜(inner membrane,IM)蛋白、膜融合蛋白(membrane fusion protein,MFP)和外膜(outer membrane,OM)蛋白,其中2对蛋白之间具有相互作用,推测分泌36个效应蛋白。T3SS包括9个保守的Hrp(hrp conserved,Hrc)蛋白和11个过敏反应与致病性(hypersensitive response and pathogenicity,Hrp)蛋白,组成88个蛋白互作对,预测分泌154个效应蛋白。T4SS包括9个毒性蛋白B(virulence B,VirB)蛋白,组成21对蛋白的相互作用关系,推测分泌11个效应蛋白。北京欧文氏菌具有2个T6SS,T6SS-1由13个T6SS蛋白和11个未知蛋白组成,形成104个蛋白互作对,T6SS-2包括11个蛋白和8个未知蛋白,组成55个蛋白互作对,SecReT6软件预测T6SS分泌6个效应蛋白。58个分泌系统基因在病原菌侵染宿主前、后均具有表达,其中34个基因具有显著的表达差异。本研究从基因组层面上解析了北京欧文氏菌分泌系统的组成、蛋白互作和分泌的效应蛋白,为该菌致病相关基因的解析提供了基础数据,为深入解析其致病机理提供理论依据。Soft rot disease which is caused by Erwinia beijingensis is the major disease in Pleurotus eryngii.Until now,the pathogenic mechanism of E.beijingensis is unknown.In the present study,the whole genome information of E.beijingensis was obtained by Nanopore and Illumina platform,the gene function annotation and secretion system composition were analyzed.The protein interactions of secretion system were predicted by string website(https://www.string-db.org/),and the effector proteins were predicted by SecRet6 and effectvet3 software.The genome size of E.beijingensis was 4.11 Mb,with GC%of 50.17%,encoding 3815genes.E.beijingensis displayed typeⅠ,typeⅢ,typeⅣand typeⅥsecretion system(T1SS,T3SS,T4SS and T6SS).T1SS was composed of 3 proteins,including inter membrane(IM)component,membrane fusion protein(MFP)and outer membrane(OM)protein channel,among which 2 pairs of proteins have interactions.It is speculated that 36 proteins were secreted through T1SS.T3SS consisted of 9 Hrc proteins and 11 Hrp proteins,with 88 pairs of protein interactions,and 154 effector proteins were predicted to be secreted by the T3SS.T4SS consisted of 9 VirB proteins with 21 pairs of protein interactions and 11 effector proteins.E.beijingensis had 2 T6SS,T6SS-1 was composed of 13 T6SS proteins and 11 hypothetical proteins,104 protein interactions were formed between T6SS proteins.T6SS-2 included 11 T6SS proteins and 8 hypothetical proteins,55 pairs of protein interactions were found in T6SS-2.It was predicted that 6effector proteins secreted into host cell by T6SS.58 secretion system genes could be expressed during infection,and 34 genes showed significant different expression.This study analyzed the composition,protein interactions and effector proteins of secretion systems in E.beijingensis from the genomic level,which provided basic data for the analysis of pathogenic genes and theoretical basis for further analysis of pathogenic mechanism.

关 键 词：软腐病 北京欧文氏菌 基因组 分泌系统 蛋白互作 

分 类 号：S6[农业科学—园艺学]