利用酵母双杂交筛选与致病疫霉菌效应蛋白PITG_07586互作的寄主蛋白  

Screening of host proteins interacting with effector PITG_07586 of Phytophthora infestans using yeast two-hybrid

作　　者：张引弟[1,2] 王荟洁 陈爱娥 刘晶 王洪洋 李灿辉[1,2] ZHANG Yindi;WANG Hujie;CHEN Aie;LIU Jing;WANG Hongyang;LI Canhui(Yunnan Key Laboratory of Potato Biology,Yunnan Normal University,Kunming 650500,China;Key Lab of Potato Biology in Universities of Yunnan Province,Yunnan Normal University,Kunming 650500,China;Teaching Affairs Department,Yunnan Normal University,Kunming 650500,China)

机构地区：[1]云南省马铃薯生物学重点实验室,云南师范大学,昆明650500 [2]云南省高校马铃薯生物学重点实验室,云南师范大学,昆明650500 [3]云南师范大学教务处,昆明650500

出　　处：《植物病理学报》2022年第4期592-600,共9页Acta Phytopathologica Sinica

基　　金：国家自然科学基金资助项目(31800134、32060499);云南省基础研究计划资助项目(2019FB023)。

摘　　要：致病疫霉是引起马铃薯晚疫病的病原菌,其分泌的效应蛋白是侵染寄主的重要毒力因子。本研究在致病疫霉转录组中筛选到一个侵染早期表达的效应蛋白基因PITG_07586(GenBank:XM_002904522.1)。该基因全长447 bp,其编码蛋白包含1个信号肽,1个核定位信号序列(RRRRKRRKKKK)。在本氏烟草中,瞬时表达该基因显著促进病原菌侵染。亚细胞定位表明GFP-PITG_07586定位在细胞核中。利用酵母双杂交技术并以PITG_07586为诱饵筛选马铃薯cDNA文库,最终获得3个靶标蛋白。经序列比对分析,这3个靶标蛋白分别是马铃薯POM30蛋白、电压依赖阴离子通道蛋白VDAC以及SRC2类蛋白。研究结果为致病疫霉菌效应蛋白PITG_07586及其靶标蛋白如何调控植物免疫提供重要依据。Phytophthora infestans is the pathogen responsible for the devastating late blight disease on potato. An array of effectors secreted by P. infestans plays an important role in the process of host infection. Based on analysis of transcriptome data of plants infected by P. infestans, we found an effector gene PITG_07586(GenBank: XM_002904522.1, 447 bp), which was up-regulated during the early stages of infection. The N-terminus of PITG_07586 contained a signal peptide(1-21 aa), and C-terminus contained a nuclear localization signal sequence(RRRRKRRKKKK). Transient expression of PITG_07586 significantly promoted P. infestans colonization in Nicotiana benthamiana. Confocal microscopic examination showed that GFP-PITG_07586 was located in the host nucleus. To further explore host proteins interacting with PITG_07586, we screened yeast cDNA library and used yeast two-hybrid assay to verify the target protein of PITG_07586 in potato. We finally got three potato target proteins that interact with PITG_07586. They were a POM30 protein, mitochondrial outer membrane protein porin VDAC, and SRC2-like protein. Our results provided a clue for further investigation of how P. infestans effector PITG_07586 associated with and modulates host immunity.

关 键 词：致病疫霉菌 PITG_07586 酵母双杂交 靶标蛋白 

分 类 号：S432.1[农业科学—植物病理学]