基于蛋白质组学的稻瘟病菌转录因子MoMsn2的调控机制研究

项目名称： 基于蛋白质组学的稻瘟病菌转录因子MoMsn2的调控机制研究

项目编号： No.31471736

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 张海峰

作者单位： 南京农业大学

项目金额： 85万元

中文摘要： HOG1介导的信号途径在稻瘟病菌的生长发育、胁迫应答及致病过程中具有重要的调控作用。申请者前期鉴定到一个与MoHog1(MoOsm1)相互作用的转录因子MoMsn2。对其进行功能分析发现，MoMsn2通过调控一系列基因的表达，参与该病菌的生长、产孢、细胞壁完整性、胁迫应答及致病性。为深入阐明MoMsn2的分子调控机制，本项目拟从蛋白质组学出发，在胁迫和非胁迫条件下，分别采用同位素相对标记与绝对定量(iTRAQ)技术和Pull-down技术鉴定受MoMsn2调控的差异表达蛋白及与其结合的蛋白，并分析这些蛋白的生物学功能及对MoMsn2定位的影响，从而提出MoMsn2介导的蛋白调控网络，最终阐明该网络在稻瘟病菌生长发育、胁迫应答及致病过程中的分子调控机制。上述工作不仅有助于深入揭示HOG1信号途径介导的分子机制，而且对设计针对该信号途径的新型杀菌剂和制定有效的病害防治策略具有重要的指导意义。

中文关键词： 转录因子；MoMsn2；蛋白质组学；调控机制；稻瘟病菌

英文摘要： The HOG1-mediated osmoregulation pathway and transcription factors play crucial roles in growth, conidiation, stress response and pathogenicity in Magnaporthe oryzae. In the latest study, we identified and characterized a transcription factor MoMsn2 that physically interacts with MoHog1. Targeted deletion of MoMSN2 revealed that MoMsn2 modulates a series of downstream genes to control aerial hyphal growth, conidiogenesis, cell wall integrity, infectious growth, stress response and virulence in M. oryzae. However, the regulatory mechanism and the targets of MoMsn2 involved in these processes need to be explored. Therefore, identification and characterization of the regulatory proteins and binding proteins of MoMsn2 will help us to better understand the molecular mechanisms of HOG1signaling pathway and MoMsn2 in the rice blast fungus. In this study, the isobaric tags for relative and absolute quantitation (iTRAQ) technique will be used to identify the differential expression proteins of MoMsn2 treatment with or without osmotic stress; and Pull-down will be further adopted to identify the binding proteins of MoMsn2 under normal or under stress conditions. The candidate binding proteins will be analyzed and their interactions with MoMsn2 will be confirmed by yeast two hybrid, co-IP or BiFC. The confirmed candidates and differential expression proteins will be further characterized, as well as their effects on the localization of MoMsn2 in M. oryzae. The results will propose a protein regulatory network mediated by MoMsn2 to address the mechanisms on growth and development, stress response and pathogenicity of M. oryzae. This study will not only better understand the regulatory mechanisms of HOG1signaling pathway, but also will help to design fungicide and develop new disease management strategies.

英文关键词： transcription factor;MoMsn2;proteomics;regulatory mechanism;Magnaporthe oryzae