稻瘟病菌转录因子MoAp1介导的基因调控网络生物学功能解析

项目名称： 稻瘟病菌转录因子MoAp1介导的基因调控网络生物学功能解析

项目编号： No.31271998

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 张正光

作者单位： 南京农业大学

项目金额： 82万元

中文摘要： 植物与病菌互作时，植物细胞产生的活性氧在植物防卫中具有非常重要的作用。已证明稻瘟病菌要成功侵染水稻，必须清除互作早期水稻细胞产生的活性氧。我们前期研究发现转录因子MoAp1作为活性氧胁迫的感受器，调控稻瘟病菌一系列基因的表达，清除活性氧，从而抑制其介导的防卫反应。因此解析MoAp1调控的基因网络有助于揭示该病菌的致病分子机制。本项目拟采用co-IP技术鉴定MoAp1的结合蛋白，并鉴定其生物学功能及其对MoAp1定位的影响。采用染色体免疫共沉淀技术鉴定MoAp1在该病菌中的DNA结合序列，结合项目组已有的转录组数据，鉴定MoAp1的靶基因，并揭示其生物学功能，从而提出MoAp1调控的基因网络，解析该网络在稻瘟病菌应答活性氧胁迫、及生长发育和致病中的分子机制。上述工作不仅对揭示MoAp1在该病菌生长发育及致病中的分子机理具有重要的理论价值，而且对设计以网络为靶标的病害控制策略具有重要指导意义。

中文关键词： 转录因子；基因调控网络；调控蛋白；靶蛋白；生物学功能

英文摘要： Reactive oxygen species (ROS) are active in the critical zone where pathogens and plants interaction happens and play important roles in defense mechanisms. Recent studies demonstrated that it is necessary for full virulence of the rice blast fungus Magnaporthe oryzae by suppressing plant-generated ROS during early stages of infection to impair ROS-mediated rice defense. Our study revealed that MoAp1, a transcription factor as a positive ROS-stress sensor and regulated gene expression to detoxify rice-generated ROS and eventually blocked ROS-triggered plant defense. Thus, research on gene regulatory network mediated by MoAp1 could help to elucidate molecular aspects of pathogenicity of M. oryzae. One of our specific objectives is to characterize biological function of MoAp1-binding proteins and evaluate the effect of these proteins on MoAp1-GFP localization after identified by co-immunoprecipitation in M. oryzae. The other specific object is to use Chromatin Immunoprecipitation followed by Illumina sequencing (ChIP-Seq) to identify the MoAp1 binding motifs and evaluate the putative MoAp1 binding motifs by electro mobility shift assays (EMSA). MoAp1 binding motifs as well as our previous RNA-sequencing data of the Moap1 mutant will used to identify potential target genes regulated by MoAp1. Finally our specific

英文关键词： transcription factor；gene regulatory network；regulatory protein；target protein；biological function