基于组织学和转录组学对甘蔗宿根矮化病的致病机理解析

项目名称： 基于组织学和转录组学对甘蔗宿根矮化病的致病机理解析

项目编号： No.31301381

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 郭莺

作者单位： 福建省亚热带植物研究所

项目金额： 22万元

中文摘要： 甘蔗宿根矮化病是世界性的甘蔗细菌病害。本项目以不带Lxx甘蔗组培苗为材料，利用表达GFP基因的Lxx突变菌株接种研究Lxx侵染的组织学过程；通过转录组测序法高通量分析与甘蔗互作过程中Lxx基因差异表达谱，通过KEGG和COG等蛋白质数据库进行比对，获得蛋白质和pathway注解；确定差异表达基因参与主要代谢途径和信号转导途径；筛选Lxx致病关键基因，通过RACE和Genome walking技术获得致病关键基因,通过生物、信息学和Southern分析基因结构，通过定量PCR和Northern杂交，对致病关键基因功能进行解析。以上研究不仅从组织学方面解析Lxx致病性，更直接深入到转录组水平，确定Lxx致病的信号转导途径和代谢途径，并获得具有自主知识产权的致病基因。这不仅为阐明甘蔗宿根矮化病的致病机理奠定基础，也为今后甘蔗宿根矮化病的防治和抗病品种选育提供技术支撑，具有重要的理论和实践意义。

中文关键词： 甘蔗宿根矮化病；LXX病原菌；病原菌与寄主互作；转录组测序；

英文摘要： Sugarcane ratoon stunting disease is the worldwide disease. In this research, tissue culture sugarcane without lxx was as study object. GFP-Lxx mutant was inoculated in sugarcane, and the infecting process was surveyed under fluorescence microscope. In effect interaction between the Lxx and sugarcane, differential gene expression profile of Lxx were acquired using the second-generation high throughput sequencing method. The sequencing results were aligned with KEGG and COG protein database, and noted the protein function and the pathway, also known differential expression gene of lxx were participated in which metabolic pathway and signal transduction pathway. The key pathogenic genes were cloned using race and genome walking method and analyzed with bioinformatics software. Function of pathogenic genes were study using the southern blot, northern blot and real-time PCR. Lxx pathogenicity were studied in this project not only in histology, but also in transcriptome. And the main metabolic pathway and signal transduction pathway were identified in connection with pathogenicity, and gained the proprietary intellectual property rights of pathogenic genes. It not only laid the foundation for clarifying the pathogenic mechanism of sugarcane ratoon stunting disease, but also provide the technical support for selectin

英文关键词： sugarcane ratoon stunting disease；LXX；Pathogen and host interaction；RNA squencing；