油菜素内酯诱导黄瓜疫病抗性的蛋白质组学研究

项目名称： 油菜素内酯诱导黄瓜疫病抗性的蛋白质组学研究

项目编号： No.31301767

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： 康云艳

作者单位： 华南农业大学

项目金额： 22万元

中文摘要： 瓜类疫霉（Phytophthora melonis）引起的疫病是危害我国黄瓜生产的毁灭性土传病害，华南地区高温高湿的气候条件尤其适宜该病的发生。油菜素甾醇类化合物是一类对植物的生长发育有重要影响的新型激素，能提高植物体对多种病害的抗性。本研究通过表油菜素内酯（EBR）及其抑制剂brassinazole预处理2d，灌根接种P.melonis游动孢子悬浮液，利用以双向电泳和质谱分析为基础的蛋白质组技术，分离和鉴定响应P.melonis侵染和EBR诱导的差异蛋白；结合生物信息学方法并采用Western blot和实时荧光定量PCR技术分析差异蛋白，明确差异蛋白的特性和生理功能，探讨P.melonis和黄瓜植株的互作机制以及EBR诱导黄瓜植株疫病抗性的分子机制。该研究可为通过EBR等外源物质的施用防治黄瓜疫病提供理论依据。

中文关键词： 油菜素内酯；黄瓜；疫病；特异蛋白；基因表达

英文摘要： Phytophthora blight, caused by Phytophthora melonis Leonian, is one of the most important soil-borne fungal diseases of cucumber plants in China.High temperature and humidity conditions in southchina are favourable for disease development.Brassinosteroids are essential steroidal plant hormones, which have shown to overcome several kinds of pathogens.In this study, cucumber plants were inoculated by adding suspension of zoospores to the soil, and 24-epibrassinolide and its inhibitor brassinazole were also injected to the soil.To study the alleviating effect of 24-epibrassinolide on phytophthora blight in cucumber plants and its mechanism of the biolgical action, we compared the high resolution two-dimensional gel electrophoresis (2-DE) protein profiles in leaves and roots among different treatments, and the main differentially expressed protein spots were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and liquid chromatography electro-spray ionization tandem mass spectrometry (LC-ESI-MS/MS). Furthermore, RT-PCR and Real-time PCR approach based on peptide sequences were used to compare transcript and protein accumulation patterns for candidate proteins. In conclusion, our study could provide the theory basis for plant-pathogen interaction model and applic

英文关键词： brassinolide；cucumis sativus；phytophthora blight；specific protein；gene expression