黄瓜韧皮部汁液响应盐胁迫的蛋白质组学及抗性相关基因功能分析

项目名称： 黄瓜韧皮部汁液响应盐胁迫的蛋白质组学及抗性相关基因功能分析

项目编号： No.31201658

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 植物保护学、园艺学与植物营养学

项目作者： 杜长霞

作者单位： 浙江农林大学

项目金额： 25万元

中文摘要： 黄瓜是世界性的重要蔬菜，也是维管系统研究的模式植物，对盐渍环境敏感。蛋白质是生理功能的执行者和生命现象的直接体现者。韧皮部作为维管系统重要组成部分，内含大量与胁迫相关蛋白。本研究以黄瓜为试材，以富含蛋白质的韧皮部汁液为研究对象，利用以双向电泳和质谱分析为基础的蛋白质组技术，分离和鉴定响应盐逆境的韧皮部汁液蛋白，研究盐胁迫对韧皮部汁液蛋白差异表达、特异蛋白合成影响；结合生物信息学方法并采用Western blot和实时荧光定量PCR技术分析盐胁迫下的差异蛋白，明确响应盐胁迫的韧皮部汁液差异蛋白特性；克隆经分析确定响应盐胁迫的韧皮部汁液差异蛋白基因，采用农杆菌介导法转入拟南芥，研究盐胁迫下转基因植株的表型和抗盐性，初步探明响应盐胁迫韧皮部汁液差异蛋白功能。该研究对解析黄瓜在盐胁迫下的分子响应和适应机理、寻找用于黄瓜耐盐性评价的特异标志物、改良黄瓜的抗盐性以及丰富植物逆境生理理论等均有重要意义。

中文关键词： 黄瓜；盐胁迫；韧皮部渗出液；蛋白质组；基因功能

英文摘要： Cucumber is a cosmopolitan important vegetable, also the model plant of vascular system research, and sensitive to salt stress. Protein is executant of physiological function and direct embodiment of life phenomenon. Plant phloem is the important constituent of vascular system, which consists of a large number of proteins related with stress. In this study, cucumber is used as experiment material. The phloem sap is used as the research object, which includes mass protein. Two-dimensional gel electrophoresis and mass spectrometry techniques based on proteomics are utilized for protein separation and identification responded to salt stress in cucumber phloem sap protein. Diversity protein and synthesis of special protein are investigated. Combining on bioinformatic approach, characteristics of function protein was analyzed by western blot and real-time fluorescent quantitative PCR techniques. The function protein in the phloem sap is confirmed. Subsequently, genes of the confirmed function protein located in phloem sap are cloned, which are introduced into arabidopsis and transgenic plants were obtained. Phenotype, salt tolerance, growth and physiological characteristics in transgenic plants are studied. The responses of molecular mechanism in phloem sap to salt stress are primarily explained. Therefore, this stud

英文关键词： cucumber；salt stress；phloem sap；proteome；gene function