番茄IL系耐旱相关基因的表达分析与功能验证

项目名称： 番茄IL系耐旱相关基因的表达分析与功能验证

项目编号： No.30871712

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 叶志彪

作者单位： 华中农业大学

项目金额： 35万元

中文摘要： 培育耐旱植物品种具有极为重要的实践价值。本项目以2个耐旱番茄IL系以及耐旱供体亲本和轮回亲本为实验材料，分别进行干旱胁迫处理，利用TOM2（Oligo）芯片杂交，初步筛选到近1800个差异表达基因。对干旱处理后，各种信号转导，生化代谢途径的转录表型进行了分析，找出了影响较大的几条代谢途径。借助生物信息学和同源序列法，初筛候选基因，利用ILs群体进行了定位，选定15个番茄耐旱候选基因，利用超量表达和抑制表达(RNAi)对其功能进行了转基因验证，发现SpWRKY，SpTR，EDR15，SpUSP等基因具有明显的抗旱功能。同时，对其组织表达谱，诱导表达谱，亚细胞定位，抗旱方面的生理生化指标进行了测定，确认了其在抗旱方面的功能。为以后的抗旱育种和抗旱的分子响应及其调控机制提供了参考。

中文关键词： 番茄; 耐旱; 基因芯片; 功能鉴定

英文摘要： The breeding of drought-resistant plant varieties has very important practical value. Two drought-tolerant tomato IL lines and drought-tolerant donor parent and the recurrent parent were suffered from drought stress treatment in this project, and hybrid using TOM2 (Oligo) microarray, above 1,800 differentially expressed genes were screened. After drought treatment, all kinds of signal transduction, biochemical metabolic pathways of transcription phenotypes were analyzed and identified several main pathways during drought resistance. Bioinformatics and homology sequence methods were employed to screen 15 candidate genes, and these genes were located using ILs. The functions of selected 15 candidate genes were confirmed with over-expression and inhibition of expression (RNAi) technology. The results indicated that SpWRKY, SpTR, EDR15, SpUSP have obvious resistance drought function. At the same time, these genes tissue expression pattern, induced expression profiles, subcellular localization, physiological and biochemical aspects of the drought were determined to confirm its function in drought areas. These results provide a reference for future drought breeding and shed light on response and molecular regulation mechanisms of drought.

英文关键词： Tomato; resistant drought; gene microarray; function identification