利用VIGS技术挖掘谷子十里香抗锈相关基因

项目名称： 利用VIGS技术挖掘谷子十里香抗锈相关基因

项目编号： No.31271787

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： 李志勇

作者单位： 河北省农林科学院谷子研究所

项目金额： 80万元

中文摘要： 谷子锈病是谷子上重要病害，流行年份不少地块植株倒伏、颗粒无收。生产中利用抗锈品种是最经济有效的防治方法。但抗锈品种应用中易出现抗性丧失现象，为此，克隆抗锈基因、揭示抗锈机理对快速培育抗锈品种，延长品种使用寿命具有重要意义。本课题组以高抗锈病资源十里香为材料，进行了重测序，锈菌诱导的转录组和数字基因表达谱测序，且标记和定位了其抗锈基因，本研究一方面对标记内基因进行生物信息学分析，找到抗锈候选基因，并利用实时荧光PCR对候选基因进行表达分析；另一方面对抗源十里香接种锈菌后不同时间叶片的表达谱中抗锈特异表达、上调表达基因进行分析，获得抗锈相关基因的信息，通过差异基因表达模式聚类分析，从分子水平上揭示谷子抗锈的分子调控途径与机理。然后，利用谷子上病毒诱导的基因沉默平台对抗锈候选基因及其抗锈相关基因进行初筛，进一步利用农杆菌介导的谷子转基因技术最终确定候选基因功能，为开展谷子抗锈分子育种奠定基础。

中文关键词： 谷子；抗锈相关基因；数字基因表达谱；实时荧光定量PCR；基因功能

英文摘要： Millet leaf rust(Uromyces setariae)is a important fungal pathogen of foxtail millet leaf disease.There were not harvest at all ,when the disease is epidemic due to millet lodging.Resistance cultivars are the most economical and environmentally-friendly way to reduce the damage caused by millet leaf rust,but the breakdown of resistance always occured.A promising measure that could simultaneously speed up millet resistance breeding and prolong working life of resistance variety is to identify the resistance mechanisms and rust resistance related gene. The rust resistance gene, derived from resistant cultivar "Shilixiang" was maped and located in a chromosome based on re-sequencing and RNA-seq﹠digital gene expression were carried out after inoculating with rust.On the one hand rust resistant candidate genes were found based on the bioinformatics analysis of candidate region and genes expression pattern were identified by real-time-PCR. On the other hand digital gene expression profiling analysis was used for estimating gene expression level after inoculating with rust and specific expressed﹠upregultate were screened for further analysis.The molecular basis of resistance mechanisms and regulate network were finaly unraveled by clustering analysis of differential gene expression pattern.Firstly the candidate gene fu

英文关键词： foxtail millet；rust related resistance gene；digital gene expression profiling；q RT-PCR；gene function