利用GWAS定位和克隆番茄灰霉病抗性基因

项目名称： 利用GWAS定位和克隆番茄灰霉病抗性基因

项目编号： No.31471887

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 王涛涛

作者单位： 华中农业大学

项目金额： 85万元

中文摘要： 番茄灰霉病是由数量位点控制的侵染性病害，目前仍没有克隆到主效的抗性基因，生产上也没有抗灰霉病的番茄品种，灰霉病已成为番茄安全生产最主要的限制因子之一。本项目以重测序的304份番茄核心种质为材料，利用全基因组关联分析（GWAS），对番茄灰霉病抗性基因进行定位，并克隆主效基因。前期我们利用GWAS定位的12个灰霉病抗性QTL位点，有10个与前人结果一致，表明定位结果是非常可靠的。特别是主效位点gw1-3和gw3-2，其SNP信号显著，是我们克隆的主要候选位点。本项目拟首先分析定位区域内序列变异与抗/感性及F2分离群体抗性的相关性，筛选候选基因；利用VIGS快速鉴定候选基因的功能，对主效基因进一步通过稳定的遗传转化进行确认；利用RNA-Seq和酵母单/双杂交技术初步研究主效基因调控的分子机理。该研究结果对揭示番茄抗灰霉病的分子机理，开发灰霉病相关的分子标记，培育抗灰霉病的番茄品种具有重要的意义。

中文关键词： 番茄；灰霉病；全基因组关联分析；基因定位；基因克隆

英文摘要： Gray mold is an important infectious disease controlled by QTL loci, the main effect resistance genes have still not been cloned, and also no resistance cultivars in the market, Gray mold has become one of the major limiting factors in tomato production. The project plans to map the locus resistant to gray mold and clone the main effect genes, using 304 re-sequencing core cultivars of tomato as materials by the method of genome-wide association analysis (GWAS). We have mapped 12 gray mold resistance loci using GWAS, ten of them are consistent with previous results, which indicated that the map is very reliable. In particular, the main effect loci gw1-3 and gw3-2, their SNP signals are significant, which are the main candidate loci for cloning. The project intends to analyze the correlation between the gene sequences in the map region and the resistance phenotype of re-sequencing materials or the F2 segregating population, then to determine the candidate genes. After that, we will identify the function of candiate genes using VIGS method, and further confirm it by stable genetic transformation. Then we study the preliminary molecular mechanisms by RNA-Seq and yeast one/two hybrid technology. The research resutls have a great significance in revealing the molecular mechanism resistant to tomato gray mold, developing gray mold associated molecular markers, cultivating tomato cultivars resistant to gray mold.

英文关键词： Tomato;gray mold;GWAS;Gene map;Gene cloning