黄瓜抗CMV主效基因CMV6.1的图位克隆及功能解析

黄瓜花叶病毒（Cucumber mosaic virus, CMV）是世界十大植物病毒之一，也是为害黄瓜生产的主要病毒。由于其遗传规律和基因调控尚不清楚，严重阻碍了黄瓜抗CMV的育种进程。我们在前期研究中定位了一个抗CMV的隐性主效位点cmv6.1，其贡献率达31.7%。通过一系列近等基因系将它精细定位在了50kb区间，包含5个候选基因。本项目拟在此基础上，对CMV6.1基因进行图位克隆及转基因验证；结合250份黄瓜核心材料的重测序数据和CMV抗性表型，分析CMV6.1基因的自然变异形式和进化模式，开发功能型分子标记；通过转录组测序技术、酵母文库构建及互作蛋白筛选，分析CMV6.1基因参与的调控网络；结合其不同等位基因对细胞超微结构和叶绿素含量等抗病毒相关表型的影响，联合分析CMV6.1的抗病功能。研究结果不仅有助于探明CMV6.1基因的抗病机制，而且为黄瓜遗传改良提供了重要的基因资源。

Cucumber mosaic virus (CMV) is one of the top ten plant viruses in the world, which is also the main virus harming cucumber production. Due to the inheritance and gene regulation for CMV resistance still unclear, the breeding process of CMV resistance in cucumber was seriously hindered. In previous studies, we mapped a recessive major locus cmv6.1 that has a 31.7% contribution for the phenotype explanation. We fine mapped the cmv6.1 into a 50 kb region using near isogenic lines, which including five candidate genes. On this basis, the project will map-based clone the cmv6.1 gene and verify the function using genetic transformation; examine the natural variation and evolutionary history of the cmv6.1 gene through the re-sequencing data and CMV resistance data of the 250 cucumber core material, develop the functional molecular marker; analyze the cmv6.1 gene regulation network using transcriptome sequencing and screening the protein interaction with cmv6.1; identify the effects of cmv6.1 different alleles for the cell ultrastructure and chlorophyll content and other virus resistant related phenotypes, analyze the cmv6.1 gene function. The results of the project will not only facilitate us to understand the resistance molecular mechanisms of cmv6.1 gene, but also provide an important genetic recourse for cucumber genetic improvement.