Rf1调控棉花细胞质雄性不育育性恢复的分子机制解析

细胞质雄性不育和恢复(CMS/Rf)系统不仅可以实现杂交种简化制种，促进作物杂种优势的利用，而且也是研究细胞核和细胞质互作的理想材料。棉花三系配套成功并用于杂交育种已多年，但CMS/Rf分子机制研究仍然滞后。申请人前期以哈克尼西棉不育胞质(CMS-D2)三系体系为实验材料，通过构建大规模遗传分离群体，组装恢复系HaiR高质量基因组，图位克隆到主效育性恢复候选基因Rf1，并利用VIGS实验对Rf1基因进行了初步的功能验证。本项目拟在此基础上，利用基因互补、过表达和CRISPR/Cas9等实验进一步验证恢复基因，深入研究Rf1调控CMS育性恢复的分子机制，阐明其起源和演化规律，为棉花三系杂交育种提供理论支持。

Cytoplasmic male sterility and fertility restorer (CMS/Rf) system provide a practical tool for facilitating hybrid seed production. It is also an ideal model for study of the interaction between nucleus and cytoplasm. The cotton three-line technology have been developed and exploited in hybrid cotton production for several years, however, compared to other plant such as rice and maize, the study of molecular basis of three line traits in cotton has significantly lagging behind. In order to cloning of Rf genes restoring CMS in cotton, we developed a large-scale of F2 segregation population from across between upland cotton varieties P30A (CMS-D2) and sea-island cotton varieties HaiR (restorer). We identified the Rf1 gene by map-based cloning based on assembling a high quality genome of HaiR, and validated its function by using virus-induced gene silencing (VIGS). This project will give detailed analysis of Rf1 biological functions be based on the previous studies by gene complementary, overexpression, and CRISPR/Cas9 experiments to determine its biology function. Furthermore, we will investigate the mechanism of Rf1 involved in CMS fertility restores by genetics, molecular biology, and biochemistry, and also explore its origins and evolution process. The results obtained will help us to get further understanding on the regulatory mechanism of CMS/Rf in cotton, and provide the theoretical support for the cotton three-line hybrid breeding.