红麻UG93 CMS恢复基因精细定位与克隆

It has a great value for kenaf industry development to utilize kenaf heterosis in Guangxi province since kenaf is a characteristic economy crop in Guangxi, China. In this grant, the UG93 cytoplasmic male sterility restorer gene (Rfu) NIL populations of Hibiscus cannabinus will be constructed by the UG93 CMS line, maintainer line and restorer line. Then molecular marker technology will be used to screen the molecular markers linked to the Rfu gene and transfer the markers to PCR markers. Furthermore, the isogenous sequences of molecular markers linked to Rfu gene in cotton genome will be found out by comparing the sequences of Rfu markers with the genome sequences of cotton. More markers that tightly linked to Rfu gene will be developped by the aid of the collinear between the kenaf and cotton. Using the results of molecular markers, we can finally fine mapping the Rfu gene by comparative genome mapping method. Functional markers will also be developped using the conservative sequence of cloned plant cytoplasmic male sterility restorer genes and PPR gene in cotton.The Rfu gene will finally cloned by positional candidate cloning and homology-based cloning method. All the marker developped in this grant will be used in kenaf hybridty test. The result of this study program will promote the breeding of UG93 CMS line and restorer line in H cannabinus by the aid of exploiting PCR markers for Rfu MAS.

红麻是广西特色经济作物，充分利用红麻杂种优势对于全国和广西农业产业发展具有重要意义。本项目以红麻UG93 CMS不育系、恢复系和保持系构建的近等基因系分离群体为材料，筛选与UG93 CMS恢复基因Rfu紧密连锁的分子标记并转化为PCR标记。同时，利用紧密连锁的分子标记序列与棉花基因组序列之间的共线性与同线性关系，将已获得的紧密连锁标记的序列定位在棉花基因组上，进而利用棉花基因组上的高度同源区甚至同源基因开发出与Rfu基因连锁更加紧密或共分离的标记；整合这些标记实现Rfu的精细定位。此外，棉花中的PPR基因以及植物中已经克隆的细胞质雄性不育恢复基因序列也将用于功能分子标记的开发并最终在本项目中结合定位候选克隆与PCR同源克隆法分离恢复基因Rfu并进行花粉管通道转基因验证。最后，尝试利用恢复基因的分子标记进行杂交种实验室内分子标记纯度鉴定。