新型油菜Ogu CMS恢复源中外源萝卜片段的序列分析及特异标记开发与利用

The Ogu CMS from radish which has a complete and stable male sterility was considered as a excellent system for utilization of heterosis in Brassica napus L., A new restorer material CLR650 for the Ogu CMS system has been created during the past few years, and could be a new type restorer for Ogu CMS in China. However, the physical location and sequence information of the introgression carrying the restorer gene Rf0 in CLR650 were still unclear, and the glucosinolate content of this restorer material need to be decreased for application. In this study, we use the method of BAC Library and DNA sequencing technique to characterize the exogenetic Raphanus fragment in CLR650, and develop specific markers linked to Rf0. After then, these markers are used for selecting the recombinant strains with low glucosinolate content deriving from CLR650×SR to improve the seed quality of CLR650. This program would clarify the originality and novelty of CLR650, and make a new scenary to the research and application of Ogu CMS in our country.

源于萝卜的Ogu CMS育性稳定、彻底，被认为是油菜杂种优势利用最具前景的细胞质不育类型。CLR650为我国自主创新培育的新型甘蓝型油菜Ogu CMS恢复材料，有望成为该系统的新型恢复源。但目前尚未对该材料中伴随恢复基因渗入的外源萝卜片段进行定位和序列研究，其硫代葡萄糖苷含量性状需要改良才能用于育种应用。本项目利用BAC文库技术和DNA测序技术，对CLR650中渗入外源萝卜片段的序列信息进行分析，然后依托甘蓝型油菜基因组的测序信息进行物理定位，开发与恢复基因连锁的特异分子标记，并利用分子标记筛选CLR650与低硫Ogu CMS恢复系SR杂交后代中的低硫苷重组株系，从而实现CLR650的高硫苷含量性状的改良。本项目的实施将在分子水平阐明项目材料的新颖性和独创性，突破国外对Ogu CMS恢复源的专利限制，开创我国Ogu CMS系统研究与应用的新局面。

本项目研究利用基因组BAC文库技术和全基因组重测序技术，明确了供试材料CLR650的外源萝卜片段的序列信息，研究表明萝卜基因组R9染色体上与Ogu CMS恢复基因连锁的60CM左右的片段渗入到油菜染色体组；根据渗入到油菜基因组的外源萝卜片段的DNA序列信息，自主开发了28个外源萝卜片段特异标记，并利用获得的分子标记与常规育种技术结合应用培育出硫代葡萄糖苷含量降低至60-70µmol/g的改良恢复系1份。通过本项目实施，在分子从面上揭示了CLR650的新颖性表现，为我国甘蓝型油菜萝卜质不育系统应用提供可利用的种质和技术基础，对突破国外Ogura CMS恢复源研究与应用的专利技术限制，提升我国Ogu-CMS系统研究与应用水平具有现实意义。