大白菜-结球甘蓝易位系外源染色体区段基因标记的开发与应用

Development and Application of Gene Markers in the Exogenous Chromosome Segment of Chinese Cabbage-Cabbage Translation Lines

为了利用基因标记对大白菜-结球甘蓝易位系进行鉴定,针对大白菜和结球甘蓝共线性基因序列差异位点设计引物,以大白菜85-1和结球甘蓝11-1为试材,进行了多态性标记的筛选,并在大白菜-结球甘蓝易位系、易位系自交后代及大白菜和结球甘蓝不同商品种间进行了可利用性鉴定。结果表明,根据大白菜和结球甘蓝263个共线性基因共设计特异引物171对,66对引物在大白菜和结球甘蓝间表现出多态性扩增,所占比例为38.60%。66个基因标记中,基于大白菜基因序列获得的2个和基于结球甘蓝基因序列获得的48个,可作为结球甘蓝相对大白菜特异的基因标记,并成功用于大白菜-结球甘蓝易位系AT1-41和AT1-47及AT1-47自交后代的鉴定。进一步利用这66个基因标记在4个大白菜和4个结球甘蓝商品种中进行了PCR扩增,有52个标记能将供试的大白菜品种和结球甘蓝品种完全鉴定出来,有11个标记在部分品种中表现出多态性。结果为深入研究大白菜-结球甘蓝易位系外源基因的表达、调控、互作和标记辅助育种奠定了基础。

In order to identify Chinese cabbage-cabbage translocation line by gene markers,the polymorphism primers were designed according to the nucleotide difference of collinear genes between Chinese cabbage and cabbage,and were screened with Chinese cabbage 85-1 and cabbage11-1 as test materials. The applicability of the primers was detected using Chinese cabbage- cabbage translocation lines and their self-crossing progenies, Chinese cabbage varieties and cabbage varieties. The results showed that among171 pairs of primers designed based on 263 collinear-gene sequences of Chinese cabbage and cabbage,66 pairs of polymorphic primers were confirmed,the ratio being 38.60%. Among the 66 gene markers,two markers from Chinese cabbage gene sequences and 48 markers from cabbage gene sequences could be used as the specific gene markers to distinguish Chinese cabbage and cabbage,which were successfully applied to identify Chinese cabbage-cabbage translocation lines AT1-41 and AT1-47,and the self-crossing progenies of AT1-47. Furthermore,PCR amplification of the 66 gene markers was carried out in four Chinese cabbage varieties and four cabbage varieties,among which 52 markers could distinguished completely the Chinese cabbage varieties and the cabbage varieties,while other11 markers presented the polymorphism in parts of the varieties. The results established the foundation for further studying the expression,regulation and interaction of exogenous genes in Chinese cabbage-cabbage translocation lines and for marker-assisted breeding.