开花时间是芸薹属蔬菜的重要性状,FLOWERING LOCUS C(FLC)是影响植物开花的关键基因.本研究利用FLCs基因外显子4的多态性区段设计正向引物,外显子7的保守区设计反向引物,建立了能够区分大白菜(Brassica rapa ssp.pekinensis,AA)BrFLC1、BrFLC2、BrFLC3和BrFLC5,与结球甘蓝(B.oleracea var.capitata,CC) BoFLC1、BoFLC2、BoFLC3和BoFLC5,共计8个基因的4对特异引物.利用该特异引物对添加结球甘蓝2号染色体的大白菜单体异附加系的FLCs基因进行鉴定,结果表明,该单体异附加系除具有大白菜的4个BrFLCs基因,同时添加了结球甘蓝的BoFLC3基因.实现了大白菜染色体组背景下结球甘蓝FLCs基因的跟踪鉴定,同时为大白菜遗传背景下添加结球甘蓝BoFLC3基因易位系的获得提供了特色材料.
Flowering time is an important trait in Brassica,and FLOWERING LOCUS C(FLCs) are key genes to flowering.In this paper,FLCs specific primers,polymorphic not only in Brassica rapa ssp.pekinensis and B.oleracea var.capitata,respectively,but also between B.rapa ssp.pekinensis and B.oleracea var.capitata,using DNA samples from 4 different type of B.oleracea var.capitata and 4 different types of B.rapa ssp.pekinensis individual plants,were screened.The 4 specific primers were obtained for genotyping of BrFLC1,BrFLC2,BrFLC3,BrFLC5,BoFLC1,BoFLC2,BoFLC3 and BoFLC5.Forward primers were designed on polymorphic region of exon 4,and reverse primers were designed on conserved region of exon 7.The corresponding FLCs between B.rapa ssp.pekinensis and B.oleracea var.capitata could be identified by 1 pair of primers.The FLCs from B.rapa ssp.pekinensis and B.oleracea var.capitata in 1 cell could be differentiated easily and quickly using 4 specific pairs of primers.The same FLCs specific bands,amplified by 4 specific primers,appeared in sesquidiploids hybrid (AAC) drived from tetraploid B.rapa ssp.pekinensis (AAAA) and diploid B.oleracea var.capitata (CC),as well as in B.rapa ssp.pekinensis and B.oleracea var.capitata.In the B.rapa ssp.pekinensis monosomic alien addition line,which introduced chromosome 2 from B.oleracea var.capitata,besides the 4 BrFLCs genes,only BoFLC3 existed in the monosomic alien addition line,detected by 4 specific primers.The results provide a new way to identify BoFLCs in the background of B.rapa ssp.pekinensis,and supply special materials for translocation lines added BoFLC3 under B.rapa ssp.pekinensis background.