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分子标记辅助选择可大大加快育种进程,然而由于大白菜抗病毒病遗传规律的复杂性,目前所定位的基因和筛选的连锁标记远不能满足育种需要,为了更好地对抗病毒病白菜(Brassica campestris ssp.pekinensis)进行分子标记辅助选择,本研究筛选了与大白菜抗芜菁花叶病毒(Turnip mosaic virus,Tu MV)抗性基因Tu RBCS01紧密连锁的分子标记。本研究在对该基因进行定位的基础上,利用回交1代(backcross1,BC_1)分离群体,采用分离群体分组分析法(bulked segregation analysis,BSA)、简单重复序列(simple sequence repeats,SSR)和序列特异引物(sequence specific primer,SSP)标记技术,筛选与该基因紧密连锁的分子标记。通过对13对SSP引物和16对SSR引物的分析表明,有8对引物在两亲本间表现多态性,有5对引物扩增出的标记与基因Tu RBCS01紧密连锁或共分离,分别为m Br4072、Bra025493-1、Bra025493-2、Bra025467-4和Bra025467-5。筛选出与大白菜Tu MV抗性基因Tu RBCS01紧密连锁的分子标记2个,分别为SAAS_m Br4072_240(1.5 c M)和Bra025493-1(1.0 c M),另有3个标记与基因Tu RBCS01共分离,分别为SAAS_Bra025493-2_749、SAAS_Bra025467-4_780和SAAS_Bra025467-5_956。上述标记丰富了大白菜抗Tu MV分子标记的数量和种类,有望用于大白菜抗病毒病分子标记辅助选择。
However, due to the complexity of inheritance of anti-virus disease in Chinese cabbage, the locus genes and screening linkage markers far can not meet the breeding needs. In order to better combat Brassica campestris ssp.pekinensis) were selected for molecular marker-assisted selection. In this study, molecular markers closely linked to the Tu MVB resistance gene Tu RBCS01 of Chinese cabbage were screened. In this study, we used backcross1 (BC1) segregation analysis (BSC), simple sequence repeats (SSR) and restriction fragment length polymorphism (SSR) Sequence specific primer (SSP) technique was used to screen the molecular markers closely linked to this gene. Through the analysis of 13 pairs of SSP primers and 16 pairs of SSR primers, 8 pairs of primers showed polymorphism among the parents, and 5 pairs of primers amplified the markers closely linked to Tu RBCS01 or co-segregated, respectively Br4072, Bra025493-1, Bra025493-2, Bra025467-4 and Bra025467-5. Two molecular markers tightly linked to Tu RBCS01 of Chinese cabbage were screened, which were SAAS_m Br4072_240 (1.5 cM) and Bra025493-1 (1.0 cM), respectively. Three other markers were co-segregated with Tu RBCS01 , SAAS_Bra025493-2_749, SAAS_Bra025467-4_780 and SAAS_Bra025467-5_956, respectively. The above markers enriched the number and variety of anti-Tu MV molecular markers of Chinese cabbage, which is expected to be used for the molecular marker-assisted selection of Chinese cabbage anti-viral disease.