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以含Me3基因的抗根结线虫辣椒自交系‘HDA149’为父本,感根结线虫辣椒自交系‘8214’为母本,构建了一个2133株的F2作图群体。采用群体分离分析法(Bulked Segregant Analysis,BSA),构建抗、感病两个DNA池,对由两亲本筛选出来的285对引物进行扩增分析,发现引物SSCP_B322和EPMS658在抗、感池间具有稳定的多态性。继续用这两对引物对F2单株进行扩增,以JoinMap 3.0软件分析发现SSCP_B322和EPMS658标记位于Me3基因两侧,遗传图距分别为0.56cM和1.33cM。运用回交群体BC1和F3群体验证实了这两个标记,可有效用于辣椒抗线虫分子标记辅助育种,也为图位克隆Me3基因奠定了基础。
A F2 population with 2133 lines was constructed using the inbred line ’HDA149’ with root-knot nematode and pepper inbred line Me3 as male parent and the root-knot nematode ’chili’ inbred line ’8214’ as female parent. Using bulked segregant analysis (BSA), two DNA pools were constructed for resistance and susceptibility. 285 pairs of primers screened from the parents were amplified and analyzed. The results showed that the primers SSCP_B322 and EPMS658 had Stable polymorphism. The two pairs of primers were used to amplify F2 plants. According to JoinMap 3.0 software analysis, SSCP_B322 and EPMS658 markers were located on both sides of Me3 gene with genetic distances of 0.56cM and 1.33cM, respectively. The two markers were confirmed by backcross BC1 and F3 populations, which could be effectively used for molecular marker-assisted breeding of pepper and nematode, and laid the foundation for the cloning of Me3 gene.