菜薹种质遗传多样性的荧光MFLP标记分析

Genetic Diversity Analysis for Germplasm of Flowering Chinese Cabbage by Using Fluorescent Microsatellite-anchored Fragment Length Polymorphism

采用M13通用接头连接锚定引物,建立了适合于菜薹的荧光微卫星锚定片段长度多态性(MFLP)技术;在此基础上,从360对选择性扩增引物组合中筛选出9对适宜的引物组合,对收集的32份菜薹种质进行等位基因多态性分析;结果显示这些引物组合的扩增产物在32份菜薹种质中的等位基因多态性在10~31之间,平均为17.7个;多态性信息量在0.61~0.98之间,平均0.81。利用获得等位基因多态性进行的遗传相似性分析表明,32份菜薹种质间的相似系数在0.277~0.836之间,平均0.624;基于多态性数据,运用除权配对法(UPGMA)进行聚类分析,将这些菜薹种质材料分为2个组群和4个亚群。这些结果显示荧光MFLP标记技术能有效地发现供试菜薹种质材料的DNA多态性,表明该技术在菜薹遗传特性分析的研究和应用领域具有可行性。

In this study,a fluorescent microsatellite-anchored fragment length polymorphism(MFLP) technique was established by adding universal M13 adapter to the anchored primers. Nine highly polymorphic primer pairs were selected from 360 selective amplification primer combinations,and were used to screen polymorphisms for 32 genotypes of flowering Chinese cabbage. The results showed that the polymorphic alleles for the nine primer pairs ranged from 10 to 31 in 32 flowering Chinese cabbage genotypes,with an average of 17.7 per primer combination. Polymorphism information content(PIC)ranged from 0.61 to 0.98 for nine primer combinations,with an average of 0.81. The genetic similarities were calculated and showed their distribution ranged from 0.277 to 0.836 among the 32 genotypes. By using polymorphic alleles data,the 32 flowering Chinese cabbage genotypes were clustered into 2 groups and 4 subgroups by UPGMA method. These results show that the developed fluorescent MFLP technique could effectively detect DNA polymorphisms in the flowering Chinese cabbage germplasm,indicating this technique is suitable for the fields of research and application of genetic analysis in flowering Chinese cabbage.