广东省野百合天然居群的随机扩增多态性DNA(RAPD)分析(英文)

Random Amplified Polymorphic DNA (RAPD) Analysis of Natural Lilium brownii from Guangdong,China

本研究选择了中国广东省7个有代表性的山区作为野百合(Lilium brownii F.E.Brown ex Miellez)天然居群的采样点,对采集到的199份样品进行了随机扩增多态性DNA(RAPD)分析,以建立野百合天然居群RAPD分析方法体系以便后续研究,并了解广东省野百合天然居群多态性情况和居群内外个体及野百合及变种百合样品的聚类情况。以新鲜叶片、硅胶干燥叶片及鳞片为材料分别提取DNA,从273条10聚寡核苷酸随机引物中筛选出20条随机引物,对7个居群共199个野百合及百合样品进行RAPD扩增。共检测到433条RAPD谱带,其中多态性条带为430条,所有个体的多态性条带百分率(PPB)为99.31%,各居群的平均多态位点比率为60.67%。通过对199个个体的RAPD数据的UPGMA聚类分析,获得了聚类图。199个样品用三种方法都成功提取出质量较高的DNA,并能扩增出清晰、明亮、稳定的RAPD条带。在聚类图遗传相似系数为0.68处发现同一居群内的所有百合个体和野百合个体在聚类图中表现了明显的分离,这是原变种野百合及变种百合的区别在分子水平上很好的反映。

Lilium brownii F.E.Brown ex Miellez,Brown lily,belongs to Lilium of Liliaeeae in planttaxonorny,is a endemic species and one of commonlly-used cross breeding parents of Lilium in China.L.brownii F.E.Brown ex Miellez var.virdulum Baker,Greenish lily,a variety of Lilium brownii,is also a well known medicinal and edible plant.Wild germplasm resources are the basic materials for genetic research and plant breeding.Howerever,due to Lilium brownii is scattered in the wild and its collection is generally only in flowering phase,it is a great difficulty for its scientific research.In the year before,researches on Lilium brownii especially molecular biology researches are still lacking in Guangdong province.In this study,total 199 Lilium brownii samples of 7 natural populations collected from representative mountain areas in Guangdong China were analyzed by RAPD markers,so as to provide good foundation for further natural germplasm resources and related studies of Lilium brownii populations,and provide good evidence of the discrimination of Brown lily and Greenish lily at molecular level.DNA from all samples was extracted from leaves and scales,which amplified clear,bright and steady RAPD bands.20 primers were screened from 273 random primers,and a total 433 DNA bands were amplified,430（99.3%） of which were polymorphic,the percentage of polymorphic bands（PPB） at the population level was 60.67% on average.The unweighted pair group method arithmetic average（UPGMA） clustery analysis of the RAPD data from 199 samples showed that all Brown lily samples and Greenish lily samples in the same population separated obviously at the value of 0.68 of genetic similarity coefficient,which was a good evidence of the discrimination of Brown lily and Greenish lily at molecular level.