论文部分内容阅读
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。
In this study, we selected seven representative mountainous areas in Guangdong province of China as the sampling points for the natural population of Lilium brownii FEBrown ex Miellez. A total of 199 collected samples were subjected to random amplified polymorphic DNA (RAPD) ) To establish RAPD analysis system of natural populations of Lilium formosana for further study and to understand the natural population polymorphism of Lilium formosana and the clustering of individual lily and wild lily varieties in different populations. DNA was extracted from fresh leaves, dried leaves and scales of silica gel, 20 random primers were selected from 273 random oligonucleotide primers, and 199 wild lily and lily samples from 7 populations were subjected to RAPD increase. A total of 433 RAPD bands were detected, of which 430 were polymorphic bands. The percentage of polymorphic bands (PPB) was 99.31% in all individuals and the average polymorphic locus ratio was 60.67% in each population. Through UPGMA cluster analysis on RAPD data of 199 individuals, the clustering graph was obtained. 199 samples successfully extracted DNA of high quality in all three ways and amplified a clear, bright, stable RAPD band. The genetic similarity coefficient of cluster graph was 0.68. It was found that all lily individuals and wild lily individuals in the same population showed obvious segregation in the clustering map, which is the difference between the original wild lily species and the mutant lily at the molecular level Good reflection.