利用SRAP标记构建不同种金腰的DNA指纹图谱

Establishment of DNA fingerprints for Chrysosplenium using SRAP Markers

金腰属植物是我国传统的民族药药材,因近年来其野生资源逐渐减少,迫切需要进行遗传资源调查和保护研究。该研究在野外考察的基础上,采用SRAP技术对24种共计36份金腰属植物进行了遗传多态性和聚类研究。结果显示利用18对SRAP引物进行扩增可获得374条多态性条带,平均每对引物扩增20.7条带型。利用生物学分析软件进行遗传参数分析表明群体的N_a为2.000 0,N_e为1.408 4,平均Nei′s指数为0.263 5,平均Shannon指数为0.419 1。UPGMA聚类分析显示在遗传相似系数约为0.70处可以将其分为3个大类群:有18个种共24份样品聚集为第Ⅰ类群,有3个种(变种)共7份样品为第Ⅱ类群,有3个种共5份样品为第Ⅲ类群。这些金腰在分子水平上呈现的差异与其分布的地理生态环境之间的差异是一致的。同时利用SRAP标记技术构建了36份金腰的DNA指纹图谱,获得每一份材料唯一的分子身份证带型。这些研究结果将为金腰属种质资源的鉴定、保护以及遗传多样性分析等提供有效方法和可靠依据,也为金腰属物种的进一步开发利用奠定了基础。

As a traditional Chinese medicinal material, Chrysosplenium is urgently needed for genetic resource investigation and protection research due to the decrease of its wild resources in recent years. After investigating the wild resources, we conducted genetic polymorphism and clustering studies of 24 species(a total of 36 samples) of Chrysosplenium using SRAP technique. The results showed that a total of 374 polymorphic bands were obtained using 18 pairs of SRAP primers to amplify these samples, on average of 20.7 bands for each primer pair. We used the biological software to analyze the population′s genetic parameter and got the Na value as 2.000 0, Ne value as 1.408 4, the average Nei′s index as 0.263 5, and the average Shannon information index as 0.419 1. UPGMA cluster analysis showed that all the samples can be divided into three major groups at the genetic similarity coefficient of 0.70: there are 18 species(24 samples) gathered for the Ⅰ groups, 3 species or variation(7 samples) for Ⅱ groups, and 3 species(5 samples) for Ⅲ groups. The differences of these Chrysosplenium species at the molecular level are consistent with that of their geographical and ecological distribution. At the same time, we used SRAP technology to construct 36 DNA digital fingerprints of Chrysosplenium and obtained the unique molecular identification band type of each material. These results will provide effective methods and reliable basis for the identification, protection and genetic diversity analysis of the germplasm resources of Chrysosplenium, and lay a foundation for the further development and utilization of them.