采用RAPD技术对人参属的亲缘关系和鉴别的分析

目的:分析人参属3种植物的亲缘关系及其可用于鉴别的特异性DNA片段。方法:采用随机扩增多态性DNA分子标记技术(RAPD)对人参属3种药材的基因组DNA进行多态性分析,筛选特异性片段并进行切胶回收、克隆及测序。结果:从120条随机引物中筛选出12条随机引物用于聚合酶链式反应(PCR)扩增,共扩增出111条扩增片段,其中多态性位点107,多态性百分率为96.40%;从聚类分析图可以看出:人参与西洋参的种间亲缘关系最近,聚为一类,而三七相对其他2种较远,单独聚为一类;将筛选出的显性标记的特异性片段进行切胶回收、克隆并测序获得其DNA序列。结论:aRAPD标记技术从分子水平提示了人参属3种植物的亲缘关系,并为后期的特异性鉴别提供基础。根据序列设计用于鉴别人参、西洋参、三七的特异性PCR引物,实现从理论研究向药材鉴别实际运用的转变。

Objective:To study the genetic relations of 3 species of Panax and specific DNA fragments for identification.Methods:Random amplified polymorphic DNA(RAPD) technology was used to analyze the polymorphism of DNA in the 3 species of Panax,and the specific fragments were selected for cuttinggel collection,cloning and sequencing.Results:Totally 12 out of 120 random primers were selected for PCR amplification and a total of 111 bands were amplified,among which 107(96.40%) bands were polymorphic. Cluster analysis indicated that Panax could be divided into 2 categories,Panax ginseng and Panax quinquefolium for one categories,Panax notoginseng for the other categories.The specific fragments of the dominant markers were selected and the DNA sequences were obtained by cloning and sequencing.Conclusion:At the molecular level,RAPD marker indicates the relationships among 3 species of Panax and provides the basis for specific identification in future.According to the acquired sequence,the specific polymerase chain reaction(PCR) primers for the identification of Panax ginseng,Panax quinquefolium or Panax notoginseng can be designed,which suggests the theoretical research could turn into the practical application to identification of medicinal materials.