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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:a RAPD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 analyze the genetic relationship and the specific DNA fragments which can be used for identification of three species of Panax species. Methods: Random amplified polymorphic DNA (RAPD) technique was used to analyze the genomic DNA of three species of Panax ginseng. The specific fragments were screened and recovered, cloned and sequenced. Results: Twelve random primers were screened from 120 random primers for polymerase chain reaction (PCR) amplification. A total of 111 amplified fragments were obtained, of which 107 were polymorphic and the percentage of polymorphism was 96.40%. From the cluster analysis, we can see that the genetic relationship between Panax ginseng and Panax quinquefolium is the most recent one, while the Panax notoginseng is more distant than the other two species and clustered together. The dominant marker The specific fragments were gel excised, cloned and sequenced to obtain the DNA sequence. Conclusion: a RAPD marker technology at the molecular level suggests the genetic relationship of the three species of Panax species, and provide the basis for the later identification. According to the sequence designed for the identification of ginseng, American ginseng, notoginseng specific PCR primers, to achieve from the theoretical study to identify the actual use of medicinal changes.