新疆特产7种乌头类药材的ISSR特征分析

Analysis on ISSR characteristics of Aconitum L.ethnic medicine in Xinjiang

目的探讨新疆特产准噶尔乌头、多根乌头、白喉乌头、那拉提乌头、空茎乌头、林地乌头、拟黄花乌头共7个种共41个居群间的遗传多样性和变异情况,为药材鉴定提供DNA特征。方法利用植物基因组提取试剂盒提取DNA,采用紫外分光光度法检测DNA的浓度和纯度,使用60种哥伦比亚引物分别对7个种共41个居群的乌头基因进行ISSR分析,应用NTSYS-PC2.10和POPGEN32对所得数据进行处理,计算不同种乌头的遗传相似系数和遗传距离,采用非加权平均法(UPGMA)进行聚类,构建亲缘关系系统图,用主坐标分析法绘制二维、三维散点图。结果从60个引物中筛选出13个条带清晰、多态性明显且重复性好的引物用于扩增,共扩增得到163个条带,其中多样性条带151个。根据ISSR聚类分析,41个居群的乌头可聚为7大类群,与种的分类相当符合;而且乌头类药材在多个引物下具有特征的DNA标记。结论乌头种质资源间具有很高的多态性,遗传变异较大;ISSR分子标记法可用于7种药材的鉴别,提供了分子水平依据,也为构建其DNA指纹图谱提供可能。

Objective To investigate the genetic diversities and variations of sevenAconitum L.species,in-cluding Aconitum soongaricum Stapf ., A.karakolicum Rapaics ., A.leucostomum Worosch ., A. leucostomum var .nalatiensis F.Zhang, A.nemorum M.Pop., A.apetalum B.Fedtsch .and A. anthoroideum DC.,and to supply DNA characteristics for accurate identification of crude drugs of Aconi-tum L.Methods Plant genome extraction kit was applied to extract DNA,and ultraviolet spectrophotom-eter was used to detect the concentrations and purity of DNA.Sixty ISSR primers were screened to analyze the DNA of total forty-one provenances of seven Aconitum L.species for ISSR amplification.Biosofteares including NTSYS-PC2.10 and POPGEN32 were used to analyze the polymorphic bands obtained.UPGMA method was used to make up the systematic diagram of genetic relationship,with two-dimension scatter-graph and three-dimension scatter-graph constructed by using the principal cordinate analysis.Results Thirteen primers selected from 60 primers were used for amplification and a total of 163 DNA bands were obtained,including 151 polymorphic bands.According to cluster analysis result of ISSR,the forty-one provenances of Aconitum L.were classified into seven groups,which accorded pretty with the classification of species;and those medicinal materials had the characteristics of DNA markers when using multiple primers.Conclusion Germplasm resources of Aconitum L.showed abundant polymorphism and higher ge-netic variation.ISSR molecular maker technology is useful for identifying species of Aconitum L.plants, supplying molecular level basis and making DNA fingerprints building possible.