不同产地哈药白喉乌头基因多态性的ISSR分析

ISSR Analysis for Genetic Polymorphism of Aconitum leucostomum from Different Habitats

目的：探讨白喉乌头种群间的遗传多样性和变异情况,并为乌头类药材之间的准确鉴定提供本质特征。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个引物分别对10个不同产地的白喉乌头野生种群样本的基因进行ISSR分析,应用POPGEN32和NTSYS-PC对所得数据进行处理,得出不同产地白喉乌头的遗传相似系数和遗传距离,采用非加权平均法（UPGMA）进行聚类分析。结果：从60个引物中筛选出11个条带清晰、多态性明显并且重复性好的引物用于扩增,共扩增得到101个条带,其中多样性条带89条,多态位点百分率（PPB）为88.1%,Shannon多样性指数（I）为0.5298,遗传相似系数（H）为0.3648,观察等位基因数（Na）和有效等位基因数（Ne）分别为1.8911和1.6555,遗传一致度在0.4950~0.6931之间,遗传距离范围在0.3666~0.7031之间;根据ISSR聚类结果可将10个不同产地的白喉乌头聚为5大类群。结论：白喉乌头种质资源间具有很高的多态性,遗传变异较大;可为种质资源遗传多样性提供分子水平的依据,为构建DNA指纹图谱奠定基础。

Objective：To investigate the genetic diversities and variations of Aconitum leucostomum,and to supply essential characteristics for identifying Aconitum crude drugs.Methods：Plant genome extraction kit was applied to extract DNA,and ultraviolet spectrophotometer was used to detect the concentrations and purity of DNA.60 ISSR primers were screened to analyze the DNA of Aconitum leucostomum from 10 habitats.Biosoftwares including POPGEN32 and NTSYS-PC were used to analyze the polymorphic bands obtained,and hence to yield the genetic similarity coefficient of the 10 habitats and map the related graphics,and cluster analysis were performed by UPGMA method.Results：11 primers selected from 60 ISSR primers were used for amplification and a total of 101 DNA bands were obtained,including 89 polymorphic bands,the average percentage of polymorphic bands（ PPB） was 88.1%.Shannon information index（ I） was 0.5298,the genetic similarity coefficient（ H） was 0.3648,observed number of alleles was 1.8911,and effective number of alleles was 1.6555.The genetic identity was from 0.4950 to 0.6931,and the genetic distances were from 0.3666 to 0.7031.According to cluster analysis result of ISSR,the 10 habitats of Aconitum leucostomum were classified into five groups.Conclusion：Germplasm resources of Aconitum leucostomum show abundant polymorphism and higher genetic variation,which might supply molecular level basis,and provide basis for building DNA fingerprint.