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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 diversity and variation of Aconite populations and to provide essential characteristics for the accurate identification of Aconitum species. Methods: The genomic DNA extraction kit was used to extract DNA. The concentration and purity of DNA were detected by ultraviolet spectrophotometry. Sixty primers were used to analyze the ISSR of 10 samples from 10 wild populations of Aconitum vulgaris. POPGEN32 and NTSYS -PC, the genetic similarity coefficient and genetic distance of Aconitum palmatum from different areas were obtained. The UPGMA was used for cluster analysis. Results: Of the 60 primers, 11 bands were screened with clear, polymorphic and reproducible primers for amplification. A total of 101 bands were amplified, of which 89 bands were polymorphic Point percentage (PPB) was 88.1%, Shannon’s diversity index (I) was 0.5298, genetic similarity coefficient (H) was 0.3648, number of observed alleles (Na) and number of effective alleles (Ne) were 1.8911 and 1.6555 , The genetic identity was between 0.4950 and 0.6931, and the genetic distance ranged between 0.3666 and 0.7031. According to the ISSR clustering results, 10 different producing areas of Aconitum vulgare were grouped into 5 groups. CONCLUSION: Aphis citricola are characterized by high polymorphism and high genetic variation, which can provide the molecular basis for the genetic diversity of germplasm resources and lay the foundation for the construction of DNA fingerprinting.