福建七叶一枝花种质资源遗传多样性的ISSR分析

Genetic diversity of Paris polyphylla Smith var.chinensis germplasm resources from Fujian based on ISSR

[目的]分析了福建七叶一枝花种质资源的遗传多样性和亲缘关系,以期快速鉴别福建省内和省外的种质资源。[方法]利用ISSR分子标记技术扩增,使用POPGENE 32软件计算平均Shannon多样性指数,并采用NTSYS计算遗传相似系数和UPGMA聚类构建亲缘关系图。[结果]从100条引物中筛选出8条扩增条带稳定且清晰、多态性高的引物进行ISSR-PCR扩增,共获得DNA条带66条,其中多态性条带62条,多态性条带比率为93.94%;27个种源平均Shannon多样性指数为0.3779,平均检测等位基因数为1.9394,平均有效等位基因数为1.3720,平均Nei′s基因多样性指数为0.2377。27个种源遗传相似系数的变异范围为0.63~0.88。UPGMA聚类表明,在相似系数为0.63时,可将福建省内与省外的七叶一枝花种源分开;在遗传相似系数为0.69时,27个种源可分为4大类群,来自南平的4份种源和来自三明的12份种源聚为一支,来自龙岩的7份种源聚为一支,来自江西、湖南、湖北的3份种源聚为一支,来自四川的1份种源单独聚为一支。[结论]福建七叶一枝花具有较高的遗传多样性,种质亲缘关系与地理环境相关,可为后续品种选育、资源鉴别和药材道地产区选择提供参考。

[Purpose]The study aimed to analyze the genetic diversity and phylogenetic relationship of Paris polyphylla Smith var.chinensis germplasm resources from Fujian Province,in order to quickly identify germplasm resources within and outside Fujian Province.[Method]Based on the result of ISSR marker amplification,the average Shannon diversity index and genetic similarity coefficient of P.polyphylla from Fujian Province were calculated by POPGENE 32 software and NTSYS,and phylogenetic relationship map was constructed using UPGMA clustering.[Result]Eight primers with stable,clear and highly polymorphic amplification bands were screened from 100 primers for ISSR-PCR amplification,and resulted in a total of 66 DNA bands,including 62 polymorphic bands,with a polymorphism rate of 93.94%.The averages of Shannon diversity index,detected allele number,effective allele number,and Nei′s gene diversity index for 27 germplasm resources were 0.3779,1.9394,1.3720 and 0.2377,respectively.The variation range of genetic similarity coefficients among 27 germplasm resources was from 0.63 to 0.88.UPGMA clustering showed that P.polyphylla populations from Fujian Province can be separated from those outside Fujian Province at a similarity coefficient of 0.63.When the genetic similarity coefficient was 0.69,27 germplasm can be divided into 4 major groups,in which group 1 was made up by 4 samples from Nanping City and 12 samples from Sanming City,group 2 was consisted of 7 samples from Longyan City,group 3 contained 3 samples from Jiangxi Province,Hunan Province and Hubei Province,and 1 sample from Sichuan Province clustered as group 4.[Conclusion]ISSR analysis revealed that P.polyphylla from Fujian Province had high genetic diversity,and the genetic relationship was related to geographical environment,which provided a certain reference basis for subsequent variety selection,resource identification,and real estate region selection.