利用ISSR及代谢产物标记检测贵州金钱草遗传多样性

目的：结合ISSR分子标记与金钱草主要代谢产物标记对贵州省共21份野生金钱草资源的遗传多样性进行分析，为其驯化种植及遗传改良提供理论依据。方法：以金钱草基因组DNA为模板，以优化的PCR体系筛选出引物进行ISSR标记扩增，应用NTsys 2.10e软件，对所得数据进行分析处理，并采用高效液相色谱法测定了主要代谢产物槲皮素和山柰素的含量，使用SPSS 19.0软件对测定数据进行统计分析与绘制聚类图。结果：用筛选出的15条ISSR引物共扩增出105条多态性条带，多态性条带占比达98.1%，各金钱草种质资源之间的遗传相似性系数范围在0.598 1~0.953 3之间，地理位置相距越近，遗传相似性越高，不同资源的槲皮素和山柰素含量差异较大，ISSR标记的聚类将21份金钱草资源分为2大类群6个亚群，代谢产物标记的聚类将其分为2大类群5个亚群。结论：贵州野生金钱草具有丰富的遗传多样性，应用ISSR及代谢产物槲皮素和山柰素标记均能较好地对金钱草资源进行聚类，但聚类结果存在差异，其亲缘关系与地理位置呈现一定的关联性。

Genetic diversity of Lysimachia christinae Hance in Guizhou by ISSR and metabolite markers

OBJECTIVE To analyze the genetic diversity of 21 wild Lysimachia christinae Hance resources in Guizhou province by using ISSR molecular markers and main metabolites of Lysimachia christinae in order to provide theoretical basis for domestication, planting and genetic improvement. METHODS Using the genomic DNA of Lysimachia christinae Hance as template, primers were screened by optimized PCR system for ISSR marker amplification, and NTsys 2.10e software was used to analyze the data. The contents of quercetin and kaempferol, the main metabolites, were determined by high performance liquid chromatography. The data were analyzed and clustered by SPSS 19.0 software. Results A total of 105 polymorphic bands were amplified with 15 ISSR primers. The polymorphic bands accounted for 98.1%. The genetic similarity coefficient ranged from 0.598 1 to 0.953 3. The closer the geographical location was, the higher the genetic similarity was. The content of quercetin and kaempferol in different resources varied greatly. The cluster of ISSR markers divided 21 resources into two groups and six subgroups, and two groups and five subgroups by the cluster of metabolite markers. CONCLUSION There are abundant genetic diversity in wild Lysimachia christinae Hance in Guizhou. ISSR and metabolite markers can be used to cluster Lysimachia christinae Hance resources, but the clustering results are different. Its genetic relationship is related to corresponding geographical location.