目的 对来自不同地域的多花黄精野生资源的核糖体内转录间隔区2（internal transcribed spacer 2，ITS2）和psbA-trnH基因间隔区序列进行系统发育分析，探索不同地理来源的多花黄精资源的遗传多样性和亲缘关系。方法 用聚合酶链式反应（polymerase chain reaction，PCR）的方法扩增获得多花黄精ITS和psbA-trnH的核酸序列，与美国国家技术信息中心（NCBI）相对应的核酸序列比对获得多花黄精ITS2和psbA-trnH核酸序列。用邻接法（Neighbor joining）构建系统发育树，用Kimura two-parameter（K2-P）模型分析遗传距离，用Mega和DNAman软件进行多重比对分析。结果 安徽青阳样品的ITS2序列为224 bp，其余24份样品的ITS2序列均为225 bp，福建泰宁的1份样品为620 bp，其余24份多花黄精样品的psbA-trnH序列均为621 bp，2个序列分别存在7个和4个变异位点。采用ITS2序列构建的系统发育树将25份资源分为2个大的类群，湖南和贵州的5份资源种群聚为一类，其他20份资源聚为一类，遗传距离显示来自贵州花溪、剑河的样品和来自福建建阳的遗传距离最大；采用psbA-trnH序列构建的系统发育树则无法将不同地域多花黄精资源区分开。结论 系统发育和变异位点分析为多花黄精资源的进化研究、道地性评价奠定了理论基础，变异位点的分析可用于相关资源的鉴定。

Objective In this work, phylogenetic analysis was used to compare the ITS2 and psbA-trnH sequences of Polygonatum cyrtonema samples from different geographical sources, so as to explore the genetic diversity and genetic relationship of these resources. Methods PCR method was used to amplify the regions of ITS and psbA-trnH, and the sequences of ITS2 and psbA-trnH were obtained after the amplified fragment sequences were blasted in NCBI database. The neighbor joining (NJ) and maximum parsimony (MP) methods were used to construct phylogenetic trees and Kimura two-parameter (K2-P) model was used to calculate the genetic distance of different samples. Mega and DNAman softwares were applied for mutiple alignment of ITS2 and psbA-trnH sequences of 25 samples of P. cyrtonema. Results The lengths of ITS2 and psbA-trnH sequences of Anhui Qingyang and Fujian Taining samples of P. cyrtonema were 224 bp and 620 bp, respectively. The lengths of ITS2 and psbA-trnH of the remaining 24 samples were 225 bp and 621 bp, respectively. ITS2 and psbA-trnH had seven and four mutation points, respectively. These 25 samples were clustered into two groups based on ITS2 sequences. Five samples in Hunan and Guizhou were clustered into one group, while the other 20 samples were clustered into another group. The genetic distance showed that the samples from Huaxi and Jianhe in Guizhou Province and Jianyang in Fujian Province had the largest genetic distance. Phylogenetic tree constructed by psbA-trnH sequences were unable to distinguish 25 samples from different geographical sources. Conclusion Phylogenetic and mutation analysis will provide the theoretic foundation to utilize the resources of P. cyrtonema, investigate their evolution, and evaluate their genuineness. The results of mutation point will also be used in the identification of related P. cyrtonema resources.

R282.12

福建省农业科学院药用植物创新团队项目（STIT2017-2-8）；热带作物生物学与遗传资源利用重点实验室开放课题（1630052017020-3）；工信部中药材生基地建设项目“多花黄精规范化种植及野生抚育产业基地建设”