《植物生理学报》 2016, 52(3): 277-284

卷柏科药用植物种间遗传多样性的ISSR分析

孙红梅¹, 谷巍^1,*, 耿超¹, 孙庆文², 曹园¹

¹南京中医药大学药学院, 南京210023; ²贵阳中医学院药学院, 贵阳550002

通信作者：谷巍；E-mail: guwei9926@126.com

摘　要：

应用ISSR分子标记技术对34种卷柏科植物(其中18种药用植物)进行遗传多样性分析。利用PopGene 32软件及NTsys 2.10e软件分析卷柏科34种植物的遗传多样性及亲缘关系, 并根据UPGMA法, 构建亲缘关系树状图。从100个随机引物中筛选出7条多态性稳定、条带清晰的引物, 共扩增出156条谱带, 多态性比例为100%。分析结果表明, 平均Shannon 信息指数为0.537, 平均 Nei’s 基因多样性指数为0.359, 平均有效等位基因数为1.617, 34种卷柏科植物种间的遗传相似性系数在0.359~0.872, 显示出该植物类群具丰富的遗传多样性。聚类分析结果表明, 34种卷柏科植物之间有较明显的分界, 其中卷柏与垫状卷柏, 剑叶卷柏与异穗卷柏, 兖州卷柏和江南卷之间的亲缘关系较近, 分别聚为一类。初步证明利用ISSR分子标记可有效地分析卷柏科药用植物的遗传多样性, 从而为卷柏科植物的分类鉴定以及资源的合理开发利用提供理论基础和科学依据。

关键词：卷柏科; ISSR; 遗传多样性; 聚类分析

收稿：2015-11-16   修定：2016-02-16

资助：江苏省中药资源产业化过程协同创新中心(ZDXMHT-1-5)、国家自然科学基金(81102772)、江苏省中药优势学科II期建设(ysxk-2014)和江苏省“青蓝工程” (2012)。 致谢 贵州省科学院生物研究所王培善教授、中国药科大学周荣汉教授、贵阳中医学院潘炉台教授和赵俊华教授对本文样品的提供与鉴定。

Genetic diversity analysis of medicinal plants of Selaginellaceae based on ISSR markers

SUN Hong-Mei¹, GU Wei^1,*, GENG Chao¹, SUN Qing-Wen², CAO Yuan¹

¹College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; ²College of Pharmacy, Guiyang College of Traditional Chinese Medicine, Guiyang 550002, China

Corresponding author: GU Wei; E-mail: guwei9926@126.com

Abstract:

ISSR (inter simple sequence repeat) markers was used to analyze the genetic diversity and relatedness of 34 species of Selaginellaceae (18 medicinal species included). The molecular marker technique ISSR was used to investigate the genetic diversity and relatedness of 34 species of Selaginellaceae. Data was analyzed by PopGene 32 and NTsys 2.10e, and a cluster diagram was presented by UPGMA. 100 primers were tested, among which seven gave reliable polymorphic banding patterns. Amplification from the accessions of the seven primers yielded 156 genetic loci with 100% polymorphism. The results from PopGene 32 software indicated that the average Shannon information index was 0.537, the average Nei’s gene diversity index was 0.359, the average effective number of alleles was 1.617, and the genetic similarity coefficient of varieties was between 0.359 and 0.872, which showed enriched genetic diversity. There are obvious boundaries between 34 species of Selaginellaceae. Interestingly, Selaginella tamariscina and S. pulvinata, S. xipholepis and S. heterostachys, S. moellendorffii and S. involvens are clustered into one group, respectively, indicating their closer relationship. The results were basically consistent with the traditional plant morphotaxonomy and indicated that the ISSR method is suitable for the genetic diversity analysis of Selaginellaceae species, thus providing a scientific foundation for their species identification and development and applications.

Key words: Selaginellaceae; ISSR; genetic diversity; cluster analysis