8份广西产绞股蓝种质的RAPD引物筛选及其遗传多样性分析

RAPD Primers Screening and Genetic Diversity Analysis of Eight Germplasms of Gynostemma Pentaphyllum from Guangxi

本研究的目的在于筛选合适的RAPD随机引物,应用RAPD技术对药用植物绞股蓝进行遗传多样性分析,并构建DNA指纹图谱。研究结果表明,我们利用生物信息学方法挑选出的20条引物中有19条引物的扩增条带清晰且多态性好;在清晰稳定出现的354条带中,294条具有多态性;其中有3条引物的扩增条带可清楚区分绞股蓝与混淆品种乌蔹莓,可建立其DNA指纹图谱。按UPGMA法进行聚类分析,计算其遗传相似系数,结果显示,8份绞股蓝供试材料聚为两类,聚类结果与其地理区域远近和生长环境一致。本研究中筛选出的19条引物适用于绞股蓝遗传多样性分析,且获得的DNA指纹图谱可用于鉴别绞股蓝。

In this research, we aim to screen the appropriate random primers, analyze the genetic diversity of Gynostemma pentaphyllum and construct DNA fingerprints with random amplified polymorphic DNA （RAPD） technique. Twenty random primers were selected by using bioinformatics methods ,of which nineteen primers can amplify clear and reproducible polymorphic bands. The PCR results showed that a total of 354 bands were amplified, including 294 polymorphic bands. Interestingly, three primers generated distinct fingerprinting patterns that could distinguish between Gynostemma pentaphyllum and Cayratia japonica. Then we constructed a dendrogram based on genetic distances by using UPGMA method. The dendogram reflected that 8 samples clustered into two taxa, and this result was in accordance with their geographic origin and growing environment. The nineteen random primers were suitable for genetic diversity analysis and the DNA fingerprints we obtained could apply to authentication of Gynos te rnma pentap hyllum.