目的采用随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)法建立川木通的分子鉴定方法,为川木通的快速鉴定奠定基础。方法从100对随机引物中筛选出5对多态性丰富的RAPD引物进行川木通10个种的扩增分析,最终得到C13为阳性序列特征性扩增区域(sequence characterized amplified regions,SCAR)标记。结果阳性SCAR标记均可以进行川木通原植物与药材的分子鉴定引物,C13可以鉴定上述2种药典品种和商品主流品种(粗齿铁线莲、钝萼铁线莲)。结论RAPD法简便易行,为亲缘关系较近的多基原药材的鉴定提供了一种新方法。
Objective To establish the identification method for Caulis Clematidis Armandii by random amplified polymorphic DNA( RAPD). Methods The fragments generated by 5 selected primers(among 100 primers) on 10 different species from seven locations. Three sequence characterized amplified region (SCAR) markers were constructed based on primer C13. Results The SCAR marker was found to identify the Chinese Pharmaceutical varieties of Caulis Clematidis Armandii in the ten species. C13 could be used to identify additional species. Conclusion RAPD is simple and operable to provide one new way to identify Chinese materia medica from multiple source.