绞股蓝属植物及其混淆品乌蔹莓的PCR-RFLP鉴别研究

Identification of Gynostemma from their adulterant Cayratia japonica by PCR-RFLP

目的:寻找简便、可重复的分子标记方法对绞股蓝属Gynostemma植物及其混淆品乌蔹莓Cayratia ja-ponica进行鉴别。方法:对7种常见药用绞股蓝属植物及其混淆品乌蔹莓的6个cpDNA片段进行PCR扩增,再利用TaqⅠ,HpaⅡ,EcoRⅠ,RsaⅠ,HhaⅠ,HindⅢ等6种限制性内切酶分别对扩增片段进行消化。结果:36种DNA片段/内切酶组合中,trnK1f-trnK2r片段与RsaⅠ组合可将乌蔹莓从绞股蓝属植物中区分出来,产物清晰、结果稳定。结论:PCR-RFLP分析方法可有效区分常见绞股蓝属植物与其混淆品乌蔹莓。

Objective： To establish a convenient and effective method for the identification of Gynostemma and Cayratia japonica . Method： Eight species, including Gynostemm pentaphyllum , G. pentagynum , G. cardiospermum , G. longipe , G. yixingense , G. laxiflontm, G. guangxiense and C. japonica were investigated through PCR - RFLP of six chloroplast DNA fragments. The six gene fragments were digested by six restriction endonuclease respectively, including Taq I , Hpa ]I , EcoR I , Rsa I , Hha ] , Hind Ill. Re- suit： Seven species of Gynostemma and their adulterant could be identified by trnKlf-trnK2r and Rsa. Conclusion： PCR -RFLP pro- vides a quick, reliable molecular marker technique for identification of Gynostemma and their adulterant Cayratia japonica.