HPLC含量测定和DNA分子标记技术对重楼属药用植物的鉴定

Identification of 10 Kinds of Paris L. by HPLC Assay and DNA Molecular Markers

目的:通过指标成分含量测定与DNA分子标记技术对重楼属药用植物样品进行鉴定,探讨DNA分子鉴定技术在中草药鉴别领域的应用前景。方法:以重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ为指标成分,利用HPLC测定指标成分含量,并用单因素方差分析对其根茎中甾体皂苷活性成分的差异进行评价;采用基于聚合酶链式反应(PCR)方法的DNA分子标记技术鉴别样品。结果:重楼属10种药用植物根茎中重楼皂苷Ⅰ、Ⅱ、Ⅵ、Ⅶ及总皂苷的含量具有极显著差异(P<0.01),且其主要化学成分种类也存在较大差异;同时ITS2序列与NJ树结果能有效区分重楼属10种药用植物。结论:DNA分子鉴定技术作为含量测定常规方法的补充,具有准确可靠、重复性高等优点,可用于重楼属药用植物的鉴定。

Objective： To identify Paris L. by determination of index ingredients and DNA molecular markers and discuss DNA molecular identification technology application in the field of herbal identification. Methods ： HPLC method was used for determina- tion of the contents of polyphyllin Ⅰ, Ⅱ, Ⅵ and Ⅶ. So as to evaluate the differences saponins active ingredients in the roots, the ANOVA analysis was used. And the samples were identified by allele - specific diagnostic polymerase chain reaction （PCR） based on sequences of internal transcribed spacer. Results：There was significant difference in the contents of polyphyllin Ⅰ, Ⅱ, Ⅵ, Ⅶ and their total contents in the roots（P 〈 0.01 ）. The species of its main chemical components were quite different. The resuits of ITS2 sequences and NJ tree can effectively distinguish genera of 10 kinds of Paris L. Conclusion ： As a supplement to con- ventional methods, for identification of Paris L. , DNA molecular technology has a great advantage in accuracy and good reliability.