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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 the samples of medicinal plant Rhizoma Paridis by means of the determination of the index components and DNA molecular markers, and to explore the application prospect of DNA molecular identification in the field of Chinese herbal medicine identification. Methods: The polysaccharide Ⅰ, Ⅱ, Ⅵ and Ⅶ were used as indexes to determine the contents of the index components by HPLC. The differences of the active ingredients of steroidal saponins in their rhizomes were evaluated by one-way analysis of variance (ANOVA) (PCR) method of DNA molecular markers to identify samples. Results: There were significant differences (P <0.01) in the contents of total saponins Ⅰ, Ⅱ, Ⅵ, Ⅶ and total saponins in the roots and stems of 10 medicinal plants of Rhodopes, and there were also significant differences in the main chemical constituents. ITS2 sequence and NJ tree results can effectively distinguish ten species of relict medicinal plants. Conclusion: DNA molecular identification technique as a supplement to conventional methods for determination of content has the advantages of accuracy, reliability and repeatability. It can be used to identify medicinal plants of Rhizoma Paridis.