基于rDNA-ITS序列及相似度分析的中药赤芍分子鉴别研究

Analysis of rDNA-ITS Sequences and Similarity of Radix Paeoniae

目的:分析中药赤芍正品及混伪品ITS序列的种内相似度及特定位点序列碱基差异,为建立中药赤芍的分子鉴别标准提供思路。方法:采用PCR扩增产物直接测序的方法对市场上购得的未知来源赤芍样品S1、S2的ITS序列(包括ITS1,ITS2,5.8S)进行测定,测定结果进行BLAST比对、相似度分析、序列特异性碱基位点差异比较;用DNAMAN对GenBank上已注册的正品赤芍和混伪品赤芍的ITS序列进行分析。结果:建立了赤芍药材分子鉴别的3个指标,即根据BLAST结果定属,种内相似度缩小属内范围并定种,再通过特异性碱基位点差异最终确定并证实样品的物种;确定了样品S1来源于芍药属植物芍药Paeonia lactiflora Pall.,样品S2来源于芍药属植物川赤芍Paeonia veitchii Lynch.。结论:本文为赤芍的分子鉴别提供较为有效的思路与方法,同时为基于rDNA-ITS相似度及序列分析进行其他中药品种鉴别提供依据。

Objective ： To establish molecular authentication standards of Radix Raeoniae. Methods ： PCR method was used to determine rDNA-ITS sequences of sample S1 and sample S2 including ITS1, 5.8S, and ITS2. The sequences were analyzed by BLAST and DNAMAN software, rDNA-ITS sequences of Paeonia lactiflora Pall. , Paeonia veitchii Lynch. and their counterfeit were analyzed by DNAMAN software. Results ： Established three molecular authentication standards on Radix Paeoniae ： （ 1 ） The genera of samples was determined by the results of BLAST ; （ 2 ） The species of samples were determined by the similarities ; （ 3 ） The specific base positions confirmed the species of samples. Based on the sequence alignment and base differences, sample S1 was identified as Paeonia lactiflora Pall. , and sample $2 was identified as Paeonia veitchii Lynch.. Conclusion ： This paper can give an effective way of Radix Paeoniae identification. What＇s more, it gives a good example for other Chinese Medicinal Materia authentication.