Identification of commercial Bupleuri Radix and its adulterants based on ITS2 barcode
Objective DNA barcoding technology,a molecular identification method,is applied to distinguishing Bupleuri Radix from its adulterants in order to ensure the quality and clinical curative effect.Methods In this study,the internal transcribel spacer 2 (ITS2) regions of 85 samples were amplified by PCR and sequenced bi-directionally.Obtained sequences were assembled using CodonCode Aligner.The genetic distances were computed by MEGA 6.0 in accordance with the kimura 2-parameter (K2P) model and the phylogenetic tree was constructed by Neighbor-joining (NJ) method.Moreover,the secondary structure of ITS2 was predicted using ITS2 database websites.Results The intra-specific genetic distances were smaller than inter-specific ones in ITS2 regions of Bupleuri Radix.NJ tree and secondary stmeture results could distinctly ditfferentiate quality product and adulterants.Only 64.7％ of the 85 samples were in accordance with the requirements of Chinese Pharmacopoeia.Conclusion ITS2 sequence can accurately and reliably identify the authenticity of Bupleuri Radix and could provide a new technique to ensure clinical safety of this traditional Chinese medicine.
NETL
NSTL
万方数据
维普
