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In this study, ITS2 barcode was used to identify Bupleurum chinense and B. longiradiatum. The ITS2 regions of 48 samples were amplified and sequenced. The sequences obtained above were aligned and the K2P distances were calculated. We used three methods, BLAST1, nearest distance and phylogenetic tree (NJ-tree), to test the identification ability. The results showed that the maximum intraspecific genetic distance of B. chinense was 0.013, and the minimum interspecific genetic distance between B. chinense and B. longiradiatum was 0.049. The NJ-tree can easily identify B. chinense and B. longiradiatum. Therefore, the ITS2 barcode is suitable to identify B. chinense and B. longiradiatum.

To determine the variation of rDNA ITS sequences in medicinal plants of the genus Bupleurum and identify corresponding DNA molecular markers associated with medicinally important members, ITS regions were amplified, sequenced, and analyzed by DNAssist version 2.2. We found that the homologous alignment of ITS sequences between members of the genus Bupleurum and associated out-groups was lower than 75%, while within-group alignment was greater than 87%. The conclusion can be drawn that ITS sequences can be used as reliable molecular markers for the identification of Radix Bupleuri.

Objective: The study the patterns of rDNA ITS sequence variation of 9 medicinal species of genus Bupleurum in China to find the DNA molecular markers for identification of these crude drugs., Method: The ITS regions of these species were amplified by PCR and sequenced, and their sequences were analyzed by DNAssist Version 2.2., Results: Homologous alignment indicated that the consanguinity of the ITS sequences between genus Bupleurum and outgroups was lower than 75%, and that within genus Bupleurum was higher than 88%. For the same species, the consanguinity was higher than 99%., Conclusion: The ITS sequences may serve as reliable molecular markers for identification of Radix Bupleuri.

Objective: To provide the basis of molecular authentication of Radix Bupleuri by the comparison of the internal transcribed spacer (ITS) sequences of five kinds of Radix Bupleuri in common use., Method: Firstly, total DNA of five kinds of Radix Bupleuri was extracted. Secondly, the ITS sequence was amplified by PCR with universal primer of ITS and PCR product was directly sequenced after purification., Result: The length of ITS1 and ITS2 sequence was 214-220 bp and 230-231 bp respectively. There were distinct variation sites between the ITS sequences of the five kinds of Radix Bupleuri., Conclusion: ITS sequence may be the evidence for the molecular authentication of Radix Bupleuri.