Murrayae Folium et Cacumen and its closely related species were identified by the ITS2 sequence which was the core barcode in the DNA barcoding system for identification of Chinese medicine.Methods:ITS2 sequence of Murrayae Folium et Cacumen,medicinal material and their mixed fakes were identified according to the standard procedure of Chinese medicine DNA barcode molecular identification method,and intra-and inter-specific variation analysis was performed.The nearest distance method,SNPs locus analysis,and tree building method are used to comprehensively evaluate the identification of the sequence ability.Results:The ITS2 sequence lengths of Murrayae Folium et Cacumen and its closely related species were ranged from 220 bp to 234 bp.The results indicated that K2P genetic distance analysis cannot effectively authenticate M.exotica,M.paniculata and M.alata.Sequence analysis found Murraya exotica and other related species have 3 stable SNPs location and in the UPGMA tree,it was shown that different species can be differentiated according to their monophy,indicating that ITS2 sequence can differentiate Murrayae Folium et Cacumen and its closely related species.A total of 44 samples of Murraya paniculata to be inspected were identified based on the DNA Barcoding System showing that that the inspected samples were all authentic.Conclusion:ITS2 sequence can be used to distinguish Murrayae Folium et Cacumen from its closely related species,which provide useful ways for market monitoring of Murrayae Folium et Cacumen and benefit to its herbgenomics research.