Identification of “Gonglao Leaf” and Its Sibling Adulterants Based on ITS2 Sequence / 世界科学技术-中医药现代化

ABSTRACT

Objective: The Phenomenon of different species with the same name has created potential threats to cinical safety of medication. Because of that, two kinds of “gonglao leaf” and their sibling adulterants have been identified by molecular methods in this study, to secure its safety in medication. Methods: Ilexcornuta, Mahoniabealei, Mahoniafortuneiand their sibling adulterants total of 9 species 45 samples were collected in this experiment,Total genomic DNA was extracted from them by the method of improved CTAB, ITS2 sequences were amplified, and the PCR products were sequenced. Sequence assembly and consensus sequence generation were performed by the CodonCode Aligner V 4.2.4.. The genetic distances were computed by MEGA 6.0 in accordance with the Kimura 2-Parameter (K2P) model and the phylogenetic tree constructed by the neighbor-joining (NJ) method. Results: The analysis results of the genetic distances, variable sites and the NJ phylogenetic tree showed that Ilexcornuta, Mahoniabealei, Mahoniafortunei were seperated from their sibling adulterants obviously. Conclusion: ITS2 sequence was able to identify two kinds of “gonglao leaf” and their sibling adulterants which can provide a basis for clinical accurate medication.