蔓荆子药材及其混伪品DNA条形码鉴定研究

Identification of Viticis Fructus and Its Adulterants by ITS2 Sequence

本研究应用ITS2序列鉴别蔓荆子及其混伪品,收集药材和基原植物样本共46份,通过提取基因组DNA、PCR扩增和双向测序获得ITS2序列,经Codon Code Aligner V4.2拼接注释获得序列,应用MEGA5.0对序列进行分析比对,计算种内和种间遗传距离(Kimura 2-Parameter,K2P),构建系统发育NJ树进行鉴定。结果表明,蔓荆子药材基于ITS2序列的种内最大K2P遗传距离均小于其与混伪品的种间最小K2P遗传距离,NJ树显示蔓荆子药材与其混伪品可明显分开,表现出良好的单系性。因此,应用ITS2序列能够准确地鉴定蔓荆子药材及其混伪品。

ITS2 sequence was used as a barcode to identify Viticis Fructus and its adulterants. The study collected 46 samples include Viticis Fructus and its adulterants. The sequences were obtained by extracting DNA, amplification, sequencing bi-direstionlly, and then assembling with CodonCode Aligner. To identify Viticis Fructus and its adulterants, the study used the methods of computing the genetic distances by Kimura 2-Parameter （K2P） model and constructing the Neighbor-joining （NJ） phylogenetic trees using MEGA5.0. The results showed that the maximum inter-specific K2P distance of Viticis Fructus was less than the minimum inter-specific K2P distance of Viticis Fructus and its adulterants and the NJ tree indicated that the Viticis Fructus and its adulterants could be distinguished clearly. Therefore, using ITS2 barcode can distinguished Viticis Fructus and its adulterants accurately.