Zhu Xunzhi, Zhang Yuxi, Liu Xia, Hou Dianyun, Gao Ting
Key Laboratory of Plant Biotechnology in Universities of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, Shandong China ; College of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, Jiangsu China.
Key Laboratory of Plant Biotechnology in Universities of Shandong Province, College of Life Sciences, Qingdao Agricultural University, Qingdao, Shandong China.
Chin Med. 2015 Nov 30;10:35. doi: 10.1186/s13020-015-0071-8. eCollection 2015.
The radix of Glehnia littoralis Fr. Schmidt ex Miq. (Beishashen), is often misidentified and adultered in Chinese medicine. Its seven common adulterants include Chuanminshen violaceum Sheh et Shan (Chuanmingshen), Changium smyrnioides Wolff (Mingdangshen), Sphallerocarpus gracilis (Bess.) K.-Pol. (Miguoqin), Adenophora polyantha Nakai (Shishashen), Silene tatarinowii Regel (Shishengyingzicao), Adenophora tetraphylla (Thunb.) Fisch (Lunyeshashen) and Adenophora stricta Miq. (Shashen). This study aims to evaluate the feasibility of the second internal transcribed spacer (ITS2) DNA barcoding to discriminate between Glehniae Radix and its common adulterants.
In this study, we collected 46 samples of G. littoralis and 59 samples of its seven common adulterants. Genomic DNA sequences were extracted from samples, including original plants and commercially processed crude drugs. The ITS2 of the ribosomal DNA sequences were amplified and sequenced bi-directionally. The sequences were assembled by CodonCode Aligner 3.5.7. The descriptive data analysis was conducted and neighbor-joining (NJ) phylogenetic tree was constructed by MEGA 5.0 in accordance with the kimura 2 -parameter (K2P) model. The identification efficiency was evaluated based on the BLAST1 methods. The ITS2 secondary structures were predicted and compared between Glehniae Radix and its adulterants by the ITS2 database.
As the 46 ITS2 sequences of G. littoralis were identical to each other, the identification efficiency of the ITS2 region was 100 %. A NJ tree based on the ITS2 sequences, and the predicted secondary structures of ITS2, distinguished Glehniae Radix from its adulterants.
DNA barcoding based on ITS2 distinguished commercial processed Glehniae Radix from common herbal adulterants.
北沙参在中药中常被误认和掺假。其七种常见掺伪品包括川明参、明党参、迷果芹、多花沙参、石生蝇子草、轮叶沙参和沙参。本研究旨在评估核糖体DNA第二内部转录间隔区（ITS2）条形码技术鉴别北沙参及其常见掺伪品的可行性。
本研究收集了46份北沙参样本及其七种常见掺伪品的59份样本。从样本中提取基因组DNA序列，包括原植物和市售加工的中药材。对核糖体DNA序列的ITS2进行双向扩增和测序。序列由CodonCode Aligner 3.5.7组装。进行描述性数据分析，并根据Kimura 2-参数（K2P）模型，用MEGA 5.0构建邻接（NJ）系统发育树。基于BLAST1方法评估鉴定效率。通过ITS2数据库预测并比较北沙参与其掺伪品的ITS2二级结构。
由于46条北沙参ITS2序列彼此相同，ITS2区域的鉴定效率为100%。基于ITS2序列构建的NJ树以及预测的ITS2二级结构，可将北沙参与其掺伪品区分开来。
基于ITS2的DNA条形码技术可区分市售加工的北沙参与常见的掺伪药材。
