Liu Chuan, Zhang Yu-Xin, Liu Yue, Chen Yi-Long, Fan Gang, Xiang Li, Xu Jiang, Zhang Yi
College of Ethnic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2016 Feb;41(4):567-571. doi: 10.4268/cjcmm20160403.
The ITS2 barcode was used toidentify Tibetan medicine "Dida", and tosecure its quality and safety in medication. A total of 13 species, 151 experimental samples for the study from the Tibetan Plateau, including Gentianaceae Swertia, Halenia, Gentianopsis, Comastoma, Lomatogonium ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner V3.7.1. The Kimura 2-Parameter (K2P) distances were calculated using MEGA 6.0. The neighbor-joining (NJ) phylogenetic trees were constructed. There are 31 haplotypes among 231 bp after alignment of all ITS2 sequence haplotypes, and the average G±C content of 61.40%. The NJ tree strongly supported that every species clustered into their own clade and high identification success rate, except that Swertia bifolia and Swertia wolfangiana could not be distinguished from each other based on the sequence divergences. DNA barcoding could be used as a fast and accurate identification method to distinguish Tibetan medicine "Dida" to ensure its safe use.
ITS2条形码被用于鉴定藏药“蒂达”，并确保其用药质量和安全。本研究从青藏高原共采集了13个物种的151份实验样本，包括龙胆科的獐牙菜属、花锚属、扁蕾属、喉毛花属、肋柱花属，对其ITS2序列进行扩增，并对纯化后的PCR产物进行测序。使用CodonCode Aligner V3.7.1进行序列组装和共有序列生成。使用MEGA 6.0计算Kimura 2-参数（K2P）距离。构建邻接法（NJ）系统发育树。所有ITS2序列单倍型比对后，231 bp中有31个单倍型，平均G±C含量为61.40%。NJ树有力地支持了每个物种都聚类到各自的分支中，识别成功率较高，只是基于序列差异无法区分双叶獐牙菜和川西獐牙菜。DNA条形码可作为一种快速准确的鉴定方法来鉴别藏药“蒂达”，以确保其安全使用。
