College of Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, 250355, People's Republic of China.
J Nat Med. 2013 Apr;67(2):296-302. doi: 10.1007/s11418-012-0681-8. Epub 2012 Jul 17.
To discriminate the cortex herbs of Chinese Pharmacopoeia, the second internal transcribed spacer (ITS2) of ribosomal DNA of 51 samples belonging to 19 kinds of cortex herbs were analyzed in this paper. Sequence assembly was performed using the CodonCode Aligner. Phylogenetic study was performed using the molecular evolutionary genetics analysis (MEGA) 4.0 software in accordance with the Kimura 2-Parameter (K2P) model. Phylogenetic tree was constructed using the neighbor-joining (NJ) method. We found that the ITS2 sequences of the studied samples ranged from 207 to 256 bp in length and easy to be amplified. The intraspecific genetic distance of the cortex herbs was between 0 and 0.073, which was lower than their interspecific genetic distance (mean, 0.868; minimum, 0.101). In the cluster dendrogram, all the studied medicinal materials with several samples, were monophyletic. It is concluded that ITS2 barcode is suitable for the identification of cortex herbs of Chinese Pharmacopoeia, and it will play an important role in the field of identification of traditional Chinese medicine (TCM).
为了鉴别《中国药典》收载的皮类药材，本研究对 19 种皮类药材的 51 个样本的核糖体 DNA 第二内转录间隔区（ITS2）进行了分析。序列组装使用 CodonCode Aligner 完成。使用 Molecular Evolutionary Genetics Analysis (MEGA) 4.0 软件，按照 Kimura 2-Parameter (K2P) 模型进行系统发育研究。采用邻接法（NJ）构建系统发育树。结果表明，所研究样本的 ITS2 序列长度为 207-256bp，易于扩增。皮类药材的种内遗传距离在 0 到 0.073 之间，低于种间遗传距离（平均值为 0.868，最小值为 0.101）。在聚类树状图中，所有具有多个样本的研究材料均为单系。结论：ITS2 条码适用于《中国药典》皮类药材的鉴定，将在中药鉴定领域发挥重要作用。
