Xing Yan-Ping, Chen Si-You, Xu Liang, Liang Yong-Man, Wang Jia-Hao, Wang Bing, Liu Tao, Kang Ting-Guo
Liaoning University of Traditional Chinese Medicine, Dalian 116600, China.
China Medical University, Shenyang 110122, China.
Zhongguo Zhong Yao Za Zhi. 2018 Oct;43(19):3862-3866. doi: 10.19540/j.cnki.cjcmm.20180703.012.
Fructus Arctii is a traditional Chinese medicine. The main counterfeit species are the seeds of Arctium tomentosum, Onopordum acanthium, Silybum marianum, Saussurea costus, Amorpha fruticosa. Traditional identification methods or molecular barcoding techniques can identify Fructus Arctii and its counterfeit species. However, the identification of the mixture of it and its spurious species is rarely reported. In this paper, we sequenced the ITS2 sequences of Fructus Arctii and 5 kinds of spurious species mix powder by high-throughput sequencing to identify the mixed powder species and providing new ideas for the identification of Fructus Arctii mix powder. The total DNA in mixed powder was extracted, and the ITS2 sequences in total DNA was amplified. Paired-end sequencing was performed on the DNA fragment of the community using the Illumina MiSeq platform. The sequence was analyzed by the software FLASH, QIIME and GraPhlAn etc. The results showed that the high quality ITS2 sequences of 39910 mix samples were obtained from the mixed samples, of which the total ITS2 sequence of the samples genus was 34 935. Phylogenetic analysis showed that the samples contained Fructus Arctii, A. tomentosum, O. acanthium, S. marianum, S. costus and A. fruticosa. Using ITS2 sequences as DNA barcodes, high-throughput sequencing technology can be used to detect the Fructus Arctii and its spurious specie in mixed powder, which can provide reference for the quality control, safe use of medicinal materials of Fructus Arctii and the identification of mixed powder of traditional Chinese medicine.
牛蒡子是一种传统中药。主要伪品有绒毛牛蒡子、刺苞菜蓟、水飞蓟、木香、紫穗槐的种子。传统鉴定方法或分子条形码技术可以鉴别牛蒡子及其伪品。然而，其与伪品混合物的鉴定鲜有报道。本文通过高通量测序对牛蒡子及其5种伪品混合粉末的ITS2序列进行测序，以鉴定混合粉末的物种，为牛蒡子混合粉末的鉴定提供新思路。提取混合粉末中的总DNA，扩增总DNA中的ITS2序列。利用Illumina MiSeq平台对群落的DNA片段进行双端测序。采用FLASH、QIIME和GraPhlAn等软件对序列进行分析。结果表明，从混合样本中获得了39910个混合样本的高质量ITS2序列，其中样本属的总ITS2序列为34935条。系统发育分析表明，样本中含有牛蒡子、绒毛牛蒡子、刺苞菜蓟、水飞蓟、木香和紫穗槐。以ITS2序列作为DNA条形码，高通量测序技术可用于检测混合粉末中的牛蒡子及其伪品，可为牛蒡子药材的质量控制、安全用药及中药混合粉末的鉴定提供参考。
