摘要
牛蒡子是我国传统中药,主要伪品为毛头牛蒡、水飞蓟、云木香、大翅蓟、紫穗槐等果实。传统鉴定方法或分子条形码技术可以对牛蒡子和其伪品进行鉴定,但是对牛蒡子与其伪品混合粉末的鉴定较困难。该文利用高通量测序技术对牛蒡子以及其5种伪品混合粉末ITS2序列进行测序,对混合粉末物种进行鉴定,为牛蒡子药材混合粉末的鉴定提供新思路。对药材混合粉末中的总DNA进行提取,并对总DNA中ITS2片段进行扩增,利用Illumina MiSeq平台对群落DNA片段进行双端(paired-end)测序,利用FLASH,QIIME以及GraPhlAn等软件对序列进行整理分析。结果显示:混合样品中共得到高质量的ITS2序列39 910条,其中,样品中的物种ITS2序列共34 935。进一步的进化分析表明样品中含牛蒡子、毛头牛蒡、水飞蓟、大翅蓟、云木香和紫穗槐。利用高通量测序技术,以ITS2序列作为DNA条形码,可以检测出混合粉末中含有牛蒡子及其伪品,为牛蒡子药材质量控制、安全用药以及中药混合粉末的鉴定提供参考。
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.
作者
邢艳萍
陈思有
许亮
梁勇满
王佳豪
王冰
刘涛
康廷国
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)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2018年第19期3862-3866,共5页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81773852)
辽宁省高校优秀青年学者成长计划项目(LJQ2014101)
