摘要
目的建立基于高通量测序技术的9种铁线莲属混合粉末分子鉴定方法。方法采用高通量测序技术,对9种铁线莲属药材混合粉末样品中的总DNA经提取,对ITS2片段进行PCR扩增,利用Illumina MiSeq平台对DNA片段进行双端测序,最后采用FLASH、QIIME、GraPhlAn及MEGA 7.0软件对序列进行整理并聚类分析,鉴定混合粉末中的物种。结果混合粉末样品中得到高质量的ITS2序列共496条,铁线莲属物种含有序列72条,比对出棉团铁线莲、女萎铁线莲、短尾铁线莲、灌木铁线莲、单叶铁线莲、黄花铁线莲、粗齿铁线莲等7个物种,未能比对出东北铁线莲和芹叶铁线莲。结论以ITS2作为DNA条形码,采用高通量测序技术,可以鉴定出铁线莲属药材混合样品中的7个物种,为混合药材的鉴定提供依据。
Objective To establish a method for identifying 9 Clematis mixed powders based on high-throughput sequencing technology.Methods The total DNA of 9 samples from Clematis was extracted and uesed ITS2 fragment for PCR amplification.The DNA fragments were separated by Illumina MiSeq platform.FLASH,QIIME,GraPhlAn and MEGA 7.0 software were used to sort and cluster DNA sequences.Results A total of 496 high-quality ITS2 sequences were obtained from the mixed powder samples,and 72 sequences were contained in the Clematis species.Seven species,including Clematis hexapetala,Clematis apiifolia,Clematis brevicaudata,Clematis fruticose,Clematis henryi,Clematis intricate,Clematis argentilucida,were compared,and Clematis manshurica and Clematis aethusifolia were not compared.Conclusion Using ITS2 as DNA barcode and high-throughput sequencing technology,seven species in the mixed samples of Clematis can be identified,which provides a basis for the identification of mixed medicinal materials.
作者
呼和
莫日根
乌雅汉
乌日汗
额尔敦呼
白明君
包桂花
许亮
HU He;MO Rigen;WU Yahan;WU Rihan;E Erdunhu;BAI Mingjun;BAO Guihua;XU Liang(Inner Mongolia Minzu University,Tongliao 028000,China;Alatan Emolle Town Community Health Service Center,New Barag West County,Hulun Buir 021300,China;International Mongolia Hospital of Inner Mongolia,Hohhot 010020,China;Key Laboratory of Mongolian Medicine and Pharmacy Research and Engineering of Ministry of Education,Tongliao 028000,China;Liaoning University of Traditional Chinese Medicine,Dalian 116000,China)
出处
《药学研究》
CAS
2024年第1期30-34,共5页
Journal of Pharmaceutical Research
基金
内蒙古卫健委蒙医药标准化研究项目(No.2023-MB030)
内蒙古自然科学基金项目(No.2023LHMS08047)
内蒙古自治区直属高校基本科研业务费项目(No.GXKY22016)。
关键词
高通量测序技术
ITS2
铁线莲属
药材
混合粉末
分子鉴定
High-throughput sequencing technology
ITS2sequence
Clematis
Mixed powder
Molecular identification
