首页|基于高通量测序技术的9种铁线莲属药材混合粉末分子鉴定

基于高通量测序技术的9种铁线莲属药材混合粉末分子鉴定

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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.
Molecular identification of mixed powder of 9 kinds of Clematis medicinal materials based on high-throughput sequencing technology
Objective To establish a method for identifying 9 Clematis mixed powders based on high-throughput se-quencing 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 argenti-lucida,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.

High-throughput sequencing technologyITS2sequenceClematisMixed powderMolecular identification

呼和、莫日根、乌雅汉、乌日汗、额尔敦呼、白明君、包桂花、许亮

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内蒙古民族大学,内蒙古通辽 028000

新巴尔虎右旗阿拉坦额莫勒镇社区卫生服务中心,内蒙古呼伦贝尔 021300

内蒙古自治区国际蒙医医院,内蒙古呼和浩特 010020

蒙医药研发工程教育部重点实验室,内蒙古通辽 028000

辽宁中医药大学,辽宁大连 116000

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高通量测序技术 ITS2 铁线莲属 药材 混合粉末 分子鉴定

内蒙古卫健委蒙医药标准化研究项目内蒙古自然科学基金内蒙古自治区直属高校基本科研业务费项目

2023-MB0302023LHMS08047GXKY22016

2024

药学研究
山东省药品检验所 山东省药学会

药学研究

CSTPCD
影响因子:0.653
ISSN:2095-5375
年,卷(期):2024.43(1)
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