成分分析

基于高通量测序技术对11种乌头属药材混合粉末鉴定研究*

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  • 1.内蒙古民族大学,通辽 028000;
    2.辽宁中医药大学,大连 116000
第一作者 撒切尔 Tel:18704792782;E-mail:1165515784@qq.com
包书茵 Tel:18647556661;E-mail:baoshuyin8881@163.com
**Tel:13948359978;E-mail:bghhtlg_123@163.com

修回日期: 2021-04-19

  网络出版日期: 2024-07-15

基金资助

*蒙医防治优势病种经典方药研发平台及质量体系建设和示范研究(2018YFC1708200); 内蒙古自治区科技重大专项(2019ZD004); 蒙医药标准化国际科技合作项目(MDKBZH2018025); 内蒙古自然科学基金项目(2018MS08124); 内蒙古自治区蒙医药重点实验室建设——蒙医验方木膝通痹饮治疗类风湿性关节炎实验研究(KJJH2001); 蒙医验方“旋覆花合剂”治疗“粘热病”性肺炎的药效学研究

Identification of mixed powder of 11 species of Aconitum Linnaeus medicinal materials based on high-throughput sequencing technology*

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  • 1. Inner Mongolia University for nationalities,Tongliao 028000,China;
    2. Liaoning University of traditional Chinese medicine,Dalian 116000,China

Revised date: 2021-04-19

  Online published: 2024-07-15

摘要

目的:建立基于高通量测序技术的11 种乌头属药材混合粉末分子鉴定方法。方法:采用高通量测序技术,对11 种乌头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IlluminaMiSeq 平台对DNA 片段进行双端测序,最后采用FLASH、QIIME、GraPhlAn 及MEGA 6.0 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:混合样品中得到高质量的ITS2 序列共82 348 条, 样品中乌头属物种含有序列69 814 条,比对出其中的8 个物种,即样品中含有草乌、川乌、阴山乌头、华北乌头、瓜叶乌头、毛叶乌头、花葶乌头、高乌头;未能比对出黄花乌头、松潘乌头、川鄂乌头3 种乌头属药材。结论:以ITS2 作为条形码,采用高通量测序技术,可以鉴定出乌头属药材混合粉末中的8 个物种,为混合药材的鉴定提供新思路。

本文引用格式

撒切尔, 包书茵, 乌仁吉如拉, 奥·乌力吉, 包桂花, 许亮, 张婷婷 . 基于高通量测序技术对11种乌头属药材混合粉末鉴定研究*[J]. 药物分析杂志, 2021 , 41(9) : 1598 -1604 . DOI: 10.16155/j.0254-1793.2021.09.13

Abstract

Objective: To establish an high-throughput sequencing method for molecular identification of mixed powders of 11 species of Aconitum medicinal materials. Method: Using high-throughput sequencing technology, total DNA from 11 kinds of Aconitum medicinal powder samples was extracted following PCR amplification of ITS2 fragments. Paired-end sequencing of the DNA fragment was performed using IlluminaMiSeq platform. Finally,using FLASH,QIIME,GraPhlAn and MEGA 6.0 software,the sequence was sorted out and the species in the mixed powder were identified by cluster analysis. Results: A total of 82 348 high-quality ITS2 sequences were obtained in the mixed samples. The Aconitum species contained 69 814 sequences. 8 species were found including Radix Aconiti kusnezoffii,Radix Aconiti Carmichaelii ,Radix Aconiti Yinschanicum,Radix Aconiti jeholense,Radix Aconiti hemsleyanum,Radix Aconiti carmichaelii,Radix Aconiti scaposum and Radix Aconiti sinomontanum. But Radix Aconiti coreanum ,Radix Aconiti sungpanense and Radix Aconiti henryi were not identified successfully. Conclusion: Using ITS2 as a barcode,the high-throughput sequencing technology can identify 8 species in the mixed powder of Aconitum medicinal materials,providing new ideas for the identification of mixed medicinal materials.

参考文献

[1] 2020 年版中华人民共和国药典. 一部[S]. 2020:39,236
ChP2020.Vol I[S]. 2020:39,236
[2] SHEN XJ,YIN TP,LI XY,et al.Diterpenoid alkaloids with chemotaxonomic significance from Aconitum spathulatum[J]. Biochem Syst Ecol,2020,91:104045.
[3] 杨武斌,王平. 乌头碱药理作用及毒性研究进展[J]. 时珍国医国药,2014,25(2):427
YANG WB,WANG P.Research progress on pharmacological effects and toxicity of aconitine[J]. Lishizhen Med Mater Med Res,2014, 25(2):427
[4] 中国科学院《中国植物志》编委会. 中国植物志英文修订版(第6 卷)[M]. 北京: 科学出版社,2001:149
Editorial Board of The Flora of China,Chinese Academy of Sciences.English Revised Edition of Flora of China (Volume 6)[M]. Beijing:Science Press ,2001:149
[5] 刘杰, 过立农, 马双成, 等. 基于DNA 条形码技术的制草乌疑似伪品基原鉴定[J]. 药物分析杂志,2020,40(2):246
LIU J,GUO LN,MA SC,et al.Based on DNA barcode technology Zhi Caowu Suspected fake origin identification[J]. Chin J Pharm Anal,2020,40(2):246
[6] 石达理. 龙胆和草乌药材的双分子鉴定及商品规格等级研究[D]. 沈阳:辽宁中医药大学,2018
SHI DL .Research on bimolecular identification and commodity specifications of Longdan and Caowu Herbs[D]. Shenyang: Liaoning University of Traditional Chinese Medicine,2018
[7] 罗群, 马丹炜, 王跃华. 川乌遗传多样性的ISSR 鉴定[J]. 中草药,2006,37(10):1554
LUO Q,MA DW,WANG YH.ISSR identification of genetic diversity of Chuanwu[J]. Chin Herb Med,2006(10):1554
[8] 张婷婷, 梁勇满, 许亮, 等. 基于高通量测序技术对五种白头翁药材混合样品的DNA 分子鉴定研究[J]. 药学学报,2018,53(11):1918
ZHANG TT,LIANG YM,XU L,et al.Based on high-throughput sequencing technology for DNA molecular identification of mixed samples of five Pulsatilla chinensis medicinal materials[J]. Acta Pharm Sin,2018,53(11):1918
[9] 邢艳萍, 陈思有, 许亮, 等. 基于高通量测序技术对牛蒡子药材与伪品药材混合粉末鉴定研究[J]. 中国中药杂志,2018,43(19):3862
XING YP,CHEN SY,XU L,et al.Based on high-throughput sequencing technology on the identification of mixed powders of Niubangzi and counterfeit medicinal materials[J]. China J Chin Mater Med,2018,43(19):3862
[10] MAGOC T,SALZBERG SL.FLASH:fast length adjustment of short reads to improve genome assemblies[J]. Bioinformatics,2011,27:2957
[11] BOKULICH NA,SUBRAMANIAN S,FAITH JJ,et al.Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing[J]. Nat Methods,2013,10:57
[12] KOLJALG U,NILSSON RH,ABARENKOV K,et al.Towards a unified paradigm for sequence-based identification offungi[J]. Mol Ecol,2013,22:5271
[13] HANDELSMAN J,RONDON MR,BRADY SF,et al.Molecular bio-logical access to the chemistry of unknown soil microbes:a newfrontier for natuml pmducts[J]. Chem Biol,1998,5(10): 245
[14] 时圣明, 潘明佳, 王洁, 等. 分子鉴定技术在中药中的应用[J]. 中草药,2016,47(17):3121
SHI SM,PAN MJ,WANG J,et al.Application of molecular identification technology in traditional Chinese medicine[J] .Chin Tradit Herb Drugs,2016,47(17):3121
[15] BINH NV,NGOC LGV,ESPINOSA WN,et al.Comprehensive comparative analysis of chloroplast genomes from seven,Panax, species,and development of an authentication system based on species-unique single nucleotide polymorphism markers[J]. J Ginseng Res,2020,44:135
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