基于高通量测序分析麻黄基原混杂  

Analysis of Ephedrae Herba-based promiscuous mixture based on high-throughput sequencing

作　　者：肖厚红 刘倩文 郭帅[1,2] 覃仕扬 徐帆[4] 曹刚 徐江 胡灏禹[2] 陈士林 XIAO Houhong;LIU Qianwen;GUO Shuai;QIN Shiyang;XU Fan;CAO Gang;XU Jiang;HU Haoyu;CHEN Shilin(School of Pharmacy,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学药学院,611137 [2]中国中医科学院中药研究所 [3]北京市公安局法医中心 [4]中国医药健康产业股份有限公司

出　　处：《环球中医药》2022年第12期2257-2265,共9页Global Traditional Chinese Medicine

基　　金：中国中医科学院优秀青年科技人才(创新类)培养专项(zz13-yq-047);科技基础资源调查专项(2018FY100700)。

摘　　要：中药麻黄来源于麻黄科植物草麻黄Ephedra sinica、中麻黄E.intermedia和木贼麻黄E.equisetina的干燥草质茎。当下麻黄栽培品已成为麻黄药材的主要来源,但由于各种原因,尤其是种质问题,导致这些品种的质量下降严重。《中华人民共和国药典(一部)》2020版收录含麻黄的中药复方制剂七十多种,可见在未来麻黄药材的需求量依旧很大,种质问题亟待解决。本文基于高通量测序,结合Sanger测序,对麻黄3个基原种的ITS2和psbA-trnH序列进行分析。结果显示:草麻黄和中麻黄在9~20 bp(CAGGAAATCCAA)这段碱基序列缺失,木贼麻黄插入了这段碱基序列;ITS2序列有7个变异位点可鉴别木贼麻黄,其中221变异位点存在嵌套峰,另外发现22位点嵌套峰可作为木贼麻黄新的鉴别特征性位点。最后,对麻黄3个基原品种的杂合度进行了考察和评价,以期为鉴别麻黄药材基原和种质混杂问题提供解决方法。The traditional Chinese medicine Ephedrae Herba is derived from the dried grass stems of Ephedra sinica,E.intermedia and E.equisetina of the family Ephedrae.At present,the cultivated products have become the main source of Ephedrae Herba medicinal materials,but due to various reasons,especially the germplasm problem,the quality of these varieties have been seriously degraded.The 2020 edition of“Chinese Pharmacopoeia”contains more than 70 kinds of traditional Chinese medicine compound preparations containing Ephedrae Herba,which shows that the demand for Ephedrae Herba is still needed in the future,and the germplasm problem needs to be solved urgently.In this paper,based on the methods of high-throughput sequencing,Sanger sequencing,ITS2 and psbA-trnH sequences,the three primitive species of Ephedrae Herba were analyzed.The results showed that the 9~20 bp(CAGGAAATCCAA)base sequence of E.sinica and E.intermedia was absent,and the E.equisetina inserted the base sequence.There were 7 variation sites in the ITS2 sequence that can identify E.equisetina.Among them,221 mutation sites had nested peaks,and 22 nested peaks can be used as new identification and characteristic sites of E.equisetina identification.Finally,the heterozygosity of three Ephedrae Herba cultivars was investigated and evaluated,in order to provide a solution for the identification of Ephedrae Herba primordia and germplasm hybridity.

关 键 词：麻黄 基原 鉴别 ITS2 种内变异 高通量测序 

分 类 号：R931[医药卫生—生药学]