基于ITS2序列的中药材苍术种苗DNA条形码鉴定

刘金欣; 李耿; 陈彩霞; 魏妙洁; 王燕; 赵春颖; 石林春

doi:10.13422/j.cnki.syfjx.2018020034

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基于ITS2序列的中药材苍术种苗DNA条形码鉴定

资源与鉴定 | 更新时间：2024-01-04

- 基于ITS2序列的中药材苍术种苗DNA条形码鉴定
- Identification of DNA Barcoding of Atractylodis Rhizoma Seedlings by ITS2 Sequence
- 中国实验方剂学杂志 2018年24卷第2期页码：34-38
- 作者机构：
- 作者简介：
- 基金信息：
  
  国家自然科学基金项目（81303158，81703659）;河北省教育厅青年拔尖人才项目（BJ201602）;河北省自然科学基金项目（H2017406031）
- DOI：10.13422/j.cnki.syfjx.2018020034
  中图分类号： S567.239
- 纸质出版日期：2018
- 稿件说明：
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刘金欣, 李耿, 陈彩霞, 等. 基于ITS2序列的中药材苍术种苗DNA条形码鉴定[J]. 中国实验方剂学杂志, 2018,24(2):34-38.

LIU Jin-xin, LI Geng, CHEN Cai-xia, et al. Identification of DNA Barcoding of Atractylodis Rhizoma Seedlings by ITS2 Sequence[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(2): 34-38.
刘金欣, 李耿, 陈彩霞, 等. 基于ITS2序列的中药材苍术种苗DNA条形码鉴定[J]. 中国实验方剂学杂志, 2018,24(2):34-38. DOI： 10.13422/j.cnki.syfjx.2018020034.

LIU Jin-xin, LI Geng, CHEN Cai-xia, et al. Identification of DNA Barcoding of Atractylodis Rhizoma Seedlings by ITS2 Sequence[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(2): 34-38. DOI： 10.13422/j.cnki.syfjx.2018020034.

摘要

目的：建立基于DNA条形码技术的中药材苍术种苗鉴定方法。方法：收集中药材苍术及其易混品原植物样品28份，构建苍术种苗鉴定参考核糖体内部转录间隔区2（ITS2）条形码数据库。从河北、山东、山西、内蒙古、辽宁及湖北等苍术主产地收集苍术种苗52份。通过提取基因组DNA，聚合酶链式反应（PCR）扩增，双向测序获得其ITS2条形码序列。应用PUAP 4.0软件计算种内、种间遗传距离，利用MEGA 7.0软件构建邻接树，对中药材苍术种苗进行DNA条形码鉴定。结果：中药材苍术及其易混品原植物ITS2条形码序列具有稳定的序列差异，邻接树呈现单系性，可以明显区分苍术及其易混品；基于标准参考数据库，42份苍术种苗为朝鲜苍术（80.8%），7份苍术种苗为苍术（13.5%），3份苍术种苗为白术（5.7%）。结论：基于ITS2序列可以准确区分中药材苍术及其混伪品种苗。苍术种苗基原物种鉴定结果以朝鲜苍术为主，提示苍术临床用药存在潜在安全风险。

Abstract

Objective:In order to guarantee the species correction of Atractylodis Rhizoma seedlings

a molecular identification method with internal transcribed spacer 2(ITS2) as DNA barcode has been tested. Method:Here

28 original plant specimens of Atractylodis Rhizoma and their common adulterants have been collected to construct its standard reference ITS2 sequence database.Fifty-two Atractylodis Rhizoma seedlings have been collected from Hebei province

Shandong province

Shanxi province

Inner Mongolia autonomous region

Liaoning province and Hubei province to verify their original species.Their ITS2 sequences have been obtained after DNA extraction

polymerase chain reaction(PCR) and bi-directional sequencing.The intra-species and inter-species genetic distances were calculated by PUAP 4.0 software.The neighbor-joining(NJ) trees were constructed by MEGA 7.0 software. Result:The stable sequence divergence has been found between ITS2 barcode sequences of Atractylodis Rhizoma and their common adulterants

their NJ trees showed monophyly.With the help of standard reference ITS2 sequence database

we found 42 Atractylodis Rhizoma seedlings were from Atractylodes koreana

7 Atractylodis Rhizoma seedlings were from A. lancea

3 Atractylodis Rhizoma seedlings were from A. macrocephala. Conclusion:Atractylodis Rhizoma seedlings are mainly come from A. koreana.This result is consistent with market survey of medicinal resources

suggesting that Atractylodis Rhizoma has safety risk.The DNA barcoding method based on the ITS2 sequence can accurately distinguish between Atractylodis Rhizoma seedlings and their adulterants seedlings.

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