凉茶药材凉粉草与混伪品的ITS2条形码鉴定

师玉华,马定乾,张景景,方广宏,徐文流,孙伟

中国药学杂志 ›› 2015, Vol. 50 ›› Issue (15) : 1282-1285.

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中国药学杂志 ›› 2015, Vol. 50 ›› Issue (15) : 1282-1285. DOI: 10.11669/cpj.2015.15.005
中药质量控制与安全性检测专栏

凉茶药材凉粉草与混伪品的ITS2条形码鉴定

  • 师玉华1,马定乾2,张景景2,方广宏3,徐文流4,孙伟1*
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Identification of Herbal Tea Ingredient Mesona chinensis and Its Adulterants Using ITS2 Barcode

  • SHI Yu-hua1, MA Ding-qian2, ZHANG Jing-jing2, FANG Guang-hong3, XU Wen-liu4, SUN Wei1*
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摘要

目的 建立一种准确、快速的凉茶药材凉粉草与其混伪品的DNA条形码鉴定方法。方法 本实验收集凉粉草及其混伪品共5个物种和4个变种的55份样本,提取基因组总DNA,通过PCR扩增和CodonCode Aligner软件拼接注释获得完整的ITS2序列,然后应用种内种间K2P遗传距离和系统进化NJ树分析进行物种鉴定。结果 凉粉草ITS2序列长度为232 bp,种内遗传距离为0,远小于其与混伪物种间的最小遗传距离0.210 2~0.310 9;NJ树结果表明,凉粉草独成一支,与其混伪品均可准确区分。结论 基于ITS2序列的DNA条形码技术能够准确有效地实现凉粉草与其混伪品的快速鉴别,保障凉茶药材凉粉草的使用安全。

Abstract

OBJECTIVE To establish an accurate and rapid method for identifying Mesona chinensis and its adulterants using the internal transcribed spacer 2 (ITS2) region as a barcode. METHODS The total genomic DNA from 55 samples representing five species and four variants including M. chinensis and its adulterants were extracted. The ITS2 regions were amplified, and the complete ITS2 sequences were annotated by CodonCode Aligner. The intra- and inter-specific genetic distances based on Kimura 2-Parameter (K2P) and the Neighbor-Joining (NJ) phylogenetic tree were analyzed to identify the species. RESULTS The lengths of all the ITS2 sequences of M. chinensis were 232 bp. The intra-specific genetic distances (0) were far shorter than the inter-specific ones between M. chinensis and its adulterants (0.210 2-0.301 9). NJ tree analysis indicated that M. chinensis were accurately differentiated from its adulterants. CONCLUSION The ITS2 region as an DNA barcode can accurately and effectively distinguish herbal tea ingredient M. chinensis from its adulterants, which provides a quick identifying method to ensure its safe use.

关键词

凉粉草 / DNA条形码鉴定 / ITS2序列 / 混伪品

Key words

Mesona chinensis / DNA barcoding / ITS2 barcode / adulterants

引用本文

导出引用
师玉华,马定乾,张景景,方广宏,徐文流,孙伟. 凉茶药材凉粉草与混伪品的ITS2条形码鉴定[J]. 中国药学杂志, 2015, 50(15): 1282-1285 https://doi.org/10.11669/cpj.2015.15.005
SHI Yu-hua, MA Ding-qian, ZHANG Jing-jing, FANG Guang-hong, XU Wen-liu, SUN Wei. Identification of Herbal Tea Ingredient Mesona chinensis and Its Adulterants Using ITS2 Barcode[J]. Chinese Pharmaceutical Journal, 2015, 50(15): 1282-1285 https://doi.org/10.11669/cpj.2015.15.005
中图分类号: R282   

参考文献

[1] LIN L S,ZHANG S,ZHAN Y L,et al. A review of chemical constituents and medicinal function of Mesona chinensis Benth. Curr Biotech (生物技术进展),2013,3(6):448-452.
[2] DENG C,LI R M. Analysis of the chemical constituents of the essential oils from Mesona chinensis Benth by gas chromatography-mass spectrometry. Chin Mod Med (中国当代医药),2012,19(13):68-69.
[3] ZHAO Z G,SHI Y P,HUANG N Z,et al. The research advances on Mesona chinensis Benth in China. J South Agr (南方农业学报),2011,42(6):657-660.
[4] CHEN Q R. Analysis of the pharmacological effects of flavonoids compounds. Chin Pract Med(中国实用医药),2012,7(21):254-255.
[5] ZHU H,LIAO Y K,SHI Q L. The microscopical identification of Mesona chinensis. Chin Med Mat (中药材),2003,26(1):16-17.
[6] ZHANG G F,GUAN J M,LIN J,et al. RAPD Fingerprint construction and genetic similarity of Mesona chinensis (Lamiaceae) in China. Genet and Mol Res,2012,11(4):3649-3657.
[7] HEBERT P D N,CYWINSKA A,BALL S L,et al. Biological identifications through DNA barcodes. P Roy Soc B-Biol Sci,2003,270(1512):313-321.
[8] KRESS W J,WURDACK K J,ZIMMER E A,et al. Use of DNA barcodes to identify flowering plants. Proc Natl Acad Sci USA,2005,102(23):8369-8374.
[9] CHEN S L,YAO H,HAN J P,et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS One,2010,5(1):e8613.
CHEN S L,YAO H,HAN J P,et al. Principles for molecular identification of traditional Chinese materia medica using DNA barcoding. China J Chin Mater Med (中国中药杂志),2013,38(2):141.
CHEN S L,PANG X H,YAO H,et al. Identification system and perspective for DNA barcoding traditional Chinese materia medica. Mod Tradit Chin Med Mater Med World Sci Tech (世界科学技术:中医药现代化),2012,13(5):747-754.
CHEN S L,PANG X H,SONG J Y,et al. A renaissance in herbal medicine identification: From morphology to DNA. Biotechnol Adv,2014,32(7):1237-1244.
LUO K,MA P,YAO H,et al. Identification of Gentianae Macrophyllae Radix using the ITS2 barcode. Acta Pharm Sin (药学学报),2012,47(12):1710-1717.
PANG X H,SONG J Y,XU H B. Using ITS2 barcode to identify ephedrae herba. China J Chin Mater Med (中国中药杂志),2012,37(8):1118-1121.
REN W,MA X X,YU J L,et al. Identification of Cimicifugae Rhizoma and its adulterants using the ITS2 sequence. China J Chin Mater Med (中国中药杂志),2014,39(12):2184-2188.
HOU D Y,SONG J Y,YANG P,et al. Application study of ITS/ITS2 barcodes in identification of Zanthoxmli Pericarpium. Chin Pharm J (中国药学杂志),2014,49(7):534-538.
SHI Y H,SUN W,XU W L,et al. Identification of herbal tea ingredient Plumeria rubra and its adulterants using DNA barcoding. China J Chin Mater Med (中国中药杂志),2014,39(12):2199-2203.
KELLER A,SCHLEICHER T,SCHULTZ J,et al. 5.8S-28S rRNA Interaction and HMM-based ITS2 annotation . Gene,2009,430(1):50-57.
TAMURA K,PETERSON D,PETERSON N,et al. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol,2011,28(10):2731-2739.
GAO D N,FAN Z G. Research on identification of Isodon lophanthoides based on psbA-trnH region. J Jiangxi Tradit Chin Med (江西中医药),2012,43(11):68-70.

基金

中国中医科学院自主课题(Z2013363-865)

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