基于ITS2序列的药用甘草亲缘关系鉴定研究

郝杰, 蔡彦朋, 刘刚, 刘育辰, 金文渊, 袁思文

中国药学杂志 ›› 2018, Vol. 53 ›› Issue (6) : 411-417.

PDF(1313 KB)
PDF(1313 KB)
中国药学杂志 ›› 2018, Vol. 53 ›› Issue (6) : 411-417. DOI: 10.11669/cpj.2018.06.002
论 著

基于ITS2序列的药用甘草亲缘关系鉴定研究

  • 郝杰1, 蔡彦朋1, 刘刚1,2,3, 刘育辰1,2,3*, 金文渊1, 袁思文1
作者信息 +

Identification of Genetic Relationships between Medicinal Licorices Based on ITS2 Sequences

  • HAO Jie1, CAI Yan-peng1, LIU Gang1,2,3, LIU Yu-chen1,2,3*, JIN Wen-yuan1, YUAN Si-wen1
Author information +
文章历史 +

摘要

目的 利用内转录间隔区序列( ITS2)分析收集的181份国内、外甘草样品,对药用甘草及其近缘种样品的亲缘关系进行研究,区分药用与非药用甘草,并推测未鉴定甘草样本与药用甘草的亲缘关系。方法 提取不同基原和产地的甘草样本基因组DNA,PCR扩增获取其ITS2序列,通过序列比对分析、最近距离法和邻接(NJ) 树分析,建立基于ITS2序列的药用甘草DNA条形码鉴定方法;并利用该方法检测国内变异植株和国外甘草样本。结果 ITS2序列能将药用甘草与其他2个近缘物种很好地区分开;国内疑似变异植株和国外甘草样本在系统树上均与药用甘草聚为一支。结论 该实验为药用甘草建立了基于ITS2条形码序列的鉴定方法;并利用该法,初步鉴定了国内变异植株和国外甘草样本与我国药用甘草的亲缘关系;为扩大药用甘草资源、保证甘草临床用药的安全性和有效性提供了参考。

Abstract

OBJECTIVE To obtain the genetic information of 181 domestic and foreign licorice samples by determining the internal transcribed spacer sequence 2 (ITS2) in order to clarify their phylogenetic relationships and differentiate the medicinal licorice species from the non-medicinal ones. METHODS Genomic DNA was first extracted from licorice samples of different origins and habitats. Then, the ITS2 sequences were amplified using PCR technique. Next, the ITS2 sequences were subjected to sequence analyses and neighbor-joining(NJ) tree analysis. Finally, the analytical method based on ITS2 DNA barcodes was established for the licorice species discrimination. The method was then applied to the phylogenetic analysis among the suspected domestic mutants, foreign licorice samples and domestic medicinal licorices, in addition to the discriminations of the medicinal and non-medicinal licorices. RESULTS The medicinal licorice could be explicitly separated from the two commonly encountered non-medicinal licorice species either by using ITS2 sequence analysis or NJ tree analysis; and the suspected domestic mutants and foreign licorice samples converged with the medicinal licorice to form one monophyletic branch in the NJ tree analysis, which indicated unambiguously conspecifics. CONCLUSION This study establishes a method for discrimination of medicinal and non-medicinal licorices based on ITS2 barcodes; and by the advantage of this method, the conspecific relationships among the suspected domestic mutants, foreign licorice samples and Chinese medicinal licorice are clarified. Thus a theoretical reference to the expansion of medicinal licorice resources and assurance of the safety and efficacy of the clinical licorice medications are provided.

关键词

甘草 / 内转录间隔区序列 / 亲缘关系 / 药用植物资源

Key words

Glycyrrhiza / ITS2 / the genetic relationship / medicinal plant resource

引用本文

导出引用
郝杰, 蔡彦朋, 刘刚, 刘育辰, 金文渊, 袁思文. 基于ITS2序列的药用甘草亲缘关系鉴定研究[J]. 中国药学杂志, 2018, 53(6): 411-417 https://doi.org/10.11669/cpj.2018.06.002
HAO Jie, CAI Yan-peng, LIU Gang, LIU Yu-chen, JIN Wen-yuan, YUAN Si-wen. Identification of Genetic Relationships between Medicinal Licorices Based on ITS2 Sequences[J]. Chinese Pharmaceutical Journal, 2018, 53(6): 411-417 https://doi.org/10.11669/cpj.2018.06.002
中图分类号: R284   

参考文献

[1] Chinese flora Commission of Chinese Academy of Sciences. Flora Reipublicae Popularis Sinicae(中国植物志)[S]. Beijing:Science and Technology Press, 1998, 42(2):167.
[2] Ch.P(2015)Vol I(中国药典2015年版.一部)[S]. 2015:86-87.
[3] XIE Z W. Theory and Application of Chinese Medicinal Herbs(中药品种理论与应用)[M]. Beijing:People′s Medical Publishing House,2008:238.
[4] ZHANG R Y, ZENG L, CAI L N, et al. Study on Variety Arrangement and Quality Control of Traditional Chinese Medicine (Volume 2) (常用中药品种整理和质量研究(第2册))[M]. Beijing:Science Press, 1995:395.
[5] LI X Y. Studies on the origin and original plants of medicinal licorice in China[J]. J Shihezi Agric Coll(石河子农学院学报),1989 (1):23-29.
[6] The State Council issued the notice on the problem about prohibiting the acquisition and sale of seaweed and stopping indiscriminate digging licorice and Ephedra[J]. Admin Ind Comm(工商行政管理), 2000, 14:49.
[7] Document [2000]13 issued by the state. The state council “The notice on the problem about prohibiting the acquisition and sales of seaweed and stopping indiscriminate digging of licorice and mahuangcao”[N]. Zhongwei Daily, 2008-11-14(3).
[8] TANG Y L, WU Y S, HUANG R S, et al. Molecular identification of Uncaria(Gouteng) through DNA barcoding[J]. Chin Med, 2016, 11:3.
[9] SHANG G F, YAN J, SHANG W. Molccubr identication of derdro bium species rchidacease[J]. Int J Mol Sci, 2015,16(9):21975-21988.
[10] HE H B, XIONG C, GUO L C, et al. Identification of Geranii Herbae using ITS2 sequence[J]. Chin Pharm J(中国药学杂志),2015, 50(1):1505-1511.
[11] ZHOU H,MA S J,SONG C, et al. Identification of Ilex asprella and its closely related species based on ITS2 barcode [J]. Chin Pharm J(中国药学杂志), 2015, 50 (19):1500-1504.
[12] HOU D Y,SONG J Y,YANG P, et al. Application study of ITS /ITS2 barcodes in identification of Zanthoxmli Pericarpiume [J]. Chin Pharm J(中国药学杂志), 2014, 49(7):534-538.
[13] LI X F,QIU T B,ZHANG H M, et al. ITS sequences and phylogenetic relationships of dandelions from different areas [J]. Chin Pharm J(中国药学杂志), 2013, 48(9):687-690.
[14] SHANG G F, MENG Y J, YU J S, et al. Application of the ribosomal DNA ITS2 region of physalis (solanaceae):DNA barcoding and phylogenetic study [J]. Front Plant Sci, 2016, 7(589):1047.
[15] XIA Z, FENG C Y, GAO Z M, et al. Authentication of DNA barcoding of scutellariabaicalensis and its related species [J]. Chin Tradit Herb Drugs(中草药), 2014, 45(1):107-112.
[16] XIA Z, GAO Z M, LI H M, et al. Molecular authentication and phylogenetic relationship of Bidenspilosa and itsrelative species [J]. Chin Tradit Herb Drugs(中草药), 2014, 45(6):828-834.
[17] YANG R, LI W D, MA Y S, et al. The molecular identification of licorice species and the qualityevaluation of licorice slices [J]. Acta Pharm Sin(药学学报), 2017, 52 (2):318-326.
[18] HERSHKOVITZ M A, ZIMMER E A. Conservation patterns in angiosperm rDNA ITS2 sequences[J]. Nucleic Acids Res, 1996, 24(15):2857-2867.
[19] CHEN S L, YAO H, HAN J P. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS One, 2010,5(1):e8613.
[20] CHEN S L. The 10th National Conference on Medicinal Plants and Phytomedicines. DNA barcode identification system of traditional Chinese Medicine[C]. 2011.
[21] HE C N. Paeonia medicinal plant genetic research [D]. Beijing:Chinese Academy of Medical Sciences, 2010.
[22] BAI Z F. Medicinal plants of Gesneriaceae genetic research [D]. Beijing:Beijing University of Chinese Medicine, 2010.
[23] ZOU X X. Study on medicinal relatives of atractylodes [D]. Beijing:Beijing University of Chinese Medicine, 2010.
[24] CHEN S L, YAO H, HAN J P, et al. Principles for molecular identification of traditional Chinese material medica using DNA barcoding[J]. Chin J Tradit Chin Med(中国中药杂志), 2013, 38 (2):141-143.
[25] KELLER A, SCHLEICHER T, SCHULTZ J, et al. 5.8S-28S rRNA interaction and HMM-based ITS2 annotation[J]. Gene, 2009, 430(1/2):50-57.
[26] CHEN S L. Standard DNA Barcodes of Chinese Metiria Medicain Chinese Pharmacopoeia(中国药典中药材DNA条形码标准序列) [M]. Beijing:Science Press,2015:148.

基金

国家自然科学基金项目资助(81473308)
PDF(1313 KB)

112

Accesses

0

Citation

Detail

段落导航
相关文章

/