基于ITS2序列的建泽泻及其混伪品鉴定研究

耿超; 谷巍; 吴启南; 巢建国; 孙红梅; 蒋玲; 韩赟

基于ITS2序列的建泽泻及其混伪品鉴定研究

耿超¹,
谷巍^1,2,,
吴启南¹,
巢建国¹,
孙红梅¹,
蒋玲¹,
韩赟¹

1.
南京中医药大学药学院，江苏南京 210023
2.
南京师范大学生命科学学院，江苏南京 210023

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出版历程
- 收稿日期: 2015-11-25
- 修回日期: 2016-01-06
- 刊出日期: 2016-03-10

Identification of Alisma orientale (Sam.) Juzep. and its adulterants using ITS2 sequence

1.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210023, China
2.
Life Sciences College, Nanjing Normal University, Nanjing, 210023, China

摘要

摘要: 目的利用ITS2条形码鉴别建泽泻及其混伪品。方法提取28份建泽泻及其混伪品基因组DNA，PCR扩增ITS2序列并进行双向测序，测序结果提交至GenBank，同时从GenBank下载泽泻科植物及其混伪品10种17条ITS2序列，对提交与下载的45条序列，应用MEGA 5.1软件进行序列比对，计算种内和种间距离，采用相似性搜索法、最近距离法进行鉴定分析，并构建NJ系统进化树直观反映鉴定结果。结果建泽泻、川泽泻ITS2序列长度均为311 bp，存在1个变异位点；建泽泻与泽泻科泽泻属物种距离较近，与泽泻科其他属及其混伪品之间遗传距离较远；NJ树结果显示建泽泻及其混伪品均可明显区分，表现出良好的单系性。结论 ITS2序列作为DNA条形码可以准确快捷地鉴别建泽泻及其混伪品，为其种质资源鉴定及临床安全用药提供了新的技术手段。
- 建泽泻 /
- 混伪品 /
- ITS2 /
- 鉴定
Abstract: OBJECTIVE The aim of this study was to identify Alisma orientale (Sam.) Juzep. and its adulterants using the ITS2 barcode. METHODS The total DNAs were extracted from twenty-eight samples of A. orientale as well as adulterants. The ITS2 sequences of these samples were amplified and sequenced. The acquired sequences were submitted to the GenBank and the other ITS2 sequences of 17 samples from ten species were downloaded from the GenBank. Total 45 ITS2 sequences were aligned and the genetic distances among them were analyzed with MEGA 5.1. The sequences analyses were performed using the BLAST1 and the nearest distance methods， and were presented intuitively by constructing Neighbor-joining (NJ) tree. RESULTS The lengths of all ITS2 sequences of A. orientale and A. plantago-aquatica were 311 bp. Only one mutation existed among them. There was significant divergence between the interspecific and intraspecific genetic distances of the ITS2 sequences. The NJ tree showed that A. orientale is obviously different from its adulterants， which showed high monophyly. CONCLUSION As a DNA barcode， ITS2 sequences can stably and accurately distinguish A. orientale from its adulterants and also provide a new clue to identify the medical germplasm resources and ensure the clinical safety in utilization of Chinese materia medica.
- Alisma orientale (Sam.) Juzep. /
- adulterants /
- ITS2 /
- identification

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参考文献(23)

[1]	陈菁瑛，苏海兰，黄玉吉，等．泽泻不同种质资源特性比较研究[J]．中国中药杂志，2009， 34（21）：2713-2717．
[2]	Chen JY， Su HL， Huang YJ， et al. Comparative studies on genetic resource of characteristics of Alisma[J]. China J Chin Mater Med， 2009， 34(21): 2713-2717.
[3]	谷巍，吴启南，巢建国，等．不同产地泽泻RAPD分析[J]．南京中医药大学学报，2009， 25（6）：452-453， Ⅳ．
[4]	Gu W， Wu QN， Chao JG， et al. RAPD analysis of rhizoma alismatis orientalitis grown in different locations[J]. J Nanjing Univ Tradit Chin Med， 2009， 25(6): 452-453，Ⅳ.
[5]	国家药典委员会．中华人民共和国药典:一部[M]．北京：化学工业出版社，2005：229．
[6]	Chinese Pharmacopoeia Commission. China Pharmacopeia: Volume A [M]. Beijing: China medical science press， 2015: 229.
[7]	Tian T， Chen H， Zhao YY. Traditional uses， phytochemistry， pharmacology， toxicology and quality control of Alisma orientale (Sam.) Juzep: a review[J]. J Ethnopharmacol， 2014， 158(Pt A): 373-387.
[8]	徐飞，吴启南，李兰，等．气质联用法分析泽泻中的挥发性成分的研究[J]．南京中医药大学学报，2011， 27（3）：277-280．
[9]	Xu F， Wu QN， Li L， et al. GC-MS analysis of volatile components in alisma orientalis[J]. J Nanjing Univ Tradit Chin Med， 2011， 27(3): 277-280.
[10]	黄琼林，马新业，梁凌玲，等．泽泻科植物DNA条形码的筛选研究[J]．中华中医药杂志，2013，28（5）：1402-1406．
[11]	Huang QL， Ma XY， Liang LL， et al. Study on screening of potential DNA barcoding in Alismataceae[J]. China J Tradit Chin Med Pharm， 2013，28(5): 1402-1406.
[12]	Gu W， Song J， Cao Y， et al. Application of the ITS2 region for barcoding medicinal plants of selaginellaceae in pteridophyta[J]. PLoS One， 2013， 8(6): e67818.
[13]	Chen S， Yao H， Han J， et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PLoS One， 2010， 5(1): e8613.
[14]	Pang XH， Song JY， Zhu YJ， et al. Applying plant DNA barcodes for Rosaceae species identification[J]. Cladistics， 2011， 27(2): 165-170.
[15]	Koetschan C， Hackl T， Müller T， et al. ITS2 database IV: interactive taxon sampling for internal transcribed spacer 2 based phylogenies[J]. Mol Phylogenet Evol， 2012， 63(3): 585-588.
[16]	赵莎，庞晓慧，宋经元，等．应用ITS2条形码鉴定中药材合欢皮、合欢花及其混伪品[J]．中国中药杂志，2014， 39（12）：2164-2168．
[17]	Zhao S， Pang XH， Song JY， et al. Identification of albiziae cortex， albiziae Flos and their adulterants using ITS2 barcoding[J]. China J Chin Mater Med， 2014， 39(12): 2164-2168.
[18]	辛天怡，姚辉，罗焜，等．羌活药材ITS/ITS2条形码鉴定及其稳定性与准确性研究[J]．药学学报，2012， 47（8）：1098-1105．
[19]	Xin TY， Yao H， Luo K， et al. Stability and accuracy of the identification of Notopterygii Rhizoma et Radix using the ITS/ITS2 barcodes[J]. Acta Pharm Sin， 2012， 47(8): 1098-1105.
[20]	沈昌明，王琳．传统中药鉴定方法的应用[J]．医药论坛杂志，2010，31（20）：205-207．
[21]	Shen CM， Wang L. Application of ttraditional identification method of Chinese materia medica[J]. J Med Forum， 2010，31(20): 205-207.
[22]	陈士林，姚辉，韩建萍，等．中药材DNA条形码分子鉴定指导原则[J]．中国中药杂志，2013， 38（2）：141-148．
[23]	Chen SL， Yao H， Han JP， et al. Principles for molecular identification of traditional Chinese materia medica using DNA barcoding[J]. China J Chin Mater Med， 2013， 38(2): 141-148.