Journal of Natural Medicines

, Volume 67, Issue 3, pp 571–575 | Cite as

Use of the potential DNA barcode ITS2 to identify herbal materials

  • Xiaohui Pang
  • Linchun Shi
  • Jingyuan Song
  • Xiaochen Chen
  • Shilin Chen
Original Paper


A potential DNA barcode, ITS2, was studied to discriminate herbal materials to confirm their identities and ensure their safe application in pharmaceuticals. Here, a total of 4385 samples of 2431 species were collected, and these samples are from 61 commonly used herbs and their closely related species or adulterants. Based on assessments of the extent of genetic divergence, the DNA barcoding gap and the ability for species discrimination, our results suggest that ITS2 is a powerful tool for distinguishing herbs. For the first dataset including 61 herbs, ITS2 correctly identified 100 % of them. For the second dataset containing 51 herbs and their 2382 closely related species, ITS2 could discriminate correctly 48 herbs from their closely related species. For the third dataset comprising 34 herbs and their 111 adulterants, ITS2 could distinguish successfully all the herbs from their adulterants. In conclusion, the ITS2 region is an efficient marker for the authentication of herbal materials, and our study will accelerate the process of the application of the DNA barcoding technique in differentiating herbs.


Herbal materials ITS2 DNA barcode Identification 



This study was supported by the Basic Scientific Research Operation Grants for State-Level Public Welfare Scientific Research Initiatives (YZ-12-08) and Beijing Municipal Science and Technology Foundation (D08080203640901).

Supplementary material

11418_2012_715_MOESM1_ESM.doc (820 kb)
Supplementary material 1 (DOC 820 kb)


  1. 1.
    Gong X, Sucher NJ (2002) Stroke therapy in traditional Chinese medicine (TCM): prospects for drug discovery and development. Phytomedicine 9:478–484PubMedCrossRefGoogle Scholar
  2. 2.
    Marian F, Widmer M, Herren S, Dönges A, Busato A (2006) Physicians’ philosophy of care: a comparison of complementary and conventional medicine. Forsch Komplementmed 13:70–77PubMedCrossRefGoogle Scholar
  3. 3.
    Hu S (1976) A contribution to our knowledge of Leonurus L., i-mu-ts’ao, the Chinese motherwort. Am J Chin Med (Gard City N Y) 4:219–237CrossRefGoogle Scholar
  4. 4.
    Kong YC, Yeung HW, Cheung YM, Hwang JC, Chan YW, Law YP, Ng KH, Yeung CH (1976) Isolation of the uterotonic principle from Leonurus Artemisia, the Chinese motherwort. Am J Chin Med (Gard City N Y) 4:373–382CrossRefGoogle Scholar
  5. 5.
    Singh B, Rastogi RP (1969) A reinvestigation of the triterpenes of Centella asiatica. Phytochemistry 8:917–921CrossRefGoogle Scholar
  6. 6.
    Cheng CL, Koo MWL (2000) Effects of Centella asiatica on ethanol induced gastric mucosal lesions in rats. Life Sci 67:2647–2653PubMedCrossRefGoogle Scholar
  7. 7.
    Ganzera M, Pöcher A, Stuppner H (2005) Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC-MS. Phytochem Anal 16:205–209PubMedCrossRefGoogle Scholar
  8. 8.
    Lee WB, Kwon HC, Cho OR, Lee KC, Choi SU, Baek NI, Lee KR (2002) Phytochemical constituents of Cirsium setidens Nakai and their cytotoxicity against human cancer cell lines. Arch Pharm Res 25:628–635PubMedCrossRefGoogle Scholar
  9. 9.
    Park MK, Rhyu MR, Yoon BK, Kwon H, Jang S, Lee YJ (2008) Modulation of the genomic estrogen receptor pathway by water extracts of Cirsium japonicum. Arch Pharm Res 31:225–230PubMedCrossRefGoogle Scholar
  10. 10.
    Kang TH, Pae HO, Yoo JC, Kim NY, Kim YC, Ko GI, Chung HT (2000) Antiproliferative effects of alkaloids from Sedum sarmentosum on murine and human hepatoma cell lines. J Ethnopharmacol 70:177–182PubMedCrossRefGoogle Scholar
  11. 11.
    Shaw PC, Ngan FN, But PPH, Wang J (1997) Authentication of Chinese medicinal materials by DNA technology. J Food Drug Anal 5:273–284Google Scholar
  12. 12.
    Tomlinson B, Chan TY, Chan JC, Critchley JA, But PP (2000) Toxicity of complementary therapies: an eastern perspective. J Clin Pharmacol 40:451–456PubMedCrossRefGoogle Scholar
  13. 13.
    Chinese Pharmacopoeia Commission (2010) The pharmacopoeia of the People’s Republic of China, vol 1. Chinese Medicine Science and Technology Press, BeijingGoogle Scholar
  14. 14.
    Yang ZY, Chao Z, Huo KK, Wu BY, Pan SL (2006) Nuclear ribosomal DNA internal transcribed spacer 1 sequences of 4 Leonurus species. Nan Fang Yi Ke Da Xue Xue Bao 26:1593–1595PubMedGoogle Scholar
  15. 15.
    Chen SL, Yao H, Han JP, Liu C, Song JY, Shi LC, Zhu YJ, Ma XY, Gao T, Pang XH, Luo K, Li Y, Li XW, Jia XC, Lin YL, Leon C (2010) Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS One 5:e8613PubMedCrossRefGoogle Scholar
  16. 16.
    Gao T, Yao H, Song JY, Liu C, Zhu YJ, Ma XY, Pang XH, Xu HX, Chen SL (2010) Identification of medicinal plants in the family Fabaceae using a potential DNA barcode ITS2. J Ethnopharmacol 130:116–121PubMedCrossRefGoogle Scholar
  17. 17.
    Gao T, Yao H, Song JY, Zhu YJ, Liu C, Chen SL (2010) Evaluating the feasibility of using candidate DNA barcodes in discriminating species of the large Asteraceae family. BMC Evol Biol 10:324PubMedGoogle Scholar
  18. 18.
    Luo K, Chen SL, Chen KL, Song JY, Yao H, Ma XY, Zhu YJ, Pang XH, Yu H, Li XW, Liu Z (2010) Assessment of candidate plant DNA barcodes using the Rutaceae family. Sci China Ser C 40:342–351Google Scholar
  19. 19.
    Pang XH, Song JY, Zhu YJ, Xie CX, Chen SL (2010) Using DNA barcoding to identify species within Euphorbiaceae. Planta Med 76:1784–1786PubMedCrossRefGoogle Scholar
  20. 20.
    Pang XH, Song JY, Zhu YJ, Xu HX, Huang LF, Chen SL (2011) Applying plant DNA barcodes for Rosaceae species identification. Cladistics 27:165–170CrossRefGoogle Scholar
  21. 21.
    Yao H, Song JY, Liu C, Luo K, Han JP, Li Y, Pang XH, Xu HX, Zhu YJ, Xiao PG, Chen SL (2010) Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS One 5:e13102PubMedCrossRefGoogle Scholar
  22. 22.
    Eddy SR (1998) Profile hidden Markov models. Bioinformatics 14:755–763PubMedCrossRefGoogle Scholar
  23. 23.
    Keller A, Schleicher T, Schultz J, Müller T, Dandekar T, Wolf M (2009) 5.8S–28S rRNA interaction and HMM-based ITS2 annotation. Gene 430:50–57PubMedCrossRefGoogle Scholar
  24. 24.
    Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792–1797PubMedCrossRefGoogle Scholar
  25. 25.
    Ross HA, Murugan S, Li WLS (2008) Testing the reliability of genetic methods of species identification via simulation. Syst Biol 57:216–230PubMedCrossRefGoogle Scholar
  26. 26.
    CBOL Plant Working Group (2009) A DNA barcode for land plants. Proc Natl Acad Sci USA 106:12794–12797CrossRefGoogle Scholar
  27. 27.
    Kress WJ, Erickson DL (2007) A two-locus global DNA barcode for land plants: the coding rbcL gene complements the non-coding trnHpsbA spacer region. PLoS One 2:e508PubMedCrossRefGoogle Scholar
  28. 28.
    Newmaster SG, Fazekas AJ, Ragupathy S (2006) DNA barcoding in land plants: evaluation of rbcL in a multigene tiered approach. Can J Bot 84:335–341CrossRefGoogle Scholar
  29. 29.
    Abu-Romman S (2011) Comparison of methods for isolating high quality DNA from sage (Salvia officinalis). J Med Plant Res 5:938–941Google Scholar
  30. 30.
    Chiou SJ, Yen JH, Fang CL, Chen HL, Lin TY (2007) Authentication of medicinal herbs using PCR-amplified ITS2 with specific primers. Planta Med 73:1421–1426PubMedCrossRefGoogle Scholar
  31. 31.
    Shaw PC, But PPH (1995) Authentication of Panax species and their adulterants by random-primed polymerase chain reaction. Planta Med 61:466–469PubMedCrossRefGoogle Scholar

Copyright information

© The Japanese Society of Pharmacognosy and Springer Japan 2012

Authors and Affiliations

  • Xiaohui Pang
    • 1
  • Linchun Shi
    • 1
  • Jingyuan Song
    • 1
  • Xiaochen Chen
    • 1
  • Shilin Chen
    • 1
  1. 1.Institute of Medicinal Plant DevelopmentChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingPeople’s Republic of China

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