Abstract
The carapace of the tortoise Chinemys reevesii is an ingredient of “Guijia”, a traditional Chinese medicine. However, C. reevesii is difficult to raise in aquaculture and is rare in the wild. Counterfeit tablets are made from carapaces of other species. In addition to C. reevesii, other species including Mauremys sinensis, Indotestudo elongate and Trachemys scripta have been used in Plastrum Testudinis as well. After processing, these carapaces are difficult to identify on the basis of morphological characteristics, which impedes law enforcement. Our study used DNA barcoding technology to identify C. reevesii and its substitutes. We extracted concentrated genomic DNA for PCR amplification. Based on the analysis of 61 full-length COI sequences, we designed four pairs of mini-barcode primers: Tu-A, Tu-B, Tu-C and Tu-D. The Tu-B primers sequenced genomic DNA with a success rate of 76.47%, and the Tu-D primers sequenced genomic DNA with a success rate of 88.24%. The identification efficiency of these two mini-barcodes was 70.59% and 64.71%, and the overall identification efficiency was approximately 76.47%. Similarly, a set of mini barcode systems was generated, which may provide an effective and low-cost method for the identification of authentic tortoise shells.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Chen, J., Jiang, Z., Li, C., Ping, X., Cui, S., Tang, S., Chu, H., Liu, B., 2015. Identification of ungulates used in a traditional Chinese medicine with DNA barcoding technology. Ecol. Evol. 5, 1818–1825.
Chinese Pharmacopoeia, 2015. China Pharmacopoeia Committee., pp. 180–181.
Dong, W., Liu, H., Xu, C., Zuo, Y., Chen, Z., Zhou, S., 2014. A chloroplast genomic strategy for designing taxon specific DNA mini-barcodes: a case study on ginsengs. BMC Genet. 15, https://doi.org/10.1186/s12863-014-0138-z.
Hajibabaei, M., Smith, M.A., Janzen, D.H., Rodriguez, J.J., Whitfiel, J.B., Hebert, P.D., 2006a. A minimalist barcode can identify a specimen whose DNA is degraded. Mol. Ecol. Notes 6, 959–964.
Hebert, P.D., Cywinska, A., Ball, S.L., deWaard, J.R., 2003. Biological identifications through DNA barcodes. Proc. R. Soc. Ser. B 270, 313–321.
Hebert, P.D., Stoeckle, M.Y., Zemlak, T.S., Francis, C.M., 2004. Identification of birds through DNA barcodes. PLoS Biol. 2, 1657–1663.
Lee, J.C., Tsai, L.C., Liao, S.P., Linacre, A., Hsieh, H.M., 2009. Species identification using the cytochrome b gene of commercial turtle shells. Forensic Sci. Int. Genet. 3, 67–73.
Hajibabaei, M., Janzen, D.H., Burns, J.M., Hallwachs, W., Hebert, P.D.N., 2006b. DNA barcodes distinguish species of tropical Lepidoptera. Proc. Natl. Acad. Sci. U. S. A. 103, 968–971.
Shokralla, S., Hellberg, R.S., Handy, S.M., King, I., Hajibabaei, M., 2015. A DNA-mini-barcoding system for authentication of processed fish products. Sci. Rep. 5, https://doi.org/10.1038/srep15894.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S., 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28, 2731–2739.
Yang, 2001. Identification of Plastrum Testudinis and its several adulterants. J. Snake 13, 77–78.
Yi, 2010. Experience of differential shell Plastrum Testudinis its adulterants. Tradit. Chin. Med. 20, 71–72.
Ward, R.D., Zemlak, T.S., Innes, B.H., Last, P.R., Hebert, P.D., 2006. DNA barcoding Australia’s fish species. Philos. Trans. R. Soc. B 360, 1847–1857.
Ward, R.D., Hanner, R., Hebert, P.D., 2009. The campaign to DNA barcode all fishes, FISH-BOL. J. Fish Biol. 74, 329–356.
Wu, 2012. Protection of endangered turtles. China Awards Sci. Technol. 151, 120–125.
Zhong, Y., Wang, K., Zhang, X., Cai, X., Chen, Y., Deng, Y., 2015. Nephrokeli, a Chinese herbal formula, may improve IgA nephropathy through regulation of the sphingosine-1-phosphate pathway. PLOS ONE 10, e0116873.
Author information
Authors and Affiliations
Corresponding author
Additional information
Author contributions
Conceived and designed the experiments: MYC and CSL. Performed the experiments: MYC, XYL, and JM. Sample collection: MYC, ML, and XNL. Wrote the paper: MYC and JM. Edited the manuscript: MYC and XH.
Rights and permissions
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Chen, M., Han, X., Ma, J. et al. Identification of Plastrum Testudinis used in traditional medicine with DNA mini-barcodes. Rev. Bras. Farmacogn. 28, 267–272 (2018). https://doi.org/10.1016/j.bjp.2018.04.008
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1016/j.bjp.2018.04.008