Morphological resemblance among Cymbopogon distans species and their adulterants which are procured from different markets in the form of dried or fresh plant tissues represents a serious problem for quality and safety of medicinal plants, as it supports frauds for substitution. In order to assure the quality control of C. distans species, DNA barcode, microscopic identification and High Performance liquid chromatography (HPLC) fingerprint were synergistically used to discriminate C. distans from its adulterants. In this work, the internal transcribed spacer 2 (ITS2) was chosen for distinguishing C. distans from their usual adulterants from 5 provinces of China. Sequences were obtained after removal of the 5.8S and 28S sections. A multiple sequence alignment was finalized. Results exhibited that ITS2 performed well, with 100% of genera being accurately distinguished. Additionally, finding indicates that the upper epidermis in leaf of C. distans was composed of one layer of wide elongated cells called Bulliform cells whereas in C. distans the upper epidermis consist of one layer cell, thus these feactures are very important for the anatomy identification. The HPLC fingerprint method was also developed, the similarities of 6 batches of C. distans samples were all more than 0.93, indicating that the samples from different geographical origins shared similar HPLC fingerprints. And the similarities between C. distans, C. citratus, C. flexuosus and Imperata cylindrica were all less than 0.93, suggesting that there was significance difference between C. distans and its adulterants. Finally, it was concluded that the DNA barcode, HPLC fingerprint and microscopic methods could effectively authenticate the quality of C. distans from their adulterants and can provide accurate and reliable information to tackle the complex quality issue of C. distans in markets. This is the first report of detailed analysis of the C. distans for effective quality and safety.
Published in | Journal of Diseases and Medicinal Plants (Volume 4, Issue 2) |
DOI | 10.11648/j.jdmp.20180402.11 |
Page(s) | 35-47 |
Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
Copyright |
Copyright © The Author(s), 2018. Published by Science Publishing Group |
Internal Transcribed Spacer (ITS2), DNA barcode, Cymbopogon distans, High Performance Liquid Chromatography (HPLC)
[1] | Baruah, J. B., Gogoi, K Das, N. M Ahmed, DK Sarmah, M. Lal, B.S. Bhau (2017). Genetic diversity study among Cymbopogon species from NE-India using RAPD and ISSR markers. Industrial Crops & Products 95: 235-243. |
[2] | Akhila A., Bertea C. M., BigheliA., CasanovaJ., Khunkitti W., Maffei M. E., Mathur A. K., Moyler D. A., Pandey A. K., Sumi H., Tiwari R., Yatagai C., (2009). Essential Oil- Bearing Grasses: The Genus Cymbopogon. Taylor & Francis Group, New York. |
[3] | Dou R, Liu Z, Yuan X, Xiangfei D, Bai R, Bi Z, Yang P, Yang Y, Dong Y, Su W, Li D, Mao C. PAMs ameliorates the imiquimod-induced psoriasis-like skin disease in mice by inhibition of translocation of NF-κB and production of inflammatory cytokines, PLoS One, 2017, 12(5): e0176823 |
[4] | Rao, B. L., (1997). Scope for development of new cultivars of Cymbopogons as a source of terpene chemicals. In: Handa, S. S., Kaul, M. K. (Eds.), Supplement to Cultivation and Utilization of Aromatic Plants. National Institute of Science Communication, Dr. KS. Krishnan Marg, New Delhi, India, pp. 71-83 |
[5] | Kool A, de Boer H. J, Krüger Å, Rydberg A, Abbad A, Björk L, et al. (2012). Molecular identification of commercialized medicinal plants in Southern Morocco. PLOS ONE. 7: e39459. pmid:22761800 |
[6] | Chen S, Pang X, Song J, Shi L, Yao H, Han J, et al. (2014). A renaissance in herbal medicine identification: from morphology to DNA. Biotechnol Adv.32: 1237–1244. pmid:25087935 |
[7] | SchlickSteiner B. C., Steiner F. M., Seifert B., Stauffer C., Christian E., Crozier RH, (2010). Integrative taxonomy: a multisource approach to exploring biodiversity. Annu Rev Entomol.55: 421-438. |
[8] | Li DZ, Gao LM, Li HT, Wang H, Ge XJ, Liu JQ, et al. Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants. ProcNatlAcad Sci. 2011; 108: 19641–19646. |
[9] | Hollingsworth ML, Clark A, Forrest LL, Richardson JR, Pennington RT, et al. (2009) Selecting barcoding loci for plants: evaluation of seven candidate loci with species-level sampling in three divergent groups of land plants. Molecular Ecology Resources 9: 439–457. |
[10] | Fazekas AJ, Burgess KS, Kesanakurti PR, Graham SW, Newmaster SG, et al. (2008). Multiple multilocus DNA barcodes from the plastid genome discriminate plant species equally well. PLoS ONE 3: e2802. |
[11] | CBOL Plant Working Group (2009) A DNA barcode for land plants. Proceedings of the National Academy of Sciences 106: 12794–12797. |
[12] | Fazekas AJ, Kesanakurti PR, Burgess KS, Percy DM, Graham SW, et al. (2009) Are plant species inherently harder to discriminate than animal species using DNA barcoding markers? Molecular Ecology Resources 9: 130–139. |
[13] | Hollingsworth PM, Graham SW, Little DP (2011) Choosing and Using a Plant DNA Barcode. PLoS ONE 6(5): e19254. https://doi.org/10.1371/journal.pone.0019254 |
[14] | Chen S, Yao H, Han J, Liu C, Song J, et al. (2010) Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species. PLoS ONE 5: e8613. |
[15] | Yao H, Song J, Liu C, Luo K, Han J, et al. (2010) Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS ONE 5: e13102. |
[16] | Li DZ, Gao LM, Li HT, Wang H, et al. China Plant BOL Group (2011) Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants. Proc Natl Acad Sci U S A 108(49): 19641–19646. |
[17] | World Health Organization: Guidelines for the Assessment of Herbal Medicines WHO, Munich, Geneva; 1991. |
[18] | FDA, (2004). Guidance for industry-botanical drug products. U.S. Food and Drug Administration. |
[19] | MEA, (2006). Guidanceon quality of herbal medicinal products/traditional herbal medicinal products. European Medicines Agency. |
[20] | Ram S., Verma, Rajendra C., Padalia, Amit Chauhan, (2013). Introduction of Cymbopogon distans (Nees ex Steud.) Wats to the sub-tropical India: Evaluation of essential-oil yield and chemical composition during annual growth. Industrial Crops and Products 49.858-863. |
[21] | Amit Chauhan, Ram S. Verma, Rajendra C. Padalia, Rakesh K. Upadhyay&Janak R. Bahl (2016): Post harvest storage effect on essential oil content and composition of Cymbopogon distans (Nees ex Steud.) Wats., Journal of Essential Oil Research, DOI: 10.1080/10412905.2016.1164762 |
[22] | Zhang JS, Zhao NN, Liu QZ, Liu ZL, Du SS, Zhou L, Deng ZW. Repellent constituents of essential oil of Cymbopogon distans aerial parts against two stored-product insects. J Agric Food Chem. (2011) Sep 28; 59(18):9910-5 |
[23] | Moore P. D., WebbJ. A, Collison M. E., (1991), Pollen analysis. Blackwell scientific publications. Google Scholar |
[24] | Johansen D. A. Plant microtechnique: Jeffrey's method, McGraw Hill Book Co., New York (NY) (1940), p. 104Google Scholar |
[25] | Chen Shouliang, Sylvia M. Phillips. ECHINOCHLOA P. Beauvois. Flora of China 22: 515–518. (2006). |
[26] | Pang X., J. Song, Y. Zhu, H. Xu, L. Huang, and S. Chen, “Applying plant DNA barcodes for Rosaceae species identification,” Cladistics, vol. 27, no. 2, pp. 165-170, (2011). |
[27] | Pang X. H., J. Y. Song, H. B. Xu, and H. Yao, (2012). “Using ITS2 barcode to identify ephedrae herba,” China Journal of Chinese MateriaMedica, vol. 37, pp. 1118-1121 |
[28] | Xin T. Y., H. Yao, K. Luo et al., (2012) “Stability and accuracy of the identification of Notopterygii Rhizoma et Radix using the ITS/ITS2 barcodes,” Acta Pharmaceutica Sinica, vol. 47, pp. 1098-1105. |
[29] | Hou D. Y., Song J. Y., Shi L. C., Ma X. C., Xin T. Y., Han J. P., et al., (2013). Stability and accuracy assessment of identification of traditional Chinese material medica using DNA barcoding: a case study on Flos Lonicerae Japonicae. Biomed Res. Int. 2013:549037. 10.1155/2013/549037 |
[30] | Xin T. Y., Yao H., Gao H. H., Zhou X. Z., Ma X. C., Xu C. Q., et al. (2013). Super food Lycium barbarum (Solanaceae) traceability via an internal transcribed spacer 2 barcode. Food Res. Int. 54, 1699–1704. |
[31] | Ben-David, T., Melamed, S., Gerson, U., Morin, S., (2007). ITS2 sequences as barcodes for identifying and analyzing spider mites (Acari: Tetranychidae). Experimental and Applied Acarology 41, 169-181 |
[32] | Li, Y., Zhou, X., Feng, G., Hu, H., Niu, L., Hebert, P. D. N., Huang, D., (2010). COI andITS2 sequences delimit species, reveal cryptic taxa and host specificity of fig-associated Sycophila (Hymenoptera, Eurytomidae). Molecular Ecology Resources 10, 31-40. |
[33] | Dou R, Bi zhen-fei, Bairui-xue, Renyao-yao, Tan rui, Li di quiang, Song liangke, Mao can quan, (2015). “Identification and analysis of corydalis boweri, Meconopsis horridula, and their closely related species of the same genus by using DNA barcode” China Journal of Chinese Materia Medica, 40(8). |
[34] | Prasad PK, Tandon V, Biswal DK, Goswami LM, Chatterjee A., (2009). Use of sequence motifs as barcodes and secondary structures of Internal Transcribed spacer 2 (ITS2, rDNA) for identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae). Bioinformation 3: 314-320. |
[35] | Qiao C, Han Q, Zhao Z, Wang Z, Xu L, Xu HX. Sequence analysis based on ITS1 region of nuclear ribosomal DNA of Amomum villosum and ten species of Alpinia. J Food Drug Anal 2009; 17:142-5. |
[36] | Gagan Shah, Maninderjit Kaur, Falgun Dhabiliya, RichaShri. Pharmacognostic Standardization of Cymbopogon citratus (dc.) stapf leaves. Pharmacognosy Journal, May-June 2012, Vol4, Issue 29. |
[37] | E. L. Kotina, B.-E. Van Wyk, P. M. Tilney, “Anatomy of the leaf and bark of Warburgia salutaris (Canellaceae), an important medicinal plant from South Africa”, South African Journal of Botany 94 (2014) 177-181 |
[38] | Ahlam Salih Eltahir and Bouran Ibrahim Abu EReish, Leaf and stem anatomy of Cymbopogon citratus and Cymbopogon schoenanthus in Sudan, J. Chem. Pharm. Res, 2010, 2(4): 766-771. |
APA Style
Diana Kavidia Muyembe, Dou Rong Kun, Song Zhilei, Fang Jin, Xue Yuan, et al. (2018). Molecular, Chemical and Microscopic Analysis of Medicinal Plant (Cymbopogon distans) from Its Adulterants. Journal of Diseases and Medicinal Plants, 4(2), 35-47. https://doi.org/10.11648/j.jdmp.20180402.11
ACS Style
Diana Kavidia Muyembe; Dou Rong Kun; Song Zhilei; Fang Jin; Xue Yuan, et al. Molecular, Chemical and Microscopic Analysis of Medicinal Plant (Cymbopogon distans) from Its Adulterants. J. Dis. Med. Plants 2018, 4(2), 35-47. doi: 10.11648/j.jdmp.20180402.11
AMA Style
Diana Kavidia Muyembe, Dou Rong Kun, Song Zhilei, Fang Jin, Xue Yuan, et al. Molecular, Chemical and Microscopic Analysis of Medicinal Plant (Cymbopogon distans) from Its Adulterants. J Dis Med Plants. 2018;4(2):35-47. doi: 10.11648/j.jdmp.20180402.11
@article{10.11648/j.jdmp.20180402.11, author = {Diana Kavidia Muyembe and Dou Rong Kun and Song Zhilei and Fang Jin and Xue Yuan and Piao Yang and Zou Jinjing and Mao Can Quan}, title = {Molecular, Chemical and Microscopic Analysis of Medicinal Plant (Cymbopogon distans) from Its Adulterants}, journal = {Journal of Diseases and Medicinal Plants}, volume = {4}, number = {2}, pages = {35-47}, doi = {10.11648/j.jdmp.20180402.11}, url = {https://doi.org/10.11648/j.jdmp.20180402.11}, eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jdmp.20180402.11}, abstract = {Morphological resemblance among Cymbopogon distans species and their adulterants which are procured from different markets in the form of dried or fresh plant tissues represents a serious problem for quality and safety of medicinal plants, as it supports frauds for substitution. In order to assure the quality control of C. distans species, DNA barcode, microscopic identification and High Performance liquid chromatography (HPLC) fingerprint were synergistically used to discriminate C. distans from its adulterants. In this work, the internal transcribed spacer 2 (ITS2) was chosen for distinguishing C. distans from their usual adulterants from 5 provinces of China. Sequences were obtained after removal of the 5.8S and 28S sections. A multiple sequence alignment was finalized. Results exhibited that ITS2 performed well, with 100% of genera being accurately distinguished. Additionally, finding indicates that the upper epidermis in leaf of C. distans was composed of one layer of wide elongated cells called Bulliform cells whereas in C. distans the upper epidermis consist of one layer cell, thus these feactures are very important for the anatomy identification. The HPLC fingerprint method was also developed, the similarities of 6 batches of C. distans samples were all more than 0.93, indicating that the samples from different geographical origins shared similar HPLC fingerprints. And the similarities between C. distans, C. citratus, C. flexuosus and Imperata cylindrica were all less than 0.93, suggesting that there was significance difference between C. distans and its adulterants. Finally, it was concluded that the DNA barcode, HPLC fingerprint and microscopic methods could effectively authenticate the quality of C. distans from their adulterants and can provide accurate and reliable information to tackle the complex quality issue of C. distans in markets. This is the first report of detailed analysis of the C. distans for effective quality and safety.}, year = {2018} }
TY - JOUR T1 - Molecular, Chemical and Microscopic Analysis of Medicinal Plant (Cymbopogon distans) from Its Adulterants AU - Diana Kavidia Muyembe AU - Dou Rong Kun AU - Song Zhilei AU - Fang Jin AU - Xue Yuan AU - Piao Yang AU - Zou Jinjing AU - Mao Can Quan Y1 - 2018/05/21 PY - 2018 N1 - https://doi.org/10.11648/j.jdmp.20180402.11 DO - 10.11648/j.jdmp.20180402.11 T2 - Journal of Diseases and Medicinal Plants JF - Journal of Diseases and Medicinal Plants JO - Journal of Diseases and Medicinal Plants SP - 35 EP - 47 PB - Science Publishing Group SN - 2469-8210 UR - https://doi.org/10.11648/j.jdmp.20180402.11 AB - Morphological resemblance among Cymbopogon distans species and their adulterants which are procured from different markets in the form of dried or fresh plant tissues represents a serious problem for quality and safety of medicinal plants, as it supports frauds for substitution. In order to assure the quality control of C. distans species, DNA barcode, microscopic identification and High Performance liquid chromatography (HPLC) fingerprint were synergistically used to discriminate C. distans from its adulterants. In this work, the internal transcribed spacer 2 (ITS2) was chosen for distinguishing C. distans from their usual adulterants from 5 provinces of China. Sequences were obtained after removal of the 5.8S and 28S sections. A multiple sequence alignment was finalized. Results exhibited that ITS2 performed well, with 100% of genera being accurately distinguished. Additionally, finding indicates that the upper epidermis in leaf of C. distans was composed of one layer of wide elongated cells called Bulliform cells whereas in C. distans the upper epidermis consist of one layer cell, thus these feactures are very important for the anatomy identification. The HPLC fingerprint method was also developed, the similarities of 6 batches of C. distans samples were all more than 0.93, indicating that the samples from different geographical origins shared similar HPLC fingerprints. And the similarities between C. distans, C. citratus, C. flexuosus and Imperata cylindrica were all less than 0.93, suggesting that there was significance difference between C. distans and its adulterants. Finally, it was concluded that the DNA barcode, HPLC fingerprint and microscopic methods could effectively authenticate the quality of C. distans from their adulterants and can provide accurate and reliable information to tackle the complex quality issue of C. distans in markets. This is the first report of detailed analysis of the C. distans for effective quality and safety. VL - 4 IS - 2 ER -