대한본초학회지 (The Korea Journal of Herbology)

  • 제33권3호
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  • Pages.25-35
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  • 2018
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  • 1229-1765(pISSN)
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  • 2288-7199(eISSN)
대한본초학회 (The Korea Association of Herbology)
권호 표지
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matK 증폭용 primer 개발 및 염기서열 분석을 통한 정력자(葶藶子) 유전자 감별

Molecular authentication of Lepidii seu Descurainiae Semen by the development of matK amplification primers and analysis of sequences

  • 문병철 (한국한의학연구원 K-herb 연구단) ;
  • 김욱진 (한국한의학연구원 K-herb 연구단) ;
  • 양선규 (한국한의학연구원 K-herb 연구단) ;
  • 박인규 (한국한의학연구원 K-herb 연구단) ;
  • 여상민 (한국한의학연구원 K-herb 연구단) ;
  • 노푸름 (한국한의학연구원 K-herb 연구단)
  • Moon, Byeong Cheol (K-herb Research Center, Korea Institute of Oriental Medicine) ;
  • Kim, Wook Jin (K-herb Research Center, Korea Institute of Oriental Medicine) ;
  • Yang, Sungyu (K-herb Research Center, Korea Institute of Oriental Medicine) ;
  • Park, Inkyu (K-herb Research Center, Korea Institute of Oriental Medicine) ;
  • Yeo, Sang Min (K-herb Research Center, Korea Institute of Oriental Medicine) ;
  • Noh, Pureum (K-herb Research Center, Korea Institute of Oriental Medicine)
  • 투고 : 2018.03.24
  • 심사 : 2018.05.25
  • 발행 : 2018.05.30
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초록

Objectives : Lepidii seu Descurainiae Semen has been frequently adulterated with the seeds of several inauthentic plant species. However, the accurate identification of these plant seeds is very difficult. To develop a reliable genetic authentication tool for Lepidii seu Descurainiae Semen, we analyzed matK sequence. Methods : To obtain the matK sequences of plant materials, genomic DNA was extracted from 24 samples and PCR amplification was carried out using matK-AF/matK-8R universal primer set and matK-LDSF/matK-LDSR primer set. For identifying species-specific nucleotides and phylogenetic analysis, matK regions were sequenced and comparatively analyzed by the ClustalW and Maximum Likelihood method. Results : We developed a new primer set to amplify matK region in Lepidii seu Descurainiae Semen and closely related plant samples. From the comparative analysis of matK sequences, we identified species-specific marker nucleotides for D. sophia, L. apetalum, L. latifolium, E. cheiranthoides, E. macilentum, and D. nemorosa, respectively. Furthermore, phylogenetic analysis revealed clear classification depending on the species. These results indicated that the matK sequence obtained a new primer set in this study was useful to identify Lepidii seu Descurainiae Semen in species level. Conclusions : We developed a primer set and identified species-specific marker nucleotides enough to distinguish authentic Lepidii seu Descurainiae Semen and adulterants at the species level based on the matK sequences. These genetic tool will be useful to prevent adulteration and to standardize the quality of Lepidii seu Descurainiae Semen.

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참고문헌

  1. Korea Institute of Oriental Medicine. Defining Dictionary for Medicinal Herbs [Korean], Retrieved March. 5, 2018, from http://boncho.kiom.re.kr/codex/.
  2. Shi P, Chao L, Wang T, Liu E, Han L, Zong Q, Li X, Zhang Y, Wang T. New boiactive flavonoid glycosides isolated from the seeds of Lepidium apetalum Willd. Fitoterapia. 2015 ; 103 : 197-205. https://doi.org/10.1016/j.fitote.2015.04.007
  3. Yang S, Kim WJ, Choi G, Yeo SM, Moon BC. Morphological authentication studies on the original plants of Lepidii seu Descurainiae Semen. Korean Herb Med Inf. 2016 ; 4(3) : 27-35.
  4. Park JC, Choi G. Review on Herbal Medicinal Materials in the Korean Pharmacopoeia and the Korean Herbal Pharmacopoeia. Korean Herb Med Inf. 2016 ; 4(2) : 9-35.
  5. Korea Food & Drug Administration (KFDA). Coloured Illustrations for Discrimination of Herbal Medicine. Homibooks, Daejeon, Korea. 2009 : 196-7.
  6. Choi G. List of Fake Korean Medicinal Herbs. Korea Institute of Oriental Medicine, Daejeon, Korea. 2016 : 75.
  7. Korea Food & Drug Administration (KFDA). The gudeline on the visual and organoleptic examination of herbal Medicine. Emun. co., Seoul, Korea. 2009 : 552-3.
  8. Zhou X, Tang L, Wu H, Zhou G, Wang T, Kou Z, Li S, Wang Z. Chemometric analyses for the characterization of raw and processed seeds for Descurainia sophia (L.) based on HPLC fingerprints. J Pharm Biomed Anal. 2015 : 111 : 1-6. https://doi.org/10.1016/j.jpba.2015.03.010
  9. Guo D, Chen NN. Identification of Descurainia sophia and Semen Plantaginis by HPCE. China Pharm. 2009 ; 20 : 683-4.
  10. Joshi K, Chavan P, Warude D, Patwardhan B. Molecular markers in herbal drug technology. Curr Sci. 2004 ; 87 : 159-65.
  11. Shucher NJ, Carles MC. Genome-based approaches to the authentication of medicinal plants. Planta Med. 2008 ; 74 : 603-23. https://doi.org/10.1055/s-2008-1074517
  12. White TJ, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes or phylogenetics. In: PCR Protocols: a guide to methods and amplifications. Academic Press, New York, USA. 1990 : 315-22.
  13. Kato H, Oginuma K, Gu Z, Hammel B, Tobe H. Phylogenetic relationships of Betulaceae based on matK sequences with particular reference at the positions of Ostryopsis . Acta Phytotax. Geobot. 1998 ; 49 : 89-97.
  14. Olmstead RG, Reeves PA. Evidence for the polyphyly of the Scropulariaceae based on choloroplast rbc L and ndh F sequences. Ann Missouri Bot Gard. 1995 ; 82 : 176-93. https://doi.org/10.2307/2399876
  15. Sang T, Crawford DN, Stuessy TF. Chloroplast DNA phylogeny, reticulate evolution and biogeography of Paeonia (Paeoniaceae). Am J Bot. 1997 ; 84 : 1120-36. https://doi.org/10.2307/2446155
  16. CBOL Plant Working Group1, A DNA barcode for land plants. Proc Natl Acad Sci USA. 2009 ; 106 : 12794-7. https://doi.org/10.1073/pnas.0905845106
  17. Hollingsworth PM, Graham SW, Little DP. Choosing and using a plant DNA barcode. PLoS ONE. 2011 ; 6(5) : e19254. https://doi.org/10.1371/journal.pone.0019254
  18. Roy S, Tyagi A, Shukla V, Kumar A, Singh UM, Chaudhary LB, Datt B, Bag SK, Singh PK, Nair NK, Husain T, Tuli R. Universal plant DNA barcode loci may not work in complex groups: a case study with Indian Berberis species. PLoS ONE. 2010 ; 5(10) : e13674. https://doi.org/10.1371/journal.pone.0013674
  19. Saddhe AA, Kumar K. DNA barcoding of plants: Selection of core markers for taxonomic groups. Plant Science Today 2018 ; 5(1) : 9-13.
  20. Zhang ZL, Song MF, Guan YH, Li HT, Niu YF, Zhang LX, Ma XJ. DNA barcoding in medicinal plants: testing the potential of a proposed barcoding marker for identification of Uncaria species from China. Biochem Syst Ecol. 2015 ; 60 : 8-14. https://doi.org/10.1016/j.bse.2015.02.017
  21. Park I, Kim WJ, Yang S, Yeo SM, Li H, Moon BC. The complete chloroplast genome sequence of Aconitum coreanum and Aconitum carmichaelii and comparative analysis with other Aconitum species. PLoS ONE. 2017 ; 12(9) : e0184257. https://doi.org/10.1371/journal.pone.0184257
  22. Kim WJ, Yang S, Choi G, Moon BC. Identification of marker nucleotides for the molecular authentication of Araliae Continentalis Radix based on the analysis of universal DNA barcode, mat K and rbcL, sequences. Kor J Herbol. 2016 ; 31(5) : 15-23. https://doi.org/10.6116/kjh.2016.31.5.15.
  23. Hall TA. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser. 1999 ; 41 : 95-8.
  24. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016 ; 33(7) : 1870-4. https://doi.org/10.1093/molbev/msw054
  25. Javinen P, Palme A, Morales LO, Lannenpaa M, Keinanen M, Sopanen T, Lascoux M. Phylogenetic relationships of Betula species (Betulaceae) based on nuclear adh and chloroplast matK sequence. Am J Bot. 2004 ; 91(11) : 1834-45. https://doi.org/10.3732/ajb.91.11.1834