Korean Journal of Medicinal Crop Science (한국약용작물학회지)

The Korean Society of Medicinal Crop Science (한국약용작물학회)
권호 표지
DOI QR코드

DOI QR Code

Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species

DNA 바코딩과 고해상 융해곡선분석에 기반한 인삼속 식물의 종 판별

  • 방경환 (농촌진흥청 국립원예특작과학원 기획조정과) ;
  • 김영창 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 임지영 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 김장욱 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 이정우 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 김동휘 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 김기홍 (농촌진흥청 국립원예특작과학원 인삼특작부) ;
  • 조익현 (농촌진흥청 국립원예특작과학원 인삼특작부)
  • Received : 2015.09.08
  • Accepted : 2015.10.07
  • Published : 2015.12.30
Download PDF
⟨ Previous Next ⟩

Abstract

Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

Keywords

References

  1. Baldwin BG. (1992). Phylogenetic utility of the internal transcribed spacers of nuclear ribosomal DNA in plants: An example from the compositae. Molecular Phylogenetics and Evolution. 1:3-16. https://doi.org/10.1016/1055-7903(92)90030-K
  2. Bang KH, Jo IH, Kim YC, Kim JU, Park HW, Shin MR, Kim YB, Kim OT, Hyun DY, Kim DH and Cha SW. (2012). Molecular identification of Korean ginseng cultivars(Panax ginseng C. A. Mey.) using peptide nucleic acid(PNA) microarray. Korean Journal of Medicinal Crop Science. 20:387-392. https://doi.org/10.7783/KJMCS.2012.20.5.387
  3. Bosmali I, Ganopoulos I, Madesis P and Tsaftaris A. (2012). Microsatellite and DNA-barcode regions typing combined with high resolution melting(HRM) analysis for food forensic uses: A case study on lentils(Lens culinaris). Food Research International. 46:141-147. https://doi.org/10.1016/j.foodres.2011.12.013
  4. Chan TW, But PP, Cheng SW, Kwok IM, Lau FW and Xu HX. (2000). Differentiation and authentication of Panax ginseng, Panax quinquefolius, and ginseng products by using HPLC/MS. Analytical Chemistry. 72:1281-1287. https://doi.org/10.1021/ac990819z
  5. Choi HK and Wen J. (2000). A phylogenetic analysis of Panax (Araliaceae): Integrating cpDNA restriction site and nuclear rDNA ITS sequence data. Plant Systematics and Evolution. 224:109-120. https://doi.org/10.1007/BF00985269
  6. Cui JF, Eneroth P and Bruhn JB. (1999). Gynostemma pentaphyllum: Identification of major sapogenins and differentiation from Panax species. European Journal of Pharmaceutical Sciences. 8:187-191. https://doi.org/10.1016/S0928-0987(99)00013-5
  7. Ganopoulos I, Bazakos C, Madesis P, Kalaitzis P and Tsaftaris A. (2013). Barcode DNA high resolution melting(Bar-HRM) analysis as a novel close-tubed and accurate tool for olive oil forensic use. Journal of the Science of Food and Agriculture. 93:2281-2286. https://doi.org/10.1002/jsfa.6040
  8. Ganopoulos I, Madesis P and Tsaftaris A. (2012). Universal ITS2 barcoding DNA region coupled with high resolution melting(HRM) analysis for seed authentication and adulteration testing in Leguminous forage and pasture species. Plant Molecular Biology Reporter. 30:1322-1328. https://doi.org/10.1007/s11105-012-0453-3
  9. Jaakola L, Suokas M and Hggman H. (2010). Novel approaches based on DNA barcoding and high resolution melting of amplicons for authenticity analyses of berry species. Food Chemistry. 123:494-500. https://doi.org/10.1016/j.foodchem.2010.04.069
  10. Jo IH, Kim YC, Kim JU, Lee SH, Lim JY, Moon JY, Noh BS, Hyun DY, Kim DH, Kim KH and Bang KH. (2014). A rapid identification of Korean ginseng cultivar, Cheonryang, using specific DNA markers. Korean Journal of Medicinal Crop Science. 22:429-434. https://doi.org/10.7783/KJMCS.2014.22.6.429
  11. Kalivas A, Ganopoulos I, Xanthopoulou A, Chatzopoulou P, Tsaftaris A and Madesis P. (2014). DNA barcode ITS2 coupled with high resolution melting(HRM) analysis for taxonomic identification of Sideritis species growing in Greece. Molecular Biology Reports. 41:5147-5155. https://doi.org/10.1007/s11033-014-3381-5
  12. Kang J, Lee S, Kang S, Kwon HN, Park JH, Kwon SW and Park S. (2008). NMR-based metabolomics approach for the differentiation of ginseng(Panax ginseng) roots from different origins. Archives of Pharmacal Research. 31:330-336. https://doi.org/10.1007/s12272-001-1160-2
  13. Kim OT, Bang KH, In DS, Lee JW, Kim YC, Shin YS, Hyun DY, Lee SS, Cha SW and Seong NS. (2007). Molecular authentication of ginseng cultivars by comparison of internal transcribed spacer and 5.8S rDNA sequences. Plant Biotechnology Reports. 1:163-167. https://doi.org/10.1007/s11816-007-0019-2
  14. Lee OR, Kim MK and Yang DC. (2012). Authentication of medicinal plants by SNP-based multiplex PCR. Methods in Molecular Biology. 862:135-147. https://doi.org/10.1007/978-1-61779-609-8_11
  15. Li WK and Fitzloff JF. (2001). A validated method for quantitative determination of saponins in notoginseng(Panax notoginseng) using high performance liquid chromatography with evaporative light scattering detection. Journal of Pharmacy and Pharmacology. 53:1637-1643. https://doi.org/10.1211/0022357011778241
  16. Madesis P, Ganopoulos I, Anagnostis A and Tsaftaris A. (2012). The application of Bar-HRM(Barcode DNA-High Resolution Melting) analysis for authenticity testing and quantitative detection of bean crops(Leguminosae) without prior DNA purification. Food Control. 25:576-582. https://doi.org/10.1016/j.foodcont.2011.11.034
  17. Mao Q, Bai M, Xu JD, Kong M, Zhu LY, Zhu H, Wang Q and Li SL. (2014). Discrimination of leaves of Panax ginseng and P. quinquefolius by ultra high performance liquid chromatography quadrupole/time of-flight mass spectrometry based metabolomics approach. Journal of Pharmaceutical and Biomedical Analysis. 97:129-140. https://doi.org/10.1016/j.jpba.2014.04.032
  18. Park MJ, Kim MK, In JG and Yang DC. (2006). Molecular identification of Korean ginseng by amplification refractory mutation system-PCR. Food Research International. 39:568-574. https://doi.org/10.1016/j.foodres.2005.11.004
  19. Shaw PC and But PP. (1995). Authentication of Panax species and their adulterants by random primed polymerase chain reaction. Planta Medica. 61:466-469. https://doi.org/10.1055/s-2006-958138
  20. Stephens AJ, Inman-Bamber J, Giffard PM and Huygens F. (2008). High resolution melting analysis of the spa repeat region of Staphylococcus aureus. Clinical Chemistry. 54:432-436. https://doi.org/10.1373/clinchem.2007.093658
  21. Wang H, Kim MK, Kwon WS, Jin H, Liang Z and Yang DC. (2011). Molecular authentication of Panax ginseng and ginseng products using robust SNP markers in ribosomal external transcribed spacer region. Journal of Pharmaceutical and Biomedical Analysis. 55:972-976. https://doi.org/10.1016/j.jpba.2011.03.037
  22. Wen J and Zimmer EA. (1996). Phylogeny and biogeography of Panax L.(the ginseng genus, Araliaceae): Inferences from ITS sequences of nuclear ribosomal DNA. Molecular Phylogenetics and Evolution. 6:167-177. https://doi.org/10.1006/mpev.1996.0069
  23. Wittwer CT, Reed GH, Gundry CN, Vandersteen JG and Pryor RJ. (2003). High resolution genotyping by amplicon melting analysis using LCGreen. Clinical Chemistry. 49:853-860. https://doi.org/10.1373/49.6.853
  24. Yang SO, Lee SW, Kim YO, Lee SW, Kim NH, Choi HK, Jung JY, Lee DH and Shin YS. (2014). Comparative analysis of metabolites in roots of Panax ginseng obtained from different sowing methods. Korean Journal of Medicinal Crop Science. 22:17-22. https://doi.org/10.7783/KJMCS.2014.22.1.17
  25. Yang WZ, Bo T, Ji S, Qiao X, Guo DA and Ye M. (2013). Rapid chemical profiling of saponins in the flower buds of Panax notoginseng by integrating MCI gel column chromatography and liquid chromatography/mass spectrometry analysis. Food Chemistry. 139:762-769. https://doi.org/10.1016/j.foodchem.2013.01.051
  26. Yao H, Song J, Liu C, Luo K, Han J, Li Y, Pang X, Xu H, Zhu Y, Xiao P and Chen S. (2010). Use of ITS2 region as the universal DNA barcode for plants and animals. PLoS ONE. 5:e13102. https://doi.org/10.1371/journal.pone.0013102
  27. Zhou L, Vandersteen J, Wang L, Fuller T, Taylor M, Palais B and Wittwer CT. (2004). High resolution DNA melting curve analysis to establish HLA genotype identity. Tissue Antigens. 64:156-164. https://doi.org/10.1111/j.1399-0039.2004.00248.x

Cited by

  1. Analysis of the chloroplast genome and SNP detection in a salt tolerant breeding line in Korean ginseng vol.43, pp.4, 2016, https://doi.org/10.5010/JPB.2016.43.4.417