Internal Transcribed Spacer Barcoding DNA Region Coupled with High Resolution Melting Analysis for Authentication of Panax Species
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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.
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.
참고문헌 (27)
- Baldwin BG., "Phylogenetic utility of the internal transcribed spacers of nuclear ribosomal DNA in plants : An example from the compositae", Molecular Phylogenetics and Evolution, 1, pp.3~16 (1992)
- 방경환, "PNA 바이오칩을 이용한 고려인삼 품종 판별", 한국약용작물학회지, 20(5), pp.387~392 (2012), 한국약용작물학회
- Bosmali I, "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, pp.141~147 (2012)
- Chan TW, "Differentiation and authentication of Panax ginseng, Panax quinquefolius, and ginseng products by using HPLC", MS, 2000, pp.72, 1281~1287
- Choi HK, "A phylogenetic analysis of Panax(Araliaceae) : Integrating cpDNA restriction site and nuclear rDNA ITS sequence data", Plant Systematics and Evolution, 224, pp.109~120 (2000)
- Cui JF, "Gynostemma pentaphyllum : Identification of major sapogenins and differentiation from Panax species", European Journal of Pharmaceutical Sciences, 8, pp.187~191 (1999)
- Ganopoulos I, "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, pp.2281~2286 (2013)
- Ganopoulos I, "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, pp.1322~1328 (2012)
- Jaakola L, "Novel approaches based on DNA barcoding and high resolution melting of amplicons for authenticity analyses of berry species", Food Chemistry, 123, pp.494~500 (2010)
- 조익현, "고려인삼 신품종 ‘천량’ 특이적 DNA 판별 마커 개발", 한국약용작물학회지, 22(6), pp.429~434 (2014), 한국약용작물학회[DOI]
- Kalivas A, "DNA barcode ITS2 coupled with high resolution melting(HRM)analysis for taxonomic identification of Sideritis species growing in Greece", Molecular Biology Reports, 41, pp.5147~5155 (2014)
- Jinho Kang, "NMR-Based Metabolomics Approach for the Differentiation of Ginseng (Panax ginseng) Roots from Different Origins", Archives of Pharmacal Research, 31(3), pp.330~336 (2008), 대한약학회
- Kim OT, "Molecular authentication of ginseng cultivars by comparison of internal transcribed spacer and 5. 8S rDNA sequences", Plant Biotechnology Reports, 1, pp.163~167 (2007)
- Lee OR, "Authentication of medicinal plants by SNP-based multiplex PCR", Methods in Molecular Biology, 862, pp.135~147 (2012)
- Li WK, "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, pp.1637~1643 (2001)
- Madesis P, "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, pp.576~582 (2012)
- Mao Q, "Discrimination of leaves of Panax ginseng and P. quinquefolius by ultra high performance liquid chromatography quadrupole", time of-flight mass spectrometry based metabolomics approach, 2014, pp.97, 129~140
- Park MJ, "Molecular identification of Korean ginseng by amplification refractory mutation system-PCR", Food Research International, 39, pp.568~574 (2006)
- Shaw PC, "Authentication of Panax species and their adulterants by random primed polymerase chain reaction", Planta Medica, 61, pp.466~469 (1995)
- Stephens AJ, "High resolution melting analysis of the spa repeat region of Staphylococcus aureus", Clinical Chemistry, 54, pp.432~436 (2008)
- Wang H, "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, pp.972~976 (2011)
- Wen J, "Phylogeny and biogeography of Panax L. (the ginseng genus, Araliaceae) : Inferences from ITS sequences of nuclear ribosomal DNA", Molecular Phylogenetics and Evolution, 6, pp.167~177 (1996)
- Wittwer CT, "High resolution genotyping by amplicon melting analysis using LCGreen", Clinical Chemistry, 49, pp.853~860 (2003)
- 양승옥, "Comparative Analysis of Metabolites in Roots of Panax ginseng Obtained from Different Sowing Methods", 한국약용작물학회지, 22(1), pp.17~22 (2014), 한국약용작물학회[DOI]
- Yang WZ, "Rapid chemical profiling of saponins in the flower buds of Panax notoginseng by integrating MCI gel column chromatography and liquid chromatography", mass spectrometry analysis, 2013, pp.139, 762~769
- Yao H, "Use of ITS2 region as the universal DNA barcode for plants and animals", PLoS ONE, 5, pp.e13102~ (2010)
- Zhou L, "High resolution DNA melting curve analysis to establish HLA genotype identity", Tissue Antigens, 64, pp.156~164 (2004)