Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Development of Primer Sets for the Detection of Polygonum multiflorum, Cynanchum wilfordii and C. auriculatum

유전자 마커를 이용한 하수오, 백수오 및 이엽우피소 종 판별법 개발

  • Kim, Kyu-Heon (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Yong-Sang (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Kim, Mi-Ra (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Ho-Yeon (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Kyu Ha (Herbal Medicine Research Division, National Institute of Food and Drug Safety Evaluation) ;
  • Kim, Jong Hwan (Herbal Medicine Research Division, National Institute of Food and Drug Safety Evaluation) ;
  • Seong, Rack Seon (Herbal Medicine Research Division, National Institute of Food and Drug Safety Evaluation) ;
  • Kang, Tae Sun (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Lee, Jin-Ha (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety) ;
  • Jang, Young-Mi (New Hazardous Substance Team, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety)
  • 김규헌 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 김용상 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 김미라 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 이호연 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 이규하 (식품의약품안전평가원 생약연구과) ;
  • 김종환 (식품의약품안전평가원 생약연구과) ;
  • 성락선 (식품의약품안전평가원 생약연구과) ;
  • 강태선 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 이진하 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀) ;
  • 장영미 (식품의약품안전처 식품의약품안전평가원 신종유해물질팀)
  • Received : 2015.04.06
  • Accepted : 2015.07.24
  • Published : 2015.09.30
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Abstract

The aim of this study was to develop rapid screening method for the identification of Chinese herbal medicine species with similar appearance, Polygonum multiflorum, Cynanchum wilfordii and C. auriculatum, by using genetic markers. As a genetic marker, psbA-trnH gene in chloroplast was selected due to differences in sequence among the three species. Species-specific primers were designed based on the sequences of the marker gene of P. multiflorum, C. wilfordii, and C. auriculatum, and the expected size of PCR products was 160, 147, and 119 bp, respectively. Under the developed conditions, cross-reaction was not detected among these three plant species. To confirm the efficiency of our species-specific primers, the optimized method was applied to a variety of processed products composed of mostly P. multiflorum and C. wilfordii, demonstrating that our method was a rapid and easy screening assay. Our findings suggest this screening method can be utilized to prevent the distribution of economically motivated adulteration food and to improve consumer's right.

본 연구에서는 건강기능성 식품원료로서 이용 빈도가 증가하고 있는 하수오, 백수오 및 이엽우피소에 대한 종 특이 프라이머를 개발하였다. 개발된 종 특이 프라이머는 하수오, 백수오 및 이엽우피소에 대해 원물뿐만 아니라 육안 확인이 어려운 가공식품 등을 대상으로 사용원료의 진위여부를 빠르고 정확하게 확인할 수 있었다. 따라서 본 연구에서 개발한 종 특이 PCR방법은 기존의 일반 프라이머를 사용하는 방법이 가지고 있는 식품 적용 한계를 극복할 수 있을 것이라고 판단하였다.

Keywords

References

  1. Chase M.W., Cowan R.S., Holling sworth P.M., Van den Berg C. and Madrinan S.: A proposal for a standardised protocol to barcode all land plants. Taxon, 56, 295-299 (2007).
  2. Chen S., Yao H., Han J., Liu C., Song J., Shi L., Zhu Y., Ma X., Gao T., Pang X., Luo K., Li Y., Li X., Jia X., Lin Y. and Leon C.: Validation of the ITS2 region as a Novel DNA barcode for identifying medicinal plant species., PLoS One, 5, e8613 (2010). https://doi.org/10.1371/journal.pone.0008613
  3. Choi H.S., Zhu M.F., Kim C.S. and Lee J.H.: Studies of name and herbal origins of Ha-Soo-Oh. Korean Journal of Oriental Medicine, 9, 81-91 (2003).
  4. Focke F., Haase I. and Fischer M.: DNA-Based identification of Spices: DNA Isolation, Whole Genome Amplification, and Polymerase Chain Reaction. Journal of Agricultural and Food Chemistry, 59, 513-520 (2011). https://doi.org/10.1021/jf103702s
  5. Han J., Luan D.H.: Sow abortion caused by feeding Cynanchum auriculatum. Anim Husb Vet Med Xumu yu Shouyi, 16(6), 266 (1984).
  6. Harmer J.E., Farrall L., Ortach M.J., Valent B. and Chunmley F.G.: Host species-specific conservation of a family of repeated sequence in the genomic of a fungal plant pathogen. Proceedings of the National Academy of Sciences of the United States of America, 86, 9981-9985 (1989). https://doi.org/10.1073/pnas.86.24.9981
  7. Hebert P.D.N., Cywinska A., Shelley L.B. and Jeremy R.: Biological identifications through DNA Barcodes. Proceedings of the Royal Society of London, Series B, 270, 313-321 (2003). https://doi.org/10.1098/rspb.2002.2218
  8. Herbert P.D.N., Retnasingham S., and DeWaard J.R.: Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species., Proceedings of the Royal Society of London, Series B, 270, 96-99 (2003). https://doi.org/10.1098/rsbl.2003.0025
  9. Hwang T.Y., Seo M.J., Lee S.K., Park H.M., Jeong K.H., Lee Y.Y., Kim S.L., Yuu H.T., Lee J.E. and Kim D.W.: Discrimination of 110 Korean soybean cultivars by sequence tagged sites(STS)-CAPS markers. Korean Journal of Breeding Science, 3, 258-272 (2012).
  10. Innocenzo M., Massimiliano P. and Enzo P.: Detection of DNA in virgin olive oils extracted from destoned fruits. European Food Research and Technology, 224, 469-475 (2007). https://doi.org/10.1007/s00217-006-0340-y
  11. Jo I.H., Bang K.H., Kim Y.C., Kim J.U., Shin M.R., Moon J.Y., Noh B.S., Hyun D.Y., Kim D.H., Cha S.W. and Kim H.S.: Analysis of mitochondrial DNA sequence and molecular marker development for identification of Panax species. Korean Journal of Medicinal Crop Sciences, 21, 91-96 (2013). https://doi.org/10.7783/KJMCS.2013.21.2.91
  12. Kim H.K., Kim Y.A., Lee A.Y. and Ko B.S.: Pattern analysis of Cynanchi wilfordi Radix and Polygoni multiflori Radix. Korea Journal of Pharmacognosy, 34(4), 278-281 (2003).
  13. Kim M.J., Song B.H., Nam S.Y, Kim I.J., Lee C.H. and Yun T.: Effects of non-supporting methods on growth and yield of Cynanchum auriculatum Royle ex Wight. Korean Journal of Medicinal Crop Sciences, 13, 268-272 (2005).
  14. Kim M.J., Kim I.J., Choi S.Y., Han D.H., Kim Y.H., Lim S.C., Kim T.J., Nam S.Y., Song B.H., Oh B.U. and Park C.G.: Comparison of Cynanchum wilfordii, C. auriculatum, Metaplexis japonica and Polygonum multiflorum by morphological characters. Korean Journal of Medicinal Crop Sciences, 22, 113-120 (2014). https://doi.org/10.7783/KJMCS.2014.22.2.113
  15. Kim M.K., Wang H., Kim Y.j., Sathiyamoorthy S., Kwon W.S. and Yang D.C.: Molecular authentication by multiplex-PCR of three similar medicinal plant species:Cynanchum wilfordii, Cynanchum auriculatum and Polygonum multiflorum (Fallopia multiflorum). Journal of Medicinal Plants Research, 7(35), 2584-2589 (2013).
  16. Kocher T.D., Thomas W.K., Meyer A., Edwards S.V., Paabo S. and Vellablanca F.X.: Dynamics of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. Proceedings of the National Academy of Sciences of the United States of America, 86, 6196-6200 (1989). https://doi.org/10.1073/pnas.86.16.6196
  17. Kress W.J., Wurdack K.J., Zimmer E.A., Weigt L.A. and Janzen D.H.: Use of DNA barcodes to identify flowering plants. Proceedings of the National Academy of Sciences of the United States of America, 102, 8369-8374 (2005). https://doi.org/10.1073/pnas.0503123102
  18. Kress W.J. and Erickson D.L.: A two-locus global DNA barcode for land plants: the coding rbcL gene complements the non-coding trnH-psbA spacer region. PLoS One, 2, e508 (2007). https://doi.org/10.1371/journal.pone.0000508
  19. Kress W.J. and Erickson D.L.: DNA barcodes : genes, genomics, and bioinformatics. Proceedings of the National Academy of Sciences of the United States of America, 105, 2761-2762 (2008). https://doi.org/10.1073/pnas.0800476105
  20. Lahaye R., Van der Bank M., Bogarin D., Warner J. and Pupulin F.: DNA barcoding the floras of biodiversity hotspots. Proceedings of the National Academy of Sciences of the United States of America, 105, 2923-2928 (2008). https://doi.org/10.1073/pnas.0709936105
  21. Moon B.C. and Choo B.G.: Gene marker for classification of Polygonum multiflorum Thungerg, Cynanchum wilforid Max. Hemsl and Cynanchum auriculatum Royle ex Wight. KR, 10-2009-0123113 (2009).
  22. Olmstead R.G., Reeves P.A.: Evidence for the polyphyly of the Scropulariaceae based in chloroplast rbcL and ndhF sequence. Ann. Missouri Bot. Gard., 82, 176-193 (1995). https://doi.org/10.2307/2399876
  23. Park Y.C., Jin S.O., Kim M.R., Kim K.H., Lee J.H., Cho T.Y., Lee H.J. and Han S.B.: Detection Method for Identification of Pueraria mirifica (Thai kudzu) in Processed Foods. Journal of Food Hygiene and Safety, 27, 466-472 (2012). https://doi.org/10.13103/JFHS.2012.27.4.466
  24. Park Y.C., Jin S.O., Lim J.Y., Kim K.H., Lee J.H., Cho T.Y., Lee H.J., Han S.B., Lee S.J., Lee K.H. and Yoon H.S.: Application for identification of food raw materials by PCR using universal primer. Journal of Food Hygiene and Safety, 27, 317-324 (2012). https://doi.org/10.13103/JFHS.2012.27.3.317
  25. Park Y.C., Lim J.Y., Kim M.R., Park Y.E., Lim J.D., Hwang C.R., Kim K.H., Lee J.H., Cho T.Y., Lee H.J., Lee S.J. and Han S.B.: Identification of faulty red pepper powder containing seasoned red-pepper sauce. Journal of Food Hygiene and Safety, 27, 182-187 (2012). https://doi.org/10.13103/JFHS.2012.27.2.182
  26. Tomoaki M., Atsushi A., Takuma T., Sumitaka Y., Yutaka O. and Yoshida M.: Identification of animal species using the partial sequences in the mitochondrial 16S rRNA Gene. Legal Medicine, 11, 449-450 (2009). https://doi.org/10.1016/j.legalmed.2009.02.002
  27. White J.J., Bruns J., Lee S. and Taylor J.: Amplification and direct sequencing of fungus ribosomal RNA genes for phylogenics. A guide to methods and application. Academic press, 315-322 (1990).