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Nested PCR을 이용한 Cowpea chlorotic mottle virus 정밀 진단 시스템 개발

Development and of Diagnostic System for Detection of Cowpea chlorotic mottle virus using by Nested PCR

  • 민병대 (국립환경과학원 환경기반연구부) ;
  • 김영석 (국립환경과학원 영산강물환경연구소) ;
  • 이시원 (국립환경과학원 환경기반연구부) ;
  • 이수헌 (경북대학교 응용생명과학부)
  • Min, Byung-Dae (Environmental Infrastructure Research Department, National Institute of Environmental Research) ;
  • Kim, Young-Suk (Yeongsan River Environment Research Center, National Institute of Environmental Research) ;
  • Lee, Siwon (Environmental Infrastructure Research Department, National Institute of Environmental Research) ;
  • Lee, Su-Heon (School of Applied Biosciences, Kyungpook National University)
  • 투고 : 2014.09.04
  • 심사 : 2014.12.31
  • 발행 : 2014.12.31

초록

Cowpea chlorotic mottle virus (CCMV)는 Group IV positive sense single strand RNA virus, Bromoviridae과, Bromovirus속으로 분류하는 식물병원성 바이러스로, 강낭콩(Phaseolus vulgaris), 나비완두(Clitoria ternatea), 담배(Nicotiana tabaccum), 대두(Glycine max), 동부(Vigna unguiculata, Vigna siensis) 및 땅콩(Arachis hypogaea)이 국내로 수입될 경우, 검사를 수행하는 관리급 검역바이러스이다. 본 연구에서는, RT-PCR, nested PCR 및 유전자-삽입 양성대조구를 개발하여, CCMV를 현장에서 신속, 정확하게 진단할 수 있는 정밀검정 시스템을 구현하였다. 본 연구에서 개발한 방법은 지속적으로 현장에서 활용되어 식물검역에 기여할 것이라고 기대된다.

Cowper chlorotic mottle virus (CCMV) is the 'controlled' quarantine virus as plant pathogenic virus that are classed as group VI (+) ssRNA virus that belongs to the genus Bromovirus and family Bromoviridae, When plants that are Phaseolus vulgaris, Clitoria ternatea, Nicotiana tabaccum, Glycine max, Vigna unguiculata and Vigna siensis, and Arachis hypogaea is imported in domestic. In this study, inspection system is implemented to analyze CCMV accurately and rapidly by developing RT-PCR, nested PCR, and gene insertion positive control. It is expected that the method developed in this study will contribute to the plant quarantine to be consistently utilized in the field.

키워드

참고문헌

  1. Animal, Plant and Fisheries Quarantine and Inspection Agency. 2013. List of Plant Quarantine Viruses in Korea in Newly Revised in 2013. Res. Plant Dis. 19:67-75.
  2. Bancroft JB, Hiebert E. 1967. Formation of an infectious nucleoprotein from protein and nucleic acid isolated from a small spherical virus. Virol. 32:354-356. https://doi.org/10.1016/0042-6822(67)90284-X
  3. Caruso P, Bertolini E, Cambra M, Lopez MM. 2003. A New and Sensitive Co-operational Polymerase Chain Reaction for Rapid Detection of Ralstonia solanacearum in Water. J. Microbiol. Methods 55:257-272. https://doi.org/10.1016/S0167-7012(03)00161-1
  4. Kim D, Hyun J, Hwang H, Lee S. 2000. RT-PCR detection of Citrus tristeza virus from early satsuma mandarin and yuzu Cheju Island. Plant Pathol. J. 16:48-51.
  5. Kim YJ, Park S, Yie SW, Kim KH. 2005. RT-PCR detection of dsRNA Mycoviruses infecting Pleurotus ostreatus and Agaricus blazei Murrill. Plant Pathol. J. 21:343-348. https://doi.org/10.5423/PPJ.2005.21.4.343
  6. Lee JS, Cho WK, Choi HS, Kim KH. 2011a. RT-PCR Detection of Five Quarantine Plant RNA Viruses Belonging to Poty and Tospoviruses. Plant Pathol. J. 27:291-296. https://doi.org/10.5423/PPJ.2011.27.3.291
  7. Lee JS, Cho WK, Lee SH, Choi HS, Kim KH. 2011b. Development of RT-PCR Based Method for Detecting Five Non-reported Quarantine Plant Viruses Infecting the Family Cucurbitaceae or Solanaceae. Plant Pathol. J. 27:93-97. https://doi.org/10.5423/PPJ.2011.27.1.093
  8. Lee S. 2013. A Study of Molecular Biological Detection Methods for Seed-transmitted Viruses in Quarantine. Ph. D. thesis. Dankook University, Cheonan, Chungcheongnam-do, Korea.
  9. Lee S, Cha M, Kim SM, Heo NY, Shin YG, Lee SH. 2014. Development of Nucleotide Primers for Dignostic RT-PCR and Nested PCR Detection of Three Seed-transmitted Viruses (CRLV, SpLV and WClMV) in Quarantine. J. Agric. & Life Sci. 48:75-83. https://doi.org/10.14397/jals.2014.48.3.75
  10. Lee S, Kang EH, Chu YM, Shin YG, Ahn TY. 2013a. Development of PCR Diagnosis System for Plant Quarantine Seed-borne Wheat streak mosaic virus. Korean J. Microbiol. 49:112-117. https://doi.org/10.7845/kjm.2013.3013
  11. Lee S, Kang EH, Heo NY, Kim SM, Kim YJ. Shin YG 2013b. Detection of Carnation necrotic fleck vrus and Carnation ringspot virus using RT-PCR. Res. Plant Dis. 19:36-44. https://doi.org/10.5423/RPD.2013.19.1.036
  12. Lee S, Kang EH, Shin YG, Lee SH. 2013c. Development of RT-PCR and Nested PCR for Detection of Four Quarantine Plant Viruses Belonging to Nepovirus. Res. Plant Dis. 19:220-225. https://doi.org/10.5423/RPD.2013.19.3.220
  13. Lee S, Shin YG. 2014. Development and Practical Use of RT-PCR for Seed-transmitted Prune dwarf virus in Quarantine. Plant Pathol. J. 30:178-182. https://doi.org/10.5423/PPJ.NT.10.2013.0099
  14. Nelson M, McClelland M. 1992. Use of DNA methyltransferase/endonuclease Enzyme Combinations for Megabase Mapping of Chromosomes. Meth. Enzymol. 216:279-303. https://doi.org/10.1016/0076-6879(92)16027-H
  15. Pan YB, Burner DM, Legendre BL. 2000. An Assessment of the Phylogenetic Relationship Among Sugarcane and Related Taxa Based on the Nucleotide Sequence of 5S rRNA Intergenic Spacers. Genetica 108:285-295. https://doi.org/10.1023/A:1004191625603
  16. Park MR, Kim KH. 2004. RT-PCR Detection of Three Non-reported Fruit Tree Viruses Useful for Quarantine Purpose in Korea. Plant Pathol. J. 20:147-154. https://doi.org/10.5423/PPJ.2004.20.2.147
  17. Priou S, Gutarra L, Aley P. 2006. An Improved Enrichment Broth for the Sensitive Detection of Ralstonia solanacearum (biovars 1 and 2A) in Soil Using DAS-ELISA. Plant Pathol. J. 55:36-45. https://doi.org/10.1111/j.1365-3059.2005.01293.x
  18. Stein A, Loebensten G, Koenic R. 1979. Detection of Cucumber mosaic virus and Bean yellow mosaic virus in gladiolus by enzymelinked immunosobent assay (ELISA) Plant Dis. Rep. 63: 185-188.