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RT-PCR Detection of Five Quarantine Plant RNA Viruses Belonging to Potyand Tospoviruses

  • Lee, Jong-Seung (Department of Agricultural Biotechnology and Plant Breeding and Genomics Institute, College of Agriculture and Life Sciences, Seoul National University) ;
  • Cho, Won-Kyong (Department of Agricultural Biotechnology and Plant Breeding and Genomics Institute, College of Agriculture and Life Sciences, Seoul National University) ;
  • Choi, Hong-Soo (Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Kook-Hyung (Department of Agricultural Biotechnology and Plant Breeding and Genomics Institute, College of Agriculture and Life Sciences, Seoul National University)
  • Received : 2011.07.13
  • Accepted : 2011.08.03
  • Published : 2011.09.01

Abstract

In order to detect quarantine plant viruses, we developed reverse transcription-polymerase chain reaction (RT-PCR) primer pairs for five single-stranded (ss) plant RNA viruses that are not currently reported in Korea but could be potential harmful plant viral pathogens. Three viruses such as Chilli veinal mottle virus (ChiVMV), Colombian datura virus (CDV), and Tobacco etch virus (TEV) belong to the genus Potyvirus while Chrysanthemum stem necrosis virus (CSNV) and Iris yellow spot virus (IYSV) are members of the genus Tospovirus. To design RT-PCR primers, we used reported gene sequences corresponding to the capsid protein and polyprotein for ChiVMV, CDV, and TEV while using nucleocapsid protein regions for CSNV and IYSV. At least two different primer pairs were designed for each virus. Fifteen out of 16 primer pairs were successfully applied in detection of individual quarantine virus with high specificity and efficiency. Taken together, this study provides a rapid and useful protocol for detection of five quarantine viruses.

Keywords

References

  1. Bezerra, I. C., Resende, R. O. de., Pozzer, L., Nagata, T., Kormelink, R. and Avila, A. C. De. 1999. Increase of tospoviral diversity in Brazil with the identification of two new tospovirus species, one from chrysanthemum and one from zucchini. Phytopathology 89:823-830. https://doi.org/10.1094/PHYTO.1999.89.9.823
  2. Carrasco, P., Daros, J. A., Agudelo-Romero, P. and Elena, S. F. 2007. A real-time RT-PCR assay for quantifying the fitness of tobacco etch virus in competition experiments. J. Virol. Methods 139:181-188. https://doi.org/10.1016/j.jviromet.2006.09.020
  3. Choi, M.-Y., Lee, S.-G., Kim, K.-H., Park, H.-H. and Kim, J. J. 2009. Recent outbreak of insect pests and climate change in Korea. International symposium on climate change and insect pest 3-304.
  4. Diaz-Montano, J., Fuchs, M., Nault, B. A., Fail, J. and Shelton, A. M. 2011. Onion thrips (Thysanoptera: Thripidae): A global pest of increasing concern in onion. J. Econ. Entomol. 104:1-13. https://doi.org/10.1603/EC10269
  5. Elena, S. F., Bedhomme, S., Carrasco, P., Cuevas, J. M., de la Iglesia, F., Lafforque, G., Lalic, J., Prosper, A., Tromas, N. and Zwart, M. P. 2011. The evolutionary genetics of emergin plant RNA viruses. Mol. Plant-Microbe. Interact. 24:287-293. https://doi.org/10.1094/MPMI-09-10-0214
  6. Garrett, K. A., Dendy, S. P., Frank, E. E., Rouse, M. N. and Travers, S. E. 2006. Climate change effects on plant disease: Genomes to ecosystems. Annu. Rev. Phytopathol. 44:489-509. https://doi.org/10.1146/annurev.phyto.44.070505.143420
  7. Harper, S. J., Delmiglio, C., Ward, L. I. and Clover, G. R. 2011. Detection of tomato black ring virus by real-time one-step RTPCR. J. Virol. Methods 171:190-194. https://doi.org/10.1016/j.jviromet.2010.10.023
  8. Hwang, J., Li, J., Liu, W. Y., An, S. J., Cho, H., Her, N. H., Yeam, I., Kim, D. and Kang, B. C. 2009. Double mutations in eIF4E and eIFiso4E confer recessive resistance to Chilli veinal mottle virus in pepper. Mol. Cells 27:329-336. https://doi.org/10.1007/s10059-009-0042-y
  9. Lee, J.-S., Cho, W. K., Lee, S.-H., Cho, H.-S. and Kim, K.-H. 2011. 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
  10. Lee, B. Y., Lim, H. R., Choi, J. Y. and Ryu, K. H. 2004. Development of molecular detection of three species of seed-transmissible viruses useful for plant quarantine. Plant Pathol. J. 20: 302-307. https://doi.org/10.5423/PPJ.2004.20.4.302
  11. Lian, S., Jonson, M. G., Cho, W. K., Choi, H.-S., Je, Y. H. and Kim, K.-H. 2011. Generation of antibodies against Rice stripe virus proteins based on recombinant proteins and synthetic polypeptides. Plant Pathol. J. 27:37-43. https://doi.org/10.5423/PPJ.2011.27.1.037
  12. Park, M.-R. and Kim, K.-H. 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
  13. Pappu, H. R., du Toit, L. J., Schwartz, H. F. and Mohan, S. K. 2006. Sequence diversity of the nucleoprotein gene of iris yellow spot virus (genus tospovirus, family bunyaviridae) isolates from the western region of the united states. Arch. Virol. 151:1015-1023. https://doi.org/10.1007/s00705-005-0681-z
  14. Salamon, P. and Palkovics, L. 2005. Occurrence of Colombian datura virus in Brugmansia hybrids, Physalis peruviana l. and Solanum muricatum Ait. in Hungary. Acta Virol. 49:117-122.
  15. Sreenivasulu, M. and Sai Gopal, D. V. R. 2010. Development of recombinant coat protein antibody based IC-RT-PCR and comparison of its sensitivity with other immunoassays for the detection of papaya ringspot virus isolates from India. Plant Pathol. J. 26:25-31. https://doi.org/10.5423/PPJ.2010.26.1.025
  16. Zhang, Y., Zhao, W., Li, M., Chen, H., Zhu, S. and Fan, Z. 2011. Real-time taqman RT-PCR for detection of maize chlorotic mottle virus in maize seeds. J. Virol. Methods 171:292-294. https://doi.org/10.1016/j.jviromet.2010.11.002

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