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DOI QR Code

Development of Multiplex RT-PCR for Simultaneous Detection of Garlic Viruses and the Incidence of Garlic Viral Disease in Garlic Genetic Resources

  • Nam, Moon (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, Yeong-Hoon (Department of Functional Crop, National Institute of Crop Science) ;
  • Park, Chung Youl (School of Applied Biosciences, Kyungpook National University) ;
  • Lee, Min-A (School of Applied Biosciences, Kyungpook National University) ;
  • Bae, Yang-Soo (School of Applied Biosciences, Kyungpook National University) ;
  • Lim, Seungmo (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Joong Hwan (Institute for Bioresources Research, Gyeongsangbuk-do Agricultural Research and Extension Services) ;
  • Moon, Jae Sun (Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Lee, Su-Heon (School of Applied Biosciences, Kyungpook National University)
  • 투고 : 2014.10.28
  • 심사 : 2014.11.16
  • 발행 : 2015.03.01

초록

Garlic generally becomes coinfected with several types of viruses belonging to the Potyvirus, Carlavirus, and Allexivirus genera. These viruses produce characteristically similar symptoms, they cannot be easily identified by electron microscopy (EM) or immunological detection methods, and they are currently widespread around the world, thereby affecting crop yields and crop quality adversely. For the early and reliable detection of garlic viruses, virus-specific sets of primers, including species-specific and genus-specific primers were designed. To effectively detect the twelve different types of garlic viruses, primer mixtures were tested and divided into two independent sets for multiplex polymerase chain reaction (PCR). The multiplex PCR assays were able to detect specific targets up to the similar dilution series with monoplex reverse transcription (RT)-PCR. Seventy-two field samples collected by the Gyeongbuk Agricultural Technology Administration were analyzed by multiplex RT-PCR. All seventy two samples were infected with at least one virus, and the coinfection rate was 78%. We conclude that the simultaneous detection system developed in this study can effectively detect and differentiate mixed viral infections in garlic.

키워드

참고문헌

  1. Ahlawat, Y. S. 1974. A masaic disease of garlic in Darjeeling hills. Sci. Cult. 40:466-467.
  2. Brierley, P. and Smith, F. F. 1944. Some virus diseases of Alliums. Phytopathology 34:990.
  3. Chen, J. et al. 2005. Characterisation of the welsh onion isolate of Shallot yellow stripe virus from China. Arch. Virol. 150:2091-2099. https://doi.org/10.1007/s00705-005-0580-3
  4. Chodorska, M., Paduch-Cichal, E., Kalinowska, E. and Szyndel, M. S. 2013. Occurrence of the viruses belonging to the Allexivirus genus on garlic plants in Poland. Prog. Plant Prot. 53:605-609.
  5. Chung, H. D. and Chang, M. U. 1979. Studies on the infection of virus in garlic in Korea. Kor. J. Hortic. Sci. Technol. 20:123-129.
  6. Dijk, P. van. 1993. Carlavirus isolates from cultivated Allium species represent three viruses. Neth. J. Plant Pathol. 99:233-257. https://doi.org/10.1007/BF01974306
  7. Dovas, C. I., Hatzibukas, E., Salomon, R., Barg, E., Shiboleth, Y. M. and Katis, N. I. 2001. Comparison of methods for virus detection in Allium spp. J. Phytopathol. 149: 731-737. https://doi.org/10.1046/j.1439-0434.2001.00705.x
  8. Keusgen, M. 2002. Health and alliums. In: Rabinowitch, H. D., Currah, L. (Eds.), Allium Crop Science: Recent Advances. CAB International, UK.
  9. Koo, B. J., Chang, M. U. and Choi, Y. D. 1998. Garlic mite-borne virus isolated from cultivated garlic in Korea. Kor. J. Plant Pathol. 14:136-144.
  10. Kumar, S., Baranwal, V. K., Joshi, S., Arya, M. and Majumder, S. 2010. Simultaneous detec-tion of mixed infection of Onion yellow dwarf virus and an Allexivirus in RT-PCRfor ensuring virus free onion bulbs. Indian J. Virol. 21:64-68. https://doi.org/10.1007/s13337-010-0008-x
  11. Majumder, S., Baranwal, V. K. and Joshi, S., 2008. Simultaneous detection of Onion yellow dwarf virus and Garlic latent virus in infected leaves and cloves of garlic by duplex RT-PCR. J. Plant Pathol. 90:369-372.
  12. Majumder, A. and Baranwal, V. K. 2014. Simultaneous detection of four garlic viruses by multiplex reverse transcription PCR and their distribution in Indian garlic accessions. J. Virol. Methods 202:34-38. https://doi.org/10.1016/j.jviromet.2014.02.019
  13. Mikaili, P., Maadirad, S., Moloudizargari, M., Aghajanshakeri, S. and Sarahroodi, S. 2013. Therapeutic uses and pharmacological properties of garlic, shallot, and their biologically active compounds. Iran J. Basic. Med. Sci. 16:1031-1048.
  14. Mohamed, N. A. and Young. B. R. 1981. Garlic yellow streak virus, a potyvirus infecting garlic in New Zwaland. Ann. Appl. Biol. 97:65-74. https://doi.org/10.1111/j.1744-7348.1981.tb02995.x
  15. Nagata, T., Lorz, H. and Widholm, J. M. 2007. Biotechnology in Agriculture and Forestry. Transgenic Crops IV pp. 428.
  16. Song, S. I., Song, J. T., Chang, M. U., Lee, J. S. and Choi, Y. D. 1997. Identification of one of the major viruses infecting garlic plants, garlic virus X. Mol. Cells 7:705-709.
  17. Sumi, S., Tsuneyoshi, T. and Furutani, H. 1993. Novel rod-shaped viruses isolated from garlic, Allium sativum, possessing a unique genome organization. J. Gen. Virol. 74:879-1885.
  18. Sward, R. J. 1991. Lettuce necrotic yellows rhabdovirus and other viruses infecting garlic. Australasian Plant Path. 19:46-51.
  19. Taskin, H., Baktemur, G., Kurul, M. and Buyukalaca, S. 2013. Use of tissue culture techniques for producing virus-free plant in garlic and their identification through real-time PCR. The Scientific World Journal Article ID 781282, 5 pages.
  20. Tsuneyoshi, T., Matsumi, T., Deng, T., Sako, I. and Sumi, S. 1998. Differentiation of Allium carlaviruses isolated from different parts of the world based on the viral coat protein sequence. Arch. Virol. 143:1093-1107. https://doi.org/10.1007/s007050050358
  21. Tsuneyoshi, T., Matsumi, T., Natsuaki, T. and Sumi, S. 1998. Nucleotide sequence analysis of virus isolates indicates the presence of three potyvirus species in Allium plants. Arch. Virol. 143:97-113. https://doi.org/10.1007/s007050050271
  22. Van der Vlugt, R. A. A., Steffens, P., Cuperus, C., Barg, E., Lesemann, D.-E., Bos, L. and Vetten, H. J. 1998. Further evidence that shallot yellow stripe virus (SYSV) is a distinct potyvirus and reidentification of welsh onion yellow stripe virus as a SYSV strain. Virology 89:148-155.
  23. Yamashita, K., Sakai, J. and Hanada, K. 1995. Leek yellow stripe virus (LYSV) isolated from garlic and its elationship to garlic mosaic virus (GMV). Ann. Phytopathol. Soc. Japan 61:273.
  24. Yamashita, K., Sakaai, J. and Hanada, K. 1996. Characterization of a new virus from garlic (Allium sativum L.), garlic miteborne mosaic virus. Ann. Phytopathol. Soc. Japan 62:483-489. https://doi.org/10.3186/jjphytopath.62.483

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