식물병연구 (Research in Plant Disease)

한국식물병리학회 (The Korean Society of Plant Pathology)
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Development of a Multiplex PCR for Simultaneous Detection of Blueberry Red Ringspot Virus and Blueberry Scorch Virus Including an Internal Control

  • Hae Min Lee (Plant Virus GenBank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University) ;
  • Eun Gyeong Song (Plant Virus GenBank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University) ;
  • Ki Hyun Ryu (Plant Virus GenBank, Department of Horticulture, Biotechnology and Landscape Architecture, Seoul Women's University)
  • 투고 : 2022.11.25
  • 심사 : 2022.12.27
  • 발행 : 2023.03.31
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초록

Blueberry red ringspot virus (BRRSV) and blueberry scorch virus (BlScV) are included in the quarantine virus list managed by the Korean Animal and Plant Quarantine Agency. A multiplex polymerase chain reaction (PCR) assay with an internal control was developed for the simultaneous detection of both viruses. The specific primers used here were designed based on the highly conserved regions of the genomic sequences of each virus, obtained from the National Center for Biotechnology Information nucleotide databases. The primers were designed to amplify a partial sequence within coat protein (CP) for detecting BRRSV and a partial sequence within the CP-16 kDa for detecting BlScV. 18S ribosomal RNA (rRNA) was used as internal control, and the primer set used in a previous study was modified in this study for detecting 18S rRNA. Each conventional PCR using the BRRSV, BlScV, and 18S rRNA primers exhibited a sensitivity of approximately 1 fg plasmid DNA. The multiplex PCR assay using the BRRSV, BlScV, and 18S rRNA primers was effective in simultaneously detecting the two viruses and 18S rRNA with a sensitivity of 1 fg plasmid DNA, similar to that of conventional PCR assays. The multiplex PCR assay developed in this study was performed using 14 blueberry cultivars grown in South Korea. BRRSV and BlScV were not detected, but 18S rRNA was all detected in all the plants tested. Therefore, our optimized multiplex PCR assay could simultaneously detect the two viruses and 18S rRNA in field samples collected from South Korea in a time-efficient manner. This approach could be valuable in crop protection and plant quarantine management.

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과제정보

This study was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through Crop Viruses and Pests Response Industry Technology Development Program, funded by the Ministry of Agriculture, Food, and Rural Affairs (MAFRA) (grant number 320044-3). This study was also supported by a sabbatical year research grant from Seoul Women's University (2021).

참고문헌

  1. Abade Dos Santos, F. A., Carvalho, C. L., Peleteiro, M. C., Parra, F. and Duarte, M. D. 2021. A versatile qPCR for diagnosis of leporid gammaherpesvirus 5 using Evagreen or Taqman technologies. Viruses 13: 715.
  2. Abarshi, M. M., Mohammed, I. U., Jeremiah, S. C., Legg, J. P., Kumar, P. L., Hillocks, R. J. et al. 2012. Multiplex RT-PCR assays for the simultaneous detection of both RNA and DNA viruses infecting cassava and the common occurrence of mixed infections by two cassava brown streak viruses in East Africa. J. Virol. Methods 179: 176-184. https://doi.org/10.1016/j.jviromet.2011.10.020
  3. Banerjee, S., Nayik, G.A., Kour, J. and Nazir, N. 2020. Blueberries. In: Antioxidants in Fruits: Properties and Health Benefits, eds. by G. A. Nayik and A. Gull, pp. 593-614. Springer, Singapore.
  4. Bristow, P. R., Martin, R. R. and Windom, G. E. 2000. Transmission, field spread, cultivar response, and impact on yield in highbush blueberry infected with blueberry scorch virus. Phytopathology 90: 474-479. https://doi.org/10.1094/PHYTO.2000.90.5.474
  5. Chang, B. S. and Mahoney, R. R. 1995. Enzyme thermostabilization by bovine serum albumin and other proteins: evidence for hydrophobic interactions. Biotechnol. Appl. Biochem. 22: 203-214. https://doi.org/10.1111/j.1470-8744.1995.tb00346.x
  6. Chang, L., Zhang, Z., Yang, H., Li, H. and Dai, H. 2007. Detection of strawberry RNA and DNA viruses by RT-PCR using total nucleic acid as a template. J. Phytopathol. 155: 431-436. https://doi.org/10.1111/j.1439-0434.2007.01254.x
  7. Cho, I. S., Chung, B. N., Cho, J. D., Choi, G. S. and Lim, H. S. 2012. First report of blueberry red ringspot virus infecting highbush blueberry in Korea. Plant Dis. 96: 1074.
  8. Ciuffo, M., Pettiti, D., Gallo, S., Masenga, V. and Turina, M. 2005. First report of blueberry scorch virus in Europe. Plant Pathol. 54: 565.
  9. Dayaram, A., Opong, A., Jaschke, A., Hadfield, J., Baschiera, M., Dobson, R. C. et al. 2012. Molecular characterisation of a novel cassava associated circular ssDNA virus. Virus Res. 166: 130-135. https://doi.org/10.1016/j.virusres.2012.03.009
  10. Fauquet, C. M., Mayo, M. A., Maniloff, J., Desselberger, U. and Ball, L. A. 2005. Virus Taxonomy: 8th Report of the International Committee on Taxonomy of Viruses. Elsevier Academic Press, San Diego, CA, USA. 1162 pp.
  11. Fuchs, M., Abawi, G. S., Marsella-Herrick, P., Cox, R., Cox, K.D., Carroll, J. E. et al. 2010. Occurrence of tomato ringspot virus and tobacco ringspot virus in highbush blueberry in New York state. J. Plant Pathol. 92: 451-459.
  12. Henderson, D. C. and Hammond, J. 2013. CKC: isolation of nucleic acids from a diversity of plants using CTAB and silica columns. Mol. Biotechnol. 53: 109-117. https://doi.org/10.1007/s12033-012-9494-y
  13. Isogai, M., Ishii, K., Umemoto, S., Watanabe, M. and Yoshikawa, N. 2009. First report of blueberry red ringspot disease caused by blueberry red ringspot virus in Japan. J. Gen. Plant Pathol. 75: 140-143. https://doi.org/10.1007/s10327-009-0145-5
  14. Jevremovic, D., Leposavic, A. and Paunovic, S. 2015. First report of blueberry mosaic-associated virus in highbush blueberry in Serbia. J. Plant Pathol. 97: 541.
  15. Kalinowska, E., Paduch-Cichal, E. and Chodorska, M. 2013. First report of Blueberry scorch virus in elderberry in Poland. Plant Dis. 97: 1515.
  16. Martin, R. R., Polashock, J. J. and Tzanetakis, I. E. 2012. New and emerging viruses of blueberry and cranberry. Viruses 4: 2831-2852. https://doi.org/10.3390/v4112831
  17. Ozay, B. and McCalla, S. E. 2021. A review of reaction enhancement strategies for isothermal nucleic acid amplification reactions. Sens. Actuator Rep. 3: 100033.
  18. Park, C. H., Song, E. G. and Ryu, K. H. 2021. A multiplex PCR assay for the simultaneous detection of five potexviruses infecting cactus plants using dual-priming oligonucleotides (DPOs) primers. J. Virol. Methods 298: 114280.
  19. Petrzik, K., Pribylova, J., Plesko, I. M. and Spak, J. 2011. Complete genome sequences of blueberry red ringspot virus (Caulimoviridae) isolates from the Czech Republic and Slovenia. Arch. Virol. 156: 1901-1903. https://doi.org/10.1007/s00705-011-1077-x
  20. Plant Quarantine Technology Center in 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. (In Korean)
  21. Saad, N., Olmstead, J. W., Jones, J. B., Varsani, A. and Harmon, P. F. 2021. Known and new emerging viruses infecting blueberry. Plants (Basel) 10: 2172.
  22. Song, H.-N., Park, M.-S., Youn, H.-S., Park, S.-J. and Hogstrand, C. 2014. Nutritional compositions and antioxidative activities of two blueberry varieties cultivated in South Korea. Korean J. Food Preserv. 21: 790-798. (In Korean) https://doi.org/10.11002/kjfp.2014.21.6.790
  23. Tang, K. F. J. and Lightner, D. V. 2000. Quantification of white spot syndrome virus DNA through a competitive polymerase chain reaction. Aquaculture 189: 11-21. https://doi.org/10.1016/S0044-8486(00)00367-7
  24. Wegener, L. A., Punja, Z. K. and Martin, R. R. 2004. First report of blueberry scorch virus in cranberry in Canada and the United States. Plant Dis. 88: 427.
  25. Xie, L. X., Zheng, S., Zhang, L. J., Zhang, X. Y. and Li, T. 2018. Blueberry scorch virus detected on blueberry plants imported into China. Plant Dis. 102: 1673.