Abstract
Wheat streak mosaic virus (WSMV), a member of the genus Tritimovirus in the family Potyviridae, has been designated as a plant quarantine pathogen in South Korea. Several diagnostic methods can be applied to diagnose viral infections in plants, but polymerase chain reaction and enzyme-linked immunosorbent assay, which can identify viral species with speed and accuracy, are mainly used in Korean plant quarantine. Many variants of different viral species with highly divergent genomic sequences are constantly being discovered by high-throughput sequencing technology. This means that previously established primers may no longer be suitable for diagnostic use. In this study, we developed a reverse transcription polymerase chain reaction assay for detecting WSMV isolates/strains using all of the WSMV sequences available in NCBI GenBank. All 13 primer sets were able to produce amplicons of the expected sizes from WSMV-infected samples. To check whether nonspecific reactions occur, some closely related viruses (one tritimovirus and five potyviruses) and target imported plants (wheat, maize, oat, and proso millet) were tested. Consequently, four primer sets, which did not produce nonspecific bands, were finally selected among the 13 primer sets. Concentration-dependent amplification tests showed that the four primer sets are adequate for use in the diagnosis of WSMV in Korean plant quarantine.
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References
Boonham N, Kreuze J, Winter S, van der Vlugt R, Bergervoet J, Tomlinson J, Mumford R. Methods in virus diagnostics: from ELISA to next generation sequencing. Virus Res. 2014;186:20–31.
Hadi BAR, Langham MAC, Osborne L, Tilmon KJ. Wheat streak mosaic virus on wheat: biology and management. J Integr Pest Manag. 2011;2(1):J1–5.
Hodge BA, Paul PA, Stewart LR. Occurrence and high-throughput sequencing of viruses in Ohio wheat. Plant Dis. 2020;104(6):1789–800.
Lim S, Igori D, Zhao F, Yoo RH, An TJ, Lim HS, Lee SH, Moon JS. Complete genome sequence of a tentative new caulimovirus from the medicinal plant Atractylodes macrocephala. Arch Virol. 2015;160(12):3127–31.
Lim S, Kim KH, Zhao F, Yoo RH, Igori D, Lee SH, Moon JS. Complete genome sequence of a novel endornavirus isolated from hot pepper. Arch Virol. 2015;160(12):3153–6.
Lim S, Yoo RH, Igori D, Zhao F, Kim KH, Moon JS. Genome sequence of a recombinant brassica yellows virus infecting Chinese cabbage. Arch Virol. 2015;160(2):597–600.
Lim S, Igori D, Zhao F, Do YS, Cho IS, Choi GS, Moon JS. Molecular detection and characterization of a divergent isolate of Plantago asiatica mosaic virus in Plantago asiatica. VirusDis. 2016;27(3):307–10.
Lim S, Kwon SY, Lee JH, Cho HS, Kim HS, Park JM, Lee SH, Moon JS. Genomic detection and molecular characterization of two distinct isolates of cycas necrotic stunt virus from Paeonia suffruticosa and Daphne odora. Virus Genes. 2019;55(5):734–7.
Nakaune R, Nakano M. Efficient methods for sample processing and cDNA synthesis by RT-PCR for the detection of grapevine viruses and viroids. J Virol Methods. 2006;134:244–9.
Novella IS, Presloid JB, Taylor RT. RNA replication errors and the evolution of virus pathogenicity and virulence. Curr Opin Virol. 2014;9:143–7.
Roossinck MJ. Deep sequencing for discovery and evolutionary analysis of plant viruses. Virus Res. 2017;239:82–6.
Roossinck MJ, Martin DP, Roumagnac P. Plant virus metagenomics: advances in virus discovery. Phytopathology. 2015;105(6):716–27.
Schubert J, Ziegler A, Rabenstein F. First detection of wheat streak mosaic virus in Germany: molecular and biological characteristics. Arch Virol. 2015;160(7):1761–6.
Singh K, Wegulo SN, Skoracka A, Kundu JK. Wheat streak mosaic virus: a century old virus with rising importance worldwide. Mol Plant Pathol. 2018;19(9):2193–206.
Stangegaard M, Dufva IH, Dufva M. Reverse transcription using random pentadecamer primers increases yield and quality of resulting cDNA. Biotechniques. 2006;40(5):649–57.
Tatineni S, Hein GL. Genetics and mechanisms underlying transmission of Wheat streak mosaic virus by the wheat curl mite. Curr Opin Virol. 2018;33:47–54.
Acknowledgements
We thank Edanz (https://en-author-services.edanzgroup.com/ac) for editing the English text of a draft of this manuscript. This work was supported by a grant (Project No. B-1543086-2019-21-03) from the Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural Affairs, Republic of Korea.
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Lee, J., Lee, K., Chun, J. et al. Development of an RT-PCR assay to detect genetically divergent wheat streak mosaic virus isolates for plant quarantine inspections in South Korea. VirusDis. 32, 150–154 (2021). https://doi.org/10.1007/s13337-020-00646-3
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DOI: https://doi.org/10.1007/s13337-020-00646-3