Cereal Research Communications

, Volume 37, Issue 4, pp 541–550 | Cite as

Discrimination of three BYDV species by one-step RT-PCR-RFLP and sequence based methods in cereal plants from the Czech Republic

  • J. K. KunduEmail author
  • J. Jarošová
  • S. Gadiou
  • G. Červená


One-step RT-PCR followed by a RFLP assay was developed for the typing of Barley yellow dwarf virus (BYDV) member of Luteovirus genera. RFLP analysis performed on 23 samples including one lab isolate showed three types of Hpa II restriction profile. A partial coat protein (CP) gene sequencing was carried out and confirmed the RFLP analysis. Both sequencing and RFLP analysis identified the presence of 3 BYDV-PAV (including the lab isolate Blatno85), 10 BYDV-MAV and 10 BYDV-PAS isolates among the field samples. One-step RT-PCR together with RFLP presents an easy and reliable assay for routine BYDV typing. These methods revealed that PAS and MAV are more dominant than PAV in the Czech Republic.




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  1. Bencharki, B., Mutterer, J., Yamani, M.E., Ziegler-Graff, V., Zaoui, D., Jonard, G. 1999. Severity of infection of Moroccan barley yellow dwarf virus PAV isolates correlates with variability in their coat protein sequences. Ann. Appl. Biol. 134:89–99.CrossRefGoogle Scholar
  2. Bisnieks, M., Kvarnheden, A., Sigvald, R., Valkonen, J.P.T. 2004. Molecular diversity of the coat protein-encoding region of Barley yellow dwarf virus -PAV and Barley yellow dwarf virus -MAV from Latvia and Sweden. Arch. Virol. 149:843–853.CrossRefGoogle Scholar
  3. Červená, G., Markytánová, J. 2005. Sledování výskytu virových zakrslostí obilnin Státní rostlinolékařskou správou v letech 2000 až 2004 (Monitoring of the dwarf viruses in cereal crops by the State Phytosanitary Administration from 2000 to 2004). Rostlinolékař 1:18–21.Google Scholar
  4. Chay, C.A., Smith, D.M., Vaughan, R., Gray, S.M. 1996. Diversity among isolates within the PAV serotype of Barley yellow dwarf virus. Phytopathology 86:370–377.CrossRefGoogle Scholar
  5. Clark, M.F., Adams, A.N. 1977. Characteristics of the microplate method of enzyme linked immunosorbent assay for detection of plant viruses. J. Gen. Virol. 34:475–483.Google Scholar
  6. D’Arcy, C.J., Domier, L L. 2005. Luteoviridae. In: C. M. Fauquet et al. (eds) Virus Taxonomy-Eighth Report of the ICTV, Springer-Verlag, NewYork, pp. 891–900.Google Scholar
  7. Gray, S., Gildow, F.E. 2003. Luteovirus-aphid interactions. Annu. Rev. Phytopathol. 41:539–566.CrossRefGoogle Scholar
  8. Haack, L., Courbon, R., Riault, G., Tanguy, S., Le Vilain, D., Henry, M., Dedryver, C.A. 1999. A plant and field study of BYDV-PAV and -MAV distribution on maize in France. Z. Pflanzenkr. Pflanzenschutz. 106:297–303.Google Scholar
  9. Henry, M., Dedryver, C.A. 1991. Occurrence of barley yellow dwarf virus in pastures of western France. Plant Pathol. 40:93–99.CrossRefGoogle Scholar
  10. Honěk, A., Jarošík, V., Dixon, A. 2006. Comparing growth patterns among field populations of cereal aphids reveals factors limiting their maximum abundance. Bull. Entomol. Res. 96:269–277.CrossRefGoogle Scholar
  11. Kroutil, P., Markytánová, J. 2005. Výskyt virových zakrslostí obilnin před sklizní v roce 2005 (The occurrence of dwarf viruses in cereal crops before the 2005 harvest). Rostlinolékař. 6:9–10.Google Scholar
  12. Kundu, J.K. 2003. A rapid and effective RNA release procedure for virus detection in woody plants by reverse transcription-polymerase chain reaction. Acta Virol. 47:147–151.PubMedGoogle Scholar
  13. Kundu, J.K. 2008. First Report of barley yellow dwarf virus-PAS in wheat and barley grown in the Czech Republic. Plant Dis. 92:1587.CrossRefGoogle Scholar
  14. Kundu, J.K. 2009. First Report of Barley yellow dwarf virus -MAV in Oat, Wheat and Barley Grown in the Czech Republic. Plant Dis. 93:964.CrossRefGoogle Scholar
  15. Kundu, J.K., Chrpová, J., Šíp, V. 2006. Biological and molecular diversities of Barley yellow dwarf virus infection in the presence/absence of resistance gene Yd2 on barley cultivars and breeding lines. In: Book of Abstracts of the “International Symposium on Management of Vector-Borne Viruses”, 7–10 February 2006, Andhra Pradesh, India, p. 35.Google Scholar
  16. Mastari, J., Lapierre, H., Dessens, J.T. 1998. Asymmetrical distribution of barley yellow dwarf virus PAV variants between host plant species. Phytopathology 88:818–821.CrossRefGoogle Scholar
  17. Miller, W.A., Liu, S., Beckett, R. 2002. Barley yellow dwarf virus: Luteoviridae or Tombusviridae? Mol. Plant Pathol. 3:177–183.CrossRefGoogle Scholar
  18. Page, D.R. 1996. TreeView: An application to display phylogenetic trees on personal computer. Comp. Appl. Biosci. 12:357–358.PubMedGoogle Scholar
  19. Paliwal, Y.C. 1982. Identification and Annual variation of variants of barley yellow dwarf virus in Ontario and Quebec. J. Plant Pathol. 4:59–64.Google Scholar
  20. Pokorny, R. 2006. Occurrence of viruses of the family Luteoviridae on maize and some annual weed grasses in the Czech Republic. Cereal Res. Commun. 34:1087–1092.CrossRefGoogle Scholar
  21. Power, A.G., Gary, S.M. 1995. Aphid transmission of barley yellow dwarf viruses: Interactions between viruses, vectors, and host plants. In: D’Arcy, C.J., Burnett, P.A. (Eds), Barley Yellow Dwarf; 40 Years of Progress. American Phytopathological Society, St. Paul, MN, pp. 259–289.Google Scholar
  22. Raus, A. 2008. Virózy obilnin: dùsledek oteplování klimatu v ochraně rostlin 21. století (Virus diseases in cereal crops: Effect of global warming in crop protection in the 21th century). Agrotip 9:8–9.Google Scholar
  23. Rizzo, T.M., Gray, S.M. 1990. Cloning and sequence analysis of a cDNA encoding the capsid protein of the MAV isolate of barley yellow dwarf virus. Nucleic Acids Res. 18 (15):4625.CrossRefGoogle Scholar
  24. Robertson, N.L., French, R. 2007a. Genetic analysis of a novel Alaska barley yellow dwarf virus in the family Luteoviridae. Arch. Virol. 152:369–382.CrossRefGoogle Scholar
  25. Robertson, N.L., French, R. 2007b. Genetic structure in natural populations of barley/cereal yellow dwarf virus isolates from Alaska. Arch. Virol. 1 52:891–902.Google Scholar
  26. Sadeghi, E., Dedryver, C.A., Riault, G., Gauthier, J.P. 1997. Variation in transmission of two BYDV-MAV isolates by multiple clones of Rhopalosiphum padi L. Eur. J. Plant Pathol. 103:515–519.CrossRefGoogle Scholar
  27. Saitou, N., Nei, M. 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4:406–425.Google Scholar
  28. Tamura, K., Dudley, J., Nei, M., Kumar, S. 2007. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24:1596–1599.CrossRefGoogle Scholar
  29. Thomson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F., Higgins, D.G. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25:4876–4882.CrossRefGoogle Scholar
  30. Vacke, J. 1964. Žlutá zakrslost ječmene v ČSSR (Barley yellow dwarf virus in CSSR). Rostl. Výr. 10:859–868.Google Scholar
  31. Vacke, J. 1991. Výskyt virù na obilninách a jejich škodlivost (The occurrence of viruses in cereal crops and their harmfulness). Sborník referátù z odborného semináře “Aktuální problémy ochrany polních plodin”, Praha, pp. 100–109.Google Scholar

Copyright information

© Akadémiai Kiadó, Budapest 2009

Authors and Affiliations

  • J. K. Kundu
    • 1
    Email author
  • J. Jarošová
    • 1
  • S. Gadiou
    • 1
  • G. Červená
    • 2
  1. 1.Department of VirologyCrop Research InstitutePragueCzech Republic
  2. 2.Division of DiagnosticsState Phytosanitary AdministrationOlomoucCzech Republic

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