Skip to main content
Log in

VPg Northern-immunoblots as a means for detection of viral RNAs in protoplasts or plants infected with grapevine fanleaf nepovirus

  • Brief Report
  • Published:
Archives of Virology Aims and scope Submit manuscript

Summary

Anti-genome-linked viral protein (anti-VPg) antibodies were produced from a synthetic peptide corresponding to the integral VPg sequence of grapevine fanleaf nepovirus-F 13. These antibodies allowed detection of viral VPg-linked proteins which occur during the processing of viral polyproteins and of viral RNAs in total RNA extracts from infected protoplasts or plants after Northern blotting. These highly specific antibodies recognised RNAs from two grapevine fanleaf virus strains but not from arabis mosaic virus.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

References

  1. Cadman C, Dias H, Harrison B (1960) Sap-transmissible viruses associated with diseases of grapevine in Europe and North America. Nature 187: 577–579

    Google Scholar 

  2. Chu PWG, Boccardo G, Francki RIB (1981) Requirement of a genome associated protein of tobacco ringspot virus for infectivity but not for in vitro translation. Virology 109: 428–430

    Google Scholar 

  3. Eggen R, Van Kammen A (1988) RNA replication in comoviruses. In: Ahlquist P, Holland L, Domingo E (eds) RNA genetics, vol 1. CRC Press, Boca Raton, pp 49–69

    Google Scholar 

  4. Frison EA, Stace-Smith R (1992) Cross-reacting and heterospecific monoclonal antibodies against arabis mosaic nepovirus. J Gen Virol 73: 2525–2530

    PubMed  Google Scholar 

  5. Gustafson GD, Milner JL, McFarland JE, Pedersen K, Larkins BA, Jackson AO (1982) Investigation of the complexity of barley strope mosaic virus RNAs with recombinant DNA clones. Virology 120: 182–193

    Google Scholar 

  6. Hans F, Fuchs M, Pinck L (1992) Replication of grapevine fanleaf virus satellite RNA transcripts inChenopodium quinoa protoplasts. J Gen Virol 73: 2517–2523

    PubMed  Google Scholar 

  7. Huss B, Walter B, Fuchs M (1989) Cross-protection between arabis mosaic virus and grapevine fanleaf virus isolates inChenopodium quinoa. Ann Appl Biol 114: 45–60

    Google Scholar 

  8. Kedzierski W, Porter JC (1991) A novel non-enzymatic procedure for removing DNA template from RNA transcription mixtures. Biotechniques 10: 210–214

    PubMed  Google Scholar 

  9. Margis R, Viry M, Pinck M, Pinck L (1991) Cloning and in vitro characterization of the grapevine fanleaf virus proteinase cistron. Virology 185: 779–787

    PubMed  Google Scholar 

  10. Margis R, Pinck L (1992) Effect of site directed mutagenesis on the presumed catalytic triad and substrate binding pocket of grapevine fanleaf virus 24-kDa proteinase. Virology 190: 884–888

    PubMed  Google Scholar 

  11. Martelli GP, Taylor CE (1989) Distribution of viruses and their nematode vectors. In: Harris KF (ed) Advances in disease vector research, vol 6. Springer, Berlin Heidelberg New York Tokyo, pp 151–189

    Google Scholar 

  12. Mayo MA, Barker H, Harrison BD (1982) Specificity and properties of the genome-linked proteins of nepoviruses. J Gen Virol 59: 149–162

    Google Scholar 

  13. Pinck L, Fuchs M, Pink M, Ravelonandro M, Walter B (1988) A satellite RNA in grapevine fanleaf virus strain F 13. J Gen Virol 69: 233–239

    Google Scholar 

  14. Pinck M, Reinbolt J, Loudes AM, Le Ret M, Pinck L (1991) Primary structure and location of the genome-linked protein (VPg) of grapevine fanleaf nepovirus. FEBS Lett 284: 117–119

    PubMed  Google Scholar 

  15. Rezaian MA, Krake LR (1987) Nucleic acid extraction and virus detection in grapevine. J Virol Methods 17: 277–285

    PubMed  Google Scholar 

  16. Ritzenthaler C, Viry M, Pinck M, Margis R, Fuchs M, Pinck L (1991) Complete nucleotide sequence and genetic organization of grapevine fanleaf nepovirus RNA 1. J Gen Virol 72: 2357–2365

    PubMed  Google Scholar 

  17. Rottier PJM, Rezelman G, Van Kammen A (1979) The inhibition of cowpea mosaic virus replication by actinomycin D. Virology 92: 299–309

    PubMed  Google Scholar 

  18. Salas M (1991) Protein-priming of DNA replication. Annu Rev Biochem 60: 39–71

    PubMed  Google Scholar 

  19. Serghini MA, Fuchs M, Pinck M, Reinbolt J, Walter B, Pinck L (1990) RNA 2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location. J Gen Virol 71: 1433–1441

    PubMed  Google Scholar 

  20. Tabor S, Richardson CC (1987) DNA sequence analysis with a modified bacteriophage T 7 DNA polymerase. Proc Natl Acad Sci USA 84: 4767–4771

    PubMed  Google Scholar 

  21. Tobin GJ, Young DC, Flanegan JB (1989) Self-catalyzed linkage of poliovirus terminal protein VPg to poliovirus RNA. Cell 59: 511–519

    PubMed  Google Scholar 

  22. Verwoerd TC, Dekker BMM, Hoekema A (1989) A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17: 2362

    PubMed  Google Scholar 

  23. Viry M, Serghini MA, Hans F, Ritzenthaler C, Pinck M, Pinck L (1993) Biologically active transcripts from cloned cDNA of genomic fanleaf nepovirus RNAs. J Gen Virol 74: 169–174

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Margis, R., Hans, F. & Pinck, L. VPg Northern-immunoblots as a means for detection of viral RNAs in protoplasts or plants infected with grapevine fanleaf nepovirus. Archives of Virology 131, 225–232 (1993). https://doi.org/10.1007/BF01379095

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01379095

Keywords

Navigation