Skip to main content
Log in

Monoclonal antibodies for detection, serological characterization and immunopurification of grapevine fleck virus

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

Ten stable hybridoma cell lines secreting monoclonal antibodies to grapevine fleck virus (GFkV) were selected after fusing spleen cells of immunized Balb/C mice with mouse myeloma cells (SP2/0-Ag 14). All MAbs reacted positively in ELISA with leaf extracts from fifty GFkV-infected grapevines from various geographical origins. MAb 2B5 was used for routine detection of GFkV and appeared to be more sensitive than polyclonal antibodies. The first attempt to purify GFkV by immunoaffinity chromatography using MAb 2B5 led to highly purified coat protein. This procedure encompassed fewer steps and allowed the use of tissues other than rootlets for satisfactory purification.

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.

Similar content being viewed by others

References

  • Bass P and Legin R (1981) Thermothérapie et multiplication in vitro d'apex de la vigne. Application à la séparation ou à l'élimination de diverses maladies de type viral et à l'évaluation des dégâts. Comptes Rendus de l'Académie agricole de France 67: 922-933

    Google Scholar 

  • Bennett B, Check IJ, Olsen MR and Hunter RL (1992) A comparison of commercially available adjuvants for use in research. Journal of Immunological Methods 153: 31-40

    Google Scholar 

  • Boscia D, Martelli GP, Savino V and Castellano MA (1991) Identi-fication of the agent of fleck disease. Vitis 30: 97-105

    Google Scholar 

  • Boscia D, Elicio V, Savino V and Martelli GP (1993) Monoclonal antibodies to grapevine fleck virus. Extended abstracts 11th Meeting ICVG, Montreux, Switzerland (Federal Agricultural Research Station of Changins, Nyon, Switzerland)

  • Boscia D, Elicio V, Savino V and Martelli GP (1995) Production of monoclonal antibodies to grapevine fleck virus. Plant Pathology 44: 160-163

    Google Scholar 

  • Boulila M, Boscia D, Di Terlizzi B, Castellano MA, Minafra A, Savino V and Martelli GP (1990) Some properties of a phloem limited non mechanically-transmissible grapevine virus. Journal of Phytopathology 129: 151-158

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Analytical Biochemistry 72: 248-254

    Google Scholar 

  • Clark MF and Adams AN (1977) Characteristics of the microplate method of enzyme linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 34: 475-483

    Google Scholar 

  • Hewitt WB, Goheen AC, Cory L and Luhn C (1972) Grapevine fleck disease, latent in many varieties, is transmitted by graft inoculation. IVth Meeting of ICVG, Colmar, France, 1970. Annales de Phytopathologie, Numéro hors série: 43-47

  • Hoogenraad NJ and Wraight CJ (1986) The effect of pristane on ascites tumor formation and monoclonal antibody production. Methods in Enzymology 121: 375-381

    Google Scholar 

  • Köhler G and Milstein C (1975) Continuous culture of fused cell secreting antibodies of predefined specificity. Nature (London) 256: 495-497

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685

    Google Scholar 

  • Martelli GP (1993) In: Graft-transmissible diseases of grapevines. Handbook for detection and diagnosis. GP Martelli (ed) FAO, Roma 137

    Google Scholar 

  • Mink GI and Parsons JL (1977) Procedures for rapid detection of virus and viruslike diseases of grapevine. Plant Disease Reporter 61: 567-571

    Google Scholar 

  • Ramel ME, Serrant P, Külling P and Gugerli P (1993) Monoclonal and polyclonal antibodies for the detection of grapevine fleck associated virus. Extended abstracts 11th Meeting ICVG, Montreux, Switzerland (Federal Agricultural Research Station of Changins, Nyon, Switzerland) 161-162

  • Schwartz Y, Boudon-Padieu E, Grange J, Meignoz R and Caudwell A (1989) Monoclonal antibodies to the mycoplasma-like organism (MLO) responsible for grapevine Flavescence dor´ee. Research in Microbiology 140: 311-324

    Google Scholar 

  • Seddas A, Meignoz R and Daire X, Boudon-Padieu E and Caudwell A (1993) Purification of grapevine Flavescence dorée MLO (mycoplasma-like-organism) by immunoaffinity. Current Microbiology 27: 229-236

    Google Scholar 

  • Seddas A, Meignoz R, Daire X and Boudon-Padieu E (1996) Generation and characterization of monoclonal antibodies to flavescence dorée phytoplasma: serological relationships and differences in electroblot immunoassay profiles of flavescence dorée and Elm yellows phytoplasmas. European Journal of Plant Pathology 102: 757-764

    Google Scholar 

  • Triolo E and Materazzi A (1986) La maculatura infettiva della vite: influenza di isolati diversi sull'attitudine alla propagazione vegetativa di Vitis rupestris St George. In: IV Symp. Int. Sélection Clonale de la Vigne, Changins. La Recherche Agronomique Suisse 3: 320-334

    Google Scholar 

  • Walter B, Bass P, Legin R, Martin C, Vernoy R, Collas Aand Vesselle G (1990) The use of a green-grafting technique for the detection of virus-like diseases of the grapevine. Journal of Phytopathology 128: 137-145

    Google Scholar 

  • Walter B and Cornuet P (1993) ELISA detection of grapevine fleck virus (GFkV). Agronomie 13: 651-657

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schieber, O., Seddas, A., Belin, C. et al. Monoclonal antibodies for detection, serological characterization and immunopurification of grapevine fleck virus. European Journal of Plant Pathology 103, 767–774 (1997). https://doi.org/10.1023/A:1008662830247

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1008662830247

Navigation