Production, characterization and use of monoclonal antibodies to grapevine virus A

Archives of Virology - Tập 127 - Trang 185-194 - 1992
D. Boscia1, E. Aslouj2, V. Elicio1, V. Savino1, M. A. Castellano1, G. P. Martelli1
1Dipartimento di Protezione delle Piante, Università degli Studi and Centro di Studio del C. N. R. sui Virus e le Virosi delle Colture Mediterranee, Bari, Italy
2Istituto Agronomico Mediterraneo, Valenzano, Italy

Tóm tắt

Four stable hybridoma cell lines secreting monoclonal antibodies to grapevine closterovirus A (GVA) were obtained by fusing spleen cells of immunized BALB/c mice with mouse myeloma cell line Sp 2/0-Ag 14. In ELISA all MAbs reacted with virus in leaf extracts fromNicotiana benthamiana, glass-house-, field-, or in vitro-grown grapevines, or with cortical shavings from mature grape canes. In IEM tests, only one of the MAbs (PA 3.F 5) decorated virus particles on the entire surface. This MAb was likely induced by a surface antigenic determinant, whereas the other three MAbs (PA 3.D 11, PA 3.C 6, and PA 3.B 9) were originated by cryptotopes. Coupling polyclonal antibodies for coating protein A-sensitized plates, and monoclonal antibody conjugates for antigen detection, gave highly efficient and reproducible results for identification of GVA in field-grown grapevines.

Tài liệu tham khảo

Boccardo G, D'Aquilio M (1981) The protein and nucleic acid of a closterovirus isolated from grapevine with stem pitting symptoms. J Gen Virol 53: 179–182 Castrovilli S, Gallitelli D (1985) A comparison of two isolates of Grapevine virus A. Phytopathol Mediterr 24: 219–220 Clark MF, Adams AN (1977) Characteristics of the microplate method of enzyme-linked immunosorbent assay for detection of plant viruses. J Gen Virol 34: 475–483 Conti M, Milne RG, Luisoni E, Boccardo G (1980) A closterovirus from stem pitting diseased grapevine. Phytopathology 70: 394–399 Engelbrecht DJ, Kasdorf GGF (1985) Association of a closterovirus with grapevines indexing positive for grapevine leafroll disease and evidence for its natural spread in grapevines. Phytopathol Mediterr 24: 101–105 Goheen HC (1988) Ruperstris stem pitting. In: Pearson RP, Goheen AC (eds) Compendium of grape diseases. American Phytopathological Society Press, St. Paul, p 53 Gugerli P, Rosciglione B, Brugger JJ, Bonnard S, Ramel ME, Tremea F (1991) Further characterization of grapevine leafroll disease. In: Proceedings of the 10th Meeting ICVG, Volos 1990, pp 59–60 Hajimorad MR, Francki RIB (1989) Preparation of soluble, biologically active alfalfa mosaic virus coat protein and its CaCl2-induced degradation. J Virol Methods 25: 49–61 Hill JH (1990) Competitive binding between monoclonal antibodies to define epitope relationships or characterize immunoglobulin idiotypes. In: Hampton R, Ball E, De Boer S (eds) Serological methods for detection and identification of viral and bacterial plant pathogens. American Phytopathological Society Press, St. Paul, pp 215–221 Hu JS, Gonsalves D, Boscia D, Namba S (1991) Use of monoclonal antibodies to characterize grapevine leafroll associated closteroviruses. Phytopathology 80: 920–925 Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T 4. Nature 227: 680–685 Martin RR, Stace-Smith R (1984) Production and characterization of monoclonal antibodies specific to potato leaf roll virus. Can J Plant Pathol 6: 206–210 Milne RG, Luisoni E (1977) Rapid immune electron microscopy of virus preparations. Methods Virol 6: 265–281 Minafra A, Russo M, Martelli GP (1992) Further studies on the use of molecular probes to grapevine closterovirus A. Vitis 31: 26–31 Monette PL, James D (1990) Detection of two strains of grapevine virus A. Plant Dis 74: 898–900 Monette PL, James D (1990) Use of in vitro cultures ofNicotiana benthamiana for the purification of grapevine virus A. Plant Cell Tissue Org Cult 23: 131–134 Rosciglione B, Castellano MA, Martelli GP, Savino V, Cannizzaro G (1983) Mealybugs transmission of grapevine virus A. Vitis 22: 331–347 Savino V, Boscia D, Martelli GP (1989) Rugose wood complex of grapevine: can grafting toVitis indicator discriminate between diseases? In: Proceedings of the 9th Meeting ICVG, Kiryat Anavim 1987, pp 91–94