1887

Abstract

A panel of monoclonal antibodies (MAbs) generated against an admixture of 12 potyvirus isolates was used to compare and differentiate diverse potyviruses. Both native and denatured virions of strains of bean yellow mosaic (BYMV), potato virus Y, tobacco etch, pea seed-borne mosaic, iris severe mosaic, iris mild mosaic and asparagus virus-1 potyviruses were used as immunogen and as antigen for screening of the hybridoma cell lines. Thirty cell lines secreting potyvirus-specific antibodies reactive in indirect antigen-coated plate (ACP-) ELISA were selected for detailed analysis. All 30 MAbs reacted with at least one strain of BYMV; 11 MAbs reacted with between one and eight of the nine BYMV strains and an additional three MAbs reacted only with isolates within the BYMV subgroup (BYMV, pea mosaic virus and clover yellow vein virus). The remaining 16 MAbs reacted with a BYMV isolate and with at least one of the other 43 potyvirus isolates tested. MAb PTY 1 reacted with all 55 potyvirus isolates tested (representing at least 33 different and distinct aphid-transmissible potyviruses). The potyvirus cross-reactive MAbs generally gave higher reactivity values in ACP-ELISA with dissociated virus than with polyclonal antibody-trapped intact virions in triple antibody sandwich ELISA (i.e. were cryptotope-specific). The BYMV strain- and virus-specific MAbs reacted strongly with both types of antigens (i.e. were metatope-specific). At least 25 distinct epitopes (12 cryptotopes and 13 metatopes) could be identified from the MAb-antigen reactivity patterns. The distribution of these epitopes between virus isolates can be used to detect and differentiate potyviruses in infected plant extracts and to examine virus architectures. Some of these epitopes are shared by potyvirus isolates not previously shown to be serologically related. The broad spectrum-reacting MAb PTY 1 recognizes a cryptotope conserved on all of the aphid-transmissible potyviruses examined and should be a valuable tool for the detection and assay of these potyviruses.

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1991-01-01
2024-10-06
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References

  1. Allison R. F., Dougherty W. G., Parks T. D., Willis L., Johnston R. E., Kelly M., Armstrong F. B. 1985; Biochemical analysis of the capsid protein gene and capsid protein of tobacco etch virus: N-terminal amino acids are located on the virion’s surface. Virology 147:309–316
    [Google Scholar]
  2. Al Moudallal Z., Altschuh D., Briand J. P., van Regenmortel M. H. V. 1984; Comparative sensitivity of different ELISA procedures for detecting monoclonal antibodies. Journal of Immunological Methods 68:35–43
    [Google Scholar]
  3. Barnett O. W., Randles J. W., Burrows P. M. 1987; Relationships among Australian and North American isolates of the bean yellow mosaic potyvirus subgroup. Phytopathology 77:791–799
    [Google Scholar]
  4. Boonekamp P. M., Pomp H. 1986; Problems concerning the production of monoclonal antibodies for plant diagnostic purposes. Acta Horticulturae 177:103–109
    [Google Scholar]
  5. Coller H. A., Coller B. S. 1986; Poisson statistical analysis of repetitive subcloning by the limiting dilution technique as a way of assessing hybridoma monoclonality. Methods in Enzymology 121:412–417
    [Google Scholar]
  6. Dougherty W. G., Willis L., Johnson R. E. 1985; Topographic analysis of tobacco etch virus capsid protein epitopes. Virology 144:66–72
    [Google Scholar]
  7. Francki R. I. B. 1983; Current problems in plant virus taxonomy. In A Critical Appraisal of Viral Taxonomy pp. 63–104 Edited by Matthews. R.E.F. Boca Raton: CRC Press;
    [Google Scholar]
  8. Francki R. I. B., McLean G. D. 1968; Purification of potato virus X and preparation of infectious ribonucleic acid by degradation with lithium chloride. Australian Journal of Biological Sciences 21:1311–1318
    [Google Scholar]
  9. Frenkel M. J., Ward C. W., Shukla D. D. 1989; The use of 3′ non-coding nucleotide sequences in the taxonomy of potyviruses: application to watermelon mosaic virus 2 and soybean mosaic virus-N. Journal of General Virology 70:2775–2783
    [Google Scholar]
  10. Gugerli P., Fries P. 1983; Characterization of monoclonal antibodies to potato virus Y and their use for virus detection. Journal of General Virology 64:2471–2477
    [Google Scholar]
  11. Hammond J., Hammond R. W. 1989; Molecular cloning, sequencing and expression in Escherichia coli of the bean yellow mosaic virus coat protein gene. Journal of General Virology 70:1961–1974
    [Google Scholar]
  12. Hammond J., Lawson R. H. 1988; An improved purification procedure for preparing potyviruses and cytoplasmic inclusions from the same tissue. Journal of Virological Methods 20:203–217
    [Google Scholar]
  13. Hill E. K., Hill J. H., Durand D. P. 1984; Production of monoclonal antibodies to viruses in the potyvirus group: use in radioimmunoassay. Journal of General Virology 65:525–532
    [Google Scholar]
  14. Hollings M., Brunt A. A. 1981; Potyviruses. In Handbook of Plant Virus Infections and Comparative Diagnosis pp 732–807 Edited by Kurstak E. Amsterdam: Elsevier;
    [Google Scholar]
  15. Hsu H. T., Franssen J. M., van der Hulst C. T. C., Derks A. F. L. M., Lawson R. H. 1988; Factors affecting selection of epitope specificity of monoclonal antibodies to tulip breaking potyviruses. Phytopathology 78:1337–1340
    [Google Scholar]
  16. Jordan R. L. 1990; Strategy and techniques for the production of monoclonal antibodies. In Serological Detection and Identification of Plant Viral and Bacterial Pathogens Edited by Hampton R. O., Ball E. St Paul, Minnesota: APS Press (in press)
    [Google Scholar]
  17. Jordan R. L., Aebig J. A. 1985; Evaluating the specific immunoreactivities of monoclonal antibodies to two plant viruses. Acta Horticulturae 164:385–393
    [Google Scholar]
  18. Jordan R. L., Hammond J. 1986; Analysis of antigenic specificity of monoclonal antibodies to several potyviruses. Phytopathology 76:1091
    [Google Scholar]
  19. Jordan R. L., Hammond J. 1988; Epitope specificity of strain-, virus-, subgroup-specific and potyvirus group cross-reactive monoclonal antibodies. Phytopathology 78:1600
    [Google Scholar]
  20. Jordan R. L., Aebig J. A., Hsu H. T. 1985; Reactivities of apple mosaic virus (ApMV)/Prunus necrotic ringspot virus (NRSV) monoclonal antibodies with ApMV, NRSV and other ilarviruses.. Phytopathology 75:1353
    [Google Scholar]
  21. Jordan R. L., Konai M., Lee I.-M., Davis R. E. 1989; Species-specific and cross-reactive monoclonal antibodies to the spiroplasmas Spiroplasma citri and S. kunkelli . Phytopathology 79:880–887
    [Google Scholar]
  22. Milne R. G. 1988; Taxonomy of the rod-shaped filamentous viruses. In The Plant Viruses vol 4 The Filamentous Plant Viruses pp. 3–50 Edited by Milne R. G. New York: Plenum Press;
    [Google Scholar]
  23. Mow at W. P. 1985; Tulip chlorotic blotch virus, a second potyvirus causing tulip flower break. Annals of Applied Biology 106:65–73
    [Google Scholar]
  24. Purcifull D. E., Zitter T. A., Hiebert E. 1975; Morphology, host range, and serological relationships of pepper mottle virus. Phytopathology 65:559–562
    [Google Scholar]
  25. Robaglia C., Durand-Tardif M., Tronchet M., Boudazin G., Astier-Manifacier S., Casse-Delbart F. 1989; Nucleotide sequence of potato virus Y (N strain) genomic RNA. Journal of General Virology 70:935–947
    [Google Scholar]
  26. Shepard J. F., Secor G. A., Purcifull D. T. 1974; Immunochemical cross-reactivity between the dissociated capsid proteins of PVY group plant viruses. Virology 58:464–475
    [Google Scholar]
  27. Sherwood J. L., Sanborn M. R., Keyser G. C. 1987; Production of monoclonal antibodies to peanut mottle virus and their use in enzyme-linked immunosorbent assay and dot-immunobinding assay. Phytopathology 77:1158–1161
    [Google Scholar]
  28. Shukla D. D., Ward C. W. 1988; Amino acid sequence homology of coat proteins as a basis for identification and classification of the potyvirus group. Journal of General Virology 69:2703–2710
    [Google Scholar]
  29. Shukla D. D., Strike P. M., Tracy S. L., Gough K. H., Ward C. W. 1988a; The N and C termini of the coat proteins of potyviruses are surface-located and the N terminus contains the major virus-specific epitopes. Journal of General Virology 69:14971508
    [Google Scholar]
  30. Shukla D. D., Thomas J. E., McKern N. M., Tracy S. C., Ward C. W. 1988b; Coat protein of potyviruses. IV. Comparison of biological properties, serological relationships, and coat protein amino acid sequences of four strains of potato virus Y. Archives of Virology 102:207–219
    [Google Scholar]
  31. Sun M. K. C., Gooding G. V., Pirone T. P., Tolin S. A. 1974; Properties of tobacco vein-mottling virus, a new pathogen of tobacco. Phytopathology 64:1133–1136
    [Google Scholar]
  32. Turpen T. 1989; Molecular cloning of a potato virus Y genome: nucleotide sequence homology in non-coding regions of potyviruses. Journal of General Virology 70:1951–1960
    [Google Scholar]
  33. Van Der Vlugt R., Allefs S., De Haan P., Goldbach R. 1989; Nucleotide sequence of the 3′-terminal region of potato virus YN RNA. Journal of General Virology 70229–233
    [Google Scholar]
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