1887

Abstract

A combination of native electrophoresis and immunodetection (Western blot) was used for the characterization of nucleoprotein particles of the potyvirus (PPV). Virus particles were electrophoresed directly from plant extracts in agarose or mixed acrylamide–agarose gels under native conditions, blotted on nitrocellulose membranes, and characterized with the aid of a coat protein-specific antibody. Using this combined methodology, called NEWeB (native electrophoresis and Western blotting), we could show that a population of particles that differ in their electrophoretic mobility can be detected in extracts of , that two different strains of PPV can be distinguished in double infections of the same plant and that virus particles from leaves contain detectable levels of helper component proteinase molecules. The potential of the NEWeB method for the study of structure and function of virus particles and similar nucleoprotein complexes in single and mixed infections is discussed.

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2000-09-01
2024-05-12
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References

  1. Blanc S., Lopez-Moya J. J., Wang R., Garcia-Lampasona S., Thornbury D. W., Pirone T. P. 1997; A specific interaction between coat protein and helper component correlates with aphid transmission of a potyvirus. Virology 231:141–147
    [Google Scholar]
  2. Hanada K. 1984; Electrophoretic analysis of virus particles of fourteen cucumovirus isolates. Annals of the Phytopathological Society of Japan 50:361–367
    [Google Scholar]
  3. Hollings M., Brunt A. A. 1981; Potyviruses. In . Handbook of Plant Virus Infections and Comparative Diagnosis pp 731–807 Edited by Kurstak E. Amsterdam: Elsevier/North Holland Biomedical Press;
    [Google Scholar]
  4. Hurtt S. S. 1987; Detection and comparison of electrophorotypes of Hibiscus chlorotic ringspot virus. Phytopathology 77:845–850
    [Google Scholar]
  5. Kasschau K. D., Cronin S., Carrington J. C. 1997; Genome amplification and long-distance movement functions associated with the central domain of tobacco etch potyvirus helper component-proteinase. Virology 228:251–262
    [Google Scholar]
  6. Katis N. 1984; Studies on aphid transmission of potato virus Y . PhD thesis University of; London, UK:
  7. Lisa V., Boccardo G., D’Agostino G., Dellavalle G., D’Aquilio M. 1981; Characterization of a potyvirus that causes zucchini yellow mosaic. Phytopathology 71:667–672
    [Google Scholar]
  8. Maiss E., Timpe U., Brisske A., Jelkmann W., Casper R., Himmler G., Mattanovich D., Katinger H. W. 1989; The complete nucleotide sequence of plum pox virus RNA. Journal of General Virology 70:513–524
    [Google Scholar]
  9. Matthews R. E. F. 1991 Plant Virology , 3rd edn. New York: Academic Press;
  10. Pirone T. P., Blanc S. 1996; Helper-dependent vector transmission of plant viruses. Annual Review of Phytopathology 34:227–247
    [Google Scholar]
  11. Riechmann J. L., Lain S., Garcia J. A. 1992; Highlights and prospects of potyvirus molecular biology. Journal of General Virology 73:1–16
    [Google Scholar]
  12. Riedel D., Lesemann D. E., Maiss E. 1998; Ultrastructural localization of nonstructural and coat proteins of 19 potyviruses using antisera to bacterially expressed proteins of plum pox potyvirus. Archives of Virology 143:2133–2158
    [Google Scholar]
  13. Roberts I. M. 1986; Immunoelectron microscopy of extracts of virus-infected plants. In Electron Microscopy of Proteins vol. 5, Viral Structure pp 293–357 Edited by Harris J. R., Horne R. W. London: Academic Press;
    [Google Scholar]
  14. Rojas M. R., Zerbini F. M., Allison R. F., Gilbertson R. L., Lucas W. J. 1997; Capsid protein and helper component proteinase function as potyvirus cell-to-cell movement proteins. Virology 237:283–295
    [Google Scholar]
  15. Tsagris M., Liu B., Tzortzakaki S., Tabler M. 1993; Abstracts of the IX International Congress of Virology . Glasgow, UK: 8–13 August 1993 p. 351
  16. Walkey D. G. A. 1991 Applied Plant Virology , 2nd edn. London & New York: Chapman and Hall;
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