1887

Abstract

In cowpea plant cells infected with cowpea mosaic virus, tubular structures containing virus particles are formed in the plasmodesmata between adjacent cells; these structures are supposedly involved in cell-to-cell spread of the virus. Here we show that similar tubular structures are also formed in cowpea protoplasts, from which the cell wall and plasmodesmata are absent. Between 12 and 21 h post-inoculation, tubule formation starts in the periphery of the protoplast at the level of the plasma membrane. Upon assembly, the virus-containing tubule is enveloped by the plasma membrane and extends into the culture medium. This suggests that the tubule has functional polarity and makes it likely that a tubule ‘grows’ into a neighbouring cell . On average, 75% of infected protoplasts were shown to possess tubular structures extending from their surface. The tubule wall was 3 to 4 nm thick and they were up to 20 µ in length, as shown by fluorescent light microscopy and negative staining electron microscopy. By analogy to infected plant cells, both the viral 58K/48K movement and capsid proteins were located in these tubules, as determined by immunofluorescent staining and immunogold labelling using specific antisera against these proteins. These results demonstrate that the formation of tubules is not necessarily dependent on the presence of plasmodesmata or the cell wall, and that they are composed, at least in part, of virus-encoded components.

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1991-11-01
2024-04-18
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References

  1. Conti G. G., Vegetti G., Bassi M., Favali M. A. 1972; Some ultrastructural and cytochemical observations on Chinese cabbage leaves infected with cauliflower mosaic virus. Virology 47:694–700
    [Google Scholar]
  2. Dawson W. O., Schlegel D. E. 1976; Synchronization of cowpea chlorotic mottle virus replication in cowpea leaves. Intervirology 7:284–291
    [Google Scholar]
  3. Di Franco A., Martelli G. P., Russo M. 1983; An ultrastructural study of olive latent ringspot virus in Gomphrena globosa. Journal of Submicroscopic Cytology 15:539–548
    [Google Scholar]
  4. Eggen R., van Kammen A. 1988; RNA replication in comoviruses. In RNA Genetics vol 1 pp 49–70 Edited by Domingo E., Holland J. J., Ahlquist P. Boca Raton: CRC Press;
    [Google Scholar]
  5. Franssen H., Goldbach R., Broekhuysen M., Moerman M., Van Kammen A. 1982; Expression of middle-component RNA of cowpea mosaic virus: in vitro generation of a precursor to both capsid proteins by a bottom-component RNA-encoded protease from infected cells. Journal of Virology 41:8–17
    [Google Scholar]
  6. Goldbach R., Rezelman G., van Kammen A. 1980; Independent replication and expression of B-component RNA of cowpea mosaic virus. Nature, London 286:297–300
    [Google Scholar]
  7. Goldbach R., Eggen R., De Jager C., Van Kammen A., Van Lent J., Rezelman G., Wellink J. 1990; Genetic organization, evolution and expression of plant viral RNA genomes. In Recognition and Response in Plant-Virus Interactions pp 147–162 Edited by Fraser R. S. S. NATO ASI Series: Cell Biology; vol 41
    [Google Scholar]
  8. Hibi T., Rezelman G., Van Kammen A. 1975; Infection of cowpea mesophyll protoplasts with cowpea mosaic virus. Virology 64:308–318
    [Google Scholar]
  9. Hibino H., Tsuchizaki T., Usugi T., Saito Y. 1977; Fine structures and developmental process of tubules induced by mulberry ringspot virus and satsuma dwarf virus infections. Annals of the Phytopathological Society of Japan 43:255–264
    [Google Scholar]
  10. Holness C. L., Lomonossoff G. P., Evans D., Maule A. J. 1989; Identification of the initiation codons for translation of cowpea mosaic virus middle component RNA using site-directed mutagenesis of an infectious cDNA clone. Virology 172:311–320
    [Google Scholar]
  11. Huang W. M., Gibson S. J., Facer P., Gu J., Polak J. M. 1983; Improved section adhesion for immunocytochemistry using high molecular weight polymers of poly-l-lysine as a slide coating. Histochemistry 77:275–279
    [Google Scholar]
  12. Hull R. 1989; The movement of viruses in plants. Annual Review of Phytopathology 27:213–240
    [Google Scholar]
  13. Jones A. T., Kinninmonth A. M., Roberts I. M. 1973; Ultrastructural changes in differentiated leaf cells infected with cherry leaf roll virus. Journal of General Virology 18:61–64
    [Google Scholar]
  14. Kim K. S. 1979; Ultrastructure of plant cells infected with the beetle-transmitted comoviruses. Fitopatologia Brasileira 4:227–239
    [Google Scholar]
  15. Kim K. S., Fulton J. P. 1971; Tubules with virus-like particles in leaf cells infected with bean pod mottle virus. Virology 43:329–337
    [Google Scholar]
  16. Kim K. S., Fulton J. P. 1973; Plant virus-induced cell wall overgrowth and associated membrane elaboration. Journal of Ultrastructure Research 45:328–342
    [Google Scholar]
  17. Kitajima E. W., Lauritis J. A. 1969; Plant virions in plasmodesmata. Virology 37:681–685
    [Google Scholar]
  18. Linstead P. J., Hills G. J., Plaskitt K. A., Wilson I. G., Harker C. L., Maule A. J. 1988; The subcellular location of the gene 1 product of cauliflower mosaic virus is consistent with a function associated with virus spread. Journal of General Virology 69:1809–1818
    [Google Scholar]
  19. Pelham H. R. B. 1979; Synthesis and proteolytic processing of cowpea mosaic virus proteins in reticulocyte lysates. Virology 96:463–477
    [Google Scholar]
  20. Rezelman G., Franssen H. J., Goldbach R. W., Ie T. S., Van Kammen A. 1982; Limits to the independence of bottom component RN A of cowpea mosaic virus. Journal of General Virology 60:335–342
    [Google Scholar]
  21. Rezelman G., van Kammen A., Wellink J. 1989; Expression of cowpea mosaic virus M RNA in cowpea protoplasts. Journal of General Virology 70:3043–3050
    [Google Scholar]
  22. Robards A. W., Lucas W. J. 1990; Plasmodesmata. Annual Review of Plant Physiology and Plant Molecular Biology 41:369–419
    [Google Scholar]
  23. Roberts I. M., Harrison B. D. 1970; Inclusion bodies and tubular structures in Chenopodium amaranticolor plants infected with strawberry latent ringspot virus. Journal of General Virology 7:47–54
    [Google Scholar]
  24. Russell W. C., Newman C., Williamson D. H. 1975; A simple cytochemical technique for demonstration of DNA in cells infected with mycoplasmas and viruses. Nature, London 253:461
    [Google Scholar]
  25. Sterk P., de Jager C. P. 1987; Interference between cowpea mosaic virus and cowpea severe mosaic virus in a cowpea host immune to cowpea mosaic virus. Journal of General Virology 68:2751–2758
    [Google Scholar]
  26. Van Beek N. A. M., Derksen A. C. G., Dijkstra J. 1985; Polyethylene glycol-mediated infection of cowpea protoplasts with Sonchus yellow net virus. Journal of General Virology 66:551–557
    [Google Scholar]
  27. Van Der Scheer C., Groenewegen J. 1971; Structure in cells of Vigna unguiculata infected with cowpea mosaic virus. Virology 46:493–497
    [Google Scholar]
  28. Van Kammen A., de Jager C. P. 1978; Cowpea mosaic virus. CMI/AAB Descriptions of Plant Viruses no 197
    [Google Scholar]
  29. Van Lent J. W. M., Verduin B. J. M. 1985; Specific gold-labelling of antibodies bound to plant viruses in mixed suspensions. Netherlands Journal of Plant Pathology 91:205–213
    [Google Scholar]
  30. Van Lent J. W. M., Verduin B. J. M. 1986; Detection of viral protein and particles in thin sections of infected plant tissue using immunogold labelling. In Developments in Applied Biology. I. Developments and Applications in Virus Testing pp 193–211 Edited by Jones R. A. C., Torrance L. Wellesboume: Association of Applied Biologists;
    [Google Scholar]
  31. Van Lent J. W. M., Groenen J. T. M., Klinge-Roode E. C., Rohrmann G. F., Zuidema D., Vlak J. M. 1990a; Localization of the 34-kDa polyhedron envelope protein in Spodoptera frugiperda cells infected with Autographa californica nuclear polyhedrosis virus. Archives of Virology 111:103–114
    [Google Scholar]
  32. van Lent J., Wellink J., Goldbach R. 1990b; Evidence for the involvement of the 58K and 48K proteins in the intercellular movement of cowpea mosaic virus. Journal of General Virology 71:219–223
    [Google Scholar]
  33. Vos P., Verver J., van Wezenbeek P., van Kammen A., Goldbach R. 1984; Study of the genetic organization of a plant viral RNA genome by in vitro expression of a full-length DNA copy. EMBO Journal 3:3049–3053
    [Google Scholar]
  34. Watanabe Y., Ooshika I., Meshi T., Okada Y. 1986; Subcellular localization of the 30K protein in TMV-inoculated tobacco protoplasts. Virology 152:414–420
    [Google Scholar]
  35. Wellink J., Van Kammen J. 1989; Cell-to-cell transport of cowpea mosaic virus requires both the 58K/48K proteins and the capsid proteins. Journal of General Virology 70:2279–2286
    [Google Scholar]
  36. Wellink J., Jaegle M., Prinz H., van Kammen A., Goldbach R. 1987; Expression of the middle component RNA of cowpea mosaic virus in vivo. Journal of General Virology 68:2577–2585
    [Google Scholar]
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