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Archives of Virology

, Volume 53, Issue 3, pp 243–254 | Cite as

Dissolution des Inclusions du Virus de la Polyédrose Nucléaire du DiptèreTipula paludosa MEIG. Etude Ultrastructurale du Virion

  • M. Bergoin
  • B. Guelpa
Article

Résume

Les inclusions de la polyédrose nucléaire deT. paludosa ont été purifiées par gradients de densité de sucrose, leur morphologie décrite au microscope électronique à balayage, et leurs conditions de solubilisation étudiées. Ces inclusions connues pour leur résistance inhabituelle ont été dissoutes dans des conditions maintenant l'intégrité des particules virales, en utilisant une solution de thioglycolate de sodium 0,125m + HCO3Na 0,125m, pH 10,5.

Les virions extraits des inclusions ont été purifiés par gradients de densité de sucrose, et leur structure étudiée au microscope électronique. En coloration négative, le virion est un bâtonnet mesurant 230 à 240 nm de long sur 100 à 110 nm de diamètre. Il est formé d'une nucléocapside entourée d'une enveloppe lâche. Sur des particules partiellement dégradées, on observe à la surface de la capside, de petites sous-unités disposées en hélice (pàs d'environ 40 Å), correspondant vraisemblablement à des capsomères. Après coloration à l'acétate d'uranyle, un filament flexueux s'observe parfois sortant d'une extrêmité de la capside.

Sur coupes fines d'hémocytes, les virions inclus dans les polyèdres ou libres dans le nucléoplasme, présentent la même morphologie mais leur diamètre n'excède pas 80–85 nm. Le virus de la polyédrose nucléaire deT. paludosa réunit tous les critères pour son incroporation dans la famille des Baculoviridae.

Dissolution of polyhedra of the nuclear polyhedrosis virus of the leatherjacketTipula paludosa MEIG. Ultrastructural study of the virion

Summary

The inclusion bodies of the nuclear polyhedrosis virus ofT. paludosa MEIG. were purified by sucrose density gradients, their morphology described using scanning electron microscopy, and the conditions for their solubilization investigated. These inclusions known to be unusually resistant to dissolution, were successfully dissolved in conditions maintaining the integrety of the virus particles by using a 0.125m sodium thioglycolate +0.125m sodium bicarbonate solution, pH 10.5.

The virions extracted from the inclusions were purified by sucrose density gradients and their fine structure studied in electron microscopy. In PTA-stained preparations the virion is rod-shaped measuring 230–240 nm in length and 100 to 110 nm in diameter. It consists of an inner nucleocapsid surrounded by a loose envelope. Partially degraded virions reveal small subunits arranged in a regular helix (pitch of approximately 40 Å) at the surface of the capsid. These subunits likely represent the virus capsomers. A flexuous ropelike structure protruding at one end of the capsid is sometimes observed in partially degraded virions stained with uranyl acetate.

In thin sections of infected hemocytes, the virions occluded on polyhedra or free in the nucleoplasm exhibit the same morphology although their diameter does not exceed 80–85 nm. The nuclear polyhedrosis virus ofT. paludosa meets all the criteria for incorporation into the Family Baculoviridae.

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Bibliographie

  1. 1.
    Bazin, F., Monsarrat, P., Bonami, J. R., Croizier, G., Meynadier, G., Quiot, J. M., Vago, C.: Particules virales de type Baculovirus observées chez le crabeCarcinus maenas. Rev. Trav. Inst. Pêches marit.38, 205–208 (1975).Google Scholar
  2. 2.
    Bergoin, M., Veyrunes, J. C., Scalla, R.: Isolation and amino acid composition of the inclusions ofMelolontha melolontha poxvirus. Virology40, 760–763 (1970).Google Scholar
  3. 3.
    Bergoin, M., Scalla, R., Duthoit, J. L., Vago, C.: Mise en évidence ed Vagoiavirus en microscopie électronique après dissolution des corps d'inclusion. Proc. Colloq. Insect Pathol. microb. Control (Wageningen, 1966), p. 63–68 (1976).Google Scholar
  4. 4.
    Bergold, G. H.: The nature of nuclear polyhedrosis viruses. In:Steinhaus, E. A. (ed.), Insect Pathology, 413–456. New York: Academic Press 1963.Google Scholar
  5. 5.
    Bird, F. T.: A virus disease of the European red mitePanonychus ulmi (Koch). Canad. J. Microbiol.13, 1131 (1967).Google Scholar
  6. 6.
    Clark, T. B., Chapman, H. C., Fukuda, T.: Nuclear-polyhedrosis and cytoplasmic-polyhedrosis virus infections inLouisana mosquitoes. (Aedes sollicitans.) J. Invertebr. Pathol.14, 284–286 (1969).Google Scholar
  7. 7.
    Couch, J. A.: Free and occluded virus, similar to Baculovirus, in hepatopancreas of pink shrimp. Nature (London)247, 229–231 (1974).Google Scholar
  8. 8.
    Dasgupta, B., Ray, H. N.: The intranuclear inclusions in the midgut of the larva ofAnopheles subpictus. Parasitology47, 194–195 (1967).Google Scholar
  9. 9.
    David, W. A. L.: The status of viruses pathogenic for insects and mites. Ann. Rev. Entomol.20, 97–117 (1975).Google Scholar
  10. 10.
    Federici, B. A., Anthony, D. W.: Formation of viron-occluding proteinic spindles in a Baculovirus disease ofAedes triseriatus. J. Invertebr. Pathol.20, 129–138 (1972).Google Scholar
  11. 11.
    Gouranton, J.: Development of an intranuclear nonocc luded rodshaped virus in some midgut cells of an adult insect,Gyrinus natator L. (Coleoptera). J. ultrastruct. Res.39, 281–294 (1972).Google Scholar
  12. 12.
    Hall, D. W., Fish, D. D.: A Baculovirus from the mosquitoWyeomyia smithii. J. Invertebr. Pathol.23, 383–388 (1974).Google Scholar
  13. 13.
    Hall, D. W., Hazard, E. I.: A nuclear-polyhedrosis virus of the caddishfly,Neophylax sp. J. Invertebr. Pathol.21, 323–324 (1973).Google Scholar
  14. 14.
    Huger, A. M.: A virus disease of the Indian rhinoceros-beetle,Oryctes rhinoceros (Linnaeus), caused by a new type of insect virus,Rhabdovirus oryctes, gen. n., sp. n. J. Invertebr. Pathol.8, 38–51 (1966).Google Scholar
  15. 15.
    Kozlov, E. A., Alexeenko, I. P.: Electron microscope investigation of the structure of the nuclear polyhedrosis virus of the silkworm,Bombyx mori. J. Invertebr. Pathol.9, 413–424 (1967).Google Scholar
  16. 16.
    Meynadier, G., Ricou, G., Bergoin, M.: Virose à corps d'inclusion chezTipula paludosa (Diptère) en France. Rev. Pathol. vég. Entomol. agr. Fr.43, 113–118 (1964).Google Scholar
  17. 17.
    Monsarrat, P., Revet, B., Gourevitch, I.: Mise en évidence, stabilisation et purification d'une strcuture nucléoprotéique intracapsidaire chez le Baculovirus d'Oryctes rhinoceros. C. R. Acad. Sci. (Paris)281, 1439–1442 (1975).Google Scholar
  18. 18.
    Monsarrat, P., Veyrunes, J. C., Meynadier, G., Croizier, G., Vago, C.: Purification et étude structurale du virus du ColéoptèreOryctes rhinoceros L. C. R. Acad. Sci. (Paris)277, 1413–1415 (1973).Google Scholar
  19. 19.
    Morgante, J. S., da Cunha, A. S., Pavan, C., Biesele, J. J., Riess, R. W., Garrido, M. C.: Development of a nuclear polyhedrosis in cells ofRynchosciara angelae (Diptera Sciaridae) and patterns of DNA synthesis in the infected cells. J. Invertebr. Pathol.24, 93–105 (1974).Google Scholar
  20. 20.
    Ponsen, M. B.: Electron microscopy of DNA-cores in nuclear polyhedral viruses. Neth. J. Plant Pathol.71, 54–56 (1965).Google Scholar
  21. 21.
    Reed, D. K., Hall, I. M.: Electron microscopy of a rod-shaped noninclusion virus infecting the citrus red mite. J. Invertebr. Pathol.20, 272–278 (1972).Google Scholar
  22. 22.
    Rennie, J.: Polyhedral disease inTipula paludosa (Meigen). Proc. roy. phys. Soc.20, 265–267 (1923).Google Scholar
  23. 23.
    Shvedchikova, N. G., Ulanov, B. P., Tarasevich, L. M.: Electron microscope investigation of DNA isolated from virus particles in the Siberian silkworm granulosis. Vop. Virusol.5, 631–632 (1968).Google Scholar
  24. 24.
    Smith, K. M.: Intranuclear changes in the polyhedrosis ofTipula paludosa. Parasitology45, 482 (1955).Google Scholar
  25. 25.
    Smith, K. M.: Insect Virology, p. 256. New York: Academic Press 1967.Google Scholar
  26. 26.
    Smith, K. M., Xeros, N.: An unusual virus disease of a dipterous larva. Nature (London)173, 866 (1954).Google Scholar
  27. 27.
    Vago, C.: Les virus libres des polyèdries nucléaires. Rev. Pathol. vég. Entomol. agr. Fr.43, 237–241 (1964).Google Scholar
  28. 28.
    Wildy, P.: In:Melnick, J. L. (ed.), Classification and nomenclature of viruses (Monographs in Virology, Vol. 5), p. 81. Basel: Karger 1971.Google Scholar

Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • M. Bergoin
    • 1
    • 2
  • B. Guelpa
    • 1
    • 2
  1. 1.Station de Recherches de CytopathologieSaint-ChristolFrance
  2. 2.Laboratoire de Pathologie ComparéeU.S.T.L.MontpellierFrance

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