Freezing in the Production of Artificial Top Component of Chicory Yellow Mottle Virus Free

Abstract

SUMMARY

Chicory yellow mottle virus (CYMV), an isometric plant virus containing RNA, is degraded when infected tissues or purified virus suspensions are frozen and thawed. Virus preparations obtained from frozen tissues contain substantial proportions of empty capsids (top component) which increase in relation to the length of time for which plant material is kept frozen before processing. Virus suspensions frozen after purification behave similarly in that, after exposure to −25 °C, they are almost totally dissociated into RNA and protein shells. Most of the dissociation takes place within the first 10 min of freezing at −25 °C. Lower temperatures (−78 °C and −196 °C) are less effective in inducing virus dissociation.

The extent of dissociation varies with the chemical composition of the suspending medium. Na-acetate and NaCl confer complete protection from freezing injury, whereas no protection is given by water and (K-Na)phosphate buffer plus NaCl. (K-Na)phosphate alone, (Na-Na)phosphate and KCl afford only partial protection.

It is possible that several factors, i.e. increased molarity and lowering of pH of solutions at the eutectic point and removal of water associated with protein macromolecules have a bearing in explaining the influence of the various chemicals.

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1972-11-01
2024-03-29
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References

  1. Dunn D. B., Hitchborn J. H. 1965; The use of bentonite in the purification of plant viruses. Virology 25:171–192
    [Google Scholar]
  2. Fraenkel-Conrat H., Williams R. C. 1955; Reconstitution of the active tobacco mosaic virus from its inactive protein and nucleic acid components. Proceedings of the National Academy of Sciences of the United States of America 41:690–698
    [Google Scholar]
  3. Francki R. I. B., Matthews R. E. F. 1961; Stimulation of turnip yellow mosaic virus protein production by 2-thiouracil. Nature, London 191:1078–1080
    [Google Scholar]
  4. Kaper J. M. 1971; Interaction in biological polyelectrolytes: studies on the dissociation and assembly of two simple RNA viruses. Journal of Colloid and Interface Science 36:1–17
    [Google Scholar]
  5. Kaper J. M., Alting Siberg R. 1969a; Degradation of turnip yellow mosaic virus by freezing and thawing in vitro: a new method for studies on the internal organization of the viral components and for isolating native RNA. Virology 38:407–413
    [Google Scholar]
  6. Kaper J. M., Alting Siberg R. 1969b; The effect of freezing on the structure of turnip yellow mosaic virus and a number of other simple plant viruses. An ultracentrifugal analysis. Cryobiology 5:366–374
    [Google Scholar]
  7. Lebeurier G., Fraenkel-Conrat H., Wurtz M., Hirth L. 1971; Self-assembly of protein subunits from alfalfa mosaic virus. Virology 43:51–61
    [Google Scholar]
  8. Matthews R. E. F. 1966; Reconstitution of TYMV-RNA with TMV proteins subunits. Virology 39:82–96
    [Google Scholar]
  9. Quacquarelli A., Vovlas C., Piazzolla P., Russo M., Martelli G. P. 1972; Characteristics of chicory yellow mottle virus (CYMV). Phytopathologia mediterrénea 11: (in the press)
    [Google Scholar]
  10. Stussi C., Lebeurier G., Hirth L. 1969; Partial reconstitution of tobacco mosaic virus. Virology 42:611–620
    [Google Scholar]
  11. Van Den Berg L. 1959; The effect of addition of sodium and potassium chloride to the reciprocal system: KH2PO4-Na2HPO4-H2O on pH and composition during freezing. Archives of Biochemistry and Biophysics 84:305–315
    [Google Scholar]
  12. Van Den Berg L., Rose D. 1959; Effect of freezing on the pH and composition of sodium and potassium phosphate solutions: The reciprocal system KH2PO4-Na2HPO4-H2O. Archives of Biochemistry and Biophysics 81:319–329
    [Google Scholar]
  13. Vovlas C., Martelli G. P., Quacquarelli A. 1971; Le virosi delle piante ortensi in Puglia. VI.II complesso delle maculature anulari della Cicoria. Phytopathologia mediterrénea 10:224–254
    [Google Scholar]
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