1887

Abstract

SUMMARY

Inocula made by mixing tobacco rattle virus (strain CAM; TRV), poly--ornithine (PLO) and either citrate buffer or phosphate buffer were compared by measuring their turbidity, by estimating the effect of centrifugation on their infectivity and by electron microscopy. Aggregates (floccules) containing virus particles were formed in the inocula but there was no evidence of any major difference in size between aggregates formed in the different buffers. The relationship between the loss of infectivity following centrifugation and the centrifugal force applied suggested that the aggregates causing infection sedimented at an average of 1340S (mol. wt. 7 × 10). Electron microscopy showed electron dense masses in most of which 10 to 20 TRV particles were embedded. Aggregates made in citrate had virus particles arranged radially whereas aggregates made in phosphate were more randomly constructed.

Preparations of sedimented aggregates, and the supernatant fraction remaining following centrifugation were poorly infective unless PLO was added. In contrast with the addition of PLO to TRV preparations, adding PLO to these centrifuged fractions restored infectivity immediately. It is suggested that PLO can have two modes of action on infection when it is added to TRV to make inocula, one involving the formation of aggregates and the other for which aggregate formation is not essential.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-40-1-19
1978-07-01
2023-02-05
Loading full text...

Full text loading...

/deliver/fulltext/jgv/40/1/JV0400010019.html?itemId=/content/journal/jgv/10.1099/0022-1317-40-1-19&mimeType=html&fmt=ahah

References

  1. Barker H., Harrison B. D. 1977; Infection of tobacco mesophyll protoplasts with raspberry ringspot virus alone and together with tobacco rattle virus. Journal of General Virology 35:125–133
    [Google Scholar]
  2. Beier H., Bruening G. 1976; Factors influencing the infection of cowpea protoplasts by cowpea mosaic virus RNA. Virology 72:363–369
    [Google Scholar]
  3. Burgess J., Motoyoshi F., Fleming E. N. 1973; The mechanism of infection of plant protoplasts by viruses. Planta (Berlin) 112:323–332
    [Google Scholar]
  4. Caspar D. L. D. 1963; Assembly and stability of the tobacco mosaic virus particle. Advances in Protein Chemistry 18:37–121
    [Google Scholar]
  5. Derrick K. S., Brlansky R. H. 1976; Assay for viruses and mycoplasmas using serologically specific electron microscopy. Phytopathology 66:815–820
    [Google Scholar]
  6. Funding L., Steensgaard J. 1973; A simple manual method for the calculation of equivalent sedimentation coefficients from rate-zonal centrifugation experiments. In MSE Application Information A8/6/73 Crawley, U.K.: MSE Scientific Instruments Ltd;
    [Google Scholar]
  7. Grout B. W. W., Coutts R. H. A. 1974; Additives for the enhancement of fusion and endocytosis in higher plant protoplasts: an electrophoretic study. Plant Science Letters 2:397–403
    [Google Scholar]
  8. Harrison B. D., Woods R. D. 1966; Serotypes and particle dimensions of tobacco rattle viruses from Europe and America. Virology 28:610–620
    [Google Scholar]
  9. Kassanis B., White R. F., Turner R. H., Woods R. D. 1977; The mechanism of virus entry during infection of tobacco protoplasts with TMV. Phytopathologische Zeitschrift 88:215–228
    [Google Scholar]
  10. Kubo S., Harrison B. D., Barker H. 1975a; Defined conditions for growth of tobacco plants as sources of protoplasts for virus infection. Journal of General Virology 28:255–257
    [Google Scholar]
  11. Kubo S., Harrison B. D., Robinson D. J. 1974; Effect of phosphate on the infection of tobacco protoplasts by tobacco rattle virus. Intervirology 3:382–387
    [Google Scholar]
  12. Kubo S., Harrison B. D., Robinson D. J., Mayo M. A. 1975b; Tobacco rattle virus in tobacco mesophyll protoplasts: infection and virus multiplication. Journal of General Virology 27:293–304
    [Google Scholar]
  13. Kubo S., Robinson D. J., Harrison B. D., Hutcheson A. M. 1976; Uptake of tobacco rattle virus by tobacco protoplasts, and the effect of phosphate on infection. Journal of General Virology 30:287–298
    [Google Scholar]
  14. Markham R. 1967; The ultracentrifuge. Methods in Virology 11:1–39
    [Google Scholar]
  15. Mayo M. A., Daney De Marcillac G. 1977; Analytical centrifugation of the disk aggregates of tobacco rattle virus protein. Virology 76:560–568
    [Google Scholar]
  16. Motoyoshi F., Oshima N. 1975; Infection with tobacco mosaic virus of leaf mesophyll protoplasts from susceptible and resistant lines of tomato. Journal of General Virology 29:81–91
    [Google Scholar]
  17. Robinson D. J. 1978; Analysis of data from boundary sedimentation experiments in the preparative ultracentrifuge. Journal of General Virology 40:31–32 (Appendix)
    [Google Scholar]
  18. Schumaker V., Rees A. 1972; Preparative centrifugation in virus research. In Principles and Techniques in Plant Virology p 336–368 Edited by Kado C. I., Agrawal H. O. New York and London: Van Nostrand Reinhold Company;
    [Google Scholar]
  19. Takebe I. 1975; The use of protoplasts in plant virology. Annual Review of Phytopathology 13:105–125
    [Google Scholar]
  20. Takebe I., Otsuki Y., Honda Y., Matsui C. 1974; Penetration of plant viruses into isolated tobacco leaf protoplasts. Proceedings of the 1st Intersectional Congress of the International Association of Microbiological Societies 355–64 Edited by Hasegawa T. Tokyo: Japanese Association for Microbiology;
    [Google Scholar]
  21. Trautman R., Hamilton M. G. 1972; Analytical ultracentrifugation. In Principles and Techniques in Plant Virology p 491–528 Edited by Kado C. I., Agrawal H. O. New York and London: Van Nostrand Reinhold Company;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-40-1-19
Loading
/content/journal/jgv/10.1099/0022-1317-40-1-19
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error