Factors Influencing the Infection of Barley Mesophyll Protoplasts with Brome Mosaic Virus RNA Free

Abstract

SUMMARY

One of the most critical factors for the infection of barley protoplasts with brome mosaic virus (BMV) RNA was the osmotic strength of the medium during protoplast isolation and inoculation. Infection was most efficient when protoplasts were isolated and inoculated in 0·5 -mannitol and then washed with 0·7 -mannitol; less infection occurred when the protoplasts were isolated in 0·5 or 0·7 -mannitol and inoculated and washed in 0·7 -mannitol. Other conditions optimal for infection were inoculum containing 1 g/ml BMV RNA or less, 1 g/ml poly--ornithine, 20 m-potassium citrate buffer at pH 4·7 and 0·5 m-CaCl, and inoculation at 0 °C. Up to 90% of protoplasts were infected in these conditions. Osmotic shock produced by increase of mannitol concentration immediately before inoculation induced invagination suggestive of pinocytosis, and this seemed to be associated with efficient infection of barley protoplasts with BMV. The discrepancy between the effect of this osmotic shock on the infection of barley protoplasts with BMV and its RNA, and the mechanism of infection are discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-41-1-63
1978-10-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/41/1/JV0410010063.html?itemId=/content/journal/jgv/10.1099/0022-1317-41-1-63&mimeType=html&fmt=ahah

References

  1. Alblas I., Bol J. F. 1977; Factors influencing the infection of cowpea mesophyll protoplasts by alfalfa mosaic virus. Journal of General Virology 36:175–185
    [Google Scholar]
  2. Aoki S., Takebe I. 1969; Infection of tobacco mesophyll protoplasts by tobacco mosaic virus ribonucleic acid. Virology 39:439–448
    [Google Scholar]
  3. Beier H., Bruening O. 1976; Factors influencing the infection of cowpea protoplasts by cowpea mosaic virus RNA. Virology 72:363–369
    [Google Scholar]
  4. Burgess I., Motoyoshi F., Fleming E. N. 1973; The mechanism of infection of plant protoplasts by viruses. Pianta, Berlin 112:323–332
    [Google Scholar]
  5. Engelhardt D. L., Zinder N. D. 1964; Host-dependent mutant of bacteriophage f2. III. Infective RNA. Virology 23:582–587
    [Google Scholar]
  6. Furusawa I., Okuno T. 1978; Infection with BMV of mesophyll protoplasts isolated from five plant species. Journal of General Virology40 in the press
    [Google Scholar]
  7. Hiruki C. 1969; Properties of single- and double-stranded ribonucleic acid from barley plants infected with bromegrass mosaic virus. Journal of Virology 3:498–505
    [Google Scholar]
  8. Koch G. 1973; Interaction of poliovirus-specific RNAs with HeLa cells and E. coli . Current Topics in Microbiology and Immunology 62:89–138
    [Google Scholar]
  9. 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]
  10. Motoyoshi F., Hull R. 1974; The infection of tobacco protoplasts with pea enation mosaic virus. Journal of General Virology 24:89–99
    [Google Scholar]
  11. Motoyoshi F., Bancroft J. B., Waits J. W., Burgess J. 1973; The infection of tobacco protoplasts with cowpea chlorotic mottle virus and its RNA. Journal of General Virology 20:177–193
    [Google Scholar]
  12. Motoyoshi F., Bancroft J. B., Watts J. W. 1974a; The infection of tobacco protoplasts with a variant of brome mosaic virus. Journal of General Virology 25:31–36
    [Google Scholar]
  13. Motoyoshi F., Watts J. W., Bancroft J. B. 1974b; Factors influencing the infection of tobacco protoplasts by cowpea chlorotic mottle virus. Journal of General Virology 25:245–256
    [Google Scholar]
  14. Okuno T., Furusawa I. 1977; A simple method for the isolation of intact mesophyll protoplasts from cereal plants. Plant & Cell Physiology 18:1357–1362
    [Google Scholar]
  15. Okuno T., Furusawa I. 1978a; Modes of infection of barley protoplasts with brome mosaic virus. Journal of General Virology 38:409–418
    [Google Scholar]
  16. Okuno T., Furusawa I. 1978b; The use of osmotic shock for the inoculation of barley protoplasts with brome mosaic virus. Journal of General Virology 39:187–190
    [Google Scholar]
  17. Okuno T., Furusawa I., Hiruki C. 1977; Infection of barley protoplasts with brome mosaic virus. Phytopathology 67:610–615
    [Google Scholar]
  18. Okuno T., Miki T., Furusawa I. 1975; Infection and multiplication of bromegrass mosaic virus in barley mesophyll protoplasts. Annals of the Phytopathological Society of Japan (Abstract in Japanese) 41:293–294
    [Google Scholar]
  19. Premecz G., Olàh T., Gulyàs A., Nytrai À., Palfi G., Farkas G. L. 1977; Is the increase in ribo- nuclease level in isolated tobacco protoplasts due to osmotic shock?. Plant Science letters 9:195–200
    [Google Scholar]
  20. Sarkar S., Upadhya M. D., Melchers G. 1974; A highly efficient method of inoculation of tobacco mesophyll protoplasts with ribonucleic acid of tobacco mosaic virus. Molecular and General Genetics 135:1–9
    [Google Scholar]
  21. Sawhney R. K., Adams W. R. J., Tsang J., Galston A. W. 1977; Leaf pretreatment with senescence retardant as a basis for oat protoplast improvement. Plant & Cell Physiology 18:1309–1317
    [Google Scholar]
  22. Takebe I. 1975; The use of protoplasts in plant virology. Annual Review of Phytopathology 13:105–125
    [Google Scholar]
  23. Takebe I. 1977; Protoplasts in the study of plant virus replication. In Comprehensive Virology xi237–283 Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-41-1-63
Loading
/content/journal/jgv/10.1099/0022-1317-41-1-63
Loading

Data & Media loading...

Most cited Most Cited RSS feed