1887

Abstract

SUMMARY

Tobacco mesophyll protoplasts were successfully inoculated with tobacco necrotic dwarf virus (TNDV), a virus closely related to potato leafroll virus. The optimum conditions for infection as assessed by staining with fluorescent antibody to virus particles were 1 µg/ml TNDV, 1 µg/ml poly--ornithine, 0.025 -phosphate buffer, pH 5.4, and 1 to 2 × 10/ml protoplasts: this combination resulted in 70 to 80% infection. Accumulation of virus antigen was first detected 12 h after inoculation, and both the number of fluorescing protoplasts and intensity of fluorescence increased markedly during the following 20 h. Virus antigen was distributed throughout the cytoplasm. Electron microscopy of thin sections revealed groups of virus-like particles in the cytoplasm and a few similar particles in the nucleus. Virus yield was estimated to be 7.8 × 10 particles per infected protoplast at 30 h after inoculation. Accumulation of virus antigen was completely inhibited by cycloheximide, but was not affected by actinomycin D or chloramphenicol, when they were applied 2 h after inoculation. Studies on the replication of aphidborne, phloem-limited viruses should be greatly facilitated by using the mesophyll protoplast system.

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1979-02-01
2024-04-27
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References

  1. Brakke M. K., Rochow W. F. 1974; Ribonucleic acid of barley yellow dwarf virus. Virology 61:240–248
    [Google Scholar]
  2. Kojima M., Murayama D. 1972; The application of polyethylene glycol precipitation for purification of potato leaf-roll virus. Annals of the Phytopathological Society of Japan 38:431–433
    [Google Scholar]
  3. Kojima M., Tamada T. 1976; Purification and serology of soybean dwarf virus. Phytopathologische Zeit-schrift 85:237–250
    [Google Scholar]
  4. Kubo S., Harrison B. D., Robinson D. J., Mayo M. A. 1975a; Tobacco rattle virus in tobacco mesophyll protoplasts: infection and virus multiplication. Journal of General Virology 27:293–304
    [Google Scholar]
  5. Kubo S., Harrison B. D., Barker H. 1975b; Defined conditions for growth of tobacco plants as sources of protoplasts for virus infection. Journal of General Virology 28:255–257
    [Google Scholar]
  6. 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 30:287–289
    [Google Scholar]
  7. Kubo S., Takanami Y. 1977; On the occurrence of tobacco necrotic dwarf, a new tobacco disease caused by a phloem-limited virus. (Abstract in Japanese) Annals of the Phytopathological Society of Japan 43:76
    [Google Scholar]
  8. Kubo S., Takanami Y. 1978a; Multiplication of tobacco necrotic dwarf virus in tobacco mesophyll protoplasts. (Abstract in Japanese) Annals of the Phytopathological Society of Japan 44:60–61
    [Google Scholar]
  9. Kubo S., Takanami Y. 1978b; Serological studies on tobacco necrotic dwarf virus. (Abstract in Japanese) Annals of the Phytopathological Society of Japan 44:398
    [Google Scholar]
  10. Mayo M. A. 1978; The effect of protoplast concentration during inoculation on the infection of tobacco protoplasts. Intervirology 9:184–188
    [Google Scholar]
  11. Motoyoshi F., Hull R. 1974; The infection of tobacco protoplasts with pea enation mosaic virus. Journal of General Virology 24:89–99
    [Google Scholar]
  12. Otsuki Y., Shimomura T., Takebe I. 1972; Tobacco mosaic virus multiplication and expression of the N gene in necrotic responding tobacco varieties. Virology 50:45–50
    [Google Scholar]
  13. Otsuki Y., Takebe I., Honda Y., Kajita S., Matsui C. 1974; Infection of tobacco mesophyll protoplasts by potato virus X. Journal of General Virology 22:375–385
    [Google Scholar]
  14. Rochow W. F. 1960; Transmission of barley yellow dwarf virus acquired from liquid extracts by aphids feeding through membranes. Virology 12:223–232
    [Google Scholar]
  15. Rochow W. F., Brakke M. K. 1964; Purification of barley yellow dwarf virus. Virology 24:310–322
    [Google Scholar]
  16. Sakai F., Takebe I. 1970; RNA and protein synthesis in protoplasts isolated from tobacco leaves. Biochimica et Biophysica Acta 244:531–540
    [Google Scholar]
  17. Sarkar S. 1976; Potato leafroll virus contains a double-stranded DNA. Virology 70:265–273
    [Google Scholar]
  18. Sarkar S., Blessing J. 1973; DNA-like properties of the nucleic acid of potato leafroll virus. Naturwissenschaften 60:480–481
    [Google Scholar]
  19. Scalla R., Rochow W. F. 1977; Protein component of two isolates of barley yellow dwarf virus. Virology 78:576–580
    [Google Scholar]
  20. Spendlove R. S. 1967; Microscopic techniques. In Methods in Virology vol III pp 475–520 Edited by Maramorosch K., Koprowski H. New York: Academic Press;
    [Google Scholar]
  21. Takanami Y., Kubo S. 1978a; Characterization of nucleic acid of tobacco necrotic dwarf and potato leafroll viruses. (Abstract in Japanese) Annals of the Phytopathological Society of Japan 44: (in the press)
    [Google Scholar]
  22. Takanami Y., Kubo S. 1978b; Enzymatic purification of two phloem-limited viruses: tobacco necrotic dwarf and potato leafroll. (Abstract in Japanese) Annals of the Phytopathological Society of Japan 44:398–399
    [Google Scholar]
  23. Takanami Y., Kubo S., Imaizumi S. 1977; Synthesis of single- and double-stranded cucumber mosaic virus RNAs in tobacco mesophyll protoplasts. Virology 80:376–389
    [Google Scholar]
  24. Takebe I. 1977; Protoplasts in the study of plant virus replication. Comprehensive Virology 11:237–283
    [Google Scholar]
  25. Takebe I., Otsuki Y. 1969; Infection of tobacco mesophyll protoplasts by tobacco mosaic virus. Proceedings of the National Academy of Sciences of the United States of America 64:843–848
    [Google Scholar]
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