Summary
Protoplasts were isolated from palisade tissue of tobacco leaves by treatment with pectinase and cellulase under aseptic conditions, and were cultured in a synthetic liquid medium. Calcofluor, a fluorescent brightener, was found to be an excellent stain for plant cell walls and was used to demonstrate regeneration of cell walls in these protoplasts. The cultured protoplasts regenerated cell walls by the 3rd day of culture, giving rise to spherical cells. The majority of the protoplasts regenerating cell walls underwent mitosis and cell division. The cycle of mitosis and cell division was repeated 2–3 times during 2 weeks of culture. Some of the nutritional conditions affecting division in the cultured protoplasts were studied.
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References
Aoki, S., Takebe, I.: Infection of tobacco mesophyll protoplasts by tobacco mosaic virus ribonucleic acid. Virology 39, 439–448 (1969).
Cocking, E. C.: Plant protoplasts. Viewpoints in biology, vol. 4, p. 170–203 (J. D. Carthy, and C. L. Duddington, eds.). London: Butterworths 1965.
—, Pojnar, E.: An electron microscopic study of the infection of isolated tomato fruit protoplasts by tobacco mosaic virus. J. gen. Virol. 4 305–312 (1969).
Cole, K.: Induced fluorescence in gametophytes of some Laminariales. Canad. J. Bot. 42, 1173–1184 (1964).
Darken, M. A.: Absorption and transport of fluorescent brighteners by microorganisms. Appl. Microbiol. 10, 387–393 (1962).
Eriksson, T.: Partial synchronization of cell division in suspension cultures of Haplopappus gracilis. Physiol. Plantarum (Cph.) 19, 900–910 (1966).
—, Jonasson, K.: Nuclear division in isolated protoplasts from cells of higher plants grown in vitro. Planta (Berl.) 89, 85–89 (1969).
Harrington, B. J., Raper, K. B.: Use of a fluorescent brightener to demonstrate cellulose in the cellular slime molds. Appl. Microbiol. 16, 106–113 (1968).
Nakazawa, S., Takamura, K., Abe, M.: Rhizoid differentiation in Fucus eggs labeled with Calcofluor White and birefringence of cell wall. Bot. Mag. (Tokyo) 82, 41–44 (1969).
Pojnar, E., Willison, J. H. M., Cocking, E. C.: Cell-wall regeneration by isolated tomato-fruit protoplasts. Protoplasma 64, 460–480 (1967).
Power, J. B., Cocking, E. C.: Isolation of leaf protoplasts: Macromolecule uptake and growth substance response. J. exp. Bot. 21, 64–70 (1970).
Takahashi, W. N.: Increasing the sensitivity of the local-lesion method of virus assay. Phytopathol. 46, 654–656 (1956).
Takebe, I., Otsuki, Y.: Infection of tobacco mesophyll protoplasts by tobacco mosaic virus. Proc. nat. Acad. Sci. (Wash.) 64, 843–850 (1969).
——, Aoki, S.: Isolation of tobacco mesophyll cells in intact and active state. Plant and Cell Physiol. 9, 115–124 (1968).
Usui, H., Takebe, I.: Division and growth of single mesophyll cells isolated enzymatically from tobacco leaves. Development, Growth and Differentiation 11, 143–151 (1969).
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Nagata, T., Takebe, I. Cell wall regeneration and cell division in isolated tobacco mesophyll protoplasts. Planta 92, 301–308 (1970). https://doi.org/10.1007/BF00385097
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DOI: https://doi.org/10.1007/BF00385097