Abstract
We have analysed the intracellular localisation of phytochrome in oat coleoptile cells by electron microscopy and confirm and extend light-microscopical findings of previous authors. We used indirect immuno-labeling with polyclonal antibodies against 60-KDa phytochrome from etiolated oat seedlings, and a gold-coupled second antibody, on ultrathin sections of LR-white-embedded material. In dark-grown seedlings, phytochrome-labeling is distributed diffusely throughout the cytoplasm. Organelles and membranes are not labeled. After photoconversion of the red-absorbing form of phytochrome to the far-red absorbing form (Pfr) (5-min red light; 660 nm), the label is sequestered uniquely in electron-dense areas within the cytoplasm. These areas are irregularly shaped, are often located in the vicinity of the vacuole, are not surrounded by a membrane, exclude cellular organelles and ribosomes and are not found in dark-grown material; an immediate 5-min farred light pulse after the red light does not cause these structures to disappear. After a dark period of 3–4 h following red-light irradiation, these electron-dense structures disappear together with any specific labeling. We suggest a Pfr-induced aggregation of an unknown, phytochrome-binding protein or proteins.
Similar content being viewed by others
Abbreviations
- Pr and Pfr :
-
phytochrome in its red and far-red absorbing form, respectively
References
Coleman, R.A., Pratt, L.H. (1974a) Electron microscopic localization of phytochrome in plants using an indirect antibody labeling method. J. Histochem. Cytochem. 22, 1039–1047
Coleman, R.A., Pratt, L.H. (1974b) Phytochrome: Immunocytochemical assay of synthesis and destruction. Planta 119, 221–231
Craig, S., Miller, C. (1984) LR White resin and improved ongrid immunogold deletion of vicilin, a pea seed storage protein. Cell Biol. Intern. Rep. 8, 879–886
Epel, B.C., Butter, W.L., Pratt, L.H., Tokuyasu, K.I. (1980) Immunofluorescence localization studies of the Pr and Pfr forms of phytochrome in the coleoptile tips of oats, corn and wheat. In: Photoreceptions and plant development, pp. 121–133, de Greef, J., ed. Antwerpen University Press, Belgium
Hendricks, S.B., Borthwick, H.A. (1967) The function of phytochrome in regulation of plant growth. Proc. Natl. Acad. Sci. USA 58, 2125–2130
Mackenzie, J.M., Coleman, R.A., Briggs, W.R., Pratt, L.H. (1975) Reversible redistribution of phytochrome within the cell upon conversion to its physiologically active form. Proc. Natl. Acad. Sci. USA 72, 799–803
Marmé, D. (1977) Phytochrome: Membranes as possible sites of primary action. Annu. Rev. Plant Physiol. 28, 173–198
Mohr, H., Shropshire, W., Jr. (1983) An introduction to photomorphogenesis for the general reader. In: Encyclopedia of plant physiology, N.S., vol. 16A: Photomorphogenesis, pp. 24–38, Shropshire, W., Jr., Mohr, H., eds. Springer, Berlin Heidelberg New York Tokyo
Mösinger, E., Batschauer, A., Schäfer, E., Apel, K. (1985) Phytochrome control of in vitro transcription of specific genes in isolated nuclei from barley (Hordeum vulgare). Eur. J. Biochem. 147, 137–142
Otto, V., Mösinger, E., Sauter, M., Schäfer, E. (1983) Phytochrome control of its own synthesis in Sorghum vulgare and Avena sativa. Photochem. Photobiol. 38, 693–700
Pratt, L.H. (1979) Phytochrome: function and properties. Photochem. Photobiol. Rev. 4, 59–124
Pratt, L.H., Coleman, R.A. (1974) Phytochrome distribution in etiolated grass seedlings as assayed by an indirect antibody-labeling method. Am. J. Bot. 61, 195–202
Quail, P.H., Briggs, W.R. (1978) Irradiation-enhanced phytochrome pelletibility. Plant Physiol. 62, 773–778
Saunders, M.J., Cordonnier, M.-M., Palevitz, B.A., Pratt, L.H. (1983) Immunofluorescence visualization of phytochrome in Pisum sativum. L. epicotyls using monoclonal antibodies. Planta 159, 545–553
Schäfer, E., Apel, K., Batschauer, A., Mösinger, E. (1986) The molecular biology of action. In: Photomorphogenesis in plants, Hendrick, R.E., Kronenberg, G.H.M., eds. Wageningen, in press
Tobin, E.M., Silverthorne, J. (1985) Light regulation of gene supression in higher plants. Annu. Rev. Plant. Physiol. 36, 569–593
Verbelen, J.-P., Pratt, L.H., Butler, W.L., Tokuyasu, K. (1982) Localization of phytochrome in oats by electron microscopy. Plant Physiol. 70, 867–871
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Speth, V., Otto, V. & Schäfer, E. Intracellular localisation of phytochrome in oat coleoptiles by electron microscopy. Planta 168, 299–304 (1986). https://doi.org/10.1007/BF00392353
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392353