Abstract
When cloned pith and leaf tissues of Nicotiana tabacum L. cv. Havana 425 are subcultured for 3 d on auxin-containing medium and labelled for 18 h with [35S]methionine, up to 10% of the labelled, soluble-protein fraction is found in a single band with an apparent molecular weight of approx. 32,000–34,000 dalton on sodium-dodecylsulfate polyacrylamide-gel electrophoretograms. The labelling of this band, designated P33, is dramatically inhibited by the cytokinin, kinetin, in some cell lines at concentrations as low as 1.4·10-8 M. P33 is a major component of the protein fraction obtained from non-habituated clones, cytokinin-habituated clones, and revertant subclones of crown-gall-transformed clones, but cannot be detected in clones habituated for both auxin and cytokinin, or crown-gall-transformed clones. The evidence supports the hypothesis that cytokinin in the presence of auxin regulates the production of a specific, major polypeptide in the soluble-protein fraction of the tissue and that this protein is not produced in tissues autotrophic for both auxin and cytokinin.
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References
Anderson, L.E., McClure, W.O. (1973) An improved scintillation cocktail of high-solubilizing power. Anal. Biochem. 51, 173–179
Axelos, M., Péaud-Lenoël, C. (1980) The apoprotein of the light-harvesting chlorophyll a/b complex of tobacco cells as a molecular marker of cytokinin activity. Plant Sci. Lett. 19, 33–41
Bevan, M., Northcote, D.H. (1981) Subculture-induced protein synthesis in tissue cultures of Glycine max and Phaseolus vulgaris. Plant 152, 24–31
Binns, A., Meins, F., Jr. (1973) Evidence that habituation of tobacco pith cells for cell division-promoting factors is heritable and potentially reversible. Proc. Natl. Acad. Sci. USA 70, 2660–2662
Bomhoff, G., Klapwijk, P.M., Kester, H.C.M., Schilperoort, R.A., Hernalsteen, I.P., Schell, J. (1976) Octopine and nopaline synthesis and breakdown genetically controlled by a plasmid of Agrobacterium tumefaciens. Mol. Gen. Genet. 145, 177–181
Bonner, W.M., Laskey, R.A. (1974) A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur. J. Biochem. 46, 83–88
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Braun, A.C. (1959) A demonstration of the recovery of the crown-gall tumor cell with the use of complex tumors of single-cell origin. Proc. Natl. Acad. Sci. USA 45, 932–938
Eschenbruch, M., Bürk, R.R. (1982) Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels. Anal. Biochem. 125, 96–99
Fairbanks, G., Steck, T.L., Wallach, D.F.H. (1971) Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10, 2606–2617
Fosket, D.E. (1980) Hormonal control of morphogenesis in cultured tissue. In: Plant growth substances 1979, pp. 363–369, Skoog, F., ed. Springer, Berlin Heidelberg New York
Fosket, D.E., Volk, M.J., Goldsmith, M.R. (1977) Polyribosome formation in relation to cytokinin-induced cell division in suspension cultures of Glycine max (L.) Merr. Plant Physiol. 60, 554–562
Jouanneau, J.-F. (1970) Renouvellement des protéines et effet spécifique de la kinétine sur des cultures de cellules de tabac. Physiol. Plant. 23, 232–244
Kaminek, M., Hadacova, V., Lustinec, J. (1981) Origin of cytokinin-and auxin-autonomy and changes in specific proteins in tobacco callus tissue. Biol. Plant. 23, 228–236
Kulaeva, O.N. (1980) Cytokinin action on enzyme activities in plants. In: Plant growth substances 1979, pp. 119–128, Skoog, F., ed. Springer, Berlin Heidelberg New York
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227, 680–685
Linsmaier, E.M., Skoog, F. (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol. Plant. 18, 100–127
Meins, F., Jr. (1982b) Tumor reversal in plants. Prog. Clin. Biol. Res. 85B, 567–576
Meins, F., Jr., Binns, A.N. (1977) Epigenetic Variation of cultured somatic cells: evidence for gradual changes in the requirement for factors promoting cell division. Proc. Natl. Acad. Sci. USA 74, 2928–2932
Meins, F., Jr., Lutz, J. (1980) The induction of cytokinin habituation in primary pith explants of tobacco. Planta 149, 402–407
Meins, F., Jr., Lutz, J., Binns, A.N. (1980) Variation in the competence of tobacco pith cells for cytokinin-habituation in culture. Differentiation 16, 71–75
Meyer, Y., Chartier, Y. (1981) Hormocal control of mitotic development in tobacco. Two-dimensional distribution of newly synthesized proteins. Plant Physiol. 68, 1273–1278
Oakley, B.R., Kirsch, D.R., Morris, N.R. (1980) A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal. Biochem. 105, 361–363
Otten, L.A.B.M., Schilperoort, R.A. (1978) A rapid microscale method for the detection of lysopine and nopaline dehydrogenase activities. Biochim. Biophys. Acta 527, 497–500
Studier, F.W. (1973) Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J. Mol. Biol. 79, 237–248
Wang, T.L., Everett, N.P., Gould, A.R., Street, H.E. (1981) Studies on the control of the cell cycle in cultured plant cells. III. The effects of cytokinin. Protoplasma 106, 23–25
Watson, B., Currier, T.C., Gordon, M.P., Chilton, M.-D., Nester, E.W. (1975) Plasmid required for virulence of Agrobacterium tumefaciens. J. Bacteriol. 123, 255–264
White, P.R. (1943) A handbook of plant tissue culture. Jacques Cattel, Lancaster, Pa.
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Eichholz, R., Harper, J., Felix, G. et al. Evidence for an abundant 33,000-dalton polypeptide regulated by cytokinins in cultured tobacco tissues. Planta 158, 410–415 (1983). https://doi.org/10.1007/BF00397733
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DOI: https://doi.org/10.1007/BF00397733