Abstract
Isolated roots of Lycopersicon esculentum Mill., cultured in axenic conditions were starved of sulphate or phosphate, and uptake capacities for the respective oxyanion-transport systems were observed for several days after sulphate or phosphate withdrawal. Sulphate-uptake capacity of the intact roots, measured in a 20-min period, increased from a control level of 100 nmol · g−1 · h−1 to 1100 nmol · g−1 · h−1 in 10 d, and phosphate-uptake capacity increased from 500 to 1400 nmol · g−1 · h−1 over 4 d. Newly synthesised polypeptides of these root cultures were pulse-labelled in vivo for 2 h, by adding [3H]leucine to the culture medium. The tissue was immediately homogenised and soluble and membrane fractions were prepared. A highly purified plasma-membrane fraction was separated from the crude microsomal membrane fraction using an aqueous two-phase partitioning technique. All fractions were analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and autoradiography. A 28-kilodalton (kDa) soluble polypeptide, and 36-, 43-, and 47-kDa plasma-membrane polypeptides were observed to have increased labelling after 4 d of sulphate deprivation. Longer periods resulted in additional polypeptides with increased [3H]leucine incorporation. The synthesis of a 25-kDa membrane polypeptide and a 65-kDa soluble polypeptide was increased after 4 d of phosphate deprivation. Two-dimensional electrophoresis afforded greater resolution of the plasmamembrane polypeptides, confirming increased synthesis of the 36-kDa polypeptide and the presence of the 28-kDa polypeptide in the plasma-membrane preparation from sulphate-starved roots. These polypeptides were also observed in protein-stained two-dimensional gels as low-abundant protein components of the plasmamembrane fraction. It is suggested that the 36-kDa polypeptide may be a component of the plasma-membrane sulphate-transport system and that the 25-kDa polypeptide may be a component of a phosphate-transport system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- kDa:
-
kilodalton(s)
- PAGE:
-
polyacrylamide gel electrophoresis
- pI:
-
isoelectric point
- SDS:
-
Sodium dodecyl sulphate
References
Butcher, D.N., Clark, J.A., Lenton, J.R. (1990) Gibberellins and the growth of excised tomato roots: comparison of gib-l mutant and wild type and responses to applied GA3] and 25,3S paclobutrazol. J. Exp. Bot. 41, 715–722
Butcher, D.N., Ingram, D.S. (1976) Plant tissue culture. Edward Arnold, London
Clarkson, D.T., Lüttge, U. (1991) Mineral nutrition: inducible and repressible nutrient transport systems. Prog. Bot. 52, 61–83
Clarkson, D.T., Saker, L.R. (1989) Sulphate influx in wheat and barley roots becomes more sensitive to specific protein-binding reagents when plants are sulphate-deficient. Planta 178, 249–257
Clarkson, D.T., Scattergood, C.B. (1982) Growth and phosphatetransport in barley and tomato plants during the development of, and recovery from, phosphate-stress. J. Exp. Bot. 33, 865–875
Clarkson, D.T., Smith, F.W., Vanden Berg, P.J. (1983) Regulation of sulphate transport in a tropical legume, Macroptilium atropurpureum, cv. Siratro. J. Exp. Bot. 34, 1463–1483
Clarkson, D., Hawkesford, M., Saker, L. (1989) The sulphate transporter in roots: physiological behaviour and attempted identification. In: Plant membrane transport: the current position, pp. 401–402, Dainty, J., De Michelis, M.I., Marrè, E., Rasi-Caldogno, F., eds, Elsevier, Amsterdam New York Oxford
Cupo, J.F., Lidgard, G.P., Lichtman, W.F. (1990) A high resolution two-dimensional gel electrophoresis and silver staining protocol demonstrated with nuclear matrix proteins. Electrophoresis 11, 500–504
Dhugga, K.S., Waines, J.G., Leonard, R.T. (1988) Correlated induction of nitrate uptake and membrane polypeptides in corn roots. Plant Physiol. 87, 120–125
Dumont, F., Loppes, R., Kremers, P. (1990) New polypeptides and in-vitro-translatable mRNAs are produced by phosphate starved cells of the unicellular algae Chlamydomonas reinhardtii. Planta 182, 610–616
Fife, C.A., Newcomb, W., Lefebvre, D.D. (1990) The effect of phosphate deprivation on protein synthesis and fixed carbon storage reserves in Brassica nigra suspension cells. Can. J. Bot. 68, 1840–1847
Goldstein, A.H., Baertlein, D.A., McDaniel, R.G. (1988a) Phosphate starvation inducible metabolism in Lycopersicon esculentum. I. Excretion of acid phosphatase by tomato plants and suspension-cultured cells. Plant Physiol. 87, 711–715
Goldstern, A.H., Danon, A., Baertlein, D.A., McDaniel, R.G. (1988b) Phosphate starvation inducible metabolism in Lycopersicon esculentum. II. Characterization of the phosphate starvation inducible-excreted acid phosphatase. Plant Physiol. 87, 716–720
Goldstein, A.H., Mayfield, S.P., Danon, A., Tibbot, B.K. (1989) Phosphate starvation inducible metabolism in Lycopersicon esculentum. III. Changes in protein secretion under nutrient stress. Plant Physiol. 91, 175–182
Hawkesford, M.J., Liddell, A.D., Leaver, C.J. (1989) Subunit composition of cytochrome c oxidase in mitochondria of Zea mays. Plant Physiol. 91, 1535–1542
Hurkman, W.J., Tanaka, C.K. (1986) Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. Plant Physiol. 81, 802–806
Larsson, C., Widell, S., Kjellbom, P. (1987) Preparation of highpurity plasma membranes. Methods Enzymol. 148, 558–568
Lee, R.B. (1982) Selectivity and kinetics of ion uptake by barley plants following nutrient deficiency. Ann. Bot. 50, 429–449
Lefebvre, D.D., Duff, S.M.G., Fife, C.A., Julien-Inalsingh, C., Plaxton, W.C. (1990) Response to phosphate deprivation in Brassica nigra suspension cells. Enhancement of intracellular, cell surface, and secreted phosphatase activities compared to increases in Pi-absorption rate. Plant Physiol. 93, 504–511
McClure, P.R., Omholt, T.E., Pace, G.M., Bouthyette, P.-Y. (1987) Nitrate-induced changes in protein synthesis and translation of RNA in maize roots. Plant Physiol. 84, 52–57
Rennenberg, H., Kemper, O., Thoene, B. (1989) Recovery of sulfate transport into heterotrophic tobacco cells from inhibition by reduced glutathione. Physiol. Plant. 76, 271–276
Reuveny, Z. (1977) Derepression of ATP sulfurylase by the sulfate analogs molybdate and selenate in cultured tobacco cells. Proc. Natl Acad. Sci. USA 74, 619–622
Ruiz-Cristin, J., Briskin, D.P. (1991) Characterization of a H+/ NO3- symport associated with plasma membrane vesicles of maize roots using 36C10 −3 as a radiotracer analog. Arch. Biochem. Biophys. 285, 74–82
Schwenn, J.D., Kemena, A. (1984) Expression of the plant sulphite reductase in cell suspension cultures from Catharanthus roseus L. Planta 160, 151–158
Sussman, M.R., Harper, J.F. (1989) Molecular biology of the plasma membrane of higher plants. Plant Cell 1, 953–960
Author information
Authors and Affiliations
Additional information
This work was supported by the Agricultural and Food Research Council via grants-in-aid to Long Ashton Research Station. We are also grateful for discussions with our colleagues D.T. Clarkson (LARS) and J.-C. Davidian (ENSA/INRA, Montpellier).
Rights and permissions
About this article
Cite this article
Hawkesford, M.J., Belcher, A.R. Differential protein synthesis in response to sulphate and phosphate deprivation: Identification of possible components of plasma-membrane transport systems in cultured tomato roots. Planta 185, 323–329 (1991). https://doi.org/10.1007/BF00201051
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00201051