Abstract
Photoheterotrophic and heterotrophic suspension cultures of tobacco (Nicotiana tabacum L.) were grown with 1 mM glutathione (reduced; GSH) as sole source of sulfur. Addition of sulfate to both cultures did not alter the rate of exponential growth, but affected the removal of GSH and sulfate in different ways. In photoheterotrophic suspensions, addition of sulfate caused a decline in the net uptake of GSH, whereas sulfate was taken up by the green cells immediately. In heterotrophic suspensions, however, addition of sulfate did not affect the net uptake of GSH and sulfate was only taken up by the cells after the GSH supply in the medium had been exhausted. Apparently, GSH uptake in photoheterotrophic cells is inhibited by sulfate, whereas sulfate uptake is inhibited by GSH in heterotrophic cells. The differences in the effect of GSH on sulfate uptake in photoheterotrophic and heterotrophic tobacco suspensions cannot be attributed to differences in the kinetic properties of sulfate carriers. In short-time transport experiments, both cultures took up sulfate almost entirely by an active-transport system as shown by experiments with metabolic inhibitors; sulfate transport of both cultures obeyed monophasic Michaelis-Menten kinetics with similar app. Km (photoheterotrophic cells: 16.0±2.0 μM; heterotrophic cells: 11.8±1.8 μM) and Vmax (photoheterotrophic cells: 323±50 nmol·min-1·g-1 dry weight; heterotrophic cells: 233±3 nmol·min-1·g-1 dry weight). Temperature- and pH-dependence of sulfate transport showed almost identical patterns. However, the cultures exhibited remarkable differences in the inhibition of sulfur influx by GSH in short-time transport experiments. Whereas 1 mM GSH inhibited sulfate transport into heterotrophic tobacco cells completely, sulfate transport into photoheterotrophic cells proceeded at more than two-thirds of its maximum velocity at this GSH concentration. The mode of action of GSH on sulfate transport in chloroplast-free tobacco cell does not appear to be direct: a 14-h exposure to 1 mM GSH was found to be necessary to completely block sulfate transport; a 4-h time of exposure did not affect this process. Consequently, glutathione does not seem to be a product of sulfur metabolism acting on sulfate-carrier entities by negative feedback control. When transferred to the whole plant, the observed differences in sulfate and glutathione influx into green and chloroplast-free cells may be interpreted as a regulatory device to prevent the uptake of excess sulfate by plants.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- DCCD:
-
N,N′-dicyclohexylcarbodiimide
- DNP:
-
dinitrophenol
- DW:
-
dry weight
- FW:
-
fresh weight
- GSH:
-
reduced glutathione
References
Anderson, J.W. (1980) Assimilation of inorganic sulfate into cysteine. In: The biochemistry of plants, vol. 5: Amino acids and derivatives, pp. 203–223, Miflin, B.J., ed. Academic Press, New York
Bergmann, L. (1960) Growth and division of single cells of higher plants in vitro. J. Gen. Physiol. 43, 841–851
Bergmann, L., Rennenberg, H. (1978) Efflux and production of glutathione in suspension cultures of Nicotiana tabacum. Z. Pflanzenphysiol. 88, 175–185
Bonas, U., Schmitz, K., Rennenberg, H., Bergmann, L. (1982) Phloem transport of sulfur in Ricinus. Planta 155, 82–88
Cacco, G., Ferrarí, G., Saccomani, M. (1980) Pattern of sulfate uptake during root elongation in maize: its correlation with productivity. Physiol. Plant. 48, 375–378
Cram, W.J. (1983) Characteristics of sulfate transport across plasmalemma and tonoplast of carrot root cells. Plant Physiol. 72, 204–211
de Kok, L., de Kan, P.L.J., Tánczos, O., Kuiper, P.J.C. (1981) Sulphate induced accumulation of glutathione and frosttolerance of spinach leaf tissue. Physiol. Plant. 53, 435–438
Hart, J.W., Filner, P. (1969) Regulation of sulfate uptake by amino acids in cultured tobacco cells. Plant Physiol. 44, 1253–1259
Holmern, K., Vange, M.S., Nissen, P. (1974) Multiphasic uptake of sulfate by barley roots. II. Effects of washing, divalent cations, inhibitors, and temperature. Physiol. Plant. 31, 302–310
Jones, S.L., Smith, I.K. (1981) Sulfate transport in cultured tobacco cells. Effect of calcium and sulfate concentration. Plant Physiol. 67, 445–448
Kahn, J.S. (1974) Physiological adaption of Euglena gracilis to uncouplers and inhibitors of oxidative phosphorylation. Arch. Biochem. Biophys. 164, 266–274
Lay, M.-M., Casida, J. (1976) Dichloroacetamide antidotes enhance thiocarbamate sulfoxide detoxification by elevating corn root glutathione content and glutathione-S-transferase activity. Pestic. Biochem. Physiol. 6, 442–456
Lineweaver, H., Burk, D. (1914) The determination of enzyme dissociation constants. J. Am. Chem. Soc. 56, 658–666
Logemann, H., Bergmann, L. (1974) Einfluß von Licht und Medium auf die „Plating Efficiency” isolierter Zellen aus Calluskulturen von Nicotiana tabacum var. „Samsun”. Planta 121, 283–292
Menon, V.K.N., Varma, A.K. (1982) Sulfate uptake in the cyanobacterium Spirulina platensis. FEMS Microbiol. Lett. 13, 141–146
Moss, M.R. (1978) Sources of sulfur in the environment; the global sulfur cycle. In: Sulfur in the environment, part I: The atmospheric cycle, pp. 23–50, Nriagu, J.O., ed. Wiley, New York
Nissen, P. (1971) Uptake of sulfate by roots and leaf slices of barley: mediated by single, multiphasic mechanisms. Physiol. Plant. 24, 315–324
Nissen, P. (1973) Multiphasic uptake in plants. I. Phosphate and sulfate. Physiol. Plant. 28, 304–316
Nissen, P. (1974) Uptake mechanisms: inorganic and organic. Annu. Rev. Plant Physiol. 25, 53–79
Pätau, K. (1943) Zur statistischen Beurteilung von Messungsreihen (Eine neue t-Tafel). Biol. Zentralbl. 63, 152–168
Rennenberg, H. (1981) Differences in the use of cysteine and glutathione as sulfur source in photoheterotrophic tobacco suspension cultures. Z. Pflanzenphysiol. 105, 31–40
Rennenberg, H. (1982) Glutathione metabolism and possible biological roles in higher plants. Phytochemistry 21, 2771–2781
Rennenberg, H. (1984) The fate of excess sulfurin higher plants. Annu. Rev. Plant Physiol. 35, 121–153
Rennenberg, H., Bergmann, L. (1979) Influences of ammonium and sulfate on the production of glutathione in suspension cultures of Nicotiana tabacum. Z. Pflanzenphysiol. 92, 133–142
Rennenberg, H., Schmitz, K., Bergmann, L. (1979) Long-distance transport of sulfur in Nicotiana tabacum. Planta 147, 57–62
Schmidt, A. (1979) Photosynthetic assimilation of sulfur compounds. In: Encyclopedia of plant physiology, N.S., vol. 6: Photosynthesis II, pp. 481–496, Gibbs, M., Latzko, E., eds. Springer, Berlin Heidelberg New York
Shargool, P.D., Ngo, T.T. (1975) The uptake of sulfate by excised roots of rape seedling (Brassica napus var. Target). Can. J. Bot. 53, 914–920
Smith, I.K. (1975) Sulfate transport in cultured tobacco cells. Plant Physiol. 55, 303–307
Smith, I.K. (1976) Characterization of sulfate transport in cultured tobacco cells. Plant Physiol. 58, 358–362
Steinkamp, R., Rennenberg, H. (1985) Degradation of glutathione in plant cells: evidence against the participation of a γ-glutamyl-transpeptidase. Z. Naturforsch. 40c, 29–33
Vange, M.S., Holmern, K., Nissen, P. (1974) Multiphasic uptake of sulfate by barley roots. I. Effects of analogues, phosphate, and pH. Physiol. Plant. 31, 292–301
Willenbrink, J. (1967) Der Transport der phosphor- und schwefelhaltigen Verbindungen. In: Handbuch der Pflanzenphysiologie, Bd. XIII, pp. 178–199, Ruhland, W., ed. Springer, Berlin Heidelberg New York
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rennenberg, H., Polle, A., Martini, N. et al. Interaction of sulfate and glutathione transport in cultured tobacco cells. Planta 176, 68–74 (1988). https://doi.org/10.1007/BF00392481
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392481