Abstract
Plasma membrane vesicles, purified by aqueous two-phase partitioning, were used to investigate the presence of sugar and amino acid carriers in cotyledons and roots of Ricinus communis L. and in roots of red beet (Beta vulgaris L.). Artificial pH and electrical gradients were generated across the plasma membrane, and [14C]acetate and [14C]tetraphenylphosphonium were used to demonstrate the presence of an internal alkaline pH gradient and an internal negative membrane potential, respectively. In Ricinus cotyledons, uptake of sucrose was more strongly inhibited than that of glutamine by p-chloromercuribenzenesulphonic acid, phlorizin and phenylglyoxal. The sucrose transport system showed a high degree of substrate specificity with only the presence of maltose and phenyl-α-glucoside significantly affecting sucrose uptake; in contrast, the glutamine transport system was inhibited by a number of other amino acids. ΔpH+gDψ-driven glutamine uptake showed saturation kinetics with a K m of 0.35 mol · m−3. Sucrose and glutamine Δψ-driven uptake was pH dependent with an optimum in the acidic range (pH 6.25) and a decrease at higher pH values. Vesicles obtained from cotyledons and roots of Ricinus showed different transport properties. In the cotyledons, gDH+gDψ-driven transport for both sucrose and glutamine were observed at similar levels; however, in the root tissue, δpH-Δψ-driven glutamine transport was the dominant uptake process. Uptake rates for glucose and fructose were low in the cotyledons whereas, in the roots, glucose and sucrose transport were slightly higher than that of fructose. In vesicles from red beet tissue there was a different uptake profile, with evidence of proton-coupled cotransport systems for sucrose and glucose, but lower uptake of glutamine and fructose. The results are discussed in relation to the reported different pathways for loading and unloading of solutes in these tissues.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CCCP:
-
carbonyl cyanide-m-chlorophyenyl hydrazone
- DEPC:
-
diethyl pyrocarbonate
- NEM:
-
N-ethylmaleimide
- PCMBS:
-
p-chloromercuribenzenesulfonic acid
- TPP:
-
tetraphenylphosphonium ion
- gDμH+ :
-
proton electrochemical potential gradient
- Δψ :
-
membrane potential
References
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
Buchout, T.J. (1989) Sucrose transport in isolated plasma membrane vesicles from sugar beet (Beta vulgaris L.). Evidence for an electrogenic sucrose-proton symport. Planta 178, 393–399
Bush, D.R. (1989) Proton-coupled sucrose transport in plasmalemma vesicles isolated from sugar beet (Beta vulgaris L. cv. Great Western) leaves. Plant Physiol. 89, 1318–1323
Bush, D.R. (1990) Electrogenicity, pH-dependence and stoichiometry of the proton-sucrose symport. Plant Physiol. 93, 1590–1596
Chapleo, S., Hall, J.L. (1989) Sugar unloading in roots of Ricinus communis L. III. The extravascular pathway of sugar transport. New Phytol. 111, 391–396
Delrot, S., Gallet, O., Li, Z-S., Lemoine, R., Gaillard, C., Laloi, M. (1991) Use of plasma membrane vesicles from sugar beet leaves for the study of sucrose transport and of sucrose transporter(s). In: Recent advances in phloem transport and assimilate compartmentation, pp. 154–166, Bonnemain, J.L., Delrot, S., Lucas, W.J. Dainty, J., eds. Ouest editions Presses Academiques, Nantes, France
Getz, H-P., Knauer, D., Willenbrink, J. (1987) Transport of sugars across the plasma membrane of beetroot protoplasts. Planta 171, 185–196
Hutchings, V. (1978) Sucrose and proton cotransport in Ricinus cotyledons. I. H+ Influx associated with sucrose uptake. Planta 138, 229–235
Komor, E. (1977) Sucrose uptake by cotyledons of Ricinus communis L: Characteristics, mechanism and regulation. Planta 137, 119–131
Komor, E., Rotter, M., Tanner, W. (1977) A proton-cotransport system in a higher plant: sucrose transport in Ricinus communis. Plant Sci. Lett. 9, 153–162
Komor, E., Orlich, G., Kohler, J., Hall, J.L., Williams, L.E. (1991) The Ricinus seedling — a model plant to study phloem transport. In: Recent advances in phloem transport and assimilate compartmentation, pp. 301–308, Bonnemain, J.L., Delrot, S., Lucas, W.J., Dainty, J. eds. Ouest editions Presses Academiques, Nantes, France
Kriedemann, P., Beevers, H. (1967) Sugar uptake and translocation in the castor bean seedling. I. Characteristics of transfer in intact and excised seedlings. Plant Physiol 42, 161–173
Larsson, C. (1983) Partition in aqueous polymer two-phase systems. In: Isolation of membranes and organelles from plant cells, pp. 277–309, Hall, J.L., Moore, A.L., eds. Academic Press, London
Lemoine, R., Delrot, S. (1989) Proton-motive-force-driven sucrose uptake in sugar beet plasma membrane vesicles. FEBS Lett. 249, 129–133
Lemoine, R., Daie, J., Wyse, R. (1988) Evidence for the presence of a sucrose carrier in immature sugar beet tap root. Plant Physiol. 86, 575–580
Li, Z-C., Bush, D.R. (1990) ΔpH-dependent amino acid transport into plasma membrane vesicles isolated from sugar beet leaves. I. Evidence for carrier-mediated, electrogenic flux through multiple transport systems. Plant Physiol. 94, 268–277
Palmgren, M.G., Sommarin, M., Ulvskov, P., Larsson, C. (1990) Effect of detergents on the H+-ATPase activity of inside-out and right-side-out plasma membrane vesicles. Biochim. Biophys. Acta 1021, 133–140
Poole, R.J. (1988) Plasma membrane and tonoplast. In: Solute transport in plant cells and tissues, pp. 83–105, Baker, D.A., Hall, J.L., eds. Longman, Harlow, UK
Ramos, S., Kaback, H.R. (1977) The electrochemical proton gradient in Escherichia coli membrane vesicles. Biochemistry 16, 848–854
Reinhold, L., Kaplan, A. (1984) Membrane transport of sugars and amino acids. Annu. Rev. Plant Physiol. 35, 45–83
Robinson, S.P., Beevers, H. (1981) Evidence for amino-acid: proton cotransport in Ricinus cotyledons. Planta 152, 527–533
Schobert, C., Komor, E. (1987) Amino acid uptake by Ricinus communis roots: characterization and physiological significance. Plant Cell Environ. 10, 493–500
Schobert, C., Komor, E. (1990) Transfer of amino acids and nitrate from the roots into the xylem of Ricinus communis seedlings. Planta 181, 85–90
Toggenburger, G., Kessler, M., Semenza, G. (1982) Phlorizin as a probe of the small intestine Na+, D-glucose cotransporter. A model. Biochim. Biophys. Acta 688, 557–571
Verstappen, R., Ranostaj, S., Rausch, T. (1991) The hexose transporters at the plasma membrane and the tonoplast of transformed plant cells: kinetic characterization of two distinct carriers. Biochim. Biophys. Acta 1073, 366–373
Williams, L.E. (1987) ATPase and proton pumping activities in cotyledons and other phloem-containing tissues of Ricinus communis and the relationship to phloem loading. Ph.D. thesis, University of Southampton, UK
Williams, L., Hall, J.L. (1987) ATPase and proton pumping activities in cotyledons and other phloem-containing tissues of Ricinus communis. J. Exp. Bot. 38, 185–202
Williams, L.E., Nelson, S.J., Hall, J.L. (1990a) Characterization of solute transport in plasma membrane vesicles isolated from cotyledons of Ricinus communis. I. Adenosine-triphosphatase and pyrophosphatase activites associated with a plasma membrane fraction isolated by phase partitioning. Planta 182, 532–539
Williams, L.E., Nelson, S.J., Hall, J.L. (1990b) Characterization of solute transport in plasma membrane vesicles isolated from cotyledons of Ricinus communis. II. Evidence for a proton-coupled mechanism for sucrose and amino acid uptake. Planta 182, 540–545
Author information
Authors and Affiliations
Additional information
We would like to thank the SERC(UK) and the Royal Society for financial support.
Rights and permissions
About this article
Cite this article
Williams, L.E., Nelson, S.J. & Hall, J.L. Characterization of solute/proton cotransport in plasma membrane vesicles from Ricinus cotyledons, and a comparison with other tissues. Planta 186, 541–550 (1992). https://doi.org/10.1007/BF00198034
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00198034