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Sucrose transport into vacuoles isolated from barley mesophyll protoplasts

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Abstract

Sucrose transport has been investigated in vacuoles isolated from barley mesophyll protoplasts. Rates of sucrose transfer across the tonoplast were even higher in vitro than in vivo indicating that the sucrose transport system had not suffered damage during isolation of the vacuoles. Sucrose transport is carrier-mediated as shown by substrate saturation of transport and sensitivity to a metabolic inhibitor and to competitive substrates. A number of sugars, in particular maltose and raffinose, decreased uptake of sucrose. Sorbitol was slowly taken up but had no effect on sucrose transport. The SH-reagent p-chloromercuribenzene sulfonate inhibited sucrose uptake completely. The apparent Km of the carrier for sucrose uptake was 21 mM. Transport was neither influenced by ATP and pyrophosphate, with or without Mg2+ present, nor by protonophores and valinomycin (with K+ present). Apparently uptake was not energy dependent. Efflux experiments with preloaded vacuoles indicated that sucrose unloading from the isolated vavuoles is mediated by the same carrier which catalyses uptake. The vacuole of mesophyll cells appears to represent an intermediary storage compartment. Uptake of photosynthetic products into the vacuole during the light apparently minimizes osmotic swelling of the small cytosolic compartment of vacuolated leaf cells when photosynthetic productivity exceeds the capacity of the phloem for translocation of sugars.

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Abbreviations

Hepes:

4-(2-hydroxyethyl)-1-piperazincethane-sulfonic acid

pCMBS:

p-chloromercuribenzene sulfonate

References

  • Arnon, D.I. (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15

    Google Scholar 

  • Bergmeyer, H.U. (1970) In: Methoden der enzymatischen Analyse, pp. 396–397, Bergmeyer, H.U., ed., 2nd edn. Verlag Chemie, Weinheim/Bergstraße

    Google Scholar 

  • Boller, T., Alibert, G. (1983) Photosynthesis in protoplasts from Melilotus alba: distribution of products between vacuole and cytosol. Z. Pflanzenphysiol. 110, 231–238

    Google Scholar 

  • Cataldo, D.A. (1974) Vein loading: the role of the symplast in intracellular transport of carbohydrate between the mesophyll and minor veins of tobacco leaves. Plant Physiol. 53, 912–917

    Google Scholar 

  • Doll, S., Rodier, F., Willenbrink, J. (1979) Accumulation of sucrose in vacuoles isolated from red beet tissue. Planta 144, 407–411

    Google Scholar 

  • Fisher, D.B., Outlaw, W.H., Jr. (1979) Sucrose compartimentation in the palisade parenchyma of Vicia faba. Plant Physiol. 64, 481–483

    Google Scholar 

  • Fliege, R., Flügge, U.I., Werdan, K., Heldt, H.W. (1978) Specific transport of inorganic phosphate, 3-phosphoglycerate and triosephosphates across the inner membrane of the envelope in spinach chloroplasts. Biochim. Biophys. Acta 502, 232–247

    PubMed  Google Scholar 

  • Gerhardt, R., Heldt, H.W. (1984) Measurement of subcellular metabolite levels in leaves by fractionation of freeze stopped material in nonaequeous media. Plant Physiol. (in press)

  • Giaquinta, R. (1977) Phloem loading of sucrose. pH dependence and selectivity. Plant Physiol. 59, 750–755

    Google Scholar 

  • Giaquinta, R. (1978) Source and sink leaf metabolism in regulation to phloem translocation. Carbon partitioning and enzymology. Plant Physiol. 61, 380–385

    Google Scholar 

  • Giersch, C., Heber, U., Kaiser, G., Walker, D.A., Robinson, S.P. (1980) Intracellular metabolite gradients and flow of carbon during photosynthesis of leaf protoplasts. Arch. Biochem. Biophys. 205, 246–259

    PubMed  Google Scholar 

  • Gordon, A.J., Ryle, G.J.A., Webb, G. (1980) The relationship between sucrose and starch during dark export from leaves of uniculm barley. J. Exp. Bot 31, 845–850

    Google Scholar 

  • Gordon, A.J., Ryle, G.J.A., Mitchell, D.F., Powell, C.E. (1982) The dynamics of carbon supply from leaves of barley plants grown in long or short days. J. Exp. Bot. 33, 241–250

    Google Scholar 

  • Guy, M., Reinhold, L., Michaeli, D. (1979) Direct evidence for a sugar transport mechanism in isolated vacuoles. Plant Physiol. 64, 61–64

    Google Scholar 

  • Heldt, H.W., Rapley, L. (1970) Specific transport of inorganic phosphate, 3-phosphoglycerate, and dihydroxyacetone phosphate and of dicarboxylates across the inner membrane of spinach chloroplasts. FEBS Lett. 10, 143–148

    PubMed  Google Scholar 

  • Kaiser, G., Martinoia, E., Wiemken, A. (1982) Rapid appearance of photosynthetic products in the vacuoles isolated from barley mesophyll protoplasts by a new fast method. Z. Pflanzenphysiol. 107, 103–113

    Google Scholar 

  • Kaiser, W.M., Kaiser, G., Prachuab, P.K., Wildman, S.G., Heber, U. (1981a) Photosynthesis under osmotic stress. Inhibition of photosynthesis of intact chloroplasts, protoplasts, and leaf slices at high osmotic potentials Planta 153, 416–422

    Google Scholar 

  • Kaiser, W.M., Kaiser, G., Schöner, S., Neimanis, S. (1981b) Photosynthesis under osmotic stress. Differential recovery of photosynthetic activities of stroma enzymes, intact chloroplasts, protoplasts, and leaf slices after exposure to high solute concentration. Planta 153, 430–435

    Google Scholar 

  • Kaiser, W.M., Paul, J.S., Bassham, J.A. (1979) Release of photosynthates from mesophyll cells in vitro and in vivo. Z. Pflanzenphysiol. 94, 377–385

    Google Scholar 

  • Leigh, R.A., Walker, R.R. (1980) ATPase and acid phosphatase activities associated with vacuoles isolated from storage roots of red beet (Beta vulgaris L.). Planta 150, 222–229

    Google Scholar 

  • Martinoia, E., Heck, U., Wiemken, A. (1981) Vacuoles as storage compartments for nitrate in barley leaves. Nature (London) 289, 292–294

    Google Scholar 

  • Smith, F.A., Raven, J.A. (1979) Intracellular pH and its regulation. Annu. Rev. Plant. Physiol. 30, 289–311

    Article  Google Scholar 

  • Sovonick, S.A., Geiger, D.A., Fellows, R.J. (1974) Evidence for active phloem loading in the minor veins of sugar beet. Plant Physiol. 54, 886–891

    Google Scholar 

  • Thom, M., Komor, E., Maretzki, A. (1982) Vacuoles from sugarcane suspension cultures. II. Characterization of sugar uptake. Plant Physiol. 69, 1320–1325

    Google Scholar 

  • Werdan, K., Heldt, H.W., Geller, G. (1972) Accumulation of bicarbonate in intact chloroplasts following a pH gradient. Biochim. Biophys. Acta 283, 430–441

    PubMed  Google Scholar 

  • Willenbrink, J., Doll, S. (1979) Characteristics of the sucrose uptake system of vacuoles isolated from red beet tissue. Kinetics and specifity of the sucrose uptake system. Planta 147, 159–162

    Google Scholar 

  • Wyn Jones, R.G., Storey, R., Leigh, R.A., Ahmad, N., Pollard, A. (1977) A hypothesis on osmotic osmoregulation. In: Regulation of cell membrane activity in plants, pp. 121–136, Marré, E., Ciferri, O., eds. Elsevier/North-Holland Biomedical Press. Amsterdam

    Google Scholar 

  • Keck, R.W., Boyer, J.S. (1974) Chloroplast response to low leaf water potentials. III. Differing inhibition of electron transport and photophosphorylation. Plant Physiol. 53, 474–479

    Google Scholar 

  • Ludlow, M.M., Björkman, O. (1984) Paraheliotropic leaf movement in Siratro as a protective mechanism against drought-induced damage to primary photosynthetic reactions: damage by excessive light and heat. Planta 161, 505–518

    Google Scholar 

  • Mohanty, P., Boyer, J.S. (1976) Chloroplast response to low leaf water potentials. IV. Quantum yield is reduced. Plant Physiol. 57, 704–709

    Google Scholar 

  • Mooney, H.A., Björkman, O., Collatz, G.J. (1977) Photosynthetic acclimation to temperature and water stress in the desert shrub Larrea divaricata. Carnegie Inst. Washington Yearb. 76, 328–335

    Google Scholar 

  • Osmond, C.B. (1981) Photorespiration and photoinhibition: some implications for the energetics of photosynthesis. Biochim Biophys. Acta 639, 77–98

    Google Scholar 

  • Osmond, C.B., Björkman, O., Anderson, D.J. (1980) Physiological processes in plant ecology: toward a synthesis with Atriplex. Springer, Berlin Heidelberg New York (Ecological Studies, vol. 36)

    Google Scholar 

  • Powles, S.B., Björkman, O. (1982) Photoinhibition of photosynthesis: effect on chlorophyll fluorescence at 77 K in intact leaves and in chloroplast membranes of Nerium oleander. Planta 156, 97–107

    Google Scholar 

  • Powles, S.B., Critchley, C. (1980) Effect of light intensity during growth on photoinhibition of intact attached bean leaflets. Plant Physiol. 65, 1181–1187

    Google Scholar 

  • Powles, S.B., Osmond, C.B., Thorne, S.W. (1979) Photoinhibition of attached, intact leaves of C3 plants illuminated in the absence of both carbon dioxide and of photorespiration. Plant Physiol. 64, 982–988

    Google Scholar 

  • Powles, S.B., Berry, J.A., Björkman, O. (1983) Interaction between light and chilling temperature on the inhibition of photosynthesis in chilling-sensitive plants. Plant Cell Environ 6, 117–123

    Google Scholar 

  • Radin, J.W., Ackerson, A.C. (1981) Water relations of cotton plants under nitrogen deficiency. III. Stomatal conductance, photosynthesis, and abcisic acid accumulation during drought. Plant Physiol. 67, 115–119

    Google Scholar 

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Dedicated to Professor Dr. W. Simonis on the occasion of his 75th birthday

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Kaiser, G., Heber, U. Sucrose transport into vacuoles isolated from barley mesophyll protoplasts. Planta 161, 562–568 (1984). https://doi.org/10.1007/BF00407090

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