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A comparative study of metabolite levels in plant leaf material in the dark

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Abstract

Metabolite levels have been compared in the dark and during photosynthesis in leaves and protoplasts from spinach, pea, wheat and barley. In protoplasts the subcellular distribution was also studied. The levels of triose phosphates and sugar bisphosphates were high in the light and low in the dark. The hexose phosphates and 3-phosphoglycerate levels in the dark were very variable depending on the plant material. In most conditions, hexose phosphates and triose phosphates were mainly in the extrachloroplast compartment, while 3-phosphoglycerate and the sugar bisphosphates were mainly in the chloroplast compartment. Leaves always had a very low triose phosphate: 3-phosphoglycerate ratio in the dark, but in protoplasts this ratio was higher. Detailed studies with spinach showed that metabolite levels were very dependent on the availability of carbohydrate in the leaf, particularly starch. Starch mobilisation is not controlled just by the availability of inorganic phosphate and accumulation of phosphorylated intermediates. Hydrolysis of starch may provide precursors for sucrose synthesis while phosphorolysis leads to provision of substrates for respiration. Starch breakdown generates high enough levels of hexose phosphate to support substantial rates of sucrose synthesis in the dark. Respiration is not greatly increased when metabolite levels are high during starch mobilisation. Higher levels of metabolites shorten the length of the induction phase of photosynthesis.

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Abbreviations

Chl:

chlorophyll

DHAP:

dihydroxyacetone phosphate

Fru2,6bisP:

fructose-2,6-bisphosphate

NMR:

nuclear magnetic resonance

PGA:

3-phosphoglyceric acid

Pi:

inorganic phosphate

RuBP:

ribulose-1,5-bisphosphate

UDPGlc:

uridine-5′-diphosphate glucose

References

  • Azcon-Bieto, J., Osmond, C.B. (1983) Relationship between photosynthesis and respiration. The effect of carbohydrate status on the rate of CO2 production by respiration in darkened and illuminated wheat leaves. Plant Physiol. 71, 574–581

    Google Scholar 

  • Azcon-Bieto, J., Lambers, D., Day, D. (1983) Effect of photosynthesis and carbohydrate status on respiratory rates and involvement of the alternative pathway in leaf respiration. Plant Physiol. 72, 598–603

    Google Scholar 

  • Chatterton, N.J., Silvius, J.E. (1980) Acclimation of photosynthetic partitioning and photosynthesis rates to changes in length of the daily photosynthetic period. Ann. Bot. 46, 739–745

    Google Scholar 

  • Delieu, T., Walker, D.A. (1981) Polarographic measurement of photosynthetic O2 evolution by leaf discs. New Phytol. 89, 165–175

    Google Scholar 

  • Dietz, K.-J., Heber, U. (1984) Rate limiting factors in leaf photosynthesis. I. Carbon fluxes in the Calvin cycle. Biochim. Biophys. Acta 767, 422–443

    Google Scholar 

  • Dietz, K.-J., Niemanis, S., Heber, U. (1984) Rate limiting factors in leaf photosynthesis. II. Electron transport and ribulose1,5bisphosphate regeneration. Biochim. Biophys. Acta 767, 444–449

    Google Scholar 

  • Doehlert, D.C., Huber, S.C. (1983) Spinach leaf sucrose phosphate synthase: activation by glucose 6-phosphate and inactivation with inorganic phosphate. FEBS Lett. 153, 293–298

    Google Scholar 

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

    Google Scholar 

  • Foyer, C., Walker, D., Spencer, C., Mann, B. (1982) Observations on the phosphate status and intracellular pH of intact cells, protoplasts and chloroplasts from photosynthetic tissue using phosphorus-31 NMR. Biochem. J. 202, 429–434

    Google Scholar 

  • Fondy, B.R., Geiger, D.A. (1982) Diurnal pattern of translocation and carbohydrate metabolism in source leaves of Beta vulgaris L. Plant Physiol. 70, 671–676

    Google Scholar 

  • Gerhardt, R., Heldt, H.W. (1984) Measurement of subcellular metabolite levels in leaves by fractionation of freeze-stopped material in non-aqueous media. Plant Physiol. 75, 542–547

    Google Scholar 

  • Giersch, Ch., 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

    Google Scholar 

  • Gordon, A.G., Ryle, G.J.A., Powell, C.E., Mitchell, D.E. (1980a) Expot mobilization and respiration of assimilates in uniculm barley during light and darkness. J. Exp. Bot. 31, 461–473

    Google Scholar 

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

    Google Scholar 

  • Hampp, R., Goller, M., Ziegler, H. (1982) Adenylate levels, energy charge and phosphorylation potential during dark-light and light-dark transitins in chloroplasts, mitochondria and cytosol of mesophyll protoplasts from Avena sativa L. Plant Physiol. 69, 448–455

    Google Scholar 

  • Herzog, B., Stitt, M., Heldt, H.W. (1984) Control of photosynthetic sucrose synthesis by fructose-2,6-bisphosphate. III. Properties of the cytosolic fructose-1,6-bisphosphatase. Plant Physiol. 75, 561–565

    Google Scholar 

  • Kaiser, G., Heber, U. (1983) Sucrose transport into vacuoles isolated from barley mesophyll protoplasts. Planta 161, 562–568

    Google Scholar 

  • Preiss, J.L. (1982) Regulation of the biosynthesis and degradation of starch. Annu. Rev. Plant Physiol. 33, 431–454

    Google Scholar 

  • Roberts, J.K.M., Wade-Jardetzky, N., Jadetzky, O. (1981) Intracellular pH measurements by 31-P nuclear magnetic resonance. Influence of factors other than pH on 31-P chemical shifts. Biochemistry 20, 5389–5394

    Google Scholar 

  • Robinson, J.M. (1984) Photosynthetic carbon metabolism in leaves and isolated chloroplasts from spinach plants grown under short and intermediate photosynthetic periods. Plant Physiol. 75, 397–409

    Google Scholar 

  • Schäfer, G., Heber, U., Heldt, H.W. (1977) Glucose transport into spinach chloroplasts. Plant Physiol. 60, 286–289

    Google Scholar 

  • Steup, M., Peavey, D.G., Gibbs, M. (1976) The regulation of starch metabolism by inorganic phosphate. Biochem. Biophys. Res. Commun. 72, 1554–1561

    Google Scholar 

  • Stitt, M., Gerhardt, R., Kürzel, B., Heldt, H.W. (1983b) A role for fructose2,6bisphosphate in the regulation of sucrose synthesis in spinach leaves. Plant Physiol. 72, 1139–1141

    Google Scholar 

  • Stitt, M., Heldt, H.W. (1981) Physiological rates of starch breakdown in isolated intact spinach chloroplast. Plant Physiol. 68, 755–761

    Google Scholar 

  • Stitt, M., Herzog, B., Heldt, H.W. (1984) Control of photosynthetic sucrose synthesis by fructose2,6bisphosphate. I. Coordination of CO2 fixation of sucrose synthesis. Plant Physiol. 75, 548–553

    Google Scholar 

  • Stitt, M., Lilley, R.McC., Heldt, H.W. (1982) Adenine nucleotide levels in the cytosol, chloroplasts and mitochondria of wheat leaf protoplasts. Plant Physiol. 70, 971–977

    Google Scholar 

  • Stitt, M., Wirtz, W., Heldt, H.W. (1980) Metabolite levels in the chloroplast and extrachloroplast compartments of spinach protoplasts. Biochim. Biophys. Acta 593, 85–102

    Google Scholar 

  • Stitt, M., Writz, W., Heldt, H.W. (1983a) Regulation of sucrose synthesis by cytoplasmic fructosebisphosphatase and sucrose phosphate synthase during photosynthesis in varying light and carbon dioxide. Plant Physiol. 72, 767–774

    Google Scholar 

  • Taussky, H.H., Shorr, E., Kutzmann, G. (1953) A microcolorimetric method for the determination of inorganic phosphorus. J. Biol. Chem. 202, 675–685

    Google Scholar 

  • Walker, D.A. (1976) CO2 fixation by intact chloroplasts: photosynthetic induction and its relation to transport phenomena and control mechanisms. In: The intact chloroplast, pp. 235–278 Barber, J., ed. Elsevier, Amsterdam

    Google Scholar 

  • Waterton, J.C., Bridges, I.G., Irving, M.P. (1983) Intercellular compartmentation detected by 31P-NMR in intact photosynthetic wheat-leaf tissue. Biochim. Biophys. Acta 763, 315–320

    Google Scholar 

  • Wirtz, W., Stitt, M., Heldt, H.W. (1980) Enzymic determination of metabolites in the subcellular compartments of spinach protoplasts. Plant Physiol. 66, 198–193

    Google Scholar 

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Stitt, M., Wirtz, W., Gerhardt, R. et al. A comparative study of metabolite levels in plant leaf material in the dark. Planta 166, 354–364 (1985). https://doi.org/10.1007/BF00401173

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