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.
Similar content being viewed by others
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
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
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
Delieu, T., Walker, D.A. (1981) Polarographic measurement of photosynthetic O2 evolution by leaf discs. New Phytol. 89, 165–175
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
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
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
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
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
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
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
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
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
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
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
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
Kaiser, G., Heber, U. (1983) Sucrose transport into vacuoles isolated from barley mesophyll protoplasts. Planta 161, 562–568
Preiss, J.L. (1982) Regulation of the biosynthesis and degradation of starch. Annu. Rev. Plant Physiol. 33, 431–454
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
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
Schäfer, G., Heber, U., Heldt, H.W. (1977) Glucose transport into spinach chloroplasts. Plant Physiol. 60, 286–289
Steup, M., Peavey, D.G., Gibbs, M. (1976) The regulation of starch metabolism by inorganic phosphate. Biochem. Biophys. Res. Commun. 72, 1554–1561
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
Stitt, M., Heldt, H.W. (1981) Physiological rates of starch breakdown in isolated intact spinach chloroplast. Plant Physiol. 68, 755–761
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
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
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
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
Taussky, H.H., Shorr, E., Kutzmann, G. (1953) A microcolorimetric method for the determination of inorganic phosphorus. J. Biol. Chem. 202, 675–685
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
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
Wirtz, W., Stitt, M., Heldt, H.W. (1980) Enzymic determination of metabolites in the subcellular compartments of spinach protoplasts. Plant Physiol. 66, 198–193
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00401173