Abstract
We have investigated the regulation of sucrose storage in cell-suspension cultures of sugarcane. When grown in batch culture, sucrose accumulation commences after about 5 d, when the nitrogen supply is exhausted. Sucrose storage is also induced by decreasing the nitrogen supply to cells growing in a chemostat. The measured activity of sucrose-phosphate synthase is high enough to account for the rate of sucrose accumulation, provided precautions are taken to avoid the hydrolysis of UDP during the assay. The cells contained high sucrose-synthase activity but pulsing experiments with [14C]glucose and unlabelled fructose indicated that this enzyme did not contribute substantially to the synthesis of sucrose, because the glucosyl and fructosyl moieties of sucrose were equally labelled. Several lines of evidence demonstrate the presence of a cycle in which sucrose is synthesized and degraded simultaneously; sucrosephosphate-synthase activity doubles during the phase when the cells are actively storing sucrose but activity is also high after storage has ceased, or when the sucrose is being remobilised; pulse experiments with [14C]fructose also showed that sucrose synthesis occurs not only during the storage phase, but also after storage has stopped and during the rapid mobilisation of sucrose; the cells contain high activities of sucrose synthase and alkaline invertase and these are both at a maximum when sucrose storage is occurring; even during the storage phase. [14C]fructose pulses lead to labelling of free glucose which is evidence for rapid synthesis and degradation of sucrose. It is proposed that the rate and extent of sucrose storage is regulated by this cycle of synthesis and degradation. Measurements of enzyme activities and metabolite levels are presented, and it is discussed which factors could contribute to the regulation of these two opposing fluxes and, hence, the rate of net sucrose storage and mobilisation.
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Abbreviations
- Fru2, 6bisP:
-
fructose-2,6-bisphosphate
- Fru6P:
-
fructose-6-phosphate
- Glc6P:
-
glucose-6-phosphate
- PFK:
-
phosphofructokinase
- PFP:
-
pyrophosphate: fructose-6-phosphate phosphotransferase
- PGA:
-
glycerate-3-phosphate
- Pi:
-
inorganic phosphate
- PPi:
-
pyrophosphate
- SPS:
-
sucrose-phosphate synthase
- UDPGlc:
-
uridinediphosphoglucose
References
Avigad, G. (1982) Sucrose and other disaccharides. In: Encyclopedia of plant physiology. N. S., vol. 13 A: Plant carbohydrates, pp. 217–347, Loewus, F. A., Tanner, W., eds. Springer, Berlin Heidelberg New York
Briskin, D. P., Thornley, W. R., Wyse, J. E. (1985) Membrane transport in isolated vesicles from sugar beet taproot II Evidence for a sucrose/H+ antiport. Plant Physiol. 79, 871–875
Dancer, J., David, M., Stitt, M. (1990a) Water stress leads to a change of partitioning in favour of sucrose in heterotrophic cell suspension cultures of Chenopodium rubrum. Plant Cell Environ., in press
Dancer, J., Veith, R., Feil, R., Komor, E., Stitt, M. (1990b) Independent changes of inorganic pyrophosphate and the ATP/ADP or UTP/UDP ratios in plant cell suspension cultures. Plant Sci 66, 59–63
Glasziou, K. T., Gaylor, K. R. (1972) Storage of sugars in stalks of sugar cane. Bot. Rev. 38, 471–490
Hargreaves, J., ap Rees, T. (1988) Turnover of starch and sucrose in roots of Pisum sativum. Phytochemistry 27, 1627–1629
Hatch, M. D., Glasziou, K. T. (1963) Sugar accumulation in sugar cane. II. Relationship of in vertase activity to sugar content and growth in storage tissue of plants grown in controlled environments. Plant Physiol. 38, 344–348
Hatzfeld, W.-D., Dancer, J., Stitt, M. (1989) Fructose-2,6-bisphosphate, metabolites and coarse control of pyrophosphate: fructose-6-phosphate phosphotransferase during triose-phosphate cycling in heterotrophic cell-suspension cultures of Chenopodium rubrum. Planta 180, 205–211
Hawker, J. S., Hatch, M. D. (1965) Mechanism of sugar storage by mature stem tissue of sugar cane. Physiol. Plant. 18, 444–453
Huber, S. C., Akazawa, T. (1986) A novel sucrose synthase pathway for sucrose degradation in cultured sycamore cells. Plant Physiol. 81, 1008–1013
Hue, L. (1980) The role of futile cycles in the regulation of carbohydrate metabolism in liver. Adv. Enzymol. 52, 247–291
Komor, E., Thom, M., Maretzki, A. (1981) The mechanism of sugar uptake by sugar cane suspension cells. Planta 153, 181–192
Komor, E., Höfner, M., Wendler, R., Thom, M., Delacruz, A. (1987) Nitrogen nutrition of sugar cane cells: regulation of amino acid uptake and the effect on cell growth and sucrose storage. Plant Physiol. Biochem. 25, 581–588
Leigh, R. A., ap Rees, T., Fuller, W. A., Banfield, J. (1979) The location of acid invertase activity and sucrose in the vacuoles of storage roots of beetroot. Biochem. J. 178, 539–547
Maretzki, A., Thom, M. (1972) Membrane transport of sugars in cell suspension of sugarcane I. Evidence of sites and specificity. Plant Physiol. 49, 172–182
Maretzki, A., Thom, M. (1988) High performance liquid chromatography-based revaluation of disaccharides produced upon incubation of sugarcane vacuoles with UDP-glucose. Plant Physiol. 88, 266–269
Nickell, L. G., Maretzki, A. (1969) Growth of suspension cultures of sugar cane cells in chemically defined media. Physiol. Plant. 22, 117–125
Preisser, J., Komor, E., (1988) Analysis of the reaction products from incubation of sugar cane vacuoles with uridine diphosphate-glucose: no evidence for the group translocator. Plant Physiol. 88, 259–265
Rufty, T. W., Huber, S. C., Volk, R. J. (1989) Alterations in leaf carbohydrate metabolism in response to nitrogen stress. Plant Physiol. 88, 725–730
Smith, R. G., Vanderbergh, G., Stitt, M., Turpin, D. M. (1989) Short term metabolite changes during transient ammonia assimilation by the N-limited alga Selanastrum minutum. Plant Physiol. 91, 749–755
Siegl, G., Stitt, M. (1990) Partial purification of two forms of sucrose-phosphate synthase from spinach leaves which differ in their kinetic properties. Plant Sci. 66, 205–210
Somogyi, M. (1952) Notes on sugar determination. J. Biol. Chem. 195, 19–23
Stitt, M. Steup, M. (1985) Starch and sucrose degradation. In: Encyclopedia of plant physiology, N. S., vol. 18: Higher plant cell respiration, pp. 347–390, Douce, R., Day, D. A., eds. Springer, Berlin Heidelberg New York
Stitt, M., Schreiber, U (1988) Interaction between sucrose synthesis and CO2 fixation. III. Response of biphasic induction kinetics and oscillations to manipulation of the relation-between electron transport, calvin cycle, and sucrose synthesis. J. Plant Physiol. 133, 263–271
Stitt, M., Huber, S. C., Kerr, P. S. (1987) Control of photosynthetic sucrose formation. In: The Biochemistry of plants, vol 10, pp. 327–409, Hatch, M. R., Boardman, N. K., eds. Academic Press, New York
Stitt, M., Wilke, I., Gerhardt, R., Heldt, H. W. (1988) Coarse control of sucrose phosphate synthase in leaves: alterations of the kinetic properties in response to the rate of photosynthesis and the accumulation of sucrose. Planta 174, 217–230
Stitt, M., Lilley, R. McC., Gerhardt, R., Heldt, H. W. (1989) Determination of metabolite levels in specific cells and subcellular compartments of plant leaves. Methods Enzymol. 174, 518–552
Thom, M., Komor, E. (1985) Electrogenic proton translocation by the ATPase of sugar cane vacuoles. Plant Physiol. 77, 329–334
Thom, M., Maretzki, A. (1985) Group translocation as a mechanism for sucrose transfer into vacuoles from sugarcane cells. Proc. Natl. Acad. Sci. USA 82, 4697–4701
Thom, M., Maretzki, A., Komor, E. (1982) Vacuoles from sugar cane suspension cultures. I. Isolation and partial characterisation. Plant Physiol. 69, 1315–1319
Walker, J. L., Huber, S. C. (1988) Regulation of sucrose phosphate synthase activity in spinach leaves by protein level and covalent modification. Planta 177, 116–120
Willenbrink, J. (1982) Storage of sugars in higher plants. In: Encyclopedia of plant physiology, N. S., vol. 13A; Plant carbohydrates I (Intracellular carbohydrates), pp. 684–699, Loewus, F. A., Tanner, W., eds. Springer, Berlin Heidelberg New York
Wilson, G. (1976) A simple and inexpensive design of chemostat enabling steady-state growth of Acer pseudoplatanus L. cells under phosphate-limiting conditions. Ann. Bot. 40, 919–932
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We are grateful to H. Schroer for technical help, to E. Bischofsberger for typing the manuscript and to L. Badewitz for the artwork. This research was supported by the Deutsche Forschungsgemeinschaft.
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Wendler, R., Veith, R., Dancer, J. et al. Sucrose storage in cell suspension cultures of Saccharum sp. (sugarcane) is regulated by a cycle of synthesis and degradation. Planta 183, 31–39 (1991). https://doi.org/10.1007/BF00197564
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DOI: https://doi.org/10.1007/BF00197564