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Sucrose enhances phosphoenolpyruvate carboxylase activity of in vitro solanum tuberosum L. under non-limiting nitrogen conditions

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Summary

The effect of sucrose on in vitro potato (ev. Kennebec) metabolism was evaluated. Plants were grown in three different media: Murashige and Skoog basal medium containing high nitrogen concentration with 0 or 20 g l−1 sucrose; or modified medium containing reduced nitrogen amount and 20 g l−1 sucrose. Plants fed with 20 g l−1 sucrose and high N exhibited higher phosphoenolpyruvate carboxylase (PEPC) and pyruvate kinase activities and high PEPC protein concentration at 7, 20 and 33 d of culture compared to those grown with 20 g l−1 sucrose and low N, or with 0 g l−1 sucrose and high nitrogen (control). The highest accumulation of starch and sucrose was found in plants grown with sucrose and low nitrogen. This accumulation occurred concomitantly with a reduced enzyme activity resulting from a low utilization of α-ketoglutarate by nitrogen assimilation, when plants were grown with reduced nitrogen. Our investigations on tricarboxylic acid cycle activity showed that sucrose led to the reduction of organic acid amounts in both leaves and roots when high nitrogen was supplied to plants. This was probably due to the intense exit of α-ketoglutarate, which was confirmed by measurements of cytosolic isocitrate dehydrogenase activity. The low leaf glutamine/glutamate ratio observed in plants grown with 20 g l−1 sucrose and high nitrogen compared to their counterparts cultivated with low nitrogen might be due to glutamine conversion into proteins when nitrogen assimilation was intense. These results demonstrate that sucrose enhanced PEPC activity by increasing protein synthesis. They also suggest that sucrose metabolism is involved in the replenishment of the tricarboxylic acid cycle by providing carbon skeletons required to sustain phosphoenolpyruvate utilization during high nitrate assimilation.

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References

  • Borland, A. M. A model for the partitioning of photosynthetically fixed carbon during the C3-CAM transition in Sedum telephium. New Phytol. 134:433–444; 1996.

    Article  CAS  Google Scholar 

  • Bradford, M. M. A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principle of protein-dye binding. Anal. Biochem. 72:248–253; 1976.

    Article  PubMed  CAS  Google Scholar 

  • Bushway, R. J.; Bureau, J. L.; McGann, D. F. Determinations of organic acids in potatoes by high performance liquid chromatography. J. Food Sci. 49:75–81; 1984.

    CAS  Google Scholar 

  • Capellades, M.; Lemeur, L.; Debergh, P. Effects of sucrose on starch accumulation and rate of photosynthesis in Rosa cultured in vitro. Plant Cell Tiss. Organ Cult. 25:21–26; 1991.

    Article  Google Scholar 

  • Champigny, M. L. Integration of photosynthetic carbon metabolism in higher plants. Photosynth. Res. 46:117–127; 1995.

    Article  CAS  Google Scholar 

  • Chen, R.; Chen, R. D. Plant NADP-dependent isocitrate dehydrogenases are predominantly localized in the cytosol. Planta 207:280–285; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Chen, R. D.; Gadal, P. Do the mitochondria provide the 2-oxoglutarate needed for glutamate synthesis in higher plant chloroplasts? Plant Physiol. Biochem. 28:141–146; 1990.

    CAS  Google Scholar 

  • Coleman, A. W.; Goff, L. J. Algae as experimental systems. New York: Alan R. Liss Inc.; 1989:33pp.

    Google Scholar 

  • Cournac, L.; Dimon, B.; Carrier, P.; Lohou, A.; Chagvardieff, P. Growth and photosynthetic characteristics of Solanum tuberosum plantlets cultivated in vitro in different conditions of aeration, sucrose supply, and CO2 enrichment. Plant Physiol. 97:112–117; 1991.

    PubMed  CAS  Google Scholar 

  • Cramer, M. D.; Lewis, O. A. M.; Lips, S. H. Inorganic carbon fixation and metabolism in maize roots as affected by nitrate and ammonium nutrition. Physiol. Plant 89:632–639; 1993.

    Article  Google Scholar 

  • Doncaster, H. D.; Leegood, R. C. Regulation of phosphoenolpyruvate carboxylase activity in maize leaves. Plant Physiol. 84:82–87; 1987.

    PubMed  CAS  Google Scholar 

  • Ehness, R.; Ecker, M.; Godtand, D.; Roitsch, T. Glucose and stress independently regulate source and sink metabolism and defense mechanisms via signal transduction pathways involving protein phosphorylation. Plant Cell 9:1825–1841; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Groß, U.; Gilles, F.; Bender, L.; Berghöfer, P.; Neumann, K. H. The influence of sucrose and an elevated CO2 concentration on photosynthesis of photoautotrophic peanut (Arachis hypogaea L.) cell cultures. Plant Cell Tiss. Organ Cult. 33:143–150; 1993.

    Article  Google Scholar 

  • Gupta, S. K.; Ku, M. S. B.; Lin, J. H.; Zhang, D.; Edwards, G. E. Light/dark modulation of phosphoenolpyruvate carboxylase in C3 and C4 species. Photosynth. Res. 42:133–143; 1994.

    Article  CAS  Google Scholar 

  • Guy, R. D.; Vanlenberghe, G. C.; Turpin, D. H. Significance of phosphoenolpyruvate carboxylase during ammonium assimilation: carbon isotope discrimination in photosynthesis and respiration by the N-limited green alga Selenastrum minutum. Plant Physiol. 89:1150–1157; 1989.

    PubMed  CAS  Google Scholar 

  • Hdider, C.; Desjardins, Y. Effects of sucrose on photosynthesis and phosphoenolpyruvate carboxylase activity of in vitro cultured strawberry plantlets. Plant Cell Tiss. Organ Cult. 36:27–33; 1994.

    Article  CAS  Google Scholar 

  • Heller, R. Relationship between carbon and nitrogen nutrition in higher plants. C. R. Acad. Agr. Fr. 78:93–104; 1997.

    Google Scholar 

  • Hoff, T.; Truong, H. N.; Caboche, M. The use of transgenic plants to study nitrate assimilation. Plant Cell Environ. 17:489–506; 1994.

    Article  CAS  Google Scholar 

  • Jiao, J. A.; Chollet, R. Light/dark regulation of maize leaf phosphoenolpyruvate carboxylase by in vivo phosphorylation. Arch. Biochem. Biophys. 261:409–417; 1988.

    Article  PubMed  CAS  Google Scholar 

  • Kotvun, T.; Daie, J. End-product control of carbon metabolism in culture-grown sugar beet plants. Plant Physiol. 108:1647–1656; 1995.

    Google Scholar 

  • Kozai, T.; Watanabe, K.; Jeong, B. R. Stem elongation and growth of Solanum tuberosum L. in vitro in response to photosynthetic photon flux, photoperiod and difference in photoperiod and dark period temperatures. Sci. Hortic. 64:1–9; 1995.

    Article  Google Scholar 

  • Krömer, S.; Gardestrom, P.; Samuelson, G. Regulation of the supply of cytosolic oxaloacetate for mitochondrial metabolism via phosphoenolpyruvate carboxylase in barley leaf protoplasts. I. The effect of covalent modification on PEPC activity, pH response, and kinetic-properties. Biochim. Biophys. Acta 1239:343–350; 1996.

    Google Scholar 

  • Kwa, S. H.; Wee, Y. C.; Lim, T. M.; Kumar, P. P. Establishment and physiological analyses of phototrophic callus of the ferm Platycerium coronarium (Koenig) Desv under CO2 enrichment. J. Exp. Bot. 46:1535–1542; 1995.

    Article  CAS  Google Scholar 

  • Lancien, M.; Ferrario-Mery, S.; Roux, Y.; Bismuth, E.; Masclaux, C.; Hirel, B. Gadal, P.; Hodges, M. Simultaneous expression of NAD-dependent isocitrate dehydrogenase and other Krebs cycle genes after nitrate resupply to short-term nitrogen-starved tobacco. Plant Physiol. 120:717–725; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Lepiniec, L.; Vidal, J.; Chollet, R.; Gadal, P.; Cretin, C. Phosphoenolpyruvate carboxylase: structure, regulation and evolution. Plant Sci. 99:111–124; 1994.

    Article  CAS  Google Scholar 

  • Leport, L.; Kandlinber, A.; Baur, B.; Kaiser, W. M. Diurnal modification of phosphoenolpyruvate carboxylase in pea leaves and roots as related to tissue malate concentration and to the nitrogen source. Planta 198:415–501; 1996.

    Article  Google Scholar 

  • Lin, M.; Turpin, D. H.; Plaxton, W. C. Pyruvate kinase isozymes from green alga, Selenastrum minutum. I. Purification and physical and immunological characterization. Arch. Biochem. Biophys. 269:219–227; 1989.

    Article  PubMed  CAS  Google Scholar 

  • Moore, B. D.; Cheng, S. H.; Sims, D.; Seemann, J. R. The biochemical and molecular basis for photosynthetic acclimation to elevated atmospheric CO2. Plant Cell Environ. 22:567–582; 1999.

    Article  CAS  Google Scholar 

  • Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15:473–479; 1962.

    Article  CAS  Google Scholar 

  • Neumann, K. H.; Groß, U.; Bender, L. Regulation of photosynthesis in Daucus carota and Arachis hypogaea cell culture by exogenous sucrose. In: Kurz, W. G. W., ed. Primary and secondary metabolism of plant cell culture II. Berlin, Heidelberg, New York: Springer-Verlag; 1989:281–291.

    Google Scholar 

  • Nimmo, G. A.; Nimmo, H. G.; Hamilton, I. D.; Fewson, C. A.; Wilkins, M. B. Purification of the phosphorylated night form and dephosphorylated day form of phosphoenolpyruvate carboxylase from Bryophyllum fedtschenkoi. Biochem. J. 239:213–220; 1986.

    PubMed  CAS  Google Scholar 

  • O'Leary, M. H. Phosphoenolpyruvate carboxylase: an enzymologist's view. Annu. Rev. Plant Physiol. 33:297–315; 1982.

    Article  Google Scholar 

  • Osmund, C. B.; Holtum, J. A. M. Crassulacean acid metabolism. In: Hatch, M. D.; Boardman, N. K., eds. Biochemistry of plants. New York: Academic Press; 1981:283–328.

    Google Scholar 

  • Paul, M.; Stitt, M. Effects of nitrogen and phosphate deficiencies on levels of carbohydrates, respiratory enzymes and metabolites in seedlings of tobacco, and their response of exogenous sucrose. Plant Cell Environ. 16:1047–1057; 1993.

    Article  CAS  Google Scholar 

  • Popova, T. N.; Podoprigora, A. Y. Catalytic properties of NAD-isocitrate dehydrogenase from pumpkin cotyledons. Biochemistry 60:1365–1371; 1995.

    Google Scholar 

  • Schäfer, C.; Simper, H.; Hofmann, B. Glucose feeding results in coordinated changes of chlorophyll content, ribulose-1,5-bisphosphate carboxylase-oxygenase activity and photosynthetic potential in photoautotrophic suspension cultured cells of Chenopoduim rubrum. Plant Cell Environ. 15:343–350; 1992.

    Article  Google Scholar 

  • Scheible, W. R.; Gonzales-Fontes, A.; Lauerer, M.; Müller-Röber, B.; Caboche, M.; Stitt, M. Nitrate acts as a signal to induce organic acid metabolism and repress starch metabolism in tobacco. Plant Cell 9:783–798; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Schuller, K. A.; Plaxton, W. C.; Turpin, D. H. Regulation of C3 phosphoenolpyruvate carboxylase from the green alga Selenastrum minutum. Properties associated with replenishment of TCA cycle intermediates during amino acids biosynthesis. Plant Physiol. 93:1303–1311; 1990.

    PubMed  CAS  Google Scholar 

  • Serret, M. D.; Trillas, M. I.; Matas, J.; Araus, J. L. Development of photoautotrophy and photoinhibition of Gardenia jasmoides plantlets during micropropagation. Plant Cell Tiss. Organ Cult. 45:1–16; 1996.

    Article  CAS  Google Scholar 

  • Shorroh, B. S.; Dixon, R. A. Molecular characterization and expression of an isocitrate dehydrogenase from alfalfa (Medicago sativa L.). Plant Mol. Biol. 20:801–807; 1992.

    Article  Google Scholar 

  • Siegel, G.; Stitt, M. Partial purification of two forms of spinach leaf sucrosephosphate synthase which differ in their kinetic properties. Plant Sci. 66:205–210; 1990.

    Article  Google Scholar 

  • Stitt, M.; Heldt, H. W. Controls of photosynthetic sucrose synthesis by fructose-2,6-bisphosphate. Intercellular metabolite distribution and properties of the cytosolic fructose bisphosphate in leaves of Zea mays L. Planta 164:179–188; 1985.

    Article  CAS  Google Scholar 

  • Sugiharto, B.; Sugiyama, T. Effects of nitrate and ammonium on gene expression of phosphoenolpyruvate carboxylase and nitrogen metabolism in maize leaf tissue during recovery from nitrogen stress. Plant Physiol. 98:1403–1408; 1992.

    PubMed  CAS  Google Scholar 

  • Sugiharto, B.; Suzuki, I.; Burnell, J. N.; Sugiyama, T. Glutamine induces the N-dependent accumulation of mRNAs encoding phosphoenolpyruvate carboxylase and carbonic anhydrase in detached maize leaf tissue. Plant Physiol. 100:2066–2070; 1992.

    PubMed  CAS  Google Scholar 

  • Tichá, I.; Cáp, F.; Pacovská, D.; Hofman, P.; Haisel, D.; Capková, V.; Schäfer, C. Culture on sugar medium enhances photosynthetic capacity and light resistance of plantlets grown in vitro. Physiol. Plant 102:155–162; 1998.

    Article  Google Scholar 

  • Turpin, D. H.; Botha, F. C.; Smith, R. G.; Feil, R.; Horsey, A. K.; Vanlerberghe, G. C. Regulation of carbon partitioning to respiration during dark ammonium assimilation by the green alga Selenastrum minutum. Plant Physiol. 93:166–175; 1990.

    PubMed  CAS  Google Scholar 

  • Vanlerberghe, G. C.; Huppe, H. C.; Vlossak, K. D. M.; Turpin, D. H. Activation of respiration to support dark nitrate and ammonium assimilation in the green alga Selenastrum minutum. Plant Physiol. 99:495–500; 1992.

    PubMed  CAS  Google Scholar 

  • Vanlerberghe, G. C.; Schuller, K. A.; Smith, R. G.; Feil, R.; Plaxton, W. C.; Turpin, D. H. Relationship between NH4 + assimilation rate and in vivo phosphoenolpyruvate carboxylase activity: regulation of anaplerotic carbon flow in the green alga Selenastrum minutum. Plant Physiol. 94:284–290; 1990.

    PubMed  CAS  Google Scholar 

  • Van Oosten, J. J.; Afif, D.; Dizengremel, P. Long-term effects of a carbon dioxide enriched atmosphere on enzymes of the primary carbon metabolism of spruce trees. Plant Physiol. Biochem. 30:541–547; 1992.

    Google Scholar 

  • Van Quy, L.; Lamaze, T.; Champigny, M. L. Effect of light and NO3− on wheat leaf phosphoenolpyruvate carboxylase activity: evidence for covalent modulation of the C3 enzyme. Plant Physiol. 97:1476–1482; 1991.

    Google Scholar 

  • Wedding, R. T.; Black, M. K.; Mayer, C. R. Activation of higher plant phosphoenolpyruvate carboxylase by glucose-6-phosphate. Plant Physiol. 90:648–652; 1989.

    PubMed  CAS  Google Scholar 

  • Weger, H. G.; Turpin, D. H. Mitochondrial respiration can support NO3 − and NO2 − reduction during photosynthesis. Interactions between photosynthesis, respiration and N assimilation in the N-limited green alga Selenastrum minutum. Plant Physiol. 89:409–415; 1989.

    Article  PubMed  CAS  Google Scholar 

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  1. Centre de Recherche en Horticulture, Département de Phytologie, Faculté des Sciences de l'Agriculture et de l'Alimentation, Université Laval, GIK 7P4, Sainte-Foy, Québec, Canada

    Boubacar Dary Sima, Yves Desjardins & Le Van Quy

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  1. Boubacar Dary Sima
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  2. Yves Desjardins
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  3. Le Van Quy
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Correspondence to Boubacar Dary Sima.

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Sima, B.D., Desjardins, Y. & Quy, L.V. Sucrose enhances phosphoenolpyruvate carboxylase activity of in vitro solanum tuberosum L. under non-limiting nitrogen conditions. In Vitro Cell.Dev.Biol.-Plant 37, 480–489 (2001). https://doi.org/10.1007/s11627-001-0085-z

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