Abstract
The aim of this work was to evaluate the influence of elevating the cytosolic activity of phosphoglucomutase (PGM; EC 5.4.2.2) on photosynthesis, growth and heterotrophic metabolism. Here we describe the generation of novel transgenic plants expressing an Escherichia coli phosphoglucomutase (EcPGM) under the control of the 35S promoter. These lines were characterised by an accumulation of leaf sucrose, despite displaying no alterations in photosynthetic carbon partitioning, and a reduced tuber starch content. Determinations of the levels of a wide range of other metabolites revealed dramatic reductions in maltose and other sugars in leaves of the transformants, as well as a modification of the pattern of organic and amino acid content in tubers of these lines. Intriguingly, the transgenics also displayed a dramatically delayed rate of sprouting and significantly enhanced rate of respiration, however, it is important to note that the severity of these traits did not always correlate with the level of transgene expression. These results are discussed in the context of current understanding of the control of respiration and the breaking of tuber dormancy.
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Abbreviations
- 3PGA:
-
3-Phosphoglycerate
- c(Ec)PGM:
-
Cytosolic (Escherichia coli) phosphoglucomutase
- Glc6P:
-
Glucose 6-phosphate
- PAR:
-
Photosynthetically active radiation
- UGPase:
-
UDPglucose pyrophosphorylase
References
Bologa KL, Fernie AR, Leisse A, Loureiro ME, Geigenberger P (2003) A bypass of sucrose synthase leads to low internal oxygen and impaired metabolic performance in growing potato tubers. Plant Physiol 132:2058–2072
von Caemmerer S, Farquhar GD (1981) Some relationships between the biochemistry of photosynthesis and the gas exchange of leaves. Planta 153:376–387
Carrera E, Bou J, Garcia-Martinez JL, Prat S (2000) Changes in GA 20-oxidase gene expression strongly affect stem length, tuber induction and tuber yield of potato plants Plant J 22:247–256
Claassens MMJ, Vreugdenhil D (2000) Is dormancy breaking of potato tubers the reverse of tuber initiation?. Potato Res 43:347–369
Dey NB, Bounelis P, Fritz TA, Bedwell DM, Marchase RB (1994) The glycosylation of phosphoglucomutase is modulated by carbon source and heat-shock in Saccharomyces-cerevisiae. J Biol Chem 269:27143–27148
Duwenig E, Steup M, Willmitzer L, Kossmann J (1997) Antisense inhibition of cytosolic phosphorylase in potato plants (Solanum tuberosum L.) affects tuber sprouting and flower formation with only little impact on carbohydrate metabolism. Plant J 12:323–333
Farre EM, Bachmann A, Willmitzer L, Trethewey RN (2001) Acceleration of potato tuber sprouting by the expression of a bacterial pyrophosphatase. Nat Biotech 19:268–272
Fernie AR, Willmitzer L (2001) Molecular and biochemical triggers of potato tuber development. Plant Physiol 127:1459–1465
Fernie AR, Roessner U, Trethewey RN, Willmitzer L (2001a) The contribution of plastidial phosphoglucomutase to the control of starch synthesis within the potato tuber. Planta 213:418–426
Fernie AR, Roscher A, Ratcliffe RG, Kruger NJ (2001b) Fructose 2,6-bisphosphatase activates pyrophosphate: fructose-6-phosphate 1-phosphotransferase and increases triose phosphate to hexose phosphate cycling in heterotrophic cells. Planta 212:250–263
Fernie AR, Willmitzer L, Trethewey RN (2002a) Sucrose to starch: a transition in molecular plant physiology. Trends Plant Sci 7:35–42
Fernie AR, Tauberger E, Lytovchenko A, Roessner U, Willmitzer L, Trethewey RN (2002b) Antisense repression of cytosolic phosphoglucomutase in potato (Solanum tuberosum) results in a severe growth retardation, reduction in tuber number and altered carbon metabolism. Planta 214:510–520
Fernie AR, Tiessen A, Stitt M, Willmitzer L, Geigenberger P (2002c) Altered metabolic fluxes result from shifts in metabolite levels in sucrose phosphorylase expressing potato tubers. Plant Cell Environ 25:1219–1232
Fernie AR, Swiedrych A, Tauberger E, Lytovchenko A, Trethewey RN, Willmitzer L (2002d) Potato plants exhibiting combined antisense repression of cytosolic and plastidial isoforms of phosphoglucomutase surprisingly approximate wild type with respect to the rate of starch synthesis. Plant Physiol Biochem 40:921–927
Fernie AR, Carrari F, Sweetlove LJ (2004) Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport. Curr Opin Plant Biol 7:1–8
Galis I, Macas J, Vlasak J, Ondrej M, Vanonckelen HA (1995) The effect of an elevated cytokinin level using the ipt gene and N-6-benzyladenine on single node and intact potato plant tuberization in vitro. J Plant Growth Reg 14:143–150
Geigenberger P, Reimholz R, Geiger M, Merlo L, Canale V, Stitt M (1997) Regulation of sucrose and starch metabolism in potato tubers in response to short-term water deficit. Planta 201:502–518
Geigenberger P, Fernie AR, Gibon Y, Christ M, Stitt M (2000) Metabolic activity decreases as an adaptive response to low internal oxygen in growing potato tubers. Biol Chem 381:723–740
Geigenberger P, Stitt M, Fernie AR (2004) Metabolic control analysis and regulation of the conversion of sucrose to starch in growing potato tubers. Plant Cell Environ 27:655–673
Gibon Y, Vigeolas H, Tiessen A, Geigenberger P, Stitt M (2002) Sensitive and high throughput metabolite assays for inorganic pyrophosphate, ADPGlc, nucleotide phosphates, and glycolytic intermediates based on a novel enzymic cycling system. Plant J 30:221–235
Gleave AP (1992) A versatile binary vector system with a T-DNA organizational-structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol Biol 20:1203–1207
Haake V, Zrenner R, Sonnewald U, Stitt M (1998) A moderate decrease of plastid aldolase activity inhibits photosynthesis, alters the levels of sugars and starch and inhibits the growth of potato plants. Plant J 14:147–157
Haake V, Geiger M, Stitt M (1999) Changes in the aldolase activity in wild type potato plants are important for acclimation to growth irradiance and carbon dioxide concentration because plastidial aldolase exerts control over ambient photosynthesis across a range of growth conditions. Plant J 17:479–489
Hajirezaei M, Sonnewald U (1999) Inhibition of potato tuber sprouting: low levels of cytosolic pyrophosphate lead to non-sprouting tubers harvested from transgenic potato plants. Potato Res 42(suppl):353–372
Hajirezaei MR, Bornke F, Peisker M, Takahata Y, Lerchl J, Kirakosyan A, Sonnewald U (2003) Decreased sucrose content triggers starch breakdown and respiration in stored potato tubers (Solanum tuberosum). J Exp Bot 54:477–488
Henkes S, Sonnewald U, Badur R, Flachmann R, Stitt M (2001) A small decrease of plastid transketolase activity in antisense tobacco transformants has dramatic effects on photosynthesis and polypropanoid metabolism. Plant Cell 13:535–551
Kammerer B, Fischer K, Hilpert B, Schubert S, Gutensohn M, Weber A, Flugge UI (1998) Molecular characterization of a carbon transporter in plastids from heterotrophic tissues: the glucose 6-phosphate phosphate antiporter. Plant Cell 10:105–117
Kruger NJ (1997) Carbohydrate synthesis and degradation. In: Dennis DT, Turpin DH, Lefebvre DD, Layzell DB (eds) Plant metabolism. Longmann, Harlow, pp 83–104
Logemann J, Schell J, Willmitzer L (1987) Improved method for the isolation of RNA from plant tissues. Ann Biochem 163:16–20
Lu M, Kleckner N (1994) Molecular cloning and characterization of the pgm gene encoding phosphoglucomutase of Escherichia coli. J Bacteriol 176:5847–5851
Lytovchenko A, Sweetlove LJ, Pauly M, Fernie AR (2002a) The influence of cytosolic phosphoglucomutase on photosynthetic carbohydrate metabolism. Planta 215:1013–1021
Lytovchenko A, Bieberich K, Willmitzer L, Fernie AR (2002b) Carbon assimilation and metabolism in potato leaves deficient in plastidial phosphoglucomutase. Planta 215:802–811
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Muschak M, Hoffmann-Benning S, Fuss H, Kossmann J, Willmitzer L, Fisahn J (1997) Gas exchange and ultrastructural analysis of transgenic potato plants expressing mRNA antisense construct targeted to the cp-fructose-1,6-bisphosphate phosphatase. Photosynthetica 33:455–465
Renz A, Merlo L, Stitt M (1993) Partial purification from potato tubers of 3 fructokinases and 3 hexokinases which show differing organ and developmental specificity. Planta 190:156–165
Rocha-Sosa M, Sonnewald U, Frommer W, Stratmann M, Schell J, Willmitzer L (1989) Both developmental and metabolic signals activate the promoter of a class-I patatin gene. EMBO J 8:23–29
Roessner U, Wagner C, Kopka J, Trethewey RN, Willmitzer L (2000) Simultaneous analysis of metabolites in potato tuber by gas chromatography—mass spectrometry. Plant J 23:131–142
Roessner U, Luedemann A, Brust D, Fiehn O, Linke T, Willmitzer L, Fernie AR (2001) Metabolic profiling allows comprehensive phenotyping of genetically and environmentally diverse systems. Plant Cell 13:11–29
Roessner-Tunali U, Urbanczyk-Wochniak E, Czechowski T, Kolbe A, Willmitzer L, Fernie AR (2003) De novo amino acid biosynthesis in potato tubers is regulated by sucrose levels. Plant Physiol 133:683–692
Rolleston FS (1972) A theoretical background to the use of measured intermediates in the study of the control of intermediary metabolism. Curr Topics Cell Regul 5:47–75
Runquist M, Kruger NJ (1999) Control of gluconeogenesis by isocitrate lyase in endosperm of germinating castor bean. Plant J 19:423–431
Sharkey TD, Savitch LV, Butz ND (1991) Photometric method for routine determination of kcat and carbamylation of rubisco. Photosynth Res 28:41–48
Sonnewald U (2001) Control of potato tuber sprouting. Trends Plant Sci 6:333–335
Sonnewald U, Hajirezaei MR, Kossmann J, Heyer A, Trethewey RN, Willmitzer L (1997) Expression of a yeast invertase in the apoplast of potato tubers increases tuber size. Nature Biotech 15:794–797
Takamiya S, Fukui T (1978) Purification and multiple isoforms of phosphoglucomutase from potato tubers. Plant J 15:697–706
Tauberger E, Fernie AR, Emmermann M, Renz A, Kossmann J, Willmitzer L, Trethewey RN (2000) Antisense inhibition of plastidial phosphoglucomutase provides compelling evidence that potato tuber amyloplasts import carbon from the cytosol in the form of glucose-6-phosphate. Plant J 23:43–53
Trethewey RN, Geigenberger P, Riedel K, Hajirezaei MR, Sonnewald U, Stitt M, Riesmeier JW, Willmitzer L (1998) Combined expression of glucokinase and invertase in potato tubers leads to a dramatic reduction in starch accumulation and a stimulation of glycolysis. Plant J 15:109–118
Trethewey RN, Fernie AR, Bachmann A, Fleischer-Notter H, Geigenberger P, Willmitzer L (2001) Expression of a bacterial sucrose phosphorylase in potato tubers results in a glucose-independent induction of glycolysis. Plant Cell Environ 24:357–365
Urbanczyk-Wochniak E, Leisse A, Roessner-Tunali U, Lytovchenko A, Reismeier J, Willmitzer L, Fernie AR (2003) Expression of a bacterial xylose isomerase in potato tubers results in an altered hexose composition and a consequent induction of metabolism. Plant Cell Physiol 44:1359–1367
Veramendi J, Fernie AR, Leisse A, Willmitzer L, Trethewey RN (2002) Potato hexokinase 2 complements transgenic Arabidopsis plants deficient in hexokinase 1 but does not play a key role in tuber carbohydrate metabolism. Plant Mol Biol 49:491–501
Acknowledgements
We are grateful to Dr. James R. Lloyd (current affiliation: Stellenbosch University, South Africa) for providing E. coli genomic DNA. We are indebted to Helga Kulka for excellent care of the plants and Frank Huhn and Dirk Zerning for help in performing sprouting experiments.
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Lytovchenko, A., Hajirezaei, M., Eickmeier, I. et al. Expression of an Escherichia coli phosphoglucomutase in potato (Solanum tuberosum L.) results in minor changes in tuber metabolism and a considerable delay in tuber sprouting. Planta 221, 915–927 (2005). https://doi.org/10.1007/s00425-005-1490-z
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DOI: https://doi.org/10.1007/s00425-005-1490-z