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Acclimation of white lupin to phosphorus deficiency involves enhanced expression of genes related to organic acid metabolism

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Abstract

White lupin (Lupinus albus L.) acclimates to phosphorus deficiency (−P) by the development of short, densely clustered lateral roots called proteoid (or cluster) roots. These specialized plant organs display increased exudation of citric and malic acid. The enhanced exudation of organic acids from P stressed white lupin roots is accompanied by increased in vitro phosphoenolpyruvate carboxylase (PEPC) and malate dehydrogenase (MDH) activity. Here we report the cloning of full-length white lupin PEPC and MDH cDNAs. RNA blot analysis indicates enhanced expression of these genes in −P proteoid roots, placing higher gene expression at the site of organic acid exudation. Correspondingly, macroarray analysis of about 1250 ESTs (expressed sequence tags) revealed induced expression of genes involved in organic acid metabolism in −P proteoid roots. In situ hybridization revealed that PEPC and MDH were both expressed in the cortex of emerging and mature proteoid rootlets. A C3 PEPC protein was partially purified from proteoid roots of P deficient white lupin. Native and subunit Mr were determined to be 440 kD and 110 kD, respectively. Citrate and malate were effective inhibitors of in vitro PEPC activity at pH 7. Addition of ATP partially relieved inhibition of PEPC by malate but had little effect on citrate inhibition. Taken together, the results presented here suggest that acclimation of white lupin to low P involves modified expression of plant genes involved in carbon metabolism.

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References

  • Aono T, Kanada N, Ijima A and Oyaizu H 2001 The response of the phosphate uptake system and the organic acid exudation system to phosphate starvation in Sesbania rostrata. Plant Cell Physiol. 15,1253–1264.

    Google Scholar 

  • Avio L, Sbrana C and Giovannetti M 1990 The response of different species of Lupinus to VAM endophytes. Symbiosis 42, 321–323.

    Google Scholar 

  • Bakrim N, Brulfert J, Vidal J and Chollet R 2001 Phosphoenolpyruvate carboxylase kinase is controlled by a similar signaling cascade in CAM and C4 plants. Biochem. Biophys. Res. Com. 286, 1158–1162.

    PubMed  Google Scholar 

  • Bradford M M 1976 A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248–254.

    PubMed  Google Scholar 

  • Braum S and Helmke P A 1995 White lupin utilizes soil phosphorus that is unavailable to soybean. Plant Soil 176, 95–100.

    Google Scholar 

  • Braun W and Abraham R 1989 Modified diffusion blotting for rapid and efficient protein transfer with PhastSystem. Electrophoresis 10, 249–253.

    PubMed  Google Scholar 

  • Chollet R, Vidal J and O'Leary M H 1996 Phosphoenolpyruvate carboxylase: a ubiquitous, highly regulated enzyme in plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47, 273–298.

    PubMed  Google Scholar 

  • Cretin C, Luchetta P, Joly C, Miginiac-Maslow M, Decottignies P, Jacquot J, Vidal J and Gadal P 1988 Identification of cDNA clone from sorghum leaf malate dehydrogenase (NADP). Eur. J. Biochem. 174, 497–501.

    PubMed  Google Scholar 

  • Devereux J, Haeberli P and Smithies O 1984 Sequence analysis programs for the VAX. Nucleic Acids Res. 12, 387–395.

    PubMed  Google Scholar 

  • Dinkelaker B, Romheld V and Marschner H 1989 Citric acid excretion and precipitation of calcium citrate in the rhizosphere of white lupin (Lupinus albus L.). Plant Cell Environ. 12, 285–292.

    Google Scholar 

  • Dinkelaker B, Hengeler C and Marschner H 1995 Distribution and function of proteoid roots and other root clusters. Botanica Acta. 108, 169–276.

    Google Scholar 

  • Dong L-Y, Masuda T, Kawaruma T, Hata S and Izui K 1998 Cloning, expression, and characterization of a root-form phosphoenolpyruvate carboxylase from Zea mays: comparison with the C4-form enzyme. Plant Cell Physiol. 39, 865–873.

    PubMed  Google Scholar 

  • Dong L, Ermolova N V and Chollet R 2001 Partial purification and biochemical characterization of a heteromeric protein phosphatase 2A holoenzyme from maize (Zea mays L.) leaves that dephosphorylates C4 phosophoenolpyruvate carboxylase. Planta 213, 379–89.

    PubMed  Google Scholar 

  • Fleming A J, Mandel T, Roth I and Kuhlemeier C 1993 The patterns of gene expression in the tomato shoot apical meristem. Plant Cell 5, 297–309.

    PubMed  Google Scholar 

  • Frank J, Claarke R J, Vater J and Holzwarth J F 2001 Influence of allosteric effectors on the kinetics and equilibrium binding of phosphoenolpyruvate (PEP) to phosphoenolpyruvate carboxylase (PEPC) from Zea mays. Biophys. Chem. 92, 53–64.

    PubMed  Google Scholar 

  • Gardner W K, Parbery D G and Barber D A 1982 The acquisition of phosphorus by Lupinus albus L. I. Some characteristics of the soil/root interface. Plant Soil 68, 19–32.

    Google Scholar 

  • Gardner W K, Barber D A and Parbery D G 1983 The acquisition of phosphorus by Lupinus albus L. III. The probable mechanism by which phosphorus movement in the soil/root interface is enhanced. Plant Soil 70, 107–124.

    Google Scholar 

  • Gietl C 1992 Malate dehydrogenase isoenzymes: Cellular location and role in the flow of metabolites between cytoplasm and cell organelles. Biochim. Biophys. Acta 110, 217–234.

    Google Scholar 

  • Gilbert G A, Knight J D, Vance C P and Allan D L 1999 Acid phosphatase activity in phosphorus-deficient white lupin roots. Plant Cell Environ. 22, 801–810.

    Google Scholar 

  • Gilbert G A, Knight J D, Vance C P and Allan D L 2000 Proteoid root development of phosphorus deficient lupin is mimicked by auxin and phosphonate. Ann. Bot. 85, 921–928.

    Google Scholar 

  • Guex N, Henry H, Flach J, Richter H and Widmer F 1995 Glyoxysomal malate dehydrogenase and malate synthase from soybean cotyledons (Glycine max L.): enzyme association, antibody production and cDNA cloning. Planta 197, 369–375.

    PubMed  Google Scholar 

  • Hartwell J, Gill A, Nimmo G A, Wilkins M B, Jenkins G I and Nimmo H G 1999 Phosphoenolpyruvate carboxylase kinase is a novel protein kinase regulated at the level of expression. Plant J 20, 333–42.

    PubMed  Google Scholar 

  • Heukeshovewn J and Dernick R 1988 Improved silver staining procedure for fast staining in PhastSystem Development Unit. I. Staining of sodium dodecyl sulfate gels. Electrophoresis 9, 28–32.

    PubMed  Google Scholar 

  • Huber S C, Huber J L and McMichael R W Jr 1994 Control of plant enzyme activity by reversible protein phosphorylation. Int. Rev. Cytol. 149, 47–98.

    Google Scholar 

  • Israel D W and Jackson W A 1982 Ion balance, uptake, and transport processes in N2-fixing and nitrate-and urea-dependent soybean plants. Plant Physiol. 112, 171–178.

    Google Scholar 

  • Jiao J, Echevarria C, Vidal J and Chollet R 1991 Protein turnover as a component in the light/dark regulation of phosphoenolpyruvate carboxylase protein-serine kinase activity in C4 plants. Proc. Natl. Acad. Sci. USA. 88, 2712–2715.

    PubMed  Google Scholar 

  • Johnson J F, Allan D L, Vance C P and Weiblen G 1996a Root carbon dioxide fixation by phosphorus-deficient Lupinus albus. Contribution to organic acid exudation by proteoid roots. Plant Physiol. 112, 19–30.

    PubMed  Google Scholar 

  • Johnson J F, Vance C P and Allan D L 1996b Phosphorus deficiency in Lupinus albus. Altered lateral root development and enhanced expression of phosphoenolpyruvate carboxylase. Plant Physiol. 112, 31–41.

    PubMed  Google Scholar 

  • Kim D J and Smith S M 1994 Expression of a single gene encoding microbody NAD-malate dehydrogenase during glyoxysome and peroxisome development in cucumber. Plant Mol. Biol. 26, 1833–1841.

    PubMed  Google Scholar 

  • Lance C and Rustin P 1984 The central role of malate in plant metabolism. Physiol. Veg. 22, 625–641.

    Google Scholar 

  • Latzko E and Kelly G J 1983 The many-faceted function of phosphoenolpyruvate carboxylase in C3 plants. Physiol. Veg. 21, 805–815.

    Google Scholar 

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

    Google Scholar 

  • Liu J, Uhde-Stone C, Li A, Vance C and Allan D 2001 A phosphate transporter with enhanced expression in proteoid roots of white lupin (Lupinus albus L.), Plant Soil 237, 257–266.

    Google Scholar 

  • Marczewski W 1989 Kinetic properties of phosphoenolpyruvate carboxylase from lupin nodules and roots. Physiol Planta. 76, 539–543.

    Google Scholar 

  • Marschner H V, Romheld V, Horst W J and Martin P 1986 Rootinduced changes in the rhizosphere: importance for the mineral nutrition of plants. Z. Pflanzenernaehr Bodenk. 149, 441–456.

    Google Scholar 

  • Marschner H, Romheld V and Cakmak I 1987 Root-induced changes of nutrient availability in the rhizosphere. J. Plant Nutr. 10, 1175–1184.

    Google Scholar 

  • Marschner H 1995 Mineral Nutrition of Higher Plants, 2nd edition. Academic Press Inc., San Diego, CA.

    Google Scholar 

  • Massonneau A, Langlade N, Leon S, Smutny J, Vogt E, Neumann G and Martinoia E 2001 Metabolic changes associated with cluster root development in white lupin (Lupinus albus L.): relationship between organic acid excretion, sucrose metabolism and energy status. Planta 213, 534–42.

    PubMed  Google Scholar 

  • Maxwell C A 1982 CO2 fixation in alfalfa and birdsfoot trefoil root nodules: synthesis, transport and partitioning of compounds from 14CO2 MS. University of Minnesota, St Paul, Minn.

    Google Scholar 

  • McCloud S A, Smith R G and Schuller K A 2001. Partial purification and characterization of pyruvate kinase from the plant fraction of soybean root nodules. Physiol. Plant.111, 283–290.

    PubMed  Google Scholar 

  • Melzer E and O'Leary M H 1987 Anaplerotic CO2 fixation by phosphoenolpyruvate carboxylase in C3 plants. Plant Physiol. 84, 58–60.

    Google Scholar 

  • Metzler M C, Rothermel B A and Nelson T 1989 Maize NADPmalate dehydrogenase: cDNA cloning, sequence and mRNA characterization. Plant Mol. Biol. 12, 712-722.

    Google Scholar 

  • Miller S S, Discroll B T, Gregerson R G, Gantt J S and Vance C P 1998 Alfalfa malate dehydrogenase (MDH): molecular cloning and characterization of five different forms reveals a unique nodule-enhanced MDH. Plant J. 15, 173–185.

    PubMed  Google Scholar 

  • Miller S S, Liu J, Allan D L, Menzhuber C J, Fedorova M and Vance C P 2001 Molecular control of acid phosphatase secreted into the rhizosphere of proteoid roots from phosphorus stressed white lupin. Plant Physiol. 127, 594–606.

    PubMed  Google Scholar 

  • Moraes T F and Plaxton W C 2000 Purification and characterization of phosphoenolpyruvate carboxylase from Brassica napus (rapeseed) suspension cell cultures: implications for phosphoenolpyruvate carboxylase regulation during phosphate starvation, and the integration of glycolysis with nitrogen assimilation. Eur. J. Biochem. 267, 4465–4476.

    PubMed  Google Scholar 

  • Musrati R A, Kollarova M, Mernik N and Mikulasova D 1998 Malate dehydrogenase: distribution, function and properties. Gen. Physiol. Biophys. 17, 193–210.

    PubMed  Google Scholar 

  • Neumann G, Massonneau A, Martinoia E and Romheld V 1999 Physiological adaptations to phosphorus deficiency during proteoid root development in white lupin. Planta 208, 373–382.

    Google Scholar 

  • Neumann G, Massonneau A, Langlade N, Dinkelaker B, Hengeler C, Romheld V and Martinoia E 2000 Physiological aspects of cluster root function and development in phosphorus-deficient white lupin (Lupinus albus L.). Ann. Bot. 85, 909–919.

    Google Scholar 

  • Pathirana S M, Vance C P, Miller S S and Gantt J S 1992 Alfalfa root nodule phosphoenolpyruvate carboxylase: characterization of the cDNA and expression in effective and plant-controlled ineffective nodules. Plant Mol. Biol. 20, 437–450

    PubMed  Google Scholar 

  • Pilbeam D J, Cakman I, Marschner H and Kirkby E A 1993 Effect of withdrawal of phosphorus on nitrate assimilation and PEP carboxylase activity in tomato. Plant Soil 154, 111–117.

    Google Scholar 

  • Raghothama K G 1999 Phosphate acquisition. Ann. Rev. Plant. Physiol. Plant. Mol. Biol. 50, 665–693.

    Google Scholar 

  • Sambrook J, Fritch E and Maniatis T 1989 Molecular Cloning. Cold Spring Harbour Laboratory Press. Cold Spring Harbour, NY.

    Google Scholar 

  • Schuller K A, Turpin D H and Plaxton W C 1990 Metabolite regulation of partially purified soybean nodule phosphoenolpyruvate carboxylase. Plant Physiol. 94, 1429–1435.

    Google Scholar 

  • Schuller K A and Werner D 1993 Phosphorylation of soybean nodule phosphoenolpyruvate carboxylase in vitro decreases sensitivity to inhibition by L-malate. Plant Physiol. 101, 1267–1273.

    PubMed  Google Scholar 

  • Scrutton M C and Fatebene F 1975 An assay system for localization of pyruvate and phosphoenolpyruvate carboxylase activity on polyacrylamide gels and its application of detection of these enzymes in tissue and extracts. Anal Biochem. 69, 247–260.

    PubMed  Google Scholar 

  • Skene K R 1998b Cluster roots: some ecological considerations. J. Ecol. 86, 1060–1064.

    Google Scholar 

  • Sugimoto T, Kawasaki T, Kato T, Whittier R F, Shibata D and Kawamura Y 1992 cDNA sequence and expression of a phosphoenolpyruvate carboxylase gene from soybean. Plant Mol. Biol. 20, 743–747.

    PubMed  Google Scholar 

  • Tadano T and Sakai H 1991 Secretion of acid phosphatase by roots of several crop species under phosphorus-deficient conditions. Soil Sci. Plant Nutr. 37, 129–140.

    Google Scholar 

  • Taybi T, Patil S, Chollet R and Cushman J C 2000 A minimal serine/threonine protein kinase circadianly regulates phosphoenolpyruvate carboxylase activity in crassulacean acid metabolism-induced leaves of the common ice plant. Plant Physiol. 123, 1471–1482.

    PubMed  Google Scholar 

  • Theodorou M E, Cornel F A, Duff S M G and Plaxton W C 1992 Phosphate starvation-inducible synthesis of the ?-subunit of the pyrophosphate-dependent phosphofructokinase in black mustard suspension cells. J. Biol. Chem. 267, 21901–21905.

    PubMed  Google Scholar 

  • Theodoru M E and Plaxton W C 1993 Metabolic adaptations of plant respiration to nutritional phosphate deprivation. Plant. Physiol. 101, 339–344.

    PubMed  Google Scholar 

  • Ting I D and Osmond C B 1973 Multiple forms of plant PEP carboxylase associated with different metabolic pathways. Plant Physiol. 51, 448–453.

    Google Scholar 

  • TrinickMJ 1977 Vesicular-arbuscular infection and soil phosphorus utilization in Lupinus spp. New Phytol. 78, 297–304.

    Google Scholar 

  • Vance C P, Stade S and Maxwell C A 1983 Alfalfa root nodule carbon dioxide fixation: I. Association with nitrogen fixation and incorporation into amino acids. Plant Physiol. 72, 469–473.

    Google Scholar 

  • Wang Y H and Chollet R 1993 In vitro phosphorylation of purified tobacco-leaf phosphoenolpyruvate carboxylase. FEBS Lett. 328, 215–218.

    PubMed  Google Scholar 

  • Watt M and Evans J R 1999a Linking development and determinacy with organic acid efflux from proteoid roots of white lupin grown with low phosphorus and ambient or elevated CO2 concentration. Plant Physiol. 120, 705–716.

    PubMed  Google Scholar 

  • Watt M and Evans J R 1999b. Proteoid roots. Physiology and development. Plant Physiol. 121, 317–323.

    PubMed  Google Scholar 

  • Zhang X Q, Li B and Chollet R 1995 In vivo regulatory phosphorylation of soybean-nodule phosphoenolpyruvate carboxylase. Plant Physiol. 108, 1561–1568.

    PubMed  Google Scholar 

  • Zhang X Q and Chollet R 1997 Phosphoenolpyruvate carboxylase protein kinase from soybean root nodules: partial purification, characterization, and up/down-regulation by photosynthate supply from the shoots. Arch Biochem Biophys. 15, 260–268.

    Google Scholar 

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Authors and Affiliations

  1. Department of Soil, Water and Climate, University of Minnesota, 1991 Upper Buford Circle, St. Paul, MN, 55108, USA

    Claudia Uhde-Stone, Glena Gilbert, Jane M-F Johnson, Kelly E. Zinn & Deborah L. Allan

  2. Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, MN, 55108, USA

    Claudia Uhde-Stone, Ralph Litjens & Stephen J. Temple

  3. Biology Dept., Viterbo University, 815 Ninth Street South, LaCrosse, WI, 54601, USA

    Glena Gilbert

  4. USDA-ARS-MWA, 803 Iowa Ave, Morris, MN, 56267, USA

    Jane M-F Johnson & Carroll P. Vance

  5. Division of Laboratory Science, Department of Technical Microbiology, Higher Institute for Professional Education Venlo, 5822 BD, Venlo, The Netherlands

    Ralph Litjens

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  1. Claudia Uhde-Stone
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  2. Glena Gilbert
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  3. Jane M-F Johnson
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Correspondence to Deborah L. Allan.

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Uhde-Stone, C., Gilbert, G., Johnson, J.MF. et al. Acclimation of white lupin to phosphorus deficiency involves enhanced expression of genes related to organic acid metabolism. Plant and Soil 248, 99–116 (2003). https://doi.org/10.1023/A:1022335519879

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