title: The metabolic flux phenotype of heterotrophic Arabidopsis cells reveals a flexible balance between the cytosolic and plastidic contributions to carbohydrate oxidation in response to phosphate limitation.
abstract: Understanding the mechanisms that allow plants to respond to variable and reduced availability of inorganic phosphate is of increasing agricultural importance because of the continuing depletion of the rock phosphate reserves that are used to combat inadequate phosphate levels in the soil. Changes in gene expression, protein levels, enzyme activities and metabolite levels all point to a reconfiguration of the central metabolic network in response to reduced availability of inorganic phosphate, but the metabolic significance of these changes can only be assessed in terms of the fluxes supported by the network. Steady-state metabolic flux analysis was used to define the metabolic phenotype of a heterotrophic Arabidopsis thaliana cell culture grown on a Murashige and Skoog medium containing 0, 1.25 or 5 mm inorganic phosphate. Fluxes through the central metabolic network were deduced from the redistribution of (13) C into metabolic intermediates and end products when cells were labelled with [1-(13) C], [2-(13) C], or [(13) C6 ]glucose, in combination with (14) C measurements of the rates of biomass accumulation. Analysis of the flux maps showed that reduced levels of phosphate in the growth medium stimulated flux through phosphoenolpyruvate carboxylase and malic enzyme, altered the balance between cytosolic and plastidic carbohydrate oxidation in favour of the plastid, and increased cell maintenance costs. We argue that plant cells respond to phosphate deprivation by reconfiguring the flux distribution through the pathways of carbohydrate oxidation to take advantage of better phosphate homeostasis in the plastid.
authors.affiliation: Department of Plant Sciences, University of Oxford, South Parks Road, Oxford, OX1 3RB, UK.
authors.forename: Shyam K
authors.initials: SK
authors.lastname: Masakapalli
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Bryant
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Kruger
authors.forename: R George
authors.initials: RG
authors.lastname: Ratcliffe
mesh_terms: D017360:Arabidopsis / Q000166:cytology / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism*; D006706:Homeostasis; D010084:Oxidation-Reduction; D010641:Phenotype; D010710:Phosphates / Q000378:metabolism*; D018087:Plastids / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000378:metabolism
chemical_list: D010710:Phosphates; D011770:Pyruvate Kinase
keywords: Arabidopsis thaliana; Pi homeostasis; carbon conversion efficiency; heterotrophic metabolism; metabolic flux analysis; phosphate availability; phosphate stress; plastid metabolism; pyruvate kinase; subcellular compartmentation