TY - JOUR A1 - Araujo, Wagner L. A1 - Nunes-Nesi, Adriano A1 - Nikoloski, Zoran A1 - Sweetlove, Lee J. A1 - Fernie, Alisdair R. T1 - Metabolic control and regulation of the tricarboxylic acid cycle in photosynthetic and heterotrophic plant tissues JF - Plant, cell & environment : cell physiology, whole-plant physiology, community physiology N2 - The tricarboxylic acid (TCA) cycle is a crucial component of respiratory metabolism in both photosynthetic and heterotrophic plant organs. All of the major genes of the tomato TCA cycle have been cloned recently, allowing the generation of a suite of transgenic plants in which the majority of the enzymes in the pathway are progressively decreased. Investigations of these plants have provided an almost complete view of the distribution of control in this important pathway. Our studies suggest that citrate synthase, aconitase, isocitrate dehydrogenase, succinyl CoA ligase, succinate dehydrogenase, fumarase and malate dehydrogenase have control coefficients flux for respiration of -0.4, 0.964, -0.123, 0.0008, 0.289, 0.601 and 1.76, respectively; while 2-oxoglutarate dehydrogenase is estimated to have a control coefficient of 0.786 in potato tubers. These results thus indicate that the control of this pathway is distributed among malate dehydrogenase, aconitase, fumarase, succinate dehydrogenase and 2-oxoglutarate dehydrogenase. The unusual distribution of control estimated here is consistent with specific non-cyclic flux mode and cytosolic bypasses that operate in illuminated leaves. These observations are discussed in the context of known regulatory properties of the enzymes and some illustrative examples of how the pathway responds to environmental change are given. KW - metabolic control analysis KW - metabolic regulation KW - respiration KW - Solanum lycopersicum (tomato) KW - TCA cycle Y1 - 2012 U6 - https://doi.org/10.1111/j.1365-3040.2011.02332.x SN - 0140-7791 VL - 35 IS - 1 SP - 1 EP - 21 PB - Wiley-Blackwell CY - Hoboken ER - TY - JOUR A1 - Fettke, Jörg A1 - Nunes-Nesi, Adriano A1 - Fernie, Alisdair R. A1 - Steup, Martin T1 - Identification of a novel heteroglycan-interacting protein, HIP 1.3, from Arabidopsis thaliana JF - Journal of plant physiology : biochemistry, physiology, molecular biology and biotechnology of plants N2 - Plastidial degradation of transitory starch yields mainly maltose and glucose. Following the export into the cytosol, maltose acts as donor for a glucosyl transfer to cytosolic heteroglycans as mediated by a cytosolic transglucosidase (DPE2; EC 2.4.1.25) and the second glucosyl residue is liberated as glucose. The cytosolic phosphorylase (Pho2/PHS2; EC 2.4.1.1) also interacts with heteroglycans using the same intramolecular sites as DPE2. Thus, the two glucosyl transferases interconnect the cytosolic pools of glucose and glucose 1-phosphate. Due to the complex monosaccharide pattern, other heteroglycan-interacting proteins (Hips) are expected to exist. Identification of those proteins was approached by using two types of affinity chromatography. Heteroglycans from leaves of Arabidopsis thaliana (Col-0) covalently bound to Sepharose served as ligands that were reacted with a complex mixture of buffer-soluble proteins from Arabidopsis leaves. Binding proteins were eluted by sodium chloride. For identification, SDS-PAGE, tryptic digestion and MALDI-TOF analyses were applied. A strongly interacting polypeptide (approximately 40 kDa; designated as HIP1.3) was observed as product of locus At1g09340. Arabidopsis mutants deficient in HIP1.3 were reduced in growth and contained heteroglycans displaying an altered monosaccharide pattern. Wild type plants express HIP1.3 most strongly in leaves. As revealed by immuno fluorescence, HIP1.3 is located in the cytosol of mesophyll cells but mostly associated with the cytosolic surface of the chloroplast envelope membranes. In an HIP1.3-deficient mutant the immunosignal was undetectable. Metabolic profiles from leaves of this mutant and wild type plants as well were determined by GC-MS. As compared to the wild type control, more than ten metabolites, such as ascorbic acid, fructose, fructose bisphosphate, glucose, glycine, were elevated in darkness but decreased in the light. Although the biochemical function of HIP1.3 has not yet been elucidated, it is likely to possess an important function in the central carbon metabolism of higher plants. KW - Arabidopsis thaliana KW - Carbohydrate binding proteins KW - Cytosolic heteroglycans KW - Maltose metabolism KW - Starch metabolism Y1 - 2011 U6 - https://doi.org/10.1016/j.jplph.2010.09.008 SN - 0176-1617 VL - 168 IS - 12 SP - 1415 EP - 1425 PB - Elsevier CY - Jena ER - TY - JOUR A1 - Nunes-Nesi, Adriano A1 - Alseekh, Saleh A1 - de Oliveira Silva, Franklin Magnum A1 - Omranian, Nooshin A1 - Lichtenstein, Gabriel A1 - Mirnezhad, Mohammad A1 - Romero Gonzalez, Roman R. A1 - Sabio y Garcia, Julia A1 - Conte, Mariana A1 - Leiss, Kirsten A. A1 - Klinkhamer, Peter Gerardus Leonardus A1 - Nikoloski, Zoran A1 - Carrari, Fernando A1 - Fernie, Alisdair R. T1 - Identification and characterization of metabolite quantitative trait loci in tomato leaves and comparison with those reported for fruits and seeds JF - Metabolomics N2 - IntroductionTo date, most studies of natural variation and metabolite quantitative trait loci (mQTL) in tomato have focused on fruit metabolism, leaving aside the identification of genomic regions involved in the regulation of leaf metabolism.ObjectiveThis study was conducted to identify leaf mQTL in tomato and to assess the association of leaf metabolites and physiological traits with the metabolite levels from other tissues.MethodsThe analysis of components of leaf metabolism was performed by phenotypying 76 tomato ILs with chromosome segments of the wild species Solanum pennellii in the genetic background of a cultivated tomato (S. lycopersicum) variety M82. The plants were cultivated in two different environments in independent years and samples were harvested from mature leaves of non-flowering plants at the middle of the light period. The non-targeted metabolite profiling was obtained by gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). With the data set obtained in this study and already published metabolomics data from seed and fruit, we performed QTL mapping, heritability and correlation analyses.ResultsChanges in metabolite contents were evident in the ILs that are potentially important with respect to stress responses and plant physiology. By analyzing the obtained data, we identified 42 positive and 76 negative mQTL involved in carbon and nitrogen metabolism.ConclusionsOverall, these findings allowed the identification of S. lycopersicum genome regions involved in the regulation of leaf primary carbon and nitrogen metabolism, as well as the association of leaf metabolites with metabolites from seeds and fruits. KW - Metabolite QTL KW - Tomato KW - Leaf metabolism KW - Metabolite network Y1 - 2019 U6 - https://doi.org/10.1007/s11306-019-1503-8 SN - 1573-3882 SN - 1573-3890 VL - 15 IS - 46 PB - Springer CY - New York ER - TY - JOUR A1 - Smith, Sarah R. A1 - Dupont, Chris L. A1 - McCarthy, James K. A1 - Broddrick, Jared T. A1 - Obornik, Miroslav A1 - Horak, Ales A1 - Füssy, Zoltán A1 - Cihlar, Jaromir A1 - Kleessen, Sabrina A1 - Zheng, Hong A1 - McCrow, John P. A1 - Hixson, Kim K. A1 - Araujo, Wagner L. A1 - Nunes-Nesi, Adriano A1 - Fernie, Alisdair R. A1 - Nikoloski, Zoran A1 - Palsson, Bernhard O. A1 - Allen, Andrew E. T1 - Evolution and regulation of nitrogen flux through compartmentalized metabolic networks in a marine diatom JF - Nature Communications N2 - Diatoms outcompete other phytoplankton for nitrate, yet little is known about the mechanisms underpinning this ability. Genomes and genome-enabled studies have shown that diatoms possess unique features of nitrogen metabolism however, the implications for nutrient utilization and growth are poorly understood. Using a combination of transcriptomics, proteomics, metabolomics, fluxomics, and flux balance analysis to examine short-term shifts in nitrogen utilization in the model pennate diatom in Phaeodactylum tricornutum, we obtained a systems-level understanding of assimilation and intracellular distribution of nitrogen. Chloroplasts and mitochondria are energetically integrated at the critical intersection of carbon and nitrogen metabolism in diatoms. Pathways involved in this integration are organelle-localized GS-GOGAT cycles, aspartate and alanine systems for amino moiety exchange, and a split-organelle arginine biosynthesis pathway that clarifies the role of the diatom urea cycle. This unique configuration allows diatoms to efficiently adjust to changing nitrogen status, conferring an ecological advantage over other phytoplankton taxa. KW - Biochemistry KW - Computational biology and bioinformatics KW - Evolution KW - Microbiology KW - Molecular biology Y1 - 2019 U6 - https://doi.org/10.1038/s41467-019-12407-y SN - 2041-1723 VL - 10 PB - Nature Publ. Group CY - London ER -