title: Quantitative Multilevel Analysis of Central Metabolism in Developing Oilseeds of Oilseed Rape during in Vitro Culture.
abstract: Seeds provide the basis for many food, feed, and fuel products. Continued increases in seed yield, composition, and quality require an improved understanding of how the developing seed converts carbon and nitrogen supplies into storage. Current knowledge of this process is often based on the premise that transcriptional regulation directly translates via enzyme concentration into flux. In an attempt to highlight metabolic control, we explore genotypic differences in carbon partitioning for in vitro cultured developing embryos of oilseed rape (Brassica napus). We determined biomass composition as well as 79 net fluxes, the levels of 77 metabolites, and 26 enzyme activities with specific focus on central metabolism in nine selected germplasm accessions. Overall, we observed a tradeoff between the biomass component fractions of lipid and starch. With increasing lipid content over the spectrum of genotypes, plastidic fatty acid synthesis and glycolytic flux increased concomitantly, while glycolytic intermediates decreased. The lipid/starch tradeoff was not reflected at the proteome level, pointing to the significance of (posttranslational) metabolic control. Enzyme activity/flux and metabolite/flux correlations suggest that plastidic pyruvate kinase exerts flux control and that the lipid/starch tradeoff is most likely mediated by allosteric feedback regulation of phosphofructokinase and ADP-glucose pyrophosphorylase. Quantitative data were also used to calculate in vivo mass action ratios, reaction equilibria, and metabolite turnover times. Compounds like cyclic 3',5'-AMP and sucrose-6-phosphate were identified to potentially be involved in so far unknown mechanisms of metabolic control. This study provides a rich source of quantitative data for those studying central metabolism.
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Jörg
authors.initials: J
authors.lastname: Schwender
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Inga
authors.initials: I
authors.lastname: Hebbelmann
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Nicolas
authors.initials: N
authors.lastname: Heinzel
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Tatjana
authors.initials: T
authors.lastname: Hildebrandt
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Alistair
authors.initials: A
authors.lastname: Rogers
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Dhiraj
authors.identifier: http://orcid.org/0000-0002-1226-2337
authors.initials: D
authors.lastname: Naik
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Matthias
authors.initials: M
authors.lastname: Klapperstück
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Hans-Peter
authors.identifier: http://orcid.org/0000-0002-4459-9727
authors.initials: HP
authors.lastname: Braun
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Falk
authors.initials: F
authors.lastname: Schreiber
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Peter
authors.identifier: http://orcid.org/0000-0003-4567-2700
authors.initials: P
authors.lastname: Denolf
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.).
authors.forename: Ljudmilla
authors.initials: L
authors.lastname: Borisjuk
authors.affiliation: Brookhaven National Laboratory, Biological, Environmental, and Climate Sciences Department, Upton, New York 11973 (J.S., I.H., A.R., D.N.);Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany (N.H., L.B., H.R.);Institut für Pflanzengenetik, Universität Hannover, 30419 Hannover, Germany (T.H., H.-P.B.);Department of Environmental Science, Indian Institute of Advanced Research, Koba, Gandhinagar 382007, Gujarat, India (D.N.);Clayton School of Information Technology, Monash University, Melbourne, Victoria 3800, Australia (M.K., F.S.);Institute of Computer Science, University Halle-Wittenberg, 06120 Halle, Germany (F.S.); andBayer CropScience, 9052 Zwijnaarde, Belgium (P.D.) rollet@ipk-gatersleben.de.
authors.forename: Hardy
authors.initials: H
authors.lastname: Rolletschek
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D055162:Biocatalysis; D018533:Biomass; D029688:Brassica napus / Q000196:embryology* / Q000378:metabolism* / Q000648:ultrastructure; D050260:Carbohydrate Metabolism; D002244:Carbon / Q000378:metabolism; D002853:Chromatography, Liquid; D006019:Glycolysis; D050356:Lipid Metabolism; D013058:Mass Spectrometry; D064688:Metabolic Flux Analysis; D008954:Models, Biological; D055361:Multilevel Analysis; D010938:Plant Oils / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism; D020543:Proteome / Q000378:metabolism; D012639:Seeds / Q000196:embryology* / Q000378:metabolism* / Q000648:ultrastructure; D013213:Starch / Q000378:metabolism; D013997:Time Factors; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D000596:Amino Acids; D010938:Plant Oils; D010940:Plant Proteins; D020543:Proteome; D002244:Carbon; D013213:Starch