title: Genetic and transgenic perturbations of carbon reserve production in Arabidopsis seeds reveal metabolic interactions of biochemical pathways.
abstract: The biosynthesis of seed oil and starch both depend on the supply of carbon from the maternal plant. The biochemical interactions between these two pathways are not fully understood. In the Arabidopsis mutant shrunken seed 1 (sse1)/pex16, a reduced rate of fatty acid synthesis leads to starch accumulation. To further understand the metabolic impact of the decrease in oil synthesis, we compared soluble metabolites in sse1 and wild type (WT) seeds. Sugars, sugar phosphates, alcohols, pyruvate, and many other organic acids accumulated in sse1 seeds as a likely consequence of the reduced carbon demand for lipid synthesis. The enlarged pool size of hexose-P, the metabolites at the crossroad of sugar metabolism, glycolysis, and starch synthesis, was likely a direct cause of the increased flow into starch. Downstream of glycolysis, more carbon entered the TCA cycle as an alternative to the fatty acid pathway, causing the total amount of TCA cycle intermediates to rise while moving the steady state of the cycle away from fumarate. To convert the excess carbon metabolites into starch, we introduced the Escherichia coli starch synthetic enzyme ADP-glucose pyrophosphorylase (AGPase) into sse1 seeds. Expression of AGPase enhanced net starch biosynthesis in the mutant, resulting in starch levels that reached 37% of seed weight. However, further increases above this level were not achieved and most of the carbon intermediates remained high in comparison with the WT, indicating that additional mechanisms limit starch deposition in Arabidopsis seeds.
authors.affiliation: Department of Crop Sciences, University of Illinois, Urbana, IL 61801, USA. yunlin@uiuc.edu
authors.forename: Yun
authors.initials: Y
authors.lastname: Lin
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Ulanov
authors.forename: Vera
authors.initials: V
authors.lastname: Lozovaya
authors.forename: Jack
authors.initials: J
authors.lastname: Widholm
authors.forename: Guirong
authors.initials: G
authors.lastname: Zhang
authors.forename: Jinhua
authors.initials: J
authors.lastname: Guo
authors.forename: Howard M
authors.initials: HM
authors.lastname: Goodman
mesh_terms: D000596:Amino Acids / Q000378:metabolism; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D050260:Carbohydrate Metabolism / Q000235:genetics / Q000502:physiology; D002244:Carbon / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005227:Fatty Acids / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D018506:Gene Expression Regulation, Plant; D051599:Glucose-1-Phosphate Adenylyltransferase / Q000235:genetics / Q000378:metabolism; D050356:Lipid Metabolism / Q000235:genetics / Q000502:physiology; D053858:Metabolic Networks and Pathways / Q000235:genetics / Q000502:physiology; D008954:Models, Biological; D010938:Plant Oils / Q000378:metabolism; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics / Q000378:metabolism*; D013213:Starch / Q000378:metabolism
chemical_list: D000596:Amino Acids; D029681:Arabidopsis Proteins; D005227:Fatty Acids; D010938:Plant Oils; D011994:Recombinant Proteins; D002244:Carbon; D013213:Starch; D051599:Glucose-1-Phosphate Adenylyltransferase