Lipid utilization, gluconeogenesis, and seedling growth in Arabidopsis mutants lacking the glyoxylate cycle enzyme malate synthase.
The aim of this research was to test the role of the glyoxylate cycle enzyme malate synthase (MLS) in lipid utilization, gluconeogenesis, and seedling growth in Arabidopsis. We hypothesized that in the absence of MLS, succinate produced by isocitrate lyase (ICL) could still feed into the tricarboxylic acid cycle, whereas glyoxylate could be converted to sugars using enzymes of the photorespiratory pathway. To test this hypothesis we isolated knock-out mls mutants and studied their growth and metabolism in comparison to wild type and icl mutant seedlings. In contrast to icl seedlings, which grow slowly and are unable to convert lipid into sugars (Eastmond, P. J., Germain, V., Lange, P. R., Bryce, J. H., Smith, S. M. & Graham, I. A. (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 5669-5674), mls seedlings grow faster, use their lipid more rapidly, and are better able to establish as plantlets. Transcriptome and metabolome analyses show that icl seedlings exhibit many features characteristic of carbohydrate starvation, whereas mls seedlings differ relatively little from wild type. In the light mls seedlings generate more sugars than icl seedlings, and when fed with [14C]acetate, 14C-labeling of sugars is three times greater than in icl seedlings and more than half that in wild type seedlings. The mls seedlings also accumulate more glycine and serine than icl or wild type seedlings, consistent with a diversion of glyoxylate into these intermediates of the photorespiratory pathway. We conclude that, in contrast to bacteria and fungi in which MLS is essential for gluconeogenesis from acetate or fatty acids, MLS is partially dispensable for lipid utilization and gluconeogenesis in Arabidopsis seedlings.
Cornah
Johanna E
JE
Institute of Cell and Molecular Biology, University of Edinburgh, King's Bldgs., Mayfield Rd., Edinburgh, EH9 3JH, United Kingdom.
Germain
Véronique
V
Ward
Jane L
JL
Beale
Michael H
MH
Smith
Steven M
SM
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D001669:Biochemical Phenomena; D001671:Biochemistry; D002241:Carbohydrates; D002845:Chromatography; D002952:Citric Acid Cycle; D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D018745:Genome, Plant; D005998:Glycine / Q000737:chemistry; D006038:Glyoxylates / Q000737:chemistry*; D007522:Isocitrate Lyase / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D008292:Malate Synthase / Q000235:genetics* / Q000502:physiology*; D008954:Models, Biological; D008957:Models, Genetic; D009154:Mutation; D010641:Phenotype; D030821:Plants, Genetically Modified; D040081:Protein Array Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000378:metabolism; D013997:Time Factors
D002241:Carbohydrates; D018076:DNA, Complementary; D006038:Glyoxylates; D008055:Lipids; D004247:DNA; D008292:Malate Synthase; D007522:Isocitrate Lyase; C031150:glyoxylic acid; D005998:Glycine