Functional Overlap of Long-Chain Acyl-CoA Synthetases in Arabidopsis.

Long-chain acyl-CoA synthetases (LACSs) play diverse and essential roles in lipid metabolism. The genomes of model eukaryotic organisms encode multiple LACS genes, and the substrate specificities of LACS homologs often overlap substantially. Homologous LACSs tend to differ in their expression patterns, localizations, and, by extension, the metabolic pathways to which they contribute. The Arabidopsis genome encodes a family of nine LACS genes, which have been characterized largely by reverse genetic analysis of mutant phenotypes. Because of redundancy, distinguishing the contributions of some Arabidopsis LACS genes has been challenging. Here, we have attempted to clarify the functions of LACSs that functionally overlap by synopsizing the results of previous work, isolating a suite of higher-order mutants that were previously lacking, and analyzing oil, wax, cutin, cuticle permeability, fertility and growth phenotypes. LACS1, LACS2, LACS4, LACS8 and LACS9 all affect cuticular lipid metabolism, but have different precise roles. Seed set, seed weight and storage oil amounts of higher-order lacs1, lacs2, lacs4, lacs8 and lacs9 mutants vary greatly, with these traits subject to different effects of fertility and oil synthesis defects. LACS4, LACS8 and LACS9 have partially redundant roles in development, as lacs4 lacs8 and lacs4 lacs9 double mutants are dwarf. lacs4 lacs8 lacs9 triple mutants were not recovered, and are assumed to be non-viable. Together, these results sketch a complex network of functions and functional interactions within the Arabidopsis LACS gene family.