RESUMEN
Plant oils in the form of triacylglycerol (TAG) are used for food, industrial feedstock and biofuel manufacture. Although TAG is typically harvested from the fruit or seeds of oil crop species, plants can also accumulate small amounts of TAG in the leaves and other vegetative tissues. Here we show that leaf TAG levels can be increased significantly (10-20 fold) by blocking fatty acid breakdown, particularly during extended dark treatments or leaf senescence in the model plant Arabidopsis. Generation of a double mutant in fatty acid breakdown and diacylglycerol acyltransferase 1 (DGAT1) resulted in a severe vegetative growth phenotype suggesting that partitioning of fatty acids to TAG in leaves is carried out predominantly by this acyltransferase. LEC2, a seed development transcription factor involved in storage product accumulation, was ectopically expressed during senescence in the fatty acid breakdown mutant COMATOSE (cts2). This resulted in accumulation of seed oil type species of TAG in senescing tissue. Our data suggests that recycled membrane fatty acids can be re-directed to TAG by expressing the seed-programme in senescing tissue or by a block in fatty acid breakdown. This work raises the possibility of producing significant amounts of oil in vegetative tissues of biomass crops such as Miscanthus.
Asunto(s)
Ácidos Grasos/metabolismo , Metabolismo de los Lípidos/fisiología , Hojas de la Planta/metabolismo , Aceites de Plantas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Cromatografía en Capa Delgada , Diacilglicerol O-Acetiltransferasa/genética , Ácidos Grasos/genética , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Metabolismo de los Lípidos/genética , Microscopía Electrónica de Transmisión , Hojas de la Planta/genética , Plantas Modificadas Genéticamente/genética , Triglicéridos/genética , Triglicéridos/metabolismoRESUMEN
The multifunctional protein (MFP) of peroxisomal beta-oxidation catalyses four separate reactions, two of which (2-trans enoyl-CoA hydratase and L-3-hydroxyacyl-CoA dehydrogenase) are core activities required for the catabolism of all fatty acids. We have isolated and characterized five Arabidopsis thaliana mutants in the MFP2 gene that is expressed predominantly in germinating seeds. Seedlings of mfp2 require an exogenous supply of sucrose for seedling establishment to occur. Analysis of mfp2-1 seedlings revealed that seed storage lipid was catabolized more slowly, long-chain acyl-CoA substrates accumulated and there was an increase in peroxisome size. Despite a reduction in the rate of beta-oxidation, mfp2 seedlings are not resistant to the herbicide 2,4-dichlorophenoxybutyric acid, which is catabolized to the auxin 2,4-dichlorophenoxyacetic acid by beta-oxidation. Acyl-CoA feeding experiments show that the MFP2 2-trans enoyl-CoA hydratase only exhibits activity against long chain (C18:0) substrates, whereas the MFP2 L-3-hydroxyacyl-CoA dehydrogenase is active on C6:0, C12:0 and C18:0 substrates. A mutation in the abnormal inflorescence meristem gene AIM1, the only homologue of MFP2, results in an abnormal inflorescence meristem phenotype in mature plants (Richmond and Bleecker, Plant Cell 11, 1999, 1911) demonstrating that the role of these genes is very different. The mfp2-1 aim1double mutant aborted during the early stages of embryo development showing that these two proteins share a common function that is essential for this key stage in the life cycle.