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 Arabidopsis acyl-CoA oxidase (ACX) family comprises isozymes with distinct fatty acid chain-length specificities that together catalyse the first step of peroxisomal fatty acid beta-oxidation. We have isolated and characterized T-DNA insertion mutants in the medium to long-chain (ACX1) and long-chain (ACX2) acyl-CoA oxidases, and show that the corresponding endogenous activities are decreased in the mutants. Lipid catabolism during germination and early post-germinative growth was unaltered in the acx1-1 mutant, but slightly delayed in the acx2-1 mutant, with 3-day-old acx2-1 seedlings accumulating long-chain acyl-CoAs. In acx1-1 and acx2-1, seedling growth and establishment in the absence of an exogenous supply of sucrose was unaffected. Seedlings of the double mutant acx1-1 acx2-1 were unable to catabolize seed storage lipid, and accumulated long-chain acyl-CoAs. The acx1-1 acx2-1 seedlings were also unable to establish photosynthetic competency in the absence of an exogenous carbon supply, a phenotype that is shared with a number of other Arabidopsis mutants disrupted in storage lipid breakdown. Germination frequency of the double mutant was significantly reduced compared with wild-type seeds. This was unaffected by the addition of exogenous sucrose, but was improved by dormancy-breaking treatments such as cold stratification and after-ripening. We show that the acx1-1, acx2-1 and acx1-2 acx2-1 double mutants and the ketoacyl-CoA thiolase-2 (kat2) mutant exhibit a sucrose-independent germination phenotype comparable with that reported for comatose (cts-2), a mutant in a peroxisomal ABC transporter which exhibits enhanced dormancy. This demonstrates an additional role beyond that of carbon provision for the beta-oxidation pathway during germination or in dormant seeds.