Increased flow of fatty acids toward beta-oxidation in developing seeds of Arabidopsis deficient in diacylglycerol acyltransferase activity or synthesizing medium-chain-length fatty acids.

Synthesis of polyhydroxyalkanoates (PHAs) from intermediates of fatty acid beta-oxidation was used as a tool to study fatty acid degradation in developing seeds of Arabidopsis. Transgenic plants expressing a peroxisomal PHA synthase under the control of a napin promoter accumulated PHA in developing seeds to a final level of 0. 06 mg g(-1) dry weight. In plants co-expressing a plastidial acyl-acyl carrier protein thioesterase from Cuphea lanceolata and a peroxisomal PHA synthase, approximately 18-fold more PHA accumulated in developing seeds. The proportion of 3-hydroxydecanoic acid monomer in the PHA was strongly increased, indicating a large flow of capric acid toward beta-oxidation. Furthermore, expression of the peroxisomal PHA synthase in an Arabidopsis mutant deficient in the enzyme diacylglycerol acyltransferase resulted in a 10-fold increase in PHA accumulation in developing seeds. These data indicate that plants can respond to the inadequate incorporation of fatty acids into triacylglycerides by recycling the fatty acids via beta-oxidation and that a considerable flow toward beta-oxidation can occur even in a plant tissue primarily devoted to the accumulation of storage lipids.

[1]  Chris Somerville,et al.  Production of Polyhydroxyalkanoates, a Family of Biodegradable Plastics and Elastomers, in Bacteria and Plants , 1995, Bio/Technology.

[2]  Steven M. L. Smith,et al.  Glycoxylate cycle enzyme activities are induced in senescent pumpkin fruits , 1996 .

[3]  A. Steinbüchel,et al.  Diversity of bacterial polyhydroxyalkanoic acids , 1995 .

[4]  J. Ohlrogge,et al.  Feedback inhibition of fatty acid synthesis in tobacco suspension cells , 1995 .

[5]  J. Ohlrogge,et al.  Expression of Lauroyl–Acyl Carrier Protein Thioesterase in Brassica napus Seeds Induces Pathways for Both Fatty Acid Oxidation and Biosynthesis and Implies a Set Point for Triacylglycerol Accumulation , 1998, Plant Cell.

[6]  Y. Poirier,et al.  Polyhydroxyalkanoate synthesis in transgenic plants as a new tool to study carbon flow through beta-oxidation. , 1999, The Plant journal : for cell and molecular biology.

[7]  Y. Poirier,et al.  Synthesis of medium-chain-length polyhydroxyalkanoates in arabidopsis thaliana using intermediates of peroxisomal fatty acid beta-oxidation. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[8]  U. K. Laemmli,et al.  Cleavage of structural proteins during , 1970 .

[9]  K. Denby,et al.  Carbon Catabolite Repression Regulates Glyoxylate Cycle Gene Expression in Cucumber. , 1994, The Plant cell.

[10]  Y. Poirier,et al.  Polyhydroxybutyrate, a Biodegradable Thermoplastic, Produced in Transgenic Plants , 1992, Science.

[11]  J. Ellis,et al.  In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants , 1993 .

[12]  W. J. Dyer,et al.  A rapid method of total lipid extraction and purification. , 1959, Canadian journal of biochemistry and physiology.

[13]  Robert V Farese,et al.  Identification of a gene encoding an acyl CoA:diacylglycerol acyltransferase, a key enzyme in triacylglycerol synthesis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  P. Benfey,et al.  Tissue‐specific expression from CaMV 35S enhancer subdomains in early stages of plant development. , 1990, The EMBO journal.

[15]  P. Covello,et al.  Alteration of Seed Fatty Acid Composition by an Ethyl Methanesulfonate-Induced Mutation in Arabidopsis thaliana Affecting Diacylglycerol Acyltransferase Activity , 1995, Plant physiology.

[16]  Y. Poirier Production of new polymeric compounds in plants. , 1999, Current opinion in biotechnology.

[17]  I. Graham,et al.  No Induction of β-oxidation in leaves of Arabidopsis that over-produce lauric acid , 1999, Planta.

[18]  A. Steinbüchel,et al.  Bacterial and other biological systems for polyester production. , 1998, Trends in biotechnology.

[19]  L. Rask,et al.  Distribution of napin and cruciferin in developing rape seed embryos. , 1992, Plant physiology.

[20]  H. S. Chae,et al.  Ethylene-Mediated Phospholipid Catabolic Pathway in Glucose-Starved Carrot Suspension Cells , 1998 .

[21]  A. Pradet,et al.  Increased Fatty Acid beta-Oxidation after Glucose Starvation in Maize Root Tips. , 1992, Plant physiology.