In characterizing the enzymes involved in the formation of very long-chain fatty acids (VLCFAs) i n the Brassicaceae, we have generated a series of mutants of Arabidopsis thaliana that have reduced VLCFA content. Here we report the characterization of a seed lipid mutant, AS1 1, which, in comparison to wild type (WT), has reduced levels of 20:l and 18:l and accumulates 18:3 as the major fatty acid in triacylglycerols. Proportions of 18:2 remain similar to WT. Cenetic analyses indicate that the fatty acid phenotype is caused by a semidominant mutation in a single nuclear gene, designated TAC1, located on chromosome 2. Biochemical analyses have shown that the AS11 phenotype is not due to a deficiency in the capacity t o elongate 18:l or to an increase in the relative A1 5 or A1 2 desaturase activities. Indeed, the ratio of desaturase/elongase activities measured in vitro is virtually identical in developing WT and AS1 1 seed homogenates. Rather, the fatty acid phenotype of AS1 1 is the result of reduced diacylglycerol acyltransferase activity throughout development, such that triacylglycerol biosynthesis is reduced. This leads to a reduction in 20:l biosynthesis during seed development, leaving more 18:l available for desaturation. Thus, we have demonstrated that changes to triacylglycerol biosynthesis can result i n dramatic changes i n fatty acid composition and, in particular, i n the accumulation of VLCFAs i n seed storage lipids.The fatty acyl composition of seed TAGs determines their physical and chemical properties and, thus, their use in edible oil or industrial applications. TAG composition depends on the interaction of several different groups of enzymes in the lipid biosynthesis pathway. The enzymes of the fatty acid synthase complex in the plastids of developing seeds are responsible for the biosynthesis of fatty acids up to and including oleic acid. Modifying enzymes, such as the extraplastidic A12 and A15 desaturases, elongases, hydroxylases, and epoxidases, yield polyunsaturated, very long-chain, hydroxy-, and epoxy-fatty acids, respectively. Acyltransferases insert specific acyl moieties onto the glyc-