To more fully understand the function of phospholipid: diacylglycerol acyltransferase (PDAT, EC 2.3.1.158) in plants we have isolated and characterized a knockout mutant of Arabidopsis thaliana L. which has a T-DNA insertion in PDAT locus At5g13640. Lipid analysis was conducted on these plants to assess the contribution of the PDAT gene to lipid composition. The fatty acid content and composition in seeds do not show significant changes in the mutant. This is a contrary situation to yeast where PDAT is a major contributor to triacylglycerol (TAG) accumulation in exponential growth phase. The results indicate that PDAT activity encoded by At5g13640 is not a major determinant of TAG synthesis in Arabidopsis seeds.