We applied metabolic control analysis to the Kennedy pathway for triacylglycerol formation in tissue cultures from the important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). When microsomal fractions were incubated at 30 °C rather than 20 °C, there was an increase in triacylglycerol labelling. This increase was accompanied by a build up of diacylglycerol (DAG) radioactivity in olive but not in oil palm, suggesting that the activity of DAG acyltransferase (DAGAT) was becoming limiting in olive. We used 2-bromooctanoate as a specific inhibitor of DAGAT and showed that the enzyme had a flux control coefficient under the experimental conditions of 0.74 in olive but only 0.12 in oil palm. These data revealed important differences in the regulation of lipid biosynthesis in cultures from different plants and suggest that changes in the endogenous activity of DAGAT is unlikely to affect oil accumulation in oil palm crops.