Olive (Olea europaea L.) callus cultures were incubated with [2-14C]ethanolamine and [Me-14C]choline in order to study phospholipid synthesis. Radioactivity from [Me-14C]choline was shown to be incorporated into the phosphatidylcholine via the CDP-base pathway. [2-14C]Ethanolamine was primarily incorporated into phosphatidylethanolamine, but significant radio-activity was also detected in phosphatidylcholine, indicating the operation of a methylation route. Incubation with [2-14C]ethanolamine indicated that phosphatidylcholine and phosphatidylethanolamine incorporated radiolabel over a similar time course. This led us to investigate the possibility that phosphatidylcholine was being synthesized by a methylation pathway distinct from the direct methylation of phosphatidylethanolamine. There was extensive incorporation of [2-14C]ethanolamine into different components of the aqueous phase of the incubations, within which phospho-base derivatives of ethanolamine were prominent. These intermediates were identified and provided evidence for the operation of an alternative methylation pathway via phosphodimethylethanolamine for the biosynthesis of phosphatidylcholine in olives.

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