Cell extracts of hyphae of Fusarium oxysporum f. sp. lycopersici rapidly transferred the methyl group of S-[methyl-3 H]adenosyl-L-methionine (Ado-Met) to endogenous phosphatidylethanolamine (PE). About 80% of the radioactivity incorporated into the phospholipid fraction was found in phosphatidylcholine (PC) while the rest of the radioactivity was present in the intermediates monomethylphosphatidylethanolamine (MePE) and dimethylphosphatidylethanolamine (DiMePE). The phospholipid methylating system had a pH optimum of 8.5, a Km of 30 μm for Ado-Met, and a Vmax of 10 nmol/h per milligram protein. The specific activity of the methylating system was highest in early log phase and lowest in the late log phase of growth.The activity of the cell-free methylating system was reduced by incubation at temperatures above 25 °C, and at 37 °C about 50% of the initial methylating activity remained after incubation for 15 min. In contrast, the activity of the in vivo methylation system almost doubled when the incubation temperature was raised from 25 to 37 °C.