In the presence of S-adenosyl-l-methionine, enzyme preparations from carrot (Daucus carota L.) cell suspension cultures specifically catalyze the methylation of caffeoyl-CoA to feruloyl-CoA. Caffeate, 5-O-caffeoyl-d-quinate, 5-O-caffeoylshikimate and methyl caffeate do not act as substrates. The caffeoyl-CoA 3-O-methyltransferase activity is stimulated in vitro by 1 mM Mg2+. The reaction is inhibited by S-adenosyl-l-homocysteine but not by coenzyme A. Extractable enzyme activity is stimulated by cell wall preparations from Phytophthora megasperma f. sp. glycinea indicating that feruloyl-CoA is an intermediate in the defence reaction of the cells.