A tomato lipase gene homologous to Arabidopsis DAD1 (lipase homologous to DAD1; LeLID1) was cloned and characterized. The corresponding transcript increased rapidly during germination of the seeds and reached a maximum level at four days after germination. Thereafter, it decreased rapidly. Little expression could be found in flowers or fruits. Immunoblot analyses showed that the gene products could be found in the cotyledons and hypocotyls, but not in the roots. In the cotyledons most LeLID1 could be recovered in a soluble fraction. The recombinant LeLID1 protein showed maximum lipase activity at pH 8.0. It showed high activity against triacylglycerols (TAGs) with long acyl chains, but little activity with phosphatidylcholine or monogalactosyldiacylglycerol. TAGs composed of short acyl chains could not be a substrate for the enzyme. A possible involvement of LeLID1 in fat mobilization during seed germination is discussed.