Lettuce (Lactuca sativa L.) is known to contain water-soluble substances that are biologically active. Aqueous or methanol extracts and residues from leaves of lettuce plants were assayed to determine their allelopathic effects, and the causative allelochemicals from fractions were quantified by means of HPLC analysis and bioassayed. Extracts from oven-dried leaf samples were more phytotoxic than those from freeze-dried samples. Leaf extracts of 40 g L$^{-1}$ were completely inhibitory on root growth of alfalfa (Medicago sativa L.), while root growths of barley (Hordeum vulgare L.) and soybean (Glycine max L.) were less sensitive. Early seedling growth of both alfalfa and barnyard grass (Echinochloa crus-galli) was significantly reduced by methanol leaf extracts. The major allelopathic substances analyzed by HPLC were coumarin, trans-cinnamic acid, o-coumaric acid, p-coumaric acid and chlorogenic acid. Of them p-coumaric acid was found as the greatest amount (8.9 mg 100 g$^{-1}$ ) in the EtOAc fraction; only coumarin was found in all the fractions. Hexane and EtOAc fractions of L. sativa reduced alfalfa root growth more than did BuOH and water fractions. These results suggest that lettuce had potent herbicidal activity, and that its activity differed depending on type and amount of causative compounds by fraction.