The allelopathic activity of Mexican sunflower [Tithonia diversifolia (Hemsl.) A. Gray] in soil under natural field conditions and the effect of water stress on the growth and allelopathic activity of this plant were investigated. Seed germination, shoot growth and root growth of tested plant species in soil collected from a field where Mexican sunflower had been grown for 5 years were less than those in soil from an area without the plant. Growth of young leaves, mature leaves, senescent leaves, stem and roots of Mexican sunflower was reduced with decrease in soil moisture level. The allelopathic activity of water extracts (per dry weight of starting material) from each part of the plants grown at low soil moisture levels was greater than that of the water extracts from the same part of the plants grown at high soil moisture levels. The allelopathic activity was found in the soil from the pots previously planted with Mexican sunflower to a similar extent (per plant) at different soil moisture levels. These results suggested that, in the field, under water stress conditions, the growth of Mexican sunflower was reduced but the plants contained a greater amount of allelopathic substance(s) per dry weight than in the absence of water stress. It was considered that allelopathic activity of Mexican sunflower in soil was maintained to a similar extent under various soil moisture conditions in natural fields.