Tithonia diversifolia (Hermsl.) A. Gray is a perennial invasive plant and spreads quickly in the invasive areas. The extracts of T. diversifolia were found to be toxic to several crop plant species such as rice, maize, sorghum, lettuce and cowpea, and several putative allelopathic substances were identified. However, there is limited information available for the effects of T. diversifolia on wild plants including weed plant species. We investigated the allelopathic potential of T. diversifolia extracts on weed plants, and searched for phytotoxic substances with allelopathic activity. An aqueous methanol extract of T. diversifolia leaves inhibited the growth of weed plants, Lolium multiflorum Lam., Phleum pretense L., Echinochloa crus-galli (L.) Beauv. The extract was then purified by several chromatographic runs and a phytotoxic substance with allelopathic activity was isolated and identified by spectral analysis as tagitinin C. The substance inhibited the growth of Lolium multiflorum, Phleum pratense and Echinochloa crus-galli at concentrations greater than 0.1 - 0.3 mM. The present results suggest that T. diversifolia may possess allelopathic potential on weed plants and tagitinin C may be responsible for the allelopathic effects of T. diversifolia. The allelopathic potential of T. diversifolia may contribute to its invasive characteristics.