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Prosopis juliflora (Sw.) DC. Is highly recognized for its invasive character and its detrimental effects on different plant species across the world. In this study, a field survey was performed to assess the impact of P. juliflora on the surrounding native plants, followed by the effect of different concentrations of its aqueous leaf extract on seed germination and seedling growth of selected Qatari flora. Our field observations indicated a high prevalence of new recruitments of P. juliflora than other native-associated plants in the study site. Germination percentage and/or seed radicle length of the test species: Acacia tortilis (Forssk.) Hayne, Prosopis cineraria (L.) Druce, Sueda aegyptica (Hasselq.) Aellen, Halopeplis perfoliata (Forssk.) Bunge ex Schweinf. & Asch, and P. juliflora were significantly (p ≤ 0.05) reduced by the extracts at different concentration levels. Moreover, the germination rate of A. tortilis was not affected; however, the seed germination of P. cineraria was significantly affected at concentrations of 6 and 8 mg mL−1. The seed radicle lengths at 2 mg mL−1 were significantly reduced for both plant species. The seed germination and radicle length of S. aegyptica and H. perfoliata were significantly decreased at 2 mg mL−1 of extract of P. juliflora. Furthermore, the height and biomass of the seedlings of Aeluropus lagopoides (L.) Trin. ex Thwaites, Caroxylon imbricatum (Forssk.), and Tetraena qatarensis (Hadidi) Beier & Thulin were significantly reduced by the aqueous extracts at 6 and 8 mg mL−1. The aqueous extract reduced the seed germination and seedling growth at 6 and 8 mg mL−1, indicating an autotoxic effect. Such allelopathic behavior of P. juliflora designates the potential investigation of using its allelochemicals as naturally based herbicides in agrosystems. [ABSTRACT FROM AUTHOR]