Phosphorus (P) is normally considered as the limited nutrient for shallow freshwater lakes and can potentially trigger eutrophication on account of high concentrations. Due to the various transportation and transformation processes, P source apportionment and management in lake ecosystems have become more and more difficult. Combining with sequential extraction of P fractions and mineralogical analysis, the isotopic compositions of oxygen in phosphate (δ18O P) of resin-extractable P from the different samples including soil, estuary sediments, pond sediments, and lake sediments in the Shijiuhu Lake catchment, China, were investigated. The results showed that δ18O P values ranged from +15.23 to +21.92‰ in agricultural soil, +16.53 to +24.10‰ in estuary sediments, +18.90 to +20.90‰ in pond sediments, and +17.42 to +19.70‰ in lake sediments. Isotopic signatures indicated that chemical fertilizers with heavier δ18O P values (+20.70 to +26.50‰) were the predominant contributors of P in the soil. The river transportation together with Fe/Al–P desorption on anaerobic condition simultaneously stimulated the enrichment of P in the lake sediments, even though the biotic activity regulated the isotope values moving toward the equilibrium. Eroded soil was the important source of P in lake and pond sediments via drainage and runoff, and conserved the source isotope signal in the samples. Stronger biotic activity in the aquatic environments dragged δ18O P values toward the equilibrium. However, conspicuous off-equilibrium isotope signature suggested the terrestrial sources in the aquatic ecosystems. The calculation of two end-member linear mixing models suggested that soils also predominantly controlled the P occurrence in the lake sediments with contribution higher than 80%, indicating that decreasing inputs from the agricultural activities is important in P reduction on catchment scale. Generally, δ18O P from different sources can provide indirect and important evidences for the identification and management of P sources in the lake catchment. [Display omitted] • Different research areas displayed the various ranges of δ18O P values. • Fertilizer with heavy δ18O P values were the predominant contributor of P in the soils. • Conspicuous off-equilibrium isotope signature suggested the terrestrial sources. • δ18O P supplied useful evidence for the identification and management of P source. [ABSTRACT FROM AUTHOR]