To maintain grassland productivity and limit resource depletion, scarce mineral P (phosphorus) fertilizers must be replaced by alternative P sources. The effect of these amendments on plant growth may depend on physicochemical soil parameters, in particular pH. The objective of this study was to investigate the effect of soil pH on biomass production, P use efficiency, and soil P forms after P amendment application (100 mg kg −1 P) using poultry manure compost (PM), rock phosphate (RP), and their combination (PMRP). We performed a growth chamber experiment with ryegrass plants (Lolium perenne) grown on two soil types with contrasting pH under controlled conditions for 7 weeks. Chemical P fractions, biomass production, and P concentrations were measured to calculate plant uptake and P use efficiency. We found a strong synergistic effect on the available soil P, while antagonistic effects were observed for ryegrass production and P uptake. We conclude that although the combination of PM and RP has positive effects in terms of soil P availability, the combined effects of the mixture must be taken into account and further evaluated for different soil types and grassland plants to maximize synergistic effects and to minimize antagonistic ones.