Phosphorus (P) is an essential nutrient element that is necessary for plant growth and development. However, most of the P exists in insoluble form. Aspergillus aculeatus has been reported to be able to solubilize insoluble forms of P. Here, to investigate the P-solubilizing effect of A. aculeatus on the performance of perennial ryegrass (Lolium perenne) under P-deficiency stress, we created four treatment groups: control [i.e., no Ca3(PO4)2 or A. aculeatus], A. aculeatus only (F), Ca3(PO4)2 and Ca3(PO4)2 + A. aculeatus [Ca3(PO4)2 + F] treatment, and Ca3(PO4)2 at concentrations of 0 and 3 g per pot (0.5 kg substrate per pot). In our results, the liquid medium inoculated with A. aculeatus exhibited enhanced soluble P and organic acid content (tartaric acid, citric acid, and aminoacetic acid) accompanied with lower pH, compared with the noninoculated regimen. Furthermore, A. aculeatus also played a primary role in increasing the soluble P content of substrate (1 sawdust: 3 sand), the growth rate, turf quality, and photosynthetic capacity of the plant exposed to Ca3(PO4)2 + F treatment, compared with other groups. Finally, in perennial ryegrass leaves, there was a dramatic increase in the valine, serine, tyrosine, and proline contents, and a remarkable decline in the glutamic acid, succinic acid, citric acid, and fumaric acid contents in the Ca3(PO4)2 + F regimen, compared with other groups. Overall, our results suggested that A. aculeatus may play a crucial role in the process of solubilizing Ca3(PO4)2 and modulating perennial ryegrass growth under P-deficiency stress.