Ectomycorrhizal fungi (ECMF) are capable of converting phosphate in the insoluble state of the soil into effective phosphorus that can be directly absorbed and used by plants, a mechanism that is essential for promoting plant growth and development.In this study, we utilized Populus davidiana × P. bolleana tissue culture seedlings as the test material. Inoculation was performed with two ectomycorrhizal fungi: Suillus luteus and Paxillus involutus. The aim was to study the activation and uptake of soil phosphorus by ectomycorrhizal. Our results showed that inoculation of both S. luteus and P. involutus significantly increased seedling height, ground diameter, biomass and tissue phosphorus content of Populus davidiana × P. bolleana seedlings. Concurrently, it reduced soil pH, increased soil acid phosphatase activity and soil effective phosphorus content. All effects on mycorrhizosphere were better than those on hyphosphere. Therefore, this study concludes that the introduction of ectomycorrhizal acidifies the soil, increases the activity of soil acid phosphatase, promotes the decomposition of phosphate in the insoluble state of the soil, increases the content of effective phosphorus in the soil, thus promoting the uptake of phosphorus by plants, and plays a role in the growth of Populus davidiana × P. bolleana. Among them, mycorrhizal affect the soil to a greater extent than their hypha.