Objective To enhance Pi absorption and utilization efficiency of soybean, a member ofPHT1gene family was isolated and characterized fromE.salsugineum, which was a homologous gene ofAtPHT1;4and consequently designated asEsPHT1;4. Results Quantitative real-time PCR (qRT-PCR) analysis showed that the transcript level ofEsPHT1;4significantly increased both in roots and leaves ofE.salsugineumunder Pi deficient conditions. Furthermore,EsPHT1;4was transferred to soybean cultivar "YD22" using anAgrobacterium-mediated cotyledonary-node transformation method. Overexpression ofEsPHT1;4in soybean not only promoted the increase of plant biomass and yield of transgenic plants upon low P stress, but also increased the accumulation and transportation of Pi from roots to leaves in the transgenic soybean lines. Conclusion EsPHT1;4was critical for controlling the accumulation and translocation of Pi in plants, and can be subsequently used as an effective foreign gene for the improvement of P use efficiency of crops by genetic manipulation.