Phosphorus (P) is one of the most important inorganic nutrients that can significantly affect plant growth and metabolism. However, 30-40% of the unavailable P in agricultural soils exists as phytate, which can not directly be absorbed by plant except for resolved by phytase. So selecting or developing new varieties that can resolve phytate-P through the modern transgenic breeding approach provided a new opportunity to improve the efficiency of phosphorus by plants. In this paper, a phytase gene (phyA), isolated from Aspergillus ficuum, was introduced into soybean by Agrobacterium-mediated transformation, and then thirty-eight soybean crosses had been made between transgenic line (JL35-phyA, male parent) and other non-transgenic varieties (female parent). PCR and RT-PCR results showed that the phyA was successfullly incorporated into soybean genome and expressed in transgenic line “JL35-phyA”. The results also showed that 427 PCR positive transgenic F1 plants from the thirty-eight cross combinations above and 377 PCR positive transgenic F2 plants from the progenies of the cross combination “JD17*L35-phyA” were obtained by the sexual hybridization transfer method.