In order to explore the most suitable culture conditions of strawberry anther in vitro regeneration system,the effects of cytokine TDZ and auxin 2,4-D on callus induction, differentiation and proliferation of strawberry anther were studied, and the most suitable hormone ratio in different stages of strawberry anther culture were selected with the strawberry cultivar "Tochiotome " anther as test material. The results showed that the combination of TDZ and 2,4-D could successfully culture Strawberry Anther regeneration plants. The most suitable induction medium was MS+2,4-D 0.2 mg/L+TDZ 1.5 mg/L. The most suitable differentiation medium was MS+2,4-D 0.05 mg/L+TDZ 2.0 mg/L. The most suitable proliferation medium was MS+TDZ 0.5 mg/L+2,4-D 0.1 mg/L.