TC25 titanium alloy plate with thickness of 6.0mm were prepared by a three-fire hot rolling process. The effects of different annealing temperatures on the microstructure, room temperature mechanical properties and high temperature mechanical properties of TC25 titanium alloy plate were studied. The results show that with the increase of annealing temperature, the primary α grows in the equiaxed microstructure formed by hot working of TC25 titanium alloy plate in the range of 760~840℃. When the annealing temperature increases to 880~900℃, the microstructure of the TC25 titanium alloy plate was transformed from the equiaxed microstructure to the duplex microstructure. And the duplex microstructure is presented with the annealing temperature increasing to 920℃. While when the annealing temperature reaches as high as 960°C, the primary α content in the duplex microstructure is reduced, and the secondary α content is increased significantly. The TC25 titanium alloy plate with duplex microstructure has better room temperature mechanical properties and high temperature mechanical properties than the TC25 titanium alloy plate with equiaxed microstructure. Annealing at 920~960℃, TC25 titanium alloy plate with duplex microstructure, excellent room temperature mechanical properties and high temperature mechanical properties can be obtained.