The oxidized Mo-50Re alloys in air at 573 K and 873 K for various times were investigated by X-Ray diffraction and positron annihilation lifetime spectroscopy. The results indicated that main orthogonal phase MoO3 together with small amount of monoclinic phase Mo8O23 were formed on on the surface of the specimens, and the oxide film of specimens oxidized at 873 K were much thicker than that of specimens oxidized at 573 K. Meanwhile, when Mo-50Re specimens oxidized in air at 873K, the defect’s size within interface layer of the specimens was larger, and the oxide film on the surface of the specimens contained much more defects. Faster oxidation process were observed occured at 873K, which was likely due to the formation of larger-size interfacial defects.