The influence of dislocation substructure of metallic materials on ultrasonic velocity has been experimentally investigated. The test materials of pure Cu, brass (Cu-35Zn), 2.25Cr-1Mo steel, and AISI 316 with different stacking fault energy (SFE) are plastically deformed in order to generate dislocation substructures. The longitudinal wave velocit $(C_L)$ decreases as a function of tensile strain in each material. The $C_L$ of Cu-35Zn and AISI 316 decreases monotonously with tensile strain, but $C_L$ of Cu and 2.25Cr-1Mo steel shows plateau phenomena due to the stable dislocation substructure. The variation of ultrasonic velocity with the extent of dislocation damping and dislocation substructures is discussed.