Ultrasonic vibration-assisted con-rod fracture splitting (UV-CFS) was used to carry out the fracture experiment of 1045 quenched and tempered steel. The effect of ultrasonic vibration on the fracture properties was studied, the fracture microstructure and the evolution of dislocations near the fracture were analyzed and the microscopic mechanism was analyzed. The results show that in the case of conventional fracture splitting without amplitude, the dimple and the fracture belong to ductile fracture. With the increase in ultrasonic amplitude, the plasticity and pore deformation of the con-rod samples decrease at first and then increase; when the amplitude reaches a certain point, the load required for cracking is reduced to a minimum and the ultrasonic hardening effect is dominant, resulting in a decrease in the plasticity of the sample, a cleavage fracture, a brittle fracture, the minimum pore deformation and high cracking quality. The research results also show that with the increase in ultrasonic amplitude, the fracture dislocation density decreases at first, then increases, and dislocation entanglement and grain breakage appear, then decrease, and multiple dislocation slip trajectories appear. The changes in the dislocation density and microstructure are consistent with the above results.