Ultrasonic assisted laser welding (ULW) is a newly developed welding method. In this paper, a new kind of ultrasonic assisted laser welding method, which is named trailing ultrasonic assisted laser welding (T-ULW) is proposed. First, the welding temperature fields as well as the weld pool temperature evolution in trailing ultrasonic assisted laser welding (T-ULW) and conventional laser welding (LW) are numerically simulated, and the shapes of the molten pools during T-ULW and LW are observed by a high-speed camera monitoring system. Both the simulation and monitoring results show that ultrasonic can reduce the temperature gradient of the weld pool and prolong the solidification time. Then, such three kinds of welding methods as T-ULW, common fixed-position ultrasonic assisted laser welding (F-ULW) and LW are used to weld SUS301 stainless steel sheet with a thickness of 0.6 mm respectively, and their performances are comparatively investigated by analyzing the experimentally-obtained macro morphologies, microstructures of welded joints. Macro morphology analysis results show that ultrasonic vibration not only elongates the molten pool, but also changes the cross-sectional shapes of the welds in such a way that decreases the top width and increases the bottom width. Microstructure analysis results indicate that the fusion zone of the welds obtained by T-ULW is uniformly distributed mainly by the refined equiaxed grains. Subsequently, the crystal refinement mechanisms of the welds in ultrasonic-assisted laser welds of SUS301 stainless steel are theoretically analyzed and experimentally verified, which show that due to ultrasonic disturbance, the temperature gradient of the molten pool is decreased, a stress field is generated in the welds and a so-called ultrasonic cavitation effect is produced in the molten pool. Such three mechanisms play very important roles in refining crystals in different ways that the decreased temperature gradient provides a good nucleation condition for the growth of equiaxed dendrite, and both the generated stress and ultrasonic cavitation cause the grain boundary to fracture, thereby form more new grain boundary, which finally lead to form fine and uniform grains in the welds. Finally, the mechanical properties of the joints are measured, and the results demonstrated that ultrasonic assisted laser welding especially T-ULW enables not only to increase the tensile strength of the welds, but also to improve the ductility of the welds.