Vibrio parahaemolyticus is a common foodborne pathogen whose motility， biofilm formation and virulence factor production are regulated by AI-2/LuxS quorum sensing system. In this study， V. parahaemolyticus with high yield of AI-2 signaling molecules was selected from nine strains Vibrio， and effects of exogenous AI-2 signaling molecules on biofilm， virulence factors and motility of V. parahaemolyticus were analyzed. The results showed that a strain of V. parahaemolyticus VP-4 with high production of AI-2 signaling molecules was obtained， and the promotion rates of biofilm， gelatinase， lecithin enzyme， amylase， motility， and hemolysis by adding exogenous AI-2 signaling molecules with different volume fractions (1%-7%) were 33.02%-67.87%， 1.67%-7.26%， 3.25%-48.15%， 1.21%-12.17%， 4.92%-35.98%， 4.15%-50.60%， and 2.06%-15.23%， respectively. Meanwhile， scanning electron microscope observation showed that the addition of exogenous AI-2 signaling molecules caused biofilm of V. parahaemolyticus to accumulate， increased in thickness and compact structure. The above results indicated that AI-2 signaling molecules could promote the formation of V. parahaemolyticus biofilm and the release of virulence factors， as well as enhance its motility. This study could provide a novel target for controlling the pathogenicity of Vibrio parahaemolyticus in seafood.