White Spot Syndrome Virus (WSSV) is one of the most common and distrous diseases for shrimp. In this study, we show that the protein VP292 that is a envelop protein of WSVV interacts with F0ATP synthase b-chain from Litopenaeus vannamei using far-western blot, ELISA, and indirect immunofluorescence analysis. Tissue distribution analysis of F0ATP synthase b-chain showed that it's transcription can be detected in muscle, hepatopancreas, intestine, hemocytes, lymphoid, and gills. Cellular localization of F0ATP synthase b-chain in shrimp hemocytes showed that F0ATP synthase b-chain was primarily located in the cytoplasm of hemocytes. The transcription levels of F0ATP synthase b-chain were significantly upregulated in intestine, hepatopancreas, hemocytes, and gills of WSSV-infected shrimp at 12 h after infection. Far-western, ELISA, and indirect immunefluorescence assay indicated that F0ATP synthase b-chain interacted with VP292. In the in vivo neutralization experiment, F0ATP synthase b-chain attained 18% protection rate of the shrimp challenged by WSSV. To the best of our knowledge, this is the first report to show that F0ATP synthase b-chain is involved in WSSV infection.