Smad family proteins are identified as intracellular signal mediators of the TGF-β superfamily.In this study,we identified two novel members of the Smad family,termed as AmphiSmad1/5/8 and AmphiSmad4,from Chinese amphioxus.Both AmphiSmad1/5/8 and AmphiSmad4 showed a typical domain structure of Smad proteins consisting of conserved MH1 and MH2 domains.Phylogenetic analysis placed AmphiSmad1/5/8 in the Smad1,5 and 8 subgroup of the R-Smad subfamily,and AmphiSmad4 in the Co-Smad subfamily.The spatial and temporal gene expression patterns of AmphiSmad1/5/8 and AmphiSmad4 showed that they may be involved in the embryonic development of notochord,myotome and alimentary canal,and may help to establish the specification of dorsal-ventral axis of amphioxus.Moreover,AmphiSmad1/5/8 and AmphiSmad4 showed extensive distribution in all adult tissues examined,suggesting that these two genes may play important roles in the morphogenesis of a variety of tissues especially notochord and gonad.