Chemokines are a superfamily of small cytokines and characterized based on their ability to induce directional migration of cells along a concentration gradient by binding to chemokine receptors, which have important roles in immunology and development. Due to the numerous and diverse members, systematic identifications of chemokine superfamily genes are difficult in many species. To that end, a comprehensive analysis of BLAST and scripting language was conducted to systematically identify and characterize chemokine system in grass carp (Ctenopharyngodon idella). Our results showed that C. idella chemokine superfamily consists of 81 chemokines and 37 receptors, in which, most genes possess typical structural features of the chemokine superfamily. Phylogenetic analyses confirmed the existence of three chemokine subfamilies (CC, CXC and XC) in C. idella and revealed their homologous relationships with other species. Chemokine receptors are transmembrane receptors and contains CCR, CXCR, XCR and ACKR subfamilies. mRNA expression analyses of chemokine superfamily genes indicated that many members are sustainably expressed in multiple tissues before and after grass carp reovirus (GCRV) or Aeromonas hydrophila infection, which provides in vivo evidence for the response patterns after viral or bacterial infection. Meanwhile, this study also explored the evolution of chemokine system from arthropod to higher vertebrates and then investigated the changes in gene number/diversification, gene organization and encoded proteins during vertebrate evolution. These results will serve the further functional and evolutional studies on chemokine superfamily.