The banana, a typical climacteric fruit, undergoes a postharvest ripening process followed by a burst in ethylene production that signals the beginning of the climacteric period. Postharvest ripening plays an important role in improving the quality of the fruit as well as limiting its shelf life. To investigate the role of glutamate decarboxylase (GAD) in climacteric ethylene biosynthesis and fruit ripening in postharvest banana, a GAD gene was isolated from banana, designated MuGAD. Coincidently with climacteric ethylene production, MuGAD expression as well as the expression of the genes encoding the Musa 1-aminocyclopropane-1-carboxylate synthase (MaACS1) and Musa 1-aminocyclopropane-1-carboxylate oxidase (MaACO1) greatly increased during natural ripening and in ethylene-treated banana. Moreover, ethylene biosynthesis, ripening progress, and MuGAD, MaACS1, and MaACO1 expression were enhanced by exogenous ethylene application and inhibited by 1-methylcyclopropene (1-MCP). Taken together, our results suggested that MuGAD is involved in the fruit ripening process in postharvest banana.