Chitin deacetylases (CDAs) are extracellular-modifying enzymes that deacetylate chitin to produce chitosan. Insect CDAs have been divided into five groups based on phylogenetic analysis. We previously reported the functional importance of group I CDAs, TcCDA1 and TcCDA2, from Tribolium castaneum in molting, morphology of cuticle as well as in movement of legs. However, ultrastructure in the cuticle after RNAi for these genes have not been investigated. In this study, we further analyzed precise localization of these proteins and ultrastructural changes/defects of the cuticles in TcCDA1- and TcCDA2-deficient insects. Loss of function of TcCDA1 and TcCDA2 causes disorganized horizontal laminae and vertical pore canals in both rigid (e.g. elytron and ventral body wall) and soft (e.g. hindwing and dorsal body wall) adult cuticles. These results indicate that TcCDA1 and TcCDA2 are critical for development and formation of the beetle cuticles