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Adaptations can endow species with complex traits that enable them to occupy new niches. To adapt to the arboreal lifestyle, treefrogs have evolved a suite of complex traits for climbing and gliding; to date, however, the molecular mechanisms underpinning these adaptations are unknown. Here, based on two de novo-assembled Asian treefrog genomes, genes related to limb development showed evidence of accelerated evolution. Furthermore, the PPL gene that regulates the keratin cytoskeleton showed conservative substitution within the genus Rhacophorus. These genetic changes contribute to limb and toe-pad morphogenesis, providing fundamental adaptations for climbing and grasping. Behavioral and morphological comparisons clarified the gliding function of interdigital webbing and the different developmental patterns between fully and weakly webbed feet. Time-ordered gene coexpression network analysis of gliding Rhacophorus kio and nongliding Rhacophorus dugritei tadpoles revealed specific coexpression patterns for fully webbed feet, which involved the Wnt signaling and vascular remodeling pathways. These findings highlight the molecular basis of phenotypes that facilitate expansion into new niches and provide insights into the importance of local adaptation in shaping phenotypes and locomotion patterns. [ABSTRACT FROM AUTHOR]