Revealing how aquatic organisms respond to dam impacts is essential for river biomonitoring and management. Traditional examinations of dam impacts on macroinvertebrate assemblages were frequently conducted within single rivers (i.e., between upstream vs. downstream locations) and based on taxonomic identities but have rarely been expanded to level of entire basins (i.e., between dammed vs. undammed rivers) and from a functional trait perspective. Here, we evaluated the effects of dams on macroinvertebrate assemblages at both the within-river and basin scales using functional traits in two comparable tropical tributaries of the Lancang-Mekong River. At different scales, maximum body size, functional feeding groups (FFG), voltinism and occurrence in drift respond significantly to dam impact. Armoring categories varied significantly between downstream sites and upstream sites, and oviposition behavior, habits and adult life span significantly differed between rivers. The key traits at the within-river scale resembled to those at the between-river scale, suggesting that within-river trait variation could further shape functional trait structure at the basin scale in dammed rivers. Furthermore, water nutrients and habitat quality induced by dams showed the most important role in shaping trait structure, although trait-environment relationships varied between the two different scales. In addition, the trait-environment relationships were stronger in the dry season than in the wet season, suggesting a more important role of environmental filtering processes in the dry season compared with the wet season. This study highlights the utility of the trait-based approach to diagnose the effects of damming and emphasizes the importance of spatial scale to examine dam impacts in riverine systems.