A hypothesis of phylogenetic relationships is presented for species of the South American catfish subfamily Centromochlinae (Auchenipteridae) based on parsimony analysis of 133 morphological characters in 47 potential ingroup taxa and one outgroup taxon. Of the 48 species previously considered valid in the subfamily, only one, Centromochlus steindachneri, was not evaluated in the present study. The phylogenetic analysis generated two most parsimonious trees, each with 202 steps, that support the monophyly of Centromochlinae composed of five valid genera: Glanidium, Gephyromochlus, Gelanoglanis, Centromochlus and Tatia. Although those five genera form a clade sister to the monotypic Pseudotatia, we exclude Pseudotatia from Centromochlinae. The parsimony analysis placed Glanidium (six species) as the sister group to all other species of Centromochlinae. Gephyromochlus contained a single species, Gephyromochlus leopardus, that is sister to the clade Gelanoglanis (five species) + Centromochlus (eight species). Based on the new taxonomic arrangement herein, Balroglanis Grant 2015 and Sauronglanis Grant 2015 are placed in the synonymy of Centromochlus. Tatia is the most diverse genus within Centromochlinae, with twenty-six species in three monophyletic clades. The species composition of Tatia differs from that of Calegari et al. (2019). Duringlanis Grant 2015 and Ferrarissoaresia Grant 2015 are considered synonyms of Tatia. All generic taxa within Centromochlinae are diagnosed based on synapomorphic morphological characters. An identification key for Centromochlinae is provided.