We examined the phylogenetic relationships among members of the Hybopsis amblops species group in a total evidence cladistic analysis using DNA sequence data from regions of three mitochondrial genes and previously published morphological data. Molecular characters consisted of 120 informative sites from 811 bases of the 12S, 82 informative sites from 571 bases of the 16S gene, and 214 informative sites from 519 bases from the ND2 protein-coding gene. Morphological characters consisted of 14 transformation series previously published by R. L. Mayden in 1989. Two most parsimonious trees were found with the topologies: (Hybopsis hypsinotus ((Hybopsis lineapunctata, Hybopsis rubrifrons) (Hybopsis amblops (Hybopsis amnis (Hybopsis winchelli, Hybopsis sp.))))) and (Hybopsis hypsinotus (Hybopsis amblops ((Hybopsis lineapunctata, Hybopsis rubrifrons) (Hybopsis amnis (Hybopsis winchelli, Hybopsis sp.))))).