Chromosomes of two Brycon species (B. lundii and B. microlepis) and Salminus hilarii were analyzed. Based on constitutive heterochromatin distribution patterns, karyotypic relationships within and between Bryconinae and Salmininae were examined. A monophyletic origin for the genus Brycon, comprising at least two chromosome synapomorphies (presence of two large submetacentric bearing paracentromeric and telomeric heterochromatin), is suggested. Moreover, Bryconinae and Salmininae may represent a monophyletic unit among Characidae, as they share several chromosome features.