By means of cloning, sequencing, and fluorescence in situ hybridization, we have determined that the EcoRI satellite DNA family is conserved in the 10 sparid species analyzed here. Its conservation, its chromosomal location at the centromere of each chromosome, and its structural features could make this satellite DNA family an important structural and/or functional element of the centromeres of these species. Monomeric units of this satellite DNA have a consensus length of 187 bp. Its sequence is characterized by a high AT content and the presence of short runs of consecutive AT base pairs. These monomeric EcoRI repeats also contain three to four copies, depending on the species, of a short sequence reflecting the repetitive duplication and subsequent divergence of an ancestral 9-bp sequence in this family. This sequence motive is conserved in some parts of the monomeric units of the different species studied at the same positions, and, precisely, surrounding the area in which the curvature of the monomeric molecule is greatest. The 9-bp sequence motive is similar to other direct-repeat sequences of the centromeric satellite DNAs of other vertebrates, including those of amphibians and mammals.