Extensive investigations on apicomplexan mitochondria, such as those of Plasmodium falciparum, revealed that ribosomal RNAs (rRNAs) are fragmented into multiple short pieces. In this study, we isolated three mitochondrial large subunit rRNA (mtLSU rRNA) fragments from the dinoflagellate Alexandrium catenella. A piece of mtLSU rRNA that possesses high sequence similarity to the P. falciparum LSU rRNA E fragment was identified in a 1.7-kbp mitochondrial (mt) DNA clone. We further confirmed that the A. catenella "E-like" fragment is indeed transcriptionally active and that the transcript could form appropriate RNA secondary structures. In addition, we identified expression of two additional rRNA fragments with sequence similarities to P. falciparum F and G fragments. Notably, the 1.7-kbp mt DNA clone contains only one of the three rRNA fragments identified in this study, suggesting that the rRNA fragments are separately encoded in the A. catenella mt genome. Given the sister relationship between apicomplexa and dinoflagellates in eukaryote phylogeny, it is most parsimonious to assume that the mt rRNA fragmentation was established prior to the separation of the two protist groups. However, current sequence data on dinoflagellate mitochondria are insufficient to reject the alternative scenario, in which the rRNA fragmentation evolved independently in apicomplexan and dinoflagellate mitochondria.