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Chromaphyosemion killifishes are a karyotypically highly diverse group of small, sexually dimorphic fishes living in rainforest rivulets in tropical West and Central Africa. In the present study, we used various chromosome banding and staining techniques to analyse the karyotypes of 13 populations representing seven described species (Chromaphyosemion loennbergii, Chromaphyosemion punctulatum, Chromaphyosemion splendopleure, Chromaphyosemion volcanum, Chromaphyosemion malumbresi, Chromaphyosemion melanogaster, Chromaphyosemion bitaeniatum) and two undescribed forms (Chromaphyosemion cf. lugens, Chromaphyosemion sp. Rio Muni GEMHS00/41). Diploid chromosome numbers (2 n) and the number of chromosome arms (NF) ranged from 2 n = 24 in C. malumbresi to 2 n = 40 in C. bitaeniatum and from NF = 40 in C. volcanum and C. cf. lugens to NF = 54 in one population of C. loennbergii. A tentative XX/XY sex chromosome system was revealed in C. loennbergii, C. melanogaster, C. malumbresi, and Chromaphyosemion sp. Rio Muni GEMHS00/41. Mapping cytogenetic data for all described Chromaphyosemion species onto a recently published mitochondrial DNA phylogeny revealed a complex pattern of chromosomal evolution with several independent reductions of 2 n and independent modifications of NF and nucleolus organizer region phenotypes. Together with the results of preliminary crossing and mate choice experiments, the cytogenetic and molecular phylogenetic data suggest that, contrary to previous hypotheses, chromosomal rearrangements are probably not the most important and certainly not the only factor driving speciation in Chromaphyosemion killifishes. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 94, 143–153.
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