ABSTRACT. Cytogenetic analyses were performed in four species of the Hypostominae subfamily, three from Hypostomus (Hypostomini) genus and Rhinelepis aspera (Rhinelepini). Three populations of Hypostomus ancistroides were analyzed, which had 2n=68 chromosomes, but presented different karyotype formulas. Hypostomus regani and H. strigaticeps, both from Ivaí river, showed 2n=72 chromosomes with two distinct cytotypes. In turn, R. aspera of the upper Paraná river basin presented 2n=54 chromosome. Multiple Nucleolar Organizer Regions (NORs) have been evidenced by silver nitrate staining in species of Hypostomus and single NOR in R. aspera. The observed variation in the chromosome number and the marked variability in karyotype formulas and NORs reveal a certain amount of karyotype variation in the genus Hypostomus suggesting the probable existence of cryptic species with independent chromosome traits. Therefore, our data can be of great value in discriminating species and understanding their chromosomal evolution.Keywords: chromosomal evolution, cryptic species, Neotropical fishes, NOR. Descrição cariotípica de possíveis novas espécies do complexo Hypostomus ancistroides e outros HypostominaeRESUMO. Foram analisadas três populações de peixes identificadas como Hypostomus ancistroides, as quais apresentaram 2n=68 cromossomos, com distintas fórmulas cariotípicas. Hypostomus regani e H. strigaticeps, ambas do rio Ivaí, apresentaram 2n=72 cromossomos com citótipos distintos. Rinelepis aspera da bacia do alto rio Paraná apresentou 2n=54 cromossomos. Nucleolar Organizer Regions (NORs) múltiplas foram evidenciadas por nitrato de Prata para as espécies do gênero Hypostomus e NOR simples para R. aspera. A variação de número cromossômico observada, como também a acentuada variação nas fórmulas cariotípicas e nas NORs, são discutidas, sugerindo a existência de possíveis espécies crípticas com caracteres cromossômicos independentes. Portanto, nossos dados podem ser de grande valia na discriminação das espécies e no entendimento de sua evolução cromossômica.
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