BackgroundNowadays, the genus Bryconamericus is placed in subfamily Stevardiinae within of Characidae, but not shows consistent evidence of monophyletism. The purpose of this work was to study the chromosomes of three species of Bryconamericus, aiming to add cytogenetic knowledge and contribute to the understanding of the chromosomal evolution of this genus.ResultsThe chromosomes of three species of Bryconamericus were analyzed using cytogenetic techniques. The karyotype of Bryconamericus stramineus contained 6 metacentric (m) + 10 submetacentric (sm) + 16 subtelocentric (st) + 20 acrocentric (a), the fundamental number (FN) of 84, one silver impregnated (Ag-NOR) pair, one pair bearing the 18S ribosomal DNA sites, another pair bearing the 5S rDNA sites, and a few positive C-bands. Bryconamericus turiuba had a karyotype containing 8 m + 10sm + 14st + 20a (FN = 84), one chromosome pair Ag-NOR, two pairs bearing the 18S rDNA sites, two pairs bearing the 5S rDNA sites, and a few C-band regions. Bryconamericus cf. iheringii had a karyotype containing 10 m + 14sm + 18st + 10a (FN = 94), including one pair with a secondary constriction Ag-NOR positive. In this karyotype the fluorescent in situ hybridization (FISH) showed the 18S and 5S rDNA probe in adjacent position.ConclusionsThe results obtained in this work showed different characteristics in the organization of two multigene families, indicating that distinct evolutionary forces acting on the diversity of rDNA sequences in the genome of three Bryconamericus species.

The species of genus Astyanax is widely distributed in freshwater neotropical zones. Astyanax is considered to be taxonomically confused, similar to other genera placed incertae sedis in Characidae. The cytogenetics of this genus is well characterized; species vary widely in diploid number, from 2n=36 chromosomes in Astyanax schubarti to 2n=50 for most species studied. The size, number, and position of different cytological markers vary among species and populations of Astyanax. We analyzed the karyotypes of individuals from three Astyanax species (Astyanax abramis, Astyanax altiparanae, and Astyanax eigenmanniorum) from populations not previously analyzed. We describe variations in several cytogenetic markers and the karyotypic relationships between them, specifically focusing on the characteristics of the conserved and divergent locations of the ribosomal genes. Our data are useful for establishing relationships between species and for investigating the karyotype evolution within the genus.

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The organization of microsatellites in B and sex chromosomes has been linked to chromosomal evolution in a number of animal groups. Here, the chromosomal organizations of (CA)₁₅, (GA)₁₅, (CG)₁₅, (GACA)₄, and (GATA)₈ microsatellites were examined in several Astyanax species with different diploid numbers: Astyanax mexicanus (2n = 50 + 1 B chromosome), A. altiparanae (2n = 50), A. marionae (2n = 48), A. fasciatus (2n = 46), and A. schubarti (2n = 36). The (CA)₁₅ and (GA)₁₅ microsatellites were dispersed across the chromosomes of A. altiparanae and A. fasciatus but were also observed as clusters (CA and GA for A. altiparanae, and CA for A. fasciatus). In A. marionae and A. schubarti, the (CA)₁₅ and (GA)₁₅ microsatellites were dispersed but were also observed as clustered signals and coincident with heterochromatic regions. In all 4 of these species, the (CG)₁₅ and (GACA)₄ microsatellites were dispersed across chromosomes, and the (GATA)₈ microsatellite was co-localized with 5S rDNA. In A. mexicanus, the (CA)₁₅, (GA)₁₅, (CG)₁₅, (GATA)₈, and (GACA)₄ microsatellites were weakly detected and dispersed across the chromosomes of the A complement. On the B chromosome, signals for the different microsatellites were weak, strong, absent, weak, and absent, respectively. The distribution of microsatellites and the locational relationship between microsatellites and 5S rDNA are discussed, and a possible evolutionary pathway is proposed for microsatellites in Astyanax.

Summary Characidae is one of the predominant fish groups in the Neotropical region with approximately 30 highly diversified subfamilies, probably not representing a monophyletic group. Three Characidae species from the Iguatemi River Basin (Mundo Novo, MS, Brazil) were analyzed: Astyanax bockmanni with 2nϭ50 chromosomes (6mϩ20smϩ8stϩ16a), Bryconamericus stramineus with 2nϭ52 chromosomes (6mϩ10smϩ16stϩ20a) and Piabina argentea with 2nϭ52 chromosomes (6mϩ24smϩ12stϩ10a), all the species showing no differences between male and female karyotypes. The Ag-NORs were located in only a single chromosome pair in the three species. The chromosomal data obtained may be useful for the taxonomy and phylogenetic studies in these fish groups.

The chromosomal location of 5S rRNA and U2 snRNA genes of Piabina argentea, Piabarchus stramineus and two Bryconamericus species from two different Brazilian river basins were investigated, in order to contribute to the understanding of evolutionary characteristics of these repetitive DNAs in the subfamily Stevardiinae. The diploid chromosome number was 2n = 52 for Bryconamericus cf. iheringii, Bryconamericus turiuba, Piabarchus stramineus and Piabina argentea. The 5S rDNA clusters were located on one chromosome pair in P. stramineus and B. cf. iheringii, and on two pairs in B. turiuba and P. argentea. The U2 snDNA clusters were located on the one pair in all species. Two-color FISH experiments showed that the co-localization between 5S rDNA and U2 snDNA in P. stramineus can represent a marker for this species. Thus, the present study demonstrated that the number of U2 snDNA clusters observed for the four species was conserved, but particular characteristics can be found in the genome of each species.A localização cromossômica dos genes de RNAr 5S e RNAsn U2 de Piabina argentea, Piabarchus stramineus e duas espécies de Bryconamericus provenientes de duas bacias hidrográficas foi investigada, com a intenção de contribuir com o entendimento de características evolutivas destes DNAs repetitivos na subfamília Stevardiinae. O número cromossômico diploide foi 2n = 52 para Bryconamericus cf. iheringii, Bryconamericus turiuba, Piabarchus stramineus e Piabina argentea. Os sítios de DNAr 5S foram localizados em um par cromossômico em P. stramineus e B. cf. iheringii, e em dois pares em B. turiuba e P. argentea. Os sítios de DNAsn U2 foram localizados em um par em todas as espécies. Experimentos de FISH com duas sondas mostraram que a co-localização entre os DNAr 5S e DNAsn U2 em P. stramineus pode representar um marcador para esta espécie. Portanto, o presente estudo demonstrou que o número de sítios de DNAsn U2 observado para as quatro espécies foi conservado, porém características particulares podem ser encontradas no genoma de cada espécie.Palavras-chave: DNA repetitivo, DNAr 5S, Cromossomo, Número diploide. Splicing.

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Repetitive sequences and their chromosomal locations have been widely studied in species of the Astyanax genus. However, the chromosomal organization of U2 snDNA remains largely unknown. The aims of this study were to examine the chromosomal contexts of U2 snRNA and 5S rRNA genes in Astyanax species and determine the degree of chromosome morphological similarity between species with different diploid numbers. Clusters of U2 snDNA and 5S rDNA were determined in nine species of Astyanax, including two karyomorphs of Astyanax fasciatus Cuvier, 1819. All species exhibited U2 snDNA clusters on two chromosome pairs, except Astyanax mexicanus De Filippi, 1853 (one pair). The 5S rDNA clusters were located on one chromosome pair in Astyanax altiparanae Garutti and Britski, 2000, and Astyanax marionae Eigenmann, 1911, two pairs in Astyanax abramis Jenyns, 1842, Astyanax asuncionensis Géry, 1972, Astyanax bockmanni Vari and Castro, 2007, Astyanax eigenmanniorum Cope, 1894, A. fasciatus (karyomorphs I and II), and Astyanax schubarti Britski, 1964, and four pairs in A. mexicanus. The relationships between the repetitive sequences in different species suggest that A. schubarti and A. mexicanus exhibit an unusual U2 snDNA chromosomal format as a result of events occurring in the evolutionary history of the Astyanax group.