Chromosome Evolution in the Genus Partamona (Apidae: Meliponini), with Comments on B Chromosome Origin.

The genus Partamona includes 33 species of stingless bees, of which 11 were studied cytogenetically. The main goal of this study was to propose a hypothesis about chromosomal evolution in Partamona by combining molecular and cytogenetic data. Cytogenetic analyses were performed on 3 Partamona species. In addition, the molecular phylogeny included mitochondrial sequences of 11 species. Although the diploid number was constant within the genus, 2n = 34, B chromosomes were reported in 7 species. Cytogenetic data showed karyotypic variations related to chromosome morphology and the amount and distribution of heterochromatin and repetitive DNA. The molecular phylogenetic reconstruction corroborated the monophyly of the genus and separated the 2 clades (A and B). This separation was also observed in the cytogenetic data, in which species within each clade shared most of the cytogenetic characteristics. Furthermore, our data suggested that the B chromosome in the genus Partamona likely originated from a common ancestor of the species that have it in clade B and, through interspecific hybridization, it appeared only in Partamona rustica from clade A. Based on the above, Partamona is an interesting genus for further investigations using molecular mapping of B chromosomes as well as for broadening phylogenetic data.

The B Chromosomes of Prochilodus lineatus (Teleostei, Characiformes) Are Highly Enriched in Satellite DNAs.

B or supernumerary chromosomes are dispensable elements that are widely present in numerous eukaryotes. Due to their non-recombining nature, there is an evident tendency for repetitive DNA accumulation in these elements. Thus, satellite DNA plays an important role in the evolution and diversification of B chromosomes and can provide clues regarding their origin. The characiform Prochilodus lineatus was one of the first discovered fish species bearing B chromosomes, with all populations analyzed so far showing one to nine micro-B chromosomes and exhibiting at least three morphological variants (Ba, Bsm, and Bm). To date, a single satellite DNA is known to be located on the B chromosomes of this species, but no information regarding the differentiation of the proposed B-types is available. Here, we characterized the satellitome of P. lineatus and mapped 35 satellite DNAs against the chromosomes of P. lineatus, of which six were equally located on all B-types and this indicates a similar genomic content. In addition, we describe, for the first time, an entire population without B chromosomes.

A genomic glimpse of B chromosomes in cichlids.

BACKGROUND: B chromosomes (Bs) are extra karyotype elements in addition to A chromosomes and are found in all major eukaryotic taxa. Among hundreds of investigated species, cichlid fishes have emerged as an interesting group of model and have contributed to unravel the complex biology of B chromosome. OBJECTIVE: We review the current state of knowledge on B chromosome investigation in cichlid fish and discuss the recent genomic advances over gene and sequences hunting on Bs and their impact on the current concept of B chromosomes. RESULTS: The Bs of cichlids have been under the subject of classical cytogenetics and high scale DNA, RNA and epigenetics analysis and a list of B chromosome genes and functional sequences has been generated. B chromosomes of cichlids are restricted to females in some species and are enriched with genes, relics of genes, transposable elements and sequences transcribing for many significant biological functions. Diverse potentially functional sequences have been described in the B chromosome of cichlids and could influence important biological characteristics as well seems to affect transcription and epigenetic modifications of the whole genome. CONCLUSION: One of the most enigmatic characteristics of Bs in cichlids is their genic content related to cell cycle and chromosome structure, and their influence over sex rates. The relationship of Bs with cell cycle and sex determination looks like to be connected with the drive of the Bs during cell divisions.

First chromosomal analysis in Deinopidae (Araneae) reveals sex Chromosome system X1X2X3X4, B chromosomes and polymorphism for centric fusion

Here we present the first cytogentic study concerning Deinopidae and their controversial phylogenetic position. This study karyologically analyzed one population of Deinopis biaculeata Simon, 1906 and five populations of Deinopis plurituberculata Mello-Leitão, 1925. The majority of specimens of D. plurituberculata exhibited 2n♂ = 40 and 2n♀ = 44 telocentric chromosomes (however some of them showed B chromosomes, belongs to Aquidauana and Botucatu population). The Deinopis biaculeata and D. plurituberculata meiosis of males showed 18 autosomal bivalents + X1X2X3X4, n = 22 and n = 18, a rare sex chromosome system (SCS) in spiders. Some individuals of D. plurituberculata from the Campo Grande population exhibited 2n♂ = 39 and 2n♀ = 43, with a metacentric chromosome (heterozygotes for centric fusion). The D. plurituberculata males with the rearrangement exhibit diplotenes with 16 autosomal bivalents + 1 autosomal trivalent + X1X2X3X4 and metaphases II with n = 22 (18 telocentric autosomes + X1X2X3X4), n = 21 (16 telocentric autosomes + a metacentric autosome + X1X2X3X4), n = 18 (18 telocentric autosomes) and n = 17 (16 telocentric autosomes + a metacentric autosome). The Ag-NORs (silver impregnation) are terminally located in a pair, coinciding with secondary constriction, which is the most common configuration for Araneae. The relatively high diploid number in Deinopis corroborates phylogenies that place it in a basal position among Entelegynes, in the UDOH grade (Uloboridae, Deinopidae, Oecobiidae and Hersiliidae). Centric fusion in only one population of D. plurituberculata suggests low dispersion capacity of this species and an absence of homozygotes for fusion suggests their low viability or a need to increase the population sampling of D. plurituberculata exhibiting the rearrangement. B chromosomes were detected in D. plurituberculata, with interpopulacional, intrapopulacional and intraindividual numerical variation, with cells presenting 0 - 3 and 0 - 6 B chromosomes in populations of Aquidauana and Botucatu, respectively.

Differential Expression of Genes Related to Sexual Determination Can Modify the Reproductive Cycle of Astyanax scabripinnis (Characiformes: Characidae) in B Chromosome Carrier Individuals.

The species complex Astyanax scabripinnis is one of the most studied with respect to origin, distribution, and frequency of B chromosomes, and is considered a model organism for evolutionary studies. Research using population inferences about the occurrence and frequency of the B chromosome shows seasonal variation between sexes, which is associated with the presence of this supernumerary element. We hypothesized that the B chromosome could influence the sex ratio of these animals. Based on this assumption, the present work aimed to investigate if differences exist among levels of gene expression with qRT-PCR of the amh (associated with testicular differentiation) and foxl2a (associated with ovarian differentiation) genes between B-carrier and non-B-carrier individuals. The results showed that for the amh gene, the difference in expression between animals with B chromosomes was not accentuated compared to that in animals without this chromosome. Expression of foxl2a in B-carrier females, however, was reduced by 73.56% compared to females that lacked the B chromosome. Males had no difference in expression of the amh and foxl2a genes between carriers and non-carriers of the B chromosome. Results indicate that the presence of B chromosomes is correlated with the differential expression of sex-associated genes. An analysis of these results integrated with data from other studies on the reproductive cycle in the same species reveals that this difference in expression may be expanding the reproductive cycle of the species.

Genetic variability in a population of Astyanax scabripinnis: recent bottleneck and the possible influence of individuals with B chromosomes

Access the genetic variability of endangered and isolated populations has become an important conservation tool. Astyanax scabripinnis is a well-known fish model for genetic studies, forming very isolated populations in headwaters. Besides that, this species frequently presents supernumerary chromosomes, which elevates the interest on genetic studies. Genetic diversity of an Astyanax scabripinnispopulation from the Atlantic Forest (Serra da Mantiqueira region, Brazil) was assessed with microsatellite markers for the first time. Since microsatellite markers are not described for this species, we tested markers described for a related species for transferability to A. scabripinnis. Six polymorphic loci were sufficiently reliable for population genetic analysis. We found that this population passed through a recent bottleneck because of the presence of an excess of heterozygotes, low allelic diversity, heterozygosity excess, and small effective population size. Individuals with and without B chromosomes were previously identified in this population and our study found private alleles in the individuals without B chromosomes. Furthermore, when individuals without B chromosomes were removed from the analysis, the population did not present heterozygosity excess, suggesting that the bottleneck event was driven by individuals with B chromosomes. Our results provide an insight into the value of microsatellite markers as molecular tools and is the first genetic study using molecular data of A. scabripinnis from this area.

Cestrum strigilatum (Ruiz & Pavón, 1799) B chromosome shares repetitive DNA sequences with A chromosomes of different Cestrum (Linnaeus, 1753) species.

Species of Cestrum (Linnaeus, 1753) have shown large diversity in the accumulation and distribution of repetitive DNA families, and B chromosomes have been described in seven species. Some types of repetitive DNA were identified in A and B chromosomes in species of this plant group, such as AT-rich SSR, 35S and 5S rDNA, C-Giemsa and C-CMA/DAPI bands and retrotransposons. To increase our understanding of the relationships of A and B chromosomes, the B of C. strigilatum Ruiz & Pavón, 1799 was microdissected, amplified and hybridized in situ against chromosomes of this species, and in six other species of this genus. FISH signals were observed in whole the B of C. strigilatum, including stretches of A chromosomes, as well as in some A chromosomes of all tested species. A strong FISH signal was seen adjacent to the 5S rDNA in the proximal region of pair 8 of all species and, due to this, we have searched for 5S rDNA fragments in the microdissected B chromosome. PCR and sequencing data evidenced 5S rDNA deletion along evolutionary pathways of the B of C. strigilatum. Although A and B chromosomes displayed redundancy in the repetitive DNA families in different species, the B of C. strigilatum seemed to differ from those Bs of other Cestrum species by the loss of rDNA fractions. A possible origin of Bs in Cestrum was discussed.

B chromosomes: from cytogenetics to systems biology.

Though hundreds to thousands of reports have described the distribution of B chromosomes among diverse eukaryote groups, a comprehensive theory of their biological role has not yet clearly emerged. B chromosomes are classically understood as a sea of repetitive DNA sequences that are poor in genes and are maintained by a parasitic-drive mechanism during cell division. Recent developments in high-throughput DNA/RNA analyses have increased the resolution of B chromosome biology beyond those of classical and molecular cytogenetic methods; B chromosomes contain many transcriptionally active sequences, including genes, and can modulate the activity of autosomal genes. Furthermore, the most recent knowledge obtained from omics analyses, which is associated with a systemic view, has demonstrated that B chromosomes can influence cell biology in a complex way, possibly favoring their own maintenance and perpetuation.

Microstructural chromosome reorganization in the genus Trichomycterus (Siluriformes: Trichomycteridae)

Trichomycterus is a specious fish genus within Trichomycterinae and displays remarkable karyotype diversity. However, knowledge about their genomic structure and location of repetitive sequence is still limited. In order to better understand the karyotype diversification, we analyzed nine species of Trichomycterus using classical and molecular cytogenetic techniques. Results revealed a conserved diploid chromosome number of 2n=54 chromosomes in all analyzed species, although remarkable differences on the constitutive heterochromatin distribution were observed. In addition, while the 18S rDNA showed a conserved distribution pattern, the 5S rDNA sites showed a quite diverse location considering the analyzed species. Remarkably, both ribosomal genes were co-located in all species, except in T . iheringi , suggesting that co-localization is probably an ancestral condition in Trichomycterus . Finally, three analyzed species showed heterochromatic B chromosomes, reinforcing the intense genomic reorganization occurring in Trichomycterus . Our results showed that chromosomal variations are not restricted to differences in karyotype formula as previously proposed, but also to modifications on the microstructural level of resolution.(AU)

Trichomycterus é um especioso gênero dentro de Trichomycterinae e exibe marcante diversidade cariotípica. No entanto, o conhecimento sobre sua estrutura genômica e localização de seqüências repetitivas ainda é restrita. Para um melhor conhecimento sobre a sua diversificação cariotípica, nós analisamos nove especies de Trichomycterus usando técnicas de citogenética clássica e molecular. Os resultados revelaram um conservado número diploide de 2n = 54 cromossomos em todas as espécies analisadas, embora diferentes marcações na distribuição da heterocromatina constitutiva tenham sido observadas. Além disso, enquanto o DNAr 18S mostrou um padrão de distribuição conservado, os sítios de DNAr 5S mostraram uma localização bastante diversa, considerando as espécies analisadas. Ambos os genes ribossomais foram co-localizados em todas as espécies, exceto em T. iheringi , sugerindo que a co-localização é provavelmente uma condição ancestral em Trichomycterus . Finalmente, três espécies analisadas mostraram cromossomos B heterocromáticos, reforçando uma intensa reoganização genômica ocorrendo em Trichomycterus . Nossos resultados mostraram que variações cromossômicas não estão restritas à diferenças na fórmula cariotípica, como proposto anteriormente, mas também às alterações a níveis de resolução estrutural.(AU)

Microstructural chromosome reorganization in the genus Trichomycterus (Siluriformes: Trichomycteridae)

Trichomycterus is a specious fish genus within Trichomycterinae and displays remarkable karyotype diversity. However, knowledge about their genomic structure and location of repetitive sequence is still limited. In order to better understand the karyotype diversification, we analyzed nine species of Trichomycterus using classical and molecular cytogenetic techniques. Results revealed a conserved diploid chromosome number of 2n=54 chromosomes in all analyzed species, although remarkable differences on the constitutive heterochromatin distribution were observed. In addition, while the 18S rDNA showed a conserved distribution pattern, the 5S rDNA sites showed a quite diverse location considering the analyzed species. Remarkably, both ribosomal genes were co-located in all species, except in T . iheringi , suggesting that co-localization is probably an ancestral condition in Trichomycterus . Finally, three analyzed species showed heterochromatic B chromosomes, reinforcing the intense genomic reorganization occurring in Trichomycterus . Our results showed that chromosomal variations are not restricted to differences in karyotype formula as previously proposed, but also to modifications on the microstructural level of resolution.

Trichomycterus é um especioso gênero dentro de Trichomycterinae e exibe marcante diversidade cariotípica. No entanto, o conhecimento sobre sua estrutura genômica e localização de seqüências repetitivas ainda é restrita. Para um melhor conhecimento sobre a sua diversificação cariotípica, nós analisamos nove especies de Trichomycterus usando técnicas de citogenética clássica e molecular. Os resultados revelaram um conservado número diploide de 2n = 54 cromossomos em todas as espécies analisadas, embora diferentes marcações na distribuição da heterocromatina constitutiva tenham sido observadas. Além disso, enquanto o DNAr 18S mostrou um padrão de distribuição conservado, os sítios de DNAr 5S mostraram uma localização bastante diversa, considerando as espécies analisadas. Ambos os genes ribossomais foram co-localizados em todas as espécies, exceto em T. iheringi , sugerindo que a co-localização é provavelmente uma condição ancestral em Trichomycterus . Finalmente, três espécies analisadas mostraram cromossomos B heterocromáticos, reforçando uma intensa reoganização genômica ocorrendo em Trichomycterus . Nossos resultados mostraram que variações cromossômicas não estão restritas à diferenças na fórmula cariotípica, como proposto anteriormente, mas também às alterações a níveis de resolução estrutural.

First description of B chromosomes in the Hyphessobrycon (Characiformes, Characidae) genus: a hypothesis for the extra element of Hyphessobryconeques Steindachner, 1882.

The Hyphessobrycon are allocated in the incertae sedis group of the Characidae family, one of the genera with more species of the group. The chromosomes of some species of Hyphessobrycon are known, and the diploid number most common for genus is 2n = 50 chromosomes. The aims of this study were to examine the karyotype macrostructure in the Hyphessobryconeques Steindachner, 1882, and show a new origin hypothesis for B chromosomes. The diploid number observed for Hyphessobryconeques was 2n = 52 chromosomes, and a karyotype formulae of 12m + 18sm + 8st + 14a, with FN (fundamental number) = 90 for both sexes. Only two females showed one B chromosome. The heterochromatin was observed mainly on centromeric regions, and in the long arm of the B chromosome. In this paper, the relationship of the B chromosome of Hyphessobryconeques with an occasional chromosome rearrangement was discussed.

B chromosomes in the species Prochilodusargenteus (Characiformes, Prochilodontidae): morphologicalidentity and dispersion.

B chromosomes have attracted the attention of Neotropical fish cytogeneticists in recent years, both for their remarkable occurrence in this group and also because of the interest in studies of the genetic structure and role played in the genome of these organisms. The aim of this study was to report the first occurrence of supernumerary chromosomes in Prochilodusargenteus (Agassiz, 1829), this being the fifth carrier species among thirteen within the genus Prochilodus (Agassiz, 1829). The extra elements identified in this species are small sized heterochromatic chromosomes characterized by a low mitotic instability index, being very similar to other supernumerary chromosomes described in the species of the genus Prochilodus. Morphology, structure and dispersion of the supernumerary genomic elements which occur in species of this genus are discussed aiming to better understand aspects involved the origin of supernumerary chromosomes and the differentiation process and relationships among species of this family.

Cytogenetic analysis of B chromosomes in one population of the fish Moenkhausia sanctaefilomenae (Steindachner, 1907) (Teleostei, Characiformes).

The aim of this study was to characterize cytogenetically one population of the fish Moenkhausia sanctaefilomenae (Steindachner, 1907), with emphasis on the analysis of B chromosomes. The nucleolar activity in the B microchromosomes was characterized, and an analysis of mitotic instability of these microchromosomes was accomplished. The results showed a diploid chromosome number of 50 chromosomes. In all individuals, we observed the presence of B microchromosomes with intra- and inter-individual variability. The analysis of the nucleolus organizing regions (NORs) by silver nitrate staining demonstrated multiple NORs. We observed active sites of ribosomal DNA in the B microchromosomes, with a frequency of 20% in the analyzed cells, which shows gene activity in these chromosomal elements. The analysis of constitutive heterochromatin patterns showed that the B microchromosomes are heterochromatic or euchromatic, which demonstrates differentiation of DNA composition between these genomic elements. The calculation of the mitotic instability index implied that B chromosomes in this species might be in a final stage of instability.

New occurrence of B chromosomes in Partamonahelleri (Friese, 1900) (Hymenoptera, Meliponini).

Cytogenetic analyses of the stingless bee Partamona helleri collected in the state of Bahia, Northeast Brazil revealed the chromosome numbers n = 18 in the haploid males and 2n = 35 in the diploid females. All karyotypes displayed one large acrocentric B chromosome, which differs from the minute B chromosomes previously described in the populations from southeastern Brazil. Giemsa staining, C-banding and DAPI/CMA(3) fluorochrome staining also revealed a remarkable interpopulational divergence regarding both the regular karyotype and the B chromosomes. The B chromosomes found in the samples from Jequié, Bahia, were entirely heterochromatic, while those found in Cravolândia, Bahia, displayed a euchromatic portion at the telomeric end of the long arm. CMA (3) labeling sites varied from seven to eight between the two localities in Bahia, due to the presence of an extra GC-rich block in the karyotype of the samples from Jequié. This is the first report of a large B chromosome in P. helleri and reveals the occurrence of a geographic differentiation within this species.

New occurrence of B chromosomes in Partamona helleri (Friese, 1900) (Hymenoptera, Meliponini)

Cytogenetic analyses of the stingless bee Partamona helleri collected in the state of Bahia, Northeast Brazil revealed the chromosome numbers n = 18 in the haploid males and 2n = 35 in the diploid females. All karyotypes displayed one large acrocentric B chromosome, which differs from the minute B chromosomes previously described in the populations from southeastern Brazil. Giemsa staining, C-banding and DAPI/CMA3 fluorochrome staining also revealed a remarkable interpopulational divergence regarding both the regular karyotype and the B chromosomes. The B chromosomes found in the samples from Jequié, Bahia, were entirely heterochromatic, while those found in Cravolândia, Bahia, displayed a euchromatic portion at the telomeric end of the long arm. CMA3 labeling sites varied from seven to eight between the two localities in Bahia, due to the presence of an extra GC-rich block in the karyotype of the samples from Jequié. This is the first report of a large B chromosome in P. helleri and reveals the occurrence of a geographic differentiation within this species.

Identification and description of distinct B chromosomes in Cyphocharax modestus (Characiformes, Curimatidae)

Cytogenetic analyses were performed in Cyphocharax modestus, collected at Paranapanema River and Tietê River (São Paulo State, Brazil). A karyotype with 2n = 54 chromosomes was observed in the animals from both Brazilian freshwater river systems. One to four B chromosomes were also detected in individuals from the Paranapanema River, which represents the probable first report of more than a single supernumerary element in a species of the Curimatidae group. C-banding revealed centromeric and telomeric heterochromatin blocks in several chromosomes of the normal karyotype complement of C. modestus. Moreover, while some B chromosomes were characterized by the complete absence of C-bands, others were totally heterochromatic. Although there was a prevalence of B chromosomes in males of C. modestus, at least one supernumerary element was found in males and/or females of several other populations of the species, which suggests that the presence of these chromosomes seems to represent a general trait of C. modestus. A possible origin of the described B chromosomes may be related to the occurrence of a chromosome non-disjunction followed by the loss of euchromatic segments, an event that should have occurred in chromosomes that present conspicuous centromeric heterochromatic blocks and even in chromosomes that lack C-bands in this region, resulting in small supernumerary elements.

Occurrence of B chromosomes in Cyphocharax modestus (Fernández-Yépez, 1948) and Steindachnerina insculpta (Fernández-Yépez, 1948)(Characiformes, Curimatidae) from the Tibagi River basin (Paraná State, Brazil)

We cytogenetically analyzed 18 individuals of Cyphocharax modestus, and 41 individuals of Steindachnerina insculpta from the Tibagi River basin (Paraná State, Brazil). All the specimens had 2n = 54 chromosomes, all meta-submetacentric (M-SM). The presence of 1 or 2 B chromosomes was seen in some individuals of C. modestus of the two sampled populations, showing inter- and intra individual variation. In S. insculpta, B chromosomes were also observed but only in one population, also showing inter- and intra individual variation. C-banding was used to show that the identified B chromosomes were totally heterochromatic in the two examined species.

Foram analisados, citogeneticamente, 18 indivíduos de Cyphocharax modestus e 41 indivíduos de Steindachnerina insculpta da bacia do rio Tibagi (Paraná, Brasil). Todos os espécimes apresentaram 2n = 54 cromossomos, todos do tipo meta-submetacêntrico (M-SM). A presença de 1 ou 2 cromossomos B foi detectada em alguns indivíduos de C. modestus de duas populações amostradas, evidenciando uma variação inter e intraindividual. Em S. insculpta, também foram observados cromossomos B, embora em somente uma população, mostrando também uma variação inter e intraindividual. Bandamento C foi usado para mostrar que os cromossomos B identificados apresentaram-se totalmente heterocromáticos nas duas espécies examinadas.

Occurrence of B chromosomes in Cyphocharax modestus (Fernández-Yépez, 1948) and Steindachnerina insculpta (Fernández-Yépez, 1948)(Characiformes, Curimatidae) from the Tibagi River basin (Paraná State, Brazil)

We cytogenetically analyzed 18 individuals of Cyphocharax modestus, and 41 individuals of Steindachnerina insculpta from the Tibagi River basin (Paraná State, Brazil). All the specimens had 2n = 54 chromosomes, all meta-submetacentric (M-SM). The presence of 1 or 2 B chromosomes was seen in some individuals of C. modestus of the two sampled populations, showing inter- and intra individual variation. In S. insculpta, B chromosomes were also observed but only in one population, also showing inter- and intra individual variation. C-banding was used to show that the identified B chromosomes were totally heterochromatic in the two examined species.

Foram analisados, citogeneticamente, 18 indivíduos de Cyphocharax modestus e 41 indivíduos de Steindachnerina insculpta da bacia do rio Tibagi (Paraná, Brasil). Todos os espécimes apresentaram 2n = 54 cromossomos, todos do tipo meta-submetacêntrico (M-SM). A presença de 1 ou 2 cromossomos B foi detectada em alguns indivíduos de C. modestus de duas populações amostradas, evidenciando uma variação inter e intraindividual. Em S. insculpta, também foram observados cromossomos B, embora em somente uma população, mostrando também uma variação inter e intraindividual. Bandamento C foi usado para mostrar que os cromossomos B identificados apresentaram-se totalmente heterocromáticos nas duas espécies examinadas.

Occurrence of B chromosomes in Cyphocharax modestus (Fernández-Yépez, 1948) and Steindachnerina insculpta (Fernández-Yépez, 1948)(Characiformes, Curimatidae) from the Tibagi River basin (Paraná State, Brazil)

We cytogenetically analyzed 18 individuals of Cyphocharax modestus, and 41 individuals of Steindachnerina insculpta from the Tibagi River basin (Paraná State, Brazil). All the specimens had 2n = 54 chromosomes, all meta-submetacentric (M-SM). The presence of 1 or 2 B chromosomes was seen in some individuals of C. modestus of the two sampled populations, showing inter- and intra individual variation. In S. insculpta, B chromosomes were also observed but only in one population, also showing inter- and intra individual variation. C-banding was used to show that the identified B chromosomes were totally heterochromatic in the two examined species.

Foram analisados, citogeneticamente, 18 indivíduos de Cyphocharax modestus e 41 indivíduos de Steindachnerina insculpta da bacia do rio Tibagi (Paraná, Brasil). Todos os espécimes apresentaram 2n = 54 cromossomos, todos do tipo meta-submetacêntrico (M-SM). A presença de 1 ou 2 cromossomos B foi detectada em alguns indivíduos de C. modestus de duas populações amostradas, evidenciando uma variação inter e intraindividual. Em S. insculpta, também foram observados cromossomos B, embora em somente uma população, mostrando também uma variação inter e intraindividual. Bandamento C foi usado para mostrar que os cromossomos B identificados apresentaram-se totalmente heterocromáticos nas duas espécies examinadas.