Karyotypic diversity in a population of Bryconamericus aff. iheringii (Characidae).

Bryconamericus comprises 56 species distributed into three groups, on the basis of the position and shape of the maxillary teeth: B. exodon, B. microcephalus and B. iheringii groups. Few cytogenetic data are available for this genus, but the diploid number of 52 chromosomes is quite common, although the karyotypic variability is extensive. This study aimed to characterize a population of B. aff. iheringii and thus contribute more cytogenetic information and better understanding of the structure and karyotypic evolution of this genus. We found 6 cytotypes with different NOR patterns: cytotype I showed a karyotype formula of 12m+10sm+16st+14a (FN = 90) and single NORs; cytotype II with 18m+14sm+10st+10a (FN = 94) and cytotype III with 20m+18sm+4st+10a (FN = 94), showing both single and multiple NORs; cytotype IV with 20m+14sm+12st+6a (NF = 98), cytotype V with 22m+18sm+8st+4a (FN = 100) and cytotype VI with 18m+24sm+6st+4a (FN = 100), all with multiple NORs. Cytotype I is the most different in relation to FN and NOR pattern, and can be regarded as belonging to another species of the genus Bryconamericus, living in sympatry in Três Bocas Stream. The remaining cytotypes may have been generated by crosses between them and by pericentric inversions. Meiotic cells were also analyzed and showed that despite the high karyotypic variability, chromosome pairing occurred normally. The great variability found in B. aff. iheringii may be related to a high degree of polymorphism; nevertheless, the possibility of occurrence of more than one species in this location is not ruled out, demonstrating the need for conservation of the Três Bocas Stream.

Physical mapping of 18S rDNA and heterochromatin in species of family Lygaeidae (Hemiptera: Heteroptera).

Analyses conducted using repetitive DNAs have contributed to better understanding the chromosome structure and evolution of several species of insects. There are few data on the organization, localization, and evolutionary behavior of repetitive DNA in the family Lygaeidae, especially in Brazilian species. To elucidate the physical mapping and evolutionary events that involve these sequences, we cytogenetically analyzed three species of Lygaeidae and found 2n (♂) = 18 (16 + XY) for Oncopeltus femoralis; 2n (♂) = 14 (12 + XY) for Ochrimnus sagax; and 2n (♂) = 12 (10 + XY) for Lygaeus peruvianus. Each species showed different quantities of heterochromatin, which also showed variation in their molecular composition by fluorochrome staining. Amplification of the 18S rDNA generated a fragment of approximately 787 bp. The alignment of the consensus sequence with sequences from other species of Heteroptera deposited in the GenBank revealed a similarity of 98% with small differences. Fluorescent in situ hybridization with the 18S rDNA fragment revealed that this ribosomal gene was located in 1 autosomal pair at different positions in the three species. No cytogenetic data are available for these Brazilian species. The basal number and the possible chromosomal changes that occurred among the different species, as well as the evolution of these DNA sequences, are discussed.

Sex chromosome differentiation in Belostoma (Insecta: Heteroptera: Belostomatidae).

Belostoma, a genus of the family Belostomatidae, includes species of great ecological importance as biocontrol agents. Few species of these species have been the subject of cytogenetic analyses. Karyotypic evolution in this genus involves agmatoploidy and simploidy; there are also different sex chromosome systems. We examined two Belostoma species (B. dilatatum and B. candidulum) collected from the Paranapanema River Basin (Brazil). Mitotic and meiotic analysis revealed 2n(♂) = 26 + X1X2X3Y for B. dilatatum and 2n(♂) = 14 + XY for B. candidulum; both karyotypes have holokinetic chromosomes. Differences in heterochromatin distribution were also observed between the species, besides variation in the localization of CMA3⁺/DAPI⁻ blocks. The existence of different types of sex chromosome systems in these species was confirmed based on arrangements of the chromosomes in different meiotic stages. We identified a new sex system in B. dilatatum, and make the first cytogenetic report on B. candidulum.

Population divergence and peculiar karyoevolutionary trends in the loricariid fish Hypostomus aff. unae from northeastern Brazil.

Loricariidae (Siluriformes, Hypostominae) is one of the most diverse catfish families. In spite of the wide distribution of loricariids in South America, cytogenetic reports are available for only a few species, mostly from southern and southeastern Brazil. We made the first chromosomal analysis of Hypostomus aff. unae from the Contas River basin in northeastern Brazil. Four populations isolated by short distances but from distinct landscapes were studied based on conventional staining, C-banding, argyrophilic nucleolar organizer regions (Ag-NOR), CMA(3)/DAPI fluorochrome staining, and fluorescent in situ hybridization with 18S rDNA probes. Although sharing the same diploid number (2n = 76) and NOR locations, each population presented exclusive karyotype formulae and specific patterns of heterochromatic and AT-rich regions. The derived karyotypes of H. aff. unae (2n >54; high number of acrocentrics bearing AT-rich interstitial heterochromatin) indicated a divergent karyoevolution, mostly driven by centric fissions, pericentric inversions and particular heterochromatin dispersion models. This finding of distinct evolutionary units in H. aff. unae will be useful for understanding the natural history of loricariids from relatively unexplored coastal basins in South America.

Karyotype diversity of four species of the incertae sedis group (Characidae) from different hydrographic basins: analysis of AgNORs, CMA3 and 18S rDNA.

A large number of genera in the tropical fish family Characidae are incertae sedis. Cytogenetic analysis was made of four of these species: Astyanax eigenmanniorum, Deuterodon stigmaturus, Hyphessobrycon luetkenii, and H. anisitsi, collected from various hydrographic basins: hydrographic system from Laguna dos Patos/RS, Tramandaí basin/RS and Tibagi River basin/PR. The first two species were collected in their type locality in the State of Rio Grande do Sul. The 2n = 48 karyotype was observed only in A. eigenmanniorum, while the other species had 2n = 50 chromosomes, with different karyotypic formulas. There was weak heterochromatin staining in the pericentromeric region of A. eigenmanniorum, D. stigmaturus and H. luetkenni chromosomes. In H. anisitsi, heterochromatin appeared to be more abundant and distributed in the pericentromeric and terminal regions of the chromosomes; three pairs showed more evident heterochromatic blocks. There were multiple Ag-NORs in all populations, visualized by FISH with an 18S rDNA probe. While D. stigmaturus and H. luetkenii had conserved AgNOR, CMA3 and 18S rDNA sites, the other two species showed intra- and interindividual variation at these sites. The karyotype variability was high, as is common in this group of fish. Different species arising from isolated hydrographic basins maintain an elevated level of karyotype differentiation, mainly with respect to chromosome structure, heterochromatin distribution and rDNA localization. This is the first report with cytogenetic data for D. stigmaturus and H. luetkenii.

Heterochromatin heterogeneity in Hypostomus prope unae (Steindachner, 1878) (Siluriformes, Loricariidae)from Northeastern Brazil.

Cytogenetic analyses using C-banding and chromosomal digestion by several restriction enzymes were carried out in four populations (named A, B, C and D) of Hypostomus prope unae (Loricariidae, Hypostominae) from Contas river basin, northeastern Brazil. These populations share 2n=76 and single NORs on the second metacentric pair but exclusive karyotype forms for each locality. Populations A and B presented conspicuous terminal and interstitial heterochromatic blocks on most of acrocentric chromosomes and equivalent to NORs with differences in both position and bearing pair. Population D showed evident marks at interstitial regions and interspersed with nucleolar region while population C presented interstitial and terminal heterochromatin segments, non-coincident with NORs. The banding pattern after digestion with the endonucleases Alu I, Bam HI, Hae III and Dde I revealed a remarkable heterogeneity within heterochromatin, allowing the identification of distinctive clusters of repeated DNA in the studied populations, besides specific patterns along euchromatic regions. The analysis using restriction enzymes has proved to be highly informative, characterizing population differences and peculiarities in the genome organization of Hypostomus prope unae.

Occurrence of natural triploidy in Rhamdia quelen (Siluriformes, Heptapteridae).

Five specimens of Rhamdia quelen collected from the Lindóia Stream, PR, Brazil, were cytogenetically analyzed. The diploid chromosome number found was 58, including 30 metacentric, 16 submetacentric, 10 subtelocentric, and 2 acrocentric chromosomes. Supernumerary or B chromosomes, frequently observed in this fish group, were not detected. One of the individuals was triploid, with 3n = 87. A silver-stained nucleolar organizer region was found on a pair of submetacentric chromosomes of the diploid specimens, and on three chromosomes of the triploid individual, confirming triploidy. Treatment with fluorochrome chromomycin A(3) revealed fluorescent bands coincident with those of the silver-stained nucleolar organizer region, in both diploid and triploid individuals, showing that this is a GC-rich region. Heterochromatin distribution was visualized by the C-banding technique, mainly in the terminal chromosome regions of the individuals and was also observed in the pericentromeric regions of some chromosomes and at both telomeres.

Cytogenetic analysis of the 18S, 5S rDNA and B chromosome of Iheringichthys labrosus (Lütken, 1874) (Siluriformes, Pimelodidae).

Cytogenetic analyses of the location of 18S and 5S ribosomal DNAs, and the base composition of B chromosomes of Iheringichthys labrosus from Tibagi River, Paraná, Brazil, are provided. AgNORs were observed in the terminal position on the long arm of a subtelocentric chromosome pair. CMA3-positive staining was observed in some chromosomes, which besides being associated with NORs, were all DAPI-negative. Chromosome B showed a strong fluorescence with CMA3. The concomitant use of 18S and 5S rDNA probes using the FISH technique revealed 18S ribosomal cistrons in a pair of subtelocentric chromosomes, on the long arm in the terminal position, coinciding with the AgNOR. The 5S sites were found in another subtelocentric pair, on the long arm in the interstitial region, near the centromere. The findings of the present study suggest that, although there are some more conserved cytogenetic characteristics, populations of I. labrosus may show their own characteristics.

Cytogenetic analysis of the 18S, 5S rDNA and B chromosome of Iheringichthys labrosus (Lütken, 1874) (Siluriformes, Pimelodidae) / Análise Citogenética de DNAr 18S, 5S e cromossomo B de Iheringichthys labrosus (Lütken, 1874) (Siluriformes, Pimelodidae)

Cytogenetic analyses of the location of 18S and 5S ribosomal DNAs, and the base composition of B chromosomes of Iheringichthys labrosus from Tibagi River, Paraná, Brazil, are provided. AgNORs were observed in the terminal position on the long arm of a subtelocentric chromosome pair. CMA3-positive staining was observed in some chromosomes, which besides being associated with NORs, were all DAPI-negative. Chromosome B showed a strong fluorescence with CMA3. The concomitant use of 18S and 5S rDNA probes using the FISH technique revealed 18S ribosomal cistrons in a pair of subtelocentric chromosomes, on the long arm in the terminal position, coinciding with the AgNOR. The 5S sites were found in another subtelocentric pair, on the long arm in the interstitial region, near the centromere. The findings of the present study suggest that, although there are some more conserved cytogenetic characteristics, populations of I. labrosus may show their own characteristics.

Foram realizadas análises citogenéticas em Iheringichthys labrosus do Rio Tibagi, Paraná, Brasil com a localização cromossômica dos DNAs ribossômicos 18S e 5S e a composição de bases de seus cromossomos B. As AgNORs foram observadas em posição terminal, no braço longo de um par de cromossomos subtelocêntricos. Marcações CMA3 positivas foram observadas em alguns cromossomos e associadas com as RONs. Porém, todas estas marcações apresentaram-se DAPI negativas. O cromossomo B mostrou-se fortemente fluorescente com CMA3. O uso concomitante das sondas de DNAr 18S e 5S, através da técnica de FISH, revelou os cístrons ribossômicos em um par de cromossomos subtelocêntricos, em posição terminal do braço longo, coincidindo com a AgNOR. Os sítios 5S foram observados em outro par subtelocêntrico, em posição intersticial do braço longo, próximo ao centrômero. Os resultados observados no presente estudo sugerem que, embora existam algumas características citogenéticas mais conservadas, as populações de I. labrosus podem mostrar suas próprias características.

Differentiation of Y chromosome in the X(1)X(1)X(2)X(2)/X(1)X(2)Y sex chromosome system of Hoplias malabaricus (Characiformes, Erythrinidae).

Six populations of Hoplias malabaricus from the Lower Paranapanema River were studied and showed 2n = 40 with metacentric/submetacentric chromosomes for females and 2n = 39 with m/sm chromosomes for males, thereby revealing a multiple sex chromosome system of the X(1)X(1)X(2)X(2)/X(1)X(2)Y type. The Y chromosome was the largest metacentric chromosome in all karyotypes. The heterochromatic blocks were located in a pericentromeric region of all pairs and in telomeric regions in some other pairs. A conspicuous pericentromeric C-band was detected in a single pair, equivalent to the X(1) chromosome. No markers on X(2) chromosomes were detected, but they could be identified by their size. The females displayed the same distribution of C and chromomycin A(3) bands and 18S rDNA sites among the populations. However, the males exhibited differences in both number and position of Giemsa C-bands on the Y chromosome, which were also evident after Chromomycin A(3) banding and FISH with the 18S rDNA probe. These cytogenetic tools allowed for the identification of possible mechanisms involved in the differentiation and evolution of the Y chromosome.

Cytotaxonomy in distinct populations of Hoplias aff. malabaricus (Characiformes, Erythrinidae) from lower Paranapanema River basin.

Cytogenetic and morphometric analyses were carried out in Hoplias aff. malabaricus specimens from six distinct populations from the lower Paranapanema River basin, located between the states of Paraná and São Paulo, Brazil. Measurements were also taken from a specimen collected in Surinam. In a population from a fish farm station at Universidade do Norte do Parana (EPUNOPAR), two sympatric cytotypes (2n = 40 and 2n = 42 chromosomes) are found. A population from a fish farm station at Universidade Estadual de Londrina (EPUEL) shows 2n = 42 meta-submetacentric chromosomes for males and females with a simple sex chromosome system of XX/XY type. Populations from the Vermelho and Rancho Alegre Rivers, Três Bocas Stream and Paranapanema River have 2n = 39 chromosomes in males and 2n = 40 chromosomes in females, showing a multiple sex chromosome system of X(1)X(1)X(2)X(2)/X(1)X(2)Y type. Twenty morphological variables were studied. These measurements were used for an analysis of the canonical variables and standard analysis of proportional measurements. The most variable measurements among the specimens are the maxilla length (MXL) and the pre-dorsal distance (PDD). Analysis of canonical variables indicates three distinct groups in the first canonical axis formed by: (1) Três Bocas Stream, (2) Rancho Alegre + Vermelho River + EPUNOPAR and (3) EPUEL + Paranapanema River. This axis retained 79.4% of information from the original matrix. Analysis of morphometrics reveals differences among populations from the Paranapanema River basin and between these and the specimen from Surinam. The morphometric and cytogenetic differences among the studied populations suggest a species complex.

Karyotype description of two species of Hypostomus (Siluriformes, Loricariidae) of the Planalto da Bodoquena, Brazil.

Hypostomus sp 3-Córrego Salobrinha NUP 4247 and Hypostomus sp 2-Rio Perdido NUP 4249, collected in the Planalto da Bodoquena, Paraguay River basin, Brazil, were characterized cytogenetically. Hypostomus sp 3-Córrego Salobrinha showed two modal numbers. This polymorphism consists of the presence of two extrachromosomes. It was not possible to define the diploid number in four specimens, where cell lineages had 2n = 83 and 2n = 84 chromosomes in one individual, and 2n = 82, 2n = 83 and 2n = 84 chromosomes in the others. These results reveal the existence of a genetic mosaic due to the occurrence of one or two extrachromosomes in this species. Hypostomus sp 2-Rio Perdido NUP 4249 showed a 2n = 84, FN = 106 with size heteromorphism in one pair of chromosomes stained with AgNO3. In both species, C banding showed a pattern of heterochromatin distribution with a few small bands in the centromeric and pericentromeric regions coinciding with chromomycin A3 staining. Until now, the major diploid number for the genus Hypostomus was 2n = 80, but the species studied here had chromosomes that in creased this number and the variation for this genus. Our results are also the first cytogenetic data on Hypostomus from the Paraguay River basin.

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.

Analyses of nucleolus organizer regions and heterochromatin of Pimelodus maculatus (Pisces, Pimelodidae).

Eighteen specimens of Pimelodus maculatus collected from Tibagi River (Sertaneja, PR, Brazil) were analyzed cytogenetically. The diploid number of 56 chromosomes was observed and karyotype was 20 M + 20 SM + 10 ST + 6 A with fundamental number (FN) of 106. Results of analyses from the nucleolus organizer regions (NORs), obtained by AgNO3, CMA3 and C-band staining showed marking in a terminal position on the long arm of a pair of subtelocentric chromosomes. The restriction enzyme AluI produced a linear differentiation similar to C-banding.