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Genetics and Molecular Biology, Volume: 31, Número: 1 Suplemento, Publicado: 2008
  • Editorial

    Foresti, Fausto; Galetti Junior, Pedro Manoel
  • Comparative cytogenetics among species of the Astyanax scabripinnis complex: evolutionary and biogeographical inferences Fish Cytogenetics

    Vicari, Marcelo Ricardo; Noleto, Rafael Bueno; Artoni, Roberto Ferreira; Moreira-Filho, Orlando; Bertollo, Luiz Antonio Carlos

    Resumo em Inglês:

    Karyotype data are presented for distinct species of the genus Astyanax from four rivers belonging to three different hydrographic basins of the State of Paraná, Brazil (Verde River - Tibagi basin, Açungui River - Ribeira basin, and Santo Antônio and Jaguariaíva Rivers - Jaguariaíva basin). Three karyotypic forms were identified, here denominated karyotype A (2n = 50 chromosomes, with 8m+18sm+10st+14a, and thirteen 18S rDNA sites); karyotype B (2n = 50 chromosomes, with 8m+18sm+10st+1f4a, and four 18S rDNA sites); and karyotype C (2n = 48 chromosomes, with 10m+16sm+10st+12a, and eight 18S rDNA sites). The pattern of constitutive heterochromatin was similar among the three karyotypic forms, with few differences. The 5S rDNA corresponds to a synapomorphic character regarding its number and chromosomal localization. The karyotypic form A occurs in the distribution center of the type locality of A. paranae, in the proximities of the town of Castro (Tibagi basin), and may have reached the headwaters of the Ribeira River by the breakdown of geographical barriers. The karyotypic forms B and C are sympatric and syntopic, occurring solely in the Jaguariaíva River basin. Our hypothesis is that the karyotypic form A corresponds to the species A. paranae and forms B and C correspond to other species of the A. scabripinnis complex.
  • First cytogenetic characterization of the sub-arctic marine fish Mallotus villosus (Müller, 1776), Osmeriformes, Osmeridae Fish Cytogenetics

    Ghigliotti, Laura; Mazzei, Federico; Ozouf-Costaz, Catherine; Christiansen, Jørgen S.; Fevolden, Svein-Erik; Pisano, Eva

    Resumo em Inglês:

    The capelin, Mallotus villosus (Osmeriformes, Osmeridae), is an ecological and commercial key component of the sub-arctic ichthyofauna. Here, we provide the first cytogenetic information on the species based on both conventional karyotyping and chromosomal mapping of 45S and 5S ribosomal genes through fluorescence in situ hybridization (FISH). The capelin genome displayed a diploid number of 54 with the karyotypic formula 26m/sm+28st/a and a fundamental number (FN) = 80. Both classes of ribosomal genes appeared to be spread out to multiple chromosomal locations, i.e. the 45S and 5S rDNA clusters were detected on six and seven chromosome pairs, respectively. A linked chromosomal organization of the major and minor ribosomal genes classes has been visualized in most of the rDNAs chromosomal locations. A comparative analysis of the available cytogenetic data for the family Osmeridae reveals diploid numbers higher than 48 and high fundamental numbers. This suggests that a rearranged karyotype is a shared feature within this family.
  • Chromosome mapping of 5S rRNA genes differentiates Brazilian populations of Leporellus vittatus (Anostomidae, Characiformes) Fish Cytogenetics

    Aguilar, Cecilia Teixeira de; Galetti Junior, Pedro Manoel

    Resumo em Inglês:

    Among the anostomid fishes, the genus Leporellus is represented by only three species: L. nattereri, endemic of the Amazon River, L. retropinnis, endemic of the Piracicaba River, and L. vittatus, widely distributed in rivers from Peru, Colombia, Guianas, and different major hydrographic basins of Brazil. A cytogenetic study carried out on specimens of Leporellus vittatus from three major Brazilian hydrographic basins evidenced a karyotype of 54 metacentric and submetacentric chromosomes. C-banding analysis revealed the presence of large pericentromeric heterochromatic segments in all chromosomes and a telomeric block coincident with the NOR sites. Ag, CMA3 or MM staining, and FISH with ribosomal probes located the 45S ribosomal genes on the terminal region of the long arm of the 12th chromosome pair of all populations. Nevertheless, in the specimens from the Paraná and São Francisco Basins the 5S rDNA clusters were interstitially located by FISH on the long arm of the 2nd chromosome pair, while in the specimens from the Tocantins-Araguaia Basin these sites were observed on the long arm of the 9th chromosome pair and on the short arm of the 17th chromosome pair. These data suggest that the species currently named Leporellus vittatus may comprise a complex of cryptic species.
  • Cytogenetic markers as diagnoses in the identification of the hybrid between Piauçu (Leporinus macrocephalus) and Piapara (Leporinus elongatus) Fish Cytogenetics

    Porto-Foresti, Fábio; Hashimoto, Diogo Teruo; Alves, Anderson Luis; Almeida, Rodrigo Braz Castilho; Senhorini, José Augusto; Bortolozzi, Jehud; Foresti, Fausto

    Resumo em Inglês:

    The genetic monitoring of interspecific hybrids involves the application of methodologies able to provide an easy and indubitable genetic characterization of both parental and hybrid individuals. In the present work, cytogenetic techniques were used to identify a hybrid lineage of "Piaupara" in order to caracterize them in relation to the parental species, Leporinus macrocephalus (piauçu) and L. elongatus (piapara). The cytogenetic analysis revealed that L. macrocephalus presented 2n = 54 chromosomes and a nucleolar organizer regions (NOR) at the telomere of the long arm of the submetacentric chromosome pair 2. Analysis of constitutive heterochromatin (C-banding) revealed a conspicuous block at the pericentromeric region on the long arm of a submetacentric chromosome pair. L. elongatus presented the same diploid number, 2n = 54, and a karyotypic formula similar to that of L. macrocephalus. The NORs were also at the telomere of the long arm of the submetacentric pair 2, which was morphologically different from that of L. macrocephalus. Heterochromatic blocks were observed at both telomeres of a submetacentric chromosome pair. The hybrid "Piaupara" presented the same diploid number (2n = 54) and karyotypic formula as the parental species and there were no visible differences between parental and hybrid individuals. Differently from the Giemsa staining, NOR- and C-banding analysis showed marked differences which allowed the identification of the hybrids by the different morphology and/or size of the chromosomes carrying the NORs and patterns of heterochromatin distribution in their chromosomes. Such genetic studies are important for fish culture since they can provide tools for monitoring natural and artificial hybridization. They are also useful in biological conservation programmes and in the proper management of natural and reared fish stocks.
  • Chromosome divergence and NOR polymorphism in Bryconamericus aff. iheringii (Teleostei, Characidae) in the hydrographic systems of the Paranapanema and Ivaí Rivers, Paraná, Brazil Fish Cytogenetics

    Capistano, Thiago Gomes; Castro, Ana Luiza de Brito Portela; Julio-Junior, Horácio Ferreira

    Resumo em Inglês:

    Cytogenetic studies were carried out in three populations of Bryconamericus aff. iheringii from two hydrographic systems of the Paranapanema and Ivaí Rivers, separated by a watershed, both belonging to the upper Paraná River basin. Specimens had a constant diploid number 2n = 52 chromosomes. However, three karyotype formulae were identified in the three populations: B. aff. iheringii from the Maringá stream had 12M+18SM+8ST+14A (FN = 90); specimens from Keller River showed 8M+28SM+6ST+10A (FN = 94) and specimens from the Tatupeba stream had 8M+20SM+8ST+16A (FN = 88). Nucleolar organizer regions (NORs) were identified by silver nitrate staining and fluorescent in situ hybridization (FISH) with an 18S rDNA probe. Specimens from Tatupeba stream had a simple NOR system located in a terminal position of the short arm of a pair of large submetacentric chromosomes. Ag-NOR and FISH methodologies revealed multiple NORs in specimens of the Maringá stream and Keller River. Differences in chromosome structure and in NOR patterns in the three populations of B. aff. iheringii revealed fixed evolutionary chromosome divergence. Aspects related to karyotypic variations and to geographic isolation of these populations are discussed.
  • Cytogenetic and morphological diversity in populations of Astyanax fasciatus (Teleostei, Characidae) from Brazilian northeastern river basins Fish Cytogenetics

    Medrado, Aline Souza; Figueiredo, Alba Vivian Amaral; Waldschmidt, Ana Maria; Affonso, Paulo Roberto Antunes de Mello; Carneiro, Paulo Luiz Souza

    Resumo em Inglês:

    In the present work, morphometric and cytogenetic analyses were carried out in populations of the fish Astyanax fasciatus (Characidae) from Contas and Recôncavo Sul River basins (State of Bahia, Brazil), providing new data on the genetic structure of this species along the region. Based on morphologic measurements, we observed that populations from the same hydrographic basin were more similar to each other (Contas and Preto do Costa Rivers), and remarkably divergent from Recôncavo Sul (Mineiro Stream), as indicated by clustering analysis. Cytogenetic data revealed a same diploid number for all populations (2n = 48), but distinct karyotype formulae (8M+24SM+12ST+4A, FN = 92 in Contas River, 8M+24SM+10ST+6A, FN = 90 in Preto do Costa River, and 8M+18SM+16ST+6A, FN = 90 in Mineiro Stream). Ag-NORs were identified at telomeres on a subtelocentric chromosome pair, although multiple ribosomal sites have been detected in some specimens from Contas River. These results show that A. fasciatus populations from northeastern river basins are well differentiated and present peculiar cytogenetic features when compared to populations from other regions. Therefore, the apparent chromosomal plasticity of this species, likely to represent a complex of cryptic forms, is corroborated. Finally, we demonstrated that morphological features can be successfully used to support other sources of genetic information.
  • Cytogenetic analyses of two endemic fish species from the São Francisco River basin: Conorhynchus conirostris and Lophiosilurus alexandri (Siluriformes) Fish Cytogenetics

    Marques, Marilza Barbosa de Almeida; Moreira-Filho, Orlando; Garcia, Caroline; Margarido, Vladimir Pavan

    Resumo em Inglês:

    Two Siluriformes species endemic to the São Francisco River basin were characterized by conventional and differential cytogenetic analyses involving C-banding, Ag-nucleolar organizer region (NOR) and chromomycin A3 (CMA3) staining, and FISH (fluorescent in situ hybridization) with 18S and 5S rDNA probes. Conorhynchus conirostris presents a higher diploid number (2n = 60) than those detected in Pimelodidae representatives, whereas Lophiosilurus alexandri, with a karyotype of 2n = 54 chromosomes, presents a chromosomal constitution similar to that found in the family Pseudopimelodidae. Plesiomorphic characteristics such as single NORs at terminal positions are found in both species, as revealed by CMA3 and silver nitrate staining, and FISH with a 18S rDNA probe. C-banding evidenced centromeric and telomeric heterochromatic blocks distributed over most of the chromosomes with a conspicuous heterochromatin segment in a pair of submetacentric chromosomes in L. alexandri. Such karyotype data, if compared to the cytogenetic pattern of other Siluriformes species, can be partially related to their degree of endemism, favorable to the occurrence and fixation of chromosomal rearrangements. The present study in representatives from these two Siluriformes families from the São Francisco River contributes to a better understanding of the karyotype evolution in species of this important order of Neotropical fishes.
  • Physical mapping of the 18S and 5S ribosomal genes in nine Characidae species (Teleostei, Characiformes) Fish Cytogenetics

    Peres, Wellington Adriano Moreira; Bertollo, Luiz Antonio Carlos; Moreira Filho, Orlando

    Resumo em Inglês:

    Characidae is one of the largest fish families of the Neotropical region, and presenting a pronounced morphological variability, certainly does not constitute a monophyletic group. The cytogenetical data also show a large chromosomal variation and can provide important information for a better understanding of the relationships between the species of this group. 18S and 5S rDNA probes were used in the present study for the chromosomal mapping in different Characidae species from the São Francisco River (Astyanax lacustris, Astyanax scabripinnis, Hasemania nana, Piabina argentea, Orthospinus franciscensis, Serrapinnus heterodon, Serrapinnus piaba and Myleus micans) and Alto Paraná (Astyanax altiparanae) basins. Species with a single pair of chromosomes bearing the nucleolar organizing regions (NORs) were identified, as well as species with multiple NORs, up to a maximum of seven 18S rDNA sites. The number of 5S rDNA site was also not constant, varying from two to eight. The mapping of the ribosomal genes was useful for the characterization and differentiation of the analyzed species.
  • Cytogenetic characterization of the strongly electric Amazonian eel, Electrophorus electricus (Teleostei, Gymnotiformes), from the Brazilian rivers Amazon and Araguaia Fish Cytogenetics

    Fonteles, Soraia B.A.; Lopes, Carlos E.; Akama, Alberto; Fernandes, Flora M.C.; Porto-Foresti, Fábio; Senhorini, José A.; Daniel-Silva, Maria de Fátima Z.; Foresti, Fausto; Almeida-Toledo, Lurdes Foresti de

    Resumo em Inglês:

    A karyotype analysis of the electric eel, Electrophorus electricus (Teleostei, Gymnotiformes), a strongly electric fish from northern South America, is presented. Two female specimens were analyzed, one from the Amazon River and one from the Araguaia River. The specimens had a chromosomal number of 2n = 52 (42M-SM + 10A). C-bands were present in a centromeric and pericentromeric position on part of the chromosomes; some interstitial C-bands were also present. Heteromorphic nucleolus organizer regions (NORs) were detected in two chromosome pairs of the specimen from the Amazon River. The chromosome number and karyotype characteristics are similar to those of other Gymnotidae species. The genera Electrophorus and Gymnotus are positioned as the basal lineages in the Gymnotiformes phylogeny.
  • Comparative cytogenetics of Carnegiella marthae and Carnegiella strigata (Characiformes, Gasteropelecidae) and description of a ZZ/ZW sex chromosome system Fish Cytogenetics

    Terencio, Maria Leandra; Schneider, Carlos Henrique; Gross, Maria Claudia; Silva, Adailton Moreira da; Feldberg, Eliana; Porto, Jorge Ivan Rebelo

    Resumo em Inglês:

    Comparative cytogenetic analyses of hatchetfishes Carnegiella marthae and Carnegiella strigata (Gasteropelecidae) from the Rio Negro basin were performed using conventional Giemsa staining, silver (Ag) -staining and C-banding. The diploid chromosome numbers of both species equaled 2n = 50 but their karyotypes were distinct. We found evidence for sex chromosomes in C. marthae since karyotype of males presented 20 M + 12 SM + 4 ST + 14 A and ZZ ST chromosomes while the females presented 20 M + 12 SM + 4 ST + 14 A and ZW ST chromosomes of distinct size. Conversely, C. strigata presented 4 M + 4 SM + 2 ST + 40 A chromosomes without sex chromosome heteromorphism. Karyotypes of both species had two NOR-bearing SM chromosomes of distinct size indicating the presence of multiple NOR phenotypes. The sex chromosome pair had specific C-banding pattern allowing identification of both Z and W. This heteromorphic system has previously been described for the gasteropelecids.
  • Karyotypic differentiation through chromosome fusion and number reduction in Imparfinis hollandi (Ostariophysi, Heptapteridae) Fish Cytogenetics

    Margarido, Vladimir Pavan; Moreira-Filho, Orlando

    Resumo em Inglês:

    The Neotropical Heptapteridae fish Imparfinis hollandi, endemic to the Iguaçu River Basin (Brazil), was cytogenetically analyzed and the diploid chromosome number of 2n = 42 chromosomes was determined (22m + 10sm + 10st), the lowest diploid number in this genus and family. Like other Heptapteridae species, only one NOR-bearing chromosome pair was detected by silver nitrate staining. Dark heterochromatic blocks were visualized in only three chromosome pairs, and chromomycin A3+ bands were coincident with Ag-NORs. Although no intercalary (TTAGGG)n sequence was observed through FISH with a telomere probe, an asymmetric karyotype showing four large chromosome pairs with diploid chromosome number reduction suggests that tandem chromosome fusions probably occurred during the karyotypic differentiation of Imparfinis hollandi.
  • Population analysis of a chromosome polymorphism in Astyanax (Teleostei, Characiformes) species endemic to the Iguaçu River Fish Cytogenetics

    Kantek, Daniel Luis Zanella; Cipriano, Roger Raupp; Noleto, Rafael Bueno; Fenocchio, Alberto Sergio; Artoni, Roberto Ferreira; Cestari, Marta Margarete

    Resumo em Inglês:

    Chromosomal analyses were performed in the fish Astyanax sp.D collected from three different points: two streams from the right bank and one from the left bank of the Upper Iguaçu River, Paraná State, Brazil. The individuals from all localities possess 2n = 50 chromosomes and a FN = 84 (4m+24sm+6st+16a). The C-banding pattern was similar in all populations. However, within each population, an interindividual variation concerning the number and localization of heterochromatic bands was observed. Some of these variations were quantified in each population, and the results indicate that the samples were not different when studying the variable frequencies. Considering that Astyanax sp.D is typical in the headwaters of the Iguaçu River, these results were not expected. The data indicate that gene flow is occurring and that the Iguaçu River is not an ecological barrier among the Astyanax sp. D populations.
  • Supernumerary chromosomes in the pufferfish Sphoeroides spengleri: first occurrence in marine Teleostean Tetraodontiformes fish Fish Cytogenetics

    Alves, Anderson Luis; Porto-Foresti, Fábio; Oliveira, Claudio; Foresti, Fausto

    Resumo em Inglês:

    Cytogenetic analyses carried out in eight specimens of Sphoeroides spengleri revealed the presence of 2n = 46 chromosomes (20 M/SM and 26 ST/A). Besides the standard karyotypical set, the presence of B microchromosomes was observed in two individuals, ranging from 0 to 2 microchromosomes per cell. A karyotype composed by 2n = 46 chromosomes with occurrence of M and SM chromosomes is considered basal for the species from the clade comprising the families Tetraodontidae, Balistidae, and Diodontidae, although it represents a derived condition for the order Tetraodontiformes, whose basal karyotype would be composed by 2n = 48 acrocentric chromosomes. The occurrence of B microchromosomes in marine Tetraodontiformes fish was not known, and this represents the first report of such a chromosomal type.
  • B chromosomes in a population of Astyanax eigenmanniorum (Characiformes, Characidae) from the Araguari River Basin (Uberlândia, MG, Brazil) Fish Cytogenetics

    Torres-Mariano, Alessandra Ribeiro; Morelli, Sandra

    Resumo em Inglês:

    A cytogenetic study was conducted on an Astyanax eigenmanniorum population from the Caetano Stream (18° 44' 56" S/ 048° 18' 39" W) - in Uberlândia, MG, Brazil - showing a modal diploid number of 48 chromosomes in the standard male and female karyotypes. However, in several specimens it was also possible to observe metaphases with one or two B chromosomes, increasing the diploid number to 49 or 50 chromosomes, respectively. The supernumerary chromosomes were totally heterochromatic and highlighted after C-banding. The silver-stained nucleolus organizing regions (Ag-NORs) were located in at least five chromosomes of the standard karyotype, thus characterizing a multiple NOR system in the species. This is the first occurrence of an A. eigenmanniorum population with 2n = 48 chromosomes, bearing supernumerary chromosomes.
  • Cytotaxonomy and karyoevolution of the genus Crenicichla (Perciformes, Cichlidae) Fish Cytogenetics

    Benzaquem, Denise Corrêa; Feldberg, Eliana; Porto, Jorge Ivan Rebelo; Gross, Maria Claudia; Zuanon, Jansen Alfredo Sampaio

    Resumo em Inglês:

    Karyotypes of six cichlid species of the genus Crenicichla were investigated. The species C. cincta, C. inpa, C. reticulata, C. lugubris, and C. cf. johanna were collected from Amazon basin, and C. britskii was collected from the Paraná-Paraguai basin. All of the analysed species showed 2n = 48 chromosomes; however, C. cincta, C. lugubris, C. cf. johanna, and C. britskii had a karyotype formula of 8M-SM+40ST-A, FN = 56, while C. inpa and C. reticulata exhibited the formula 6M-SM+42ST-A, FN = 54. Analysis of active Ag-NORs revealed two NOR-bearing chromosomes in all species; however, theses cistrons were located on different chromosome pairs and/or in different chromosome locations in each species. This condition bears evolutionary significance, since it is the main chromosome marker of the process of karyotypic evolution among the species of the genus Crenicichla. In general, C-banding revealed a similar constitutive heterochromatin pattern in all species, although it was possible to detect some features that led us to infer that Crenicichla also presents a species-specific heterochromatin pattern.
  • Cytogenetic studies in fishes of the genera Hassar, Platydoras and Opsodoras (Doradidae, Siluriformes) from Jarí and Xingú Rivers, Brazil Fish Cytogenetics

    Milhomem, Susana Suely Rodrigues; Souza, Augusto Cesar Paes de; Nascimento, Aline Lira do; Carvalho Jr., Jaime Ribeiro; Feldberg, Eliana; Pieczarka, Julio Cesar; Nagamachi, Cleusa Yoshiko

    Resumo em Inglês:

    We studied the karyotypes of Hassar cf. orestis and an undescribed Hassar species from the Jarí River and Opsodoras ternetzi, H. orestis and Platydoras cf. costatus from the Xingú River, all with 2n = 58. Constitutive heterochromatin is located in the centromere in most metacentric pairs; in some chromosomes this banding is not present, or it is located on the whole chromosome arm or in the distal regions. The NOR is located on a single biarmed pair at a distal region of the short arm in H. cf. orestis, H. orestis and P. cf. costatus at a distal region of the long arm in O. ternetzi and at a proximal region of the long arm in the Hassar species. In all species (except for Hassar sp.) the CMA3 analysis revealed a rich G-C region coincident with the NOR. Probably inversions occurred in the NOR chromosome during the chromosomal differentiation of the Doradidae species here described.
  • Localization of ribosomal genes in three Pimelodus species (Siluriformes, Pimelodidae) of the São Francisco River: 5S genes as species markers and conservation of the 18S rDNA sites Fish Cytogenetics

    Garcia, Caroline; Moreira Filho, Orlando

    Resumo em Inglês:

    Pimelodidae is one of the most representative of Neotropical catfish families. However, these fish are still poorly studied in terms of cytogenetics, especially regarding the application of more accurate techniques such as the chromosomal localization of ribosomal genes. In the present work, fluorescent in situ hybridization with 5S and 18S rDNA probes was employed for rDNA site mapping in Pimelodus sp., P. fur and P. maculatus from the São Francisco River in the Três Marias municipality - MG. The results from the application of the 18S probe confirmed the previous data obtained by silver nitrate staining, identifying a simple nucleolar organizing region system for these species. However, the labeling results from the 5S rDNA probe demonstrated a difference in the number and localization of these sites between the analyzed species. The obtained data allowed inferences on the possible processes involved in the karyotypic evolution of this genus.
  • Identification and description of distinct B chromosomes in Cyphocharax modestus (Characiformes, Curimatidae) Fish Cytogenetics

    Santos, Lessandra Viviane De Rosa; Foresti, Fausto; Martins, Cesar; Oliveira, Claudio; Wasko, Adriane Pinto

    Resumo em Inglês:

    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.
  • Chromosomal evidence of population subdivision in the freshwater fish Leporinus elongatus in the Upper Paraná River basin Fish Cytogenetics

    Molina, Wagner Franco; Shibatta, Oscar; Galetti Jr., Pedro Manoel

    Resumo em Inglês:

    Cytogenetic analyses performed in populations of the Neotropical freshwater fish Leporinus elongatus of the Upper Paraná River basin showed a chromosome polymorphism involving heterochromatic segments of the nucleolar organizing pair in this species. The NOR-bearing chromosome pair was characterized by two phenotypes identified by the absence (C¹) or presence (C²) of a heterochromatic segment located in an interstitial position of the long arm in this pair. The meiotic segregation of these variations results in three distinct cytotypes, C¹C¹, C¹C² and C²C². Both populations were in Hardy-Weinberg equilibrium. Sex-related cytotype divergence was identified and multivariate analyses revealed that the C²C² individuals presented morphometric differentiation relative to body height in relation to the other cytotypes. A chromosome differentiation and a cytotype proportion difference observed between both populations suggest the occurrence of population subdivision within this species along the Upper Paraná River basin. These findings might contribute to the knowledge of the population biology of this fish and for its conservation.
  • First report of a B chromosome in a natural population of Astyanax altiparanae (Characiformes, Characidae) Fish Cytogenetics

    Hashimoto, Diogo Teruo; Gonçalves, Vanessa Regina; Bortolozzi, Jehud; Foresti, Fausto; Porto-Foresti, Fábio

    Resumo em Inglês:

    Several species of the genus Astyanax have already been genetically studied, and B-chromosomes have been considered to be an interesting feature in some species of this group. In the present paper we report, for the first time, the occurrence of a B microchromosome in a natural population of A. altiparanae. This additional genomic element was identified as an acrocentric chromosome, similar in size to the smallest chromosomal pairs of the standard karyotype. Analysis of the constitutive heterochromatin pattern by C-banding evidenced heterochromatic blocks located on centromeric, pericentromeric, and interstitial regions of some chromosomes, and also positive marks in a subtelocentric chromosomal pair that presented the short arms entirely heterochromatic. The application of this methodology also revealed a heterochromatic pattern in the extra chromosome, a typical feature of supernumerary chromosomes.
  • Use of chromosome microdissection in fish molecular cytogenetics Fish Cytogenetics

    Henning, Frederico; Trifonov, Vladimir; Almeida-Toledo, Lurdes Foresti de

    Resumo em Inglês:

    Chromosome microdissection is a technique in which whole chromosomes or chromosomal segments are dissected under an inverted microscope yielding chromosome-specific sequences. Several protocol modifications introduced during the past 15 years reduced the number of chromosomes required for most applications. This is of particular interest to fish molecular cytogenetics, since most species present highly uniform karyotypes which make impossible the collection of multiple copies of the same chromosome. Probes developed in this manner can be used to investigate chromosome homologies in closely related species. Here we describe a protocol recently used in the gymnotiform species group Eigenmannia and review the major steps involved in the generation of these markers focusing on protocol modifications aiming to reduce the number of required chromosomes.
  • Phylogenetic analysis of the order Pleuronectiformes (Teleostei) based on sequences of 12S and 16S mitochondrial genes Fish Molecular Genetics

    Azevedo, Marisa F.C.; Oliveira, Claudio; Pardo, Belén G.; Martínez, Paulino; Foresti, Fausto

    Resumo em Inglês:

    The fish order Pleuronectiformes, composed of 14 families, has two suborders: Psettodoidei (with one family) and Pleuronectoidei (with thirteen families). The relationships among families of Pleuronectoidei and among the genera of their families have extensively been debated and a consensus has not yet been reached. In the present study, partial sequences of the 12S and 16S mitochondrial rRNA genes were obtained from 19 species belonging to the families Achiridae, Bothidae, Cynoglossidae, Paralichthyidae, Pleuronectidae, Scophthalmidae, and Soleidae. Additional sequences of 42 pleuronectiform species were obtained from GenBank. Phylogenetic analyses were conducted by the methods of maximum-parsimony, maximum-likelihood and Bayesian inference. Our results corroborate the monophyletic status of all families, excluding Paralichthyidae. In the family Achiridae, the genus Catathyridium (freshwater) was the sister group of Trinectes (saltwater), and Hypoclinemus (freshwater) was the sister group of Achirus (saltwater). Assuming that the putative ancestor of achirids lived in saltwater, it is suggested that the freshwater habitats in South America were colonized independently by different achirid lineages.
  • The complete mitochondrial genome of the pirarucu (Arapaima gigas, Arapaimidae, Osteoglossiformes) Fish Molecular Genetics

    Hrbek, Tomas; Farias, Izeni Pires

    Resumo em Inglês:

    We sequenced the complete mitochondrial genome of the pirarucu, Arapaima gigas, the largest fish of the Amazon basin, and economically one of the most important species of the region. The total length of the Arapaima gigas mitochondrial genome is 16,433 bp. The mitochondrial genome contains 13 protein-coding genes, two rRNA genes and 22 tRNA genes. Twelve of the thirteen protein-coding genes are coded on the heavy strand, while nad6 is coded on the light strand. The Arapaima gene order and content is identical to the common vertebrate form, as is codon usage and base composition. Its control region is atypical in being short at 767 bp. The control region also contains a conserved ATGTA motif recently identified in the Asian arowana, three conserved sequence blocks (CSB-1, CBS-2 and CBS-3) and its 3' end contains long series of di- and mono-nucleotide microsatellite repeats. Other osteoglossiform species for which control region sequences have been published show similar control region characteristics.
  • 5S rDNA characterization in twelve Sciaenidae fish species (Teleostei, Perciformes): depicting gene diversity and molecular markers Fish Molecular Genetics

    Alves-Costa, Fernanda A.; Martins, Cesar; Matos, Fernanda Del Campos de; Foresti, Fausto; Oliveira, Claudio; Wasko, Adriane P.

    Resumo em Inglês:

    In order to extend the genetic data on the Sciaenidae fish family, the present study had the purpose to characterize PCR-generated 5S rDNA repeats of twelve species of this group through PAGE (Polyacrylamide Gel Electrophoresis) analysis. The results showed the occurrence of at least two different 5S rDNA size classes in all the species. Moreover, 5S rDNA repeats of one of the studied species - Isopisthus parvipinnis - were cloned and subjected to nucleotide sequencing and Southern blot membrane hybridization analyses, which permitted to confirm the existence of two major 5S rDNA classes. Phylogenetic analysis based on the nucleotide sequences of different 5S rDNA repeats of I. parvipinnis lead to their separation into two major clusters. These results may reflect the high dynamism that rules the evolution rate of 5S rDNA repeats. The obtained data suggest that 5S rDNA can be useful in genetic analyses to identify species-specific markers and determine relationships among species of the Sciaenidae group.
  • Genetic and morphometric differences between yellowtail snapper (Ocyurus chrysurus, Lutjanidae) populations of the tropical West Atlantic Fish Molecular Genetics

    Vasconcellos, Anderson V.; Vianna, Paulo; Paiva, Paulo C.; Schama, Renata; Solé-Cava, Antonio

    Resumo em Inglês:

    Populations of Ocyurus chrysurus were compared genetically and morphometrically along the West Atlantic coast to test the null hypothesis of population homogeneity in the area. Brazilian populations were found to be differentiated in shape (canonical variates analysis; F[48,515] = 10.84, p < 0.0001). Analyses of mitochondrial DNA sequences (663 bp of the control region) did not show any differences between Brazilian populations but could detect differences between Brazilian and Caribbean (Belize) populations. The samples from Pernambuco differed significantly from the other Brazilian populations in allozyme frequencies (11 loci; F ST = 0.167; p < 0.05), but this may have resulted from the small number of samples analysed for that population. Sequence variation of Belize samples departed from neutral expectations (Fu's FS = -8.88; p < 0.001). A mismatch distribution analysis points to an ancient population expansion in that area. We conclude that the genetic data do not allow the rejection of the null hypothesis of panmixia for Brazilian yellowtail snapper populations which should be treated as a single genetic stock, with a latitudinal gradient on their morphology which probably results from phenotypic plasticity. On the other hand, there is a severe restriction to gene flow between O. chrysurus populations from the Caribbean and from the southwestern Atlantic.
  • Genetic diversity of Hypostomus ancistroides (Teleostei, Loricariidae) from an urban stream Fish Molecular Genetics

    Sofia, Silvia H.; Galindo, Bruno A.; Paula, Francine M.; Sodré, Leda M.K.; Martinez, Cláudia B.R.

    Resumo em Inglês:

    In this study, random amplified polymorphic DNA (RAPD) markers were applied to analyze the genetic diversity of samples of the Neotropical catfish Hypostomus ancistroides, collected from four sites (S1, S2, S3 and S4) along an urban stream in Southern Brazil. The 11 primers used in RAPD analysis amplified 147 loci, 76 (51.7%) of which were polymorphic. The proportions of polymorphic loci observed in the four samples were: 29.93% (S1), 31.97% (S2), 23.81% (S3) and 38.77% (S4). The average heterozygosity within sampling localities ranged from 0.1230 to 0.1526 and unbiased genetic distances ranged from 0.0253 to 0.0445. AMOVA partitioned 90.85% of the total variation within samples and 9.15% among samples. Excepting for the sample pair S1-S2 (phiST = 0.02784; p > 0.05), all others pairwise phiST values were significantly greater than zero, indicating moderate genetic differentiation among catfish samples from four localities. The relative low levels of genetic variation detected in all groups studied could be related to different factors, including the sedentary habit of these fish, which can be eroding the genetic variation of H. ancistroides from each locality.
  • A test of the utility of DNA barcoding in the radiation of the freshwater stingray genus Potamotrygon (Potamotrygonidae, Myliobatiformes) Fish Molecular Genetics

    Toffoli, Daniel; Hrbek, Tomas; Araújo, Maria Lúcia Góes de; Almeida, Maurício Pinto de; Charvet-Almeida, Patricia; Farias, Izeni Pires

    Resumo em Inglês:

    DNA barcoding is a recently proposed global standard in taxonomy based on DNA sequences. The two main goals of DNA barcoding methodology are assignment of specimens to a species and discovery of new species. There are two main underlying assumptions: i) reciprocal monophyly of species, and ii) intraspecific divergence is always less than interspecific divergence. Here we present a phylogenetic analysis of the family Potamotrygonidae based on mitochondrial cytochrome c oxidase I gene, sampling 10 out of the 18 to 20 valid species including two non-described species. Potamotrygonidae systematics is still not fully resolved with several still-to-be-described species while some other species are difficult to delimit due to overlap in morphological characters and because of sharing a complex color patterns. Our results suggest that the family passed through a process of rapid speciation and that the species Potamotrygon motoro, P. scobina, and P. orbignyi share haplotypes extensively. Our results suggest that systems of identification of specimens based on DNA sequences, together with morphological and/or ecological characters, can aid taxonomic studies, but delimitation of new species based on threshold values of genetic distances are overly simplistic and misleading.
  • Soluble malate dehydrogenase of Geophagus brasiliensis (Cichlidae, Perciformes): isolated isoforms and kinetics properties Fish Molecular Genetics

    Aquino-Silva, Maria Regina de; Schwantes, Maria Luiza Barcellos; Munin, Flavia Simone; Schwantes, Arno Rudi; Santos, Silvana Pereira dos

    Resumo em Inglês:

    Kinetic properties and thermal stabilities of Geophagus brasiliensis skeletal muscle unfractionated malate dehydrogenase (MDH, EC 1.1.1.37) and its isolated isoforms were analyzed to examine a possible sMDH-B* locus duplication in a fixation process influenced by genetic drift. Two optimal pHs were detected: 7.5 for AB1 unfractionated muscle phenotype and its B1 isoform, and 8.0 for AB1B2 unfractionated muscle phenotype, A and B2 isoforms. While G. brasiliensis A isoform could be characterized as thermostable, the duplicated B isoform cannot be assumed as thermolabile. Km values for isolated B2 isoforms were 1.6 times lower than for B1. A duplication event in progress best explains the electrophoretic six-band pattern detected in G. brasiliensis, which would be caused by genetic drift.
  • Phylogeny of the Serrasalmidae (Characiformes) based on mitochondrial DNA sequences Fish Molecular Genetics

    Ortí, Guillermo; Sivasundar, Arjun; Dietz, Kelly; Jégu, Michel

    Resumo em Inglês:

    Previous studies based on DNA sequences of mitochondrial (mt) rRNA genes showed three main groups within the subfamily Serrasalminae: (1) a "pacu" clade of herbivores (Colossoma, Mylossoma, Piaractus); (2) the "Myleus" clade (Myleus, Mylesinus, Tometes, Ossubtus); and (3) the "piranha" clade (Serrasalmus, Pygocentrus, Pygopristis, Pristobrycon, Catoprion, Metynnis). The genus Acnodon was placed as the sister taxon of clade (2+3). However, poor resolution within each clade was obtained due to low levels of variation among rRNA gene sequences. Complete sequences of the hypervariable mtDNA control region for a total of 45 taxa, and additional sequences of 12S and 16S rRNA from a total of 74 taxa representing all genera in the family are now presented to address intragroup relationships. Control region sequences of several serrasalmid species exhibit tandem repeats of short motifs (12 to 33 bp) in the 3' end of this region, accounting for substantial length variation. Bayesian inference and maximum parsimony analyses of these sequences identify the same groupings as before and provide further evidence to support the following observations: (a) Serrasalmus gouldingi and species of Pristobrycon (non-striolatus) form a monophyletic group that is the sister group to other species of Serrasalmus and Pygocentrus; (b) Catoprion, Pygopristis, and Pristobrycon striolatus form a well supported clade, sister to the group described above; (c) some taxa assigned to the genus Myloplus (M. asterias, M tiete, M ternetzi, and M rubripinnis) form a well supported group whereas other Myloplus species remain with uncertain affinities (d) Mylesinus, Tometes and Myleus setiger form a monophyletic group.
  • Identities among actin-encoding cDNAs of the Nile tilapia (Oreochromis niloticus) and other eukaryote species revealed by nucleotide and amino acid sequence analyses Fish Molecular Genetics

    Poletto, Andréia B.; Wasko, Adriane P.; Oliveira, Claudio; Azevedo, Alexandre; Carvalho, Robson F.; Silva, Maeli Dal Pai; Foresti, Fausto; Martins, Cesar

    Resumo em Inglês:

    Actin-encoding cDNAs of Nile tilapia (Oreochromis niloticus) were isolated by RT-PCR using total RNA samples of different tissues and further characterized by nucleotide sequencing and in silico amino acid (aa) sequence analysis. Comparisons among the actin gene sequences of O. niloticus and those of other species evidenced that the isolated genes present a high similarity to other fish and other vertebrate actin genes. The highest nucleotide resemblance was observed between O. niloticus and O. mossambicus a-actin and b-actin genes. Analysis of the predicted aa sequences revealed two distinct types of cytoplasmic actins, one cardiac muscle actin type and one skeletal muscle actin type that were expressed in different tissues of Nile tilapia. The evolutionary relationships between the Nile tilapia actin genes and diverse other organisms is discussed.
  • Description of novel microsatellite loci in the Neotropical fish Prochilodus argenteus and cross-amplification in P. costatus and P. lineatus Fish Molecular Genetics

    Barbosa, Anna C.D.R.; Galzerani, Felipe; Corrêa, Thaís C.; Galetti Jr., Pedro M.; Hatanaka, Terumi

    Resumo em Inglês:

    Prochilodus is one of the most important fish resources of South America, in addition to the important role it plays in nutrient cycling of Neotropical rivers. In the present study, we describe the isolation and characterization of nine novel microsatellite loci in Prochilodus argenteus. The number of alleles per polymorphic locus varied from 5 (Par76) to 21 (Par85), revealing a total of 116 alleles. The values of observed and expected heterozygosities ranged from 0.629 (Par69) to 0.926 (Par85 and Par86) and from 0.643 (Par66) to 0.931 (Par80), respectively. Furthermore, the ability of these and other previously described microsatellite markers to amplify orthologous loci was tested in two related species, Prochilodus costatus and Prochilodus lineatus. These loci will be useful for studies of population genetic structure in this group of fishes, and in aiding future genetic mapping studies of P. argenteus.
  • Discrimination of Shark species by simple PCR of 5S rDNA repeats Fish Molecular Genetics

    Pinhal, Danillo; Gadig, Otto BF; Wasko, Adriane P; Oliveira, Claudio; Ron, Ernesto; Foresti, Fausto; Martins, Cesar

    Resumo em Inglês:

    Sharks are suffering from intensive exploitation by worldwide fisheries leading to a severe decline in several populations in the last decades. The lack of biological data on a species-specific basis, associated with a k-strategist life history make it difficult to correctly manage and conserve these animals. The aim of the present study was to develop a DNA-based procedure to discriminate shark species by means of a rapid, low cost and easily applicable PCR analysis based on 5S rDNA repeat units amplification, in order to contribute conservation management of these animals. The generated agarose electrophoresis band patterns allowed to unequivocally distinguish eight shark species. The data showed for the first time that a simple PCR is able to discriminate elasmobranch species. The described 5S rDNA PCR approach generated species-specific genetic markers that should find broad application in fishery management and trade of sharks and their subproducts.
  • A phylogenetic analysis of Brycon and Henochilus (Characiformes, Characidae, Bryconinae) based on the mitochondrial gene 16S rRNA Fish Molecular Genetics

    Hilsdorf, Silva; Oliveira, Claudio; Lima, Flávio César Thadeo de; Matsumoto, Cristianne Kayoko

    Resumo em Inglês:

    The genus Brycon, the largest subunit of the Bryconinae, has 42 valid species distributed from southern Mexico to the La Plata River in Argentina. Henochilus is a monotypic genus, comprising a single species (H. wheatlandii) found in the upper Rio Doce basin. In the present study, partial sequences of the mitochondrial gene 16S were obtained for fifteen species of Brycon and for Henochilus wheatlandii. The results showed that the genus Brycon is paraphyletic, since Henochilus is the sister-group of B. ferox and B. insignis. The most basal species analyzed were the trans-Andean species B. henni, B. petrosus, and B. chagrensis.
  • Can Lutjanus purpureus (South red snapper) be "legally" considered a red snapper (Lutjanus campechanus)? Fish Molecular Genetics

    Gomes, Grazielle; Schneider, Horacio; Vallinoto, Marcelo; Santos, Simoni; Orti, Guillermo; Sampaio, Iracilda

    Resumo em Inglês:

    Red snappers (Lutjanus purpureus in Brazil and Lutjanus campechanus in USA and Gulf of Mexico) are both under clear effect of overfishing. Because of their high morphological similarity it has already been suggested that they could possibly be considered as a single species. To investigate the degree of similarity and the genetic structure of red snapper populations we constructed a common dataset of partial D-loop mtDNA sequences of L. purpureus from Brazil (Amapá, Pará and Maranhão) and L. campechanus from the Atlantic coast of the USA (Florida, Louisiana and Mississippi). Phylogenetic and population genetic analyses surprisingly depicted high similarity between L. campechanus and L. purpureus, compatible with the hypothesis of a single species of red snapper for the Western Atlantic Ocean. These preliminary but very curious findings open an important discussion regarding the legislation involved on the capture of this overexploited fish resources as well as regarding their taxonomy.
  • Evidence of lack of population substructuring in the Brazilian freshwater fish Prochilodus costatus Fish Molecular Genetics

    Carvalho-Costa, Luis F.; Hatanaka, Terumi; Galetti Jr., Pedro M.

    Resumo em Inglês:

    Curimbatá-pióa (Prochilodus costatus Valenciennes, 1850) is an endemic species from the São Francisco River Basin with migratory habits, important ecological role in the ecosystem and relevant fishing importance. The present study aimed to assess the genetic variation in P. costatus in order to investigate its population genetic structure. Genetic variation was studied at three sites downstream the Três Marias dam (Upper-middle São Francisco River, State of Minas Gerais) through six specific microsatellite loci. Fish from the three sites had quite similar genetic diversity levels and no genetic differentiation was detected, suggesting that P. costatus might represent a single reproductive unit in the studied area. Alternatively, the present study was not able to detect putative coexistent and comigrating populations along the main channel river. Our findings could be very helpful for the management and conservation of this fish.
  • Molecular cloning and sequence analysis of growth hormone cDNA of Neotropical freshwater fish Pacu (Piaractus mesopotamicus) Fish Molecular Genetics

    Pinheiro, Janeth Silva; Wolff, José Luiz Caldas; Araújo, Ronaldo de Carvalho; Hilsdorf, Alexandre Wagner Silva

    Resumo em Inglês:

    RT-PCR was used for amplifying Piaractus mesopotamicus growth hormone (GH) cDNA obtained from mRNA extracted from pituitary cells. The amplified fragment was cloned and the complete cDNA sequence was determined. The cloned cDNA encompassed a sequence of 543 nucleotides that encoded a polypeptide of 178 amino acids corresponding to mature P. mesopotamicus GH. Comparison with other GH sequences showed a gap of 10 amino acids localized in the N terminus of the putative polypeptide of P. mesopotamicus. This same gap was also observed in other members of the family. Neighbor-joining tree analysis with GH sequences from fishes belonging to different taxonomic groups placed the P. mesopotamicus GH within the Otophysi group. To our knowledge, this is the first GH sequence of a Neotropical characiform fish deposited in GenBank.
  • A brief history of fish genetics in Brazil History, Story And Memories

    Foresti, Fausto
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