RESUMO
Abstract Genetic and phylogenetic relationships among seven piranha species of the genera Serrasalmus and Pygocentrus from the Paraná-Paraguay, São Francisco and Tocantins River basins were evaluated in the present study by partial sequences of two mitochondrial genes, Cytochrome b and Cytochrome c Oxidase I. Phylogenetic analysis of Maximum-Likelihood and Bayesian inference were performed. Results indicated, in general, greater genetic similarity between the two species of Pygocentrus (P. nattereri and P. piraya), between Serrasalmus rhombeus and S. marginatus and between S. maculatus, S. brandtii and S. eigenmanni. Pygocentrus nattereri, S. rhombeus and S. maculatus showed high intraspecific genetic variability. These species have each one, at least two different mitochondrial lineages that, currently, occur in sympatry (S. rhombeus) or in allopatry (P. nattereri and S. maculatus). Species delimitation analysis and the high values of genetic distances observed between populations of S. rhombeus and of S. maculatus indicated that each species may corresponds to a complex of cryptic species. The non-monophyletic condition of S. rhombeus and S. maculatus reinforces the hypothesis. The geographic distribution and the genetic differentiation pattern observed for the piranha species analyzed herein are discussed regarding the geological and hydrological events that occurred in the hydrographic basins.
Resumo Relações genéticas e filogenéticas de sete espécies de piranhas dos gêneros Serrasalmus e Pygocentrus das bacias hidrográficas Paraná-Paraguai, São Francisco e Tocantins foram avaliadas com base em sequências parciais dos genes mitocondriais Citocromo b e Citocromo c Oxidase I. Foram realizadas análises filogenéticas de Máxima Verossimilhança e de inferência Bayesiana. Os resultados indicaram, em geral, maior similaridade genética entre as duas espécies de Pygocentrus (P. nattereri e P. piraya), entre Serrasalmus rhombeus e S. marginatus e entre S. maculatus, S. brandtii e S. eigenmanni. Pygocentrus nattereri, S. rhombeus e S. maculatus revelaram ter alta variabilidade genética intraespecífica. Essas espécies têm, cada uma, pelo menos duas linhagens mitocondriais que, atualmente, ocorrem em simpatria (S. rhombeus) ou alopatria (P. nattereri e S. maculatus). Análises de delimitação de espécies e os altos valores de distância genética observados entre as populações de S. rhombeus e de S. maculatus indicam que cada espécie pode, na verdade, corresponder a um complexo de espécies crípticas. A condição não-monofilética de S. rhombeus e S. maculatus reforça essa hipótese. A distribuição geográfica e o padrão de diferenciação genética observados para as espécies de piranhas analisadas são discutidos com relação aos eventos geológicos e hidrológicos que ocorreram nas bacias hidrográficas.
Assuntos
Animais , Caraciformes , Paraguai , Filogenia , Brasil , Teorema de Bayes , RiosRESUMO
Genetic and phylogenetic relationships among seven piranha species of the genera Serrasalmus and Pygocentrus from the Paraná-Paraguay, São Francisco and Tocantins River basins were evaluated in the present study by partial sequences of two mitochondrial genes, Cytochrome b and Cytochrome c Oxidase I. Phylogenetic analysis of Maximum-Likelihood and Bayesian inference were performed. Results indicated, in general, greater genetic similarity between the two species of Pygocentrus (P. nattereri and P. piraya), between Serrasalmus rhombeus and S. marginatus and between S. maculatus, S. brandtii and S. eigenmanni. Pygocentrus nattereri, S. rhombeus and S. maculatus showed high intraspecific genetic variability. These species have each one, at least two different mitochondrial lineages that, currently, occur in sympatry (S. rhombeus) or in allopatry (P. nattereri and S. maculatus). Species delimitation analysis and the high values of genetic distances observed between populations of S. rhombeus and of S. maculatus indicated that each species may corresponds to a complex of cryptic species. The non-monophyletic condition of S. rhombeus and S. maculatus reinforces the hypothesis. The geographic distribution and the genetic differentiation pattern observed for the piranha species analyzed herein are discussed regarding the geological and hydrological events that occurred in the hydrographic basins.
Assuntos
Caraciformes , Animais , Teorema de Bayes , Brasil , Paraguai , Filogenia , RiosRESUMO
Abstract Genetic and phylogenetic relationships among seven piranha species of the genera Serrasalmus and Pygocentrus from the Paraná-Paraguay, São Francisco and Tocantins River basins were evaluated in the present study by partial sequences of two mitochondrial genes, Cytochrome b and Cytochrome c Oxidase I. Phylogenetic analysis of Maximum-Likelihood and Bayesian inference were performed. Results indicated, in general, greater genetic similarity between the two species of Pygocentrus (P. nattereri and P. piraya), between Serrasalmus rhombeus and S. marginatus and between S. maculatus, S. brandtii and S. eigenmanni. Pygocentrus nattereri, S. rhombeus and S. maculatus showed high intraspecific genetic variability. These species have each one, at least two different mitochondrial lineages that, currently, occur in sympatry (S. rhombeus) or in allopatry (P. nattereri and S. maculatus). Species delimitation analysis and the high values of genetic distances observed between populations of S. rhombeus and of S. maculatus indicated that each species may corresponds to a complex of cryptic species. The non-monophyletic condition of S. rhombeus and S. maculatus reinforces the hypothesis. The geographic distribution and the genetic differentiation pattern observed for the piranha species analyzed herein are discussed regarding the geological and hydrological events that occurred in the hydrographic basins.
Resumo Relações genéticas e filogenéticas de sete espécies de piranhas dos gêneros Serrasalmus e Pygocentrus das bacias hidrográficas Paraná-Paraguai, São Francisco e Tocantins foram avaliadas com base em sequências parciais dos genes mitocondriais Citocromo b e Citocromo c Oxidase I. Foram realizadas análises filogenéticas de Máxima Verossimilhança e de inferência Bayesiana. Os resultados indicaram, em geral, maior similaridade genética entre as duas espécies de Pygocentrus (P. nattereri e P. piraya), entre Serrasalmus rhombeus e S. marginatus e entre S. maculatus, S. brandtii e S. eigenmanni. Pygocentrus nattereri, S. rhombeus e S. maculatus revelaram ter alta variabilidade genética intraespecífica. Essas espécies têm, cada uma, pelo menos duas linhagens mitocondriais que, atualmente, ocorrem em simpatria (S. rhombeus) ou alopatria (P. nattereri e S. maculatus). Análises de delimitação de espécies e os altos valores de distância genética observados entre as populações de S. rhombeus e de S. maculatus indicam que cada espécie pode, na verdade, corresponder a um complexo de espécies crípticas. A condição não-monofilética de S. rhombeus e S. maculatus reforça essa hipótese. A distribuição geográfica e o padrão de diferenciação genética observados para as espécies de piranhas analisadas são discutidos com relação aos eventos geológicos e hidrológicos que ocorreram nas bacias hidrográficas.
RESUMO
Astyanax altiparanae, belonging to the bimaculatus group, which includes species with similar colors and morphology, occurs in the upper Paraná River basin. As the use of mitochondrial DNA has made great strides in the diagnosis of species, in previous researches, two strains were detected in A. altiparanae with a high divergence in the D-loop region, provisionally called AltoPR and AltoPR-D. Evidence led to the hypothesis that the two strains did not belong to the same species. Phylogenetic hypotheses were produced by maximum-likelihood. Mean internal distances of the AltoPR and AltoPR-D groups were respectively 0.002 and 0.003, with the distance between them being 0.037. Sequences from GenBank of specimens collected from the Paraíba do Sul River basin were also divided into two groups, of which one may be identified as AltoPR. Since the other group provided an intermediate distance when compared to AltoPR-D, an in-depth investigation was required. The other species analyzed showed a greater distance and was revealed to be a monophyletic taxon. The results suggested that they are really two species and that neither corresponds to the other species used in the current study.
Assuntos
Characidae/classificação , Characidae/genética , Animais , Sequência de Bases , Brasil , DNA Mitocondrial , Enganação , Evolução Molecular , Geografia , Haplótipos , Mutação INDEL , Dados de Sequência Molecular , Filogenia , Polimorfismo de Nucleotídeo Único , RiosRESUMO
The objectives of this study were to standardize a PCR-RFLP genotyping method for the AY_731081:g.1900T>C SNP of the equine CD14 gene, and to characterize this SNP and two other polymorphisms (AY_005808: c.1530A>G of the TLR4 gene and AX_463789: g.133T>C of the Cε gene) in Mangalarga horses, in order to contribute to future studies investigating the association between DNA markers and traits related to immune system physiology in this breed. A total of 151 Mangalarga horses of both sexes and variable ages, representative of the population of São Paulo State, were used. PCR-RFLP was found to be adequate for genotyping of the AY_731081: g.1900T>C SNP of the equine CD14 gene. However, this polymorphism is probably not present in Mangalarga horses, thus impairing association studies using this marker in the breed. The population genetic parameters obtained for the TLR4 AY_005808:c.1530A>G and Cε AX_463789:g.133T>C polymorphisms suggest the use of these markers in association studies with immune system-related traits in Mangalarga horses.
Os objetivos deste trabalho foram a padronização da metodologia PCR-RFLP para genotipagem do SNP AY_731081:g.1900T>C do gene CD14 equino, bem como a caracterização em equinos da raça brasileira Mangalarga deste e de outros dois polimorfismos, o AY_005808: c.1530A>G do TLR4 e o AX_463789: g.133T>C do Cε, a fim de promover o embasamento necessário para futuras pesquisas visando à associação entre marcadores de DNA e características relacionadas à fisiologia do sistema imune na raça. Para tanto, foram utilizados 151 animais Mangalarga, de ambos os sexos e de idades variadas, representativos da população do estado de São Paulo. O método de PCR-RFLP mostrou-se adequado para a genotipagem do SNP AY_731081: g.1900T>C do gene CD14 equino. Entretanto, tal polimorfismo provavelmente não ocorre em equinos Mangalarga, impossibilitando estudos de associação com o marcador na raça. Os parâmetros genético-populacionais obtidos para os polimorfismos AY_005808:c.1530A>G do gene TLR4 e o AX_463789:g.133T>C do gene Cε demonstraram a possibilidade de realização de pesquisas.
Assuntos
Animais , Cavalos/genética , Polimorfismo Genético , Técnicas de Genotipagem/veterinária , Anotação de Sequência Molecular/métodos , Reação em Cadeia da Polimerase/veterináriaRESUMO
The genetic divergence among invasive and native populations of Plagioscion squamosissimus from four Neotropical hydrographic basins was assessed using the hypervariable domain of the mitochondrial DNA (mtDNA) control region. Plagioscion squamosissimus is native to the neighbouring hydrographic basins of the Parnaíba and Amazon Rivers, and the latter includes the Araguaia-Tocantins drainage, but it is invasive in other basins due to introductions. The mtDNA nucleotide polymorphism supported the hypothesis that the Amazon and Parnaíba populations constitute the same species and are separated into two independent evolutionary lineages. Absence of nucleotide polymorphism was observed within and among P. squamosissimus populations invasive to the uppper and middle Paraná River basins. Nucleotide divergence was null or low comparing the Paraná invasive populations with the populations native to the Parnaíba River basin, whereas it was significantly high compared to Tocantins populations. These results ascertain that P. squamosissimus populations invasive to the upper Paraná River basin and to the middle Paraná River basin downstream of the Itaipu dam are derived from the Parnaíba River basin. The genetic data presented are potentially useful to assist further studies on P. squamosissimus taxonomic and geographic distribution, development of ecological guidelines for managing populations invasive to the upper Paraná River basin and for preservation of native fish diversity.
Assuntos
Variação Genética , Espécies Introduzidas , Perciformes/genética , Animais , DNA Mitocondrial/genética , Deriva Genética , Haplótipos , Perciformes/classificação , Filogenia , Rios , América do Sul , Clima TropicalRESUMO
The morphological discrimination between the species Astyanax altiparanae and A. asuncionensis of the upper Paraná River and Paraguay River basins, respectively, has always been difficult. Two D-loop haplogroups of A. altiparanae are known, one with the presence (AltoPR) or the absence (AltoPR-D) of a 32-bp block similar to that in A. asuncionensis. We examined these samples to characterize A. altiparanae and verify whether A. asuncionensis occurred in the upper Paraná River prior to the submergence of the Sete Quedas Falls when Itaipu reservoir was impounded. D-loop sequences were analyzed in A. altiparanae of the upper Paraná and Iguaçu Rivers and those of A. asuncionensis of the upper Paraguay River. The haplogroup AltoPR was found at all sites of the upper Paraná and Iguaçu Rivers, whereas AltoPR-D occurred in the Itaipu reservoir, floodplain and in the Tietê and Grande Rivers. Two haplogroups of A. asuncionensis were identified and both did not have the 32-bp block. However, AltoPR and AltoPR-D differed from one another in 5.1% of their bases and between 8.9 and 12.5% with regard to the haplogroups of the upper Paraguay basin. Further, AltoPR-D occurred in the Grande River upstream the Marimbondo Falls and other older reservoirs than Itaipu. The results reject the hypothesis of the establishment of A. asuncionensis and suggest that the haplogroup AltoPR-D existed in the upper Paraná River before the impounding of the Itaipu reservoir. Moreover, morphological similarity and high genetic variation within the altiparanae/asuncionensis group suggest the existence of a cryptic species complex.
Assuntos
Characidae/genética , DNA Mitocondrial/genética , Animais , Sequência de Bases , Brasil , Primers do DNA , Evolução Molecular , Haplótipos , Dados de Sequência Molecular , Polimorfismo Genético , Homologia de Sequência do Ácido NucleicoRESUMO
Brycon is one of the main genera of Neotropical freshwater fish. In Brazil, Brycon species have been found in many hydrographic basins, such as the Amazon, Paraná, Paraguay, and Araguaia-Tocantins basins. We examined the phylogenetic relationships among the species Brycon orbignyanus, B. hilarii, B. cf. pesu, B. cephalus, B. falcatus, and B. gouldingi, using mitochondrial and nuclear molecular markers. Specimens of B. orbignyanus were collected in the Paraná River. Specimens of B. hilarii were collected in the Manso River. Specimens of B. cephalus were obtained from a fish farm, and specimens of B. cf. pesu, B. falcatus and B. gouldingi were sampled in the Araguaia-Tocantins basin. DNA extraction was carried out using the phenol/chloroform method. Molecular polymorphism studies of Brycon species were carried out with the inter-simple sequence repeat (ISSR) technique, using the total DNA of six specimens of each species. In DNA amplification of B. cf. pesu, eight specimens were used. The partial sequence of mitochondrial cytochrome b was amplified by PCR. The PCR products were used directly in sequencing reactions. Each ISSR primer produced from 7 to 14 scorable and reproducible bands. The (GGAC)(3)A and (GGAC)(3)C primers produced the greatest number of species-specific bands. A 264-bp fragment, corresponding to the partial region of mitochondrial DNA cytochrome b, was sequenced and used for analysis. According to the phylogenetic tree obtained from the data, these Brycon species can be divided into two clades: one comprised only B. cf. pesu, and the second comprised the remaining Brycon species. We conclude that ISSR primers can be used for the identification of species-specific bands in fish, such as Brycon spp.
Assuntos
Ecossistema , Peixes/genética , Água Doce , Filogenia , Animais , Sequência de Bases , Brasil , Citocromos b/genética , DNA Mitocondrial/genética , Eletroforese em Gel de Ágar , Geografia , Repetições Minissatélites/genética , Dados de Sequência Molecular , Análise de Componente Principal , Especificidade da EspécieRESUMO
Pseudoplatystoma corruscans (Spix and Agassiz, 1829) and Pseudoplatystoma reticulatum (Eingenmann and Eigenmann, 1889) are large migratory catfishes of high biological importance and great commercial value in South America. Because fertile crossbreeds can be artificially produced in hatcheries, a high genetic proximity between these two Pimelodidae species is conceivable. Possible escape of crossbred specimens from pisciculture stations is a serious environmental concern. Despite their importance, knowledge of P. corruscans and P. reticulatum biology, ecology, population diversity and genetics is limited. In the present work, the genetic divergence between P. corruscans and P. reticulatum populations from the Paraná River Basin was analyzed on the basis of polymorphisms in ISSR fragments and in the hypervariable sequence of the mitochondrial DNA (mtDNA) control region. Estimates of intraspecific haplotype (h > 0.5) and nucleotide diversities (pi < 0.01) indicate that P. corruscans and P. reticulatum have survived a historical population decline, followed by a demographic expansion. The interspecific polymorphisms within the mtDNA control region and ISSR fragments were suitable as diagnostic molecular markers and could be used to discriminate the two species. A unique Pseudoplatystoma specimen, captured in the Upper Paraná River Floodplain, was identified by these DNA diagnostic markers as a hybrid P. reticulatum x P. corruscans, which possibly escaped from pisciculture. The integrity of the natural population of P. corruscans in the Upper Paraná River is at risk of genetic introgression or homogenization due to the presence of hybrids and the transposition of P. reticulatum upstream through the Canal da Piracema at Itaipu Dam. Data presented herein improve the understanding of the genetic relatedness between P. corruscans and P. reticulatum and represent potential tools for future programs of conservation and surveillance of genetic introgression events and the genetic integrity of these populations.
Assuntos
Peixes-Gato/genética , DNA Mitocondrial/genética , Animais , Genética Populacional , Reação em Cadeia da Polimerase , Polimorfismo Genético/genética , RiosRESUMO
Pseudoplatystoma corruscans (Spix and Agassiz, 1829) and Pseudoplatystoma reticulatum (Eingenmann and Eigenmann, 1889) are large migratory catfishes of high biological importance and great commercial value in South America. Because fertile crossbreeds can be artificially produced in hatcheries, a high genetic proximity between these two Pimelodidae species is conceivable. Possible escape of crossbred specimens from pisciculture stations is a serious environmental concern. Despite their importance, knowledge of P. corruscans and P. reticulatum biology, ecology, population diversity and genetics is limited. In the present work, the genetic divergence between P. corruscans and P. reticulatum populations from the Paraná River Basin was analyzed on the basis of polymorphisms in ISSR fragments and in the hypervariable sequence of the mitochondrial DNA (mtDNA) control region. Estimates of intraspecific haplotype (h > 0.5) and nucleotide diversities (π < 0.01) indicate that P. corruscans and P. reticulatum have survived a historical population decline, followed by a demographic expansion. The interspecific polymorphisms within the mtDNA control region and ISSR fragments were suitable as diagnostic molecular markers and could be used to discriminate the two species. A unique Pseudoplatystoma specimen, captured in the Upper Paraná River Floodplain, was identified by these DNA diagnostic markers as a hybrid P. reticulatum x P. corruscans, which possibly escaped from pisciculture. The integrity of the natural population of P. corruscans in the Upper Paraná River is at risk of genetic introgression or homogenization due to the presence of hybrids and the transposition of P. reticulatum upstream through the Canal da Piracema at Itaipu Dam. Data presented herein improve the understanding of the genetic relatedness between P. corruscans and P. reticulatum and represent potential tools for future programs of conservation and surveillance of genetic ...
Pseudoplatystoma corruscans Spix e Agassiz, 1829 e Pseudoplatystoma reticulatum Eigenmann e Eigenmann, 1889 são peixes migratórios de grande porte, com alta importância biológica e elevado valor comercial na América do Sul. Híbridos férteis são obtidos em cativeiro e, portanto, é esperada alta proximidade genética entre essas duas espécies de Pimelodidae. Escapes de espécimes híbridos a partir de estações de piscicultura representam um sério problema ambiental. Apesar da sua importância, conhecimentos sobre a biologia, ecologia, diversidade de populações e genética de P. corruscans e P. reticulatum são escassos. No presente trabalho, foi avaliada a divergência genética entre P. corruscans e P. reticulatum da Bacia do Rio Paraná, com base em fragmentos ISSR e na seqüência D-loop do DNA mitocondrial (mtDNA). As estimativas das diversidades intra-específicas haplotípica (h > 0,5) e nucleotídica (π < 0,01) evidenciaram que P. corruscans e P. reticulatum sobreviveram a um declínio populacional histórico, seguido de expansão demográfica. Os polimorfismos interespecíficos no mtDNA e nos fragmentos ISSR foram eficientes para diagnósticos e discriminaram as duas espécies. Um espécime de Pseudoplatystoma capturado na planície de inundação do Alto Rio Paraná foi identificado com esses marcadores moleculares como híbrido P. reticulatum x P. corruscans, que possivelmente escapou de psicicultura. A integridade da população de P. corruscans no Alto Rio Paraná está ameaçada, por introgressão ou homogeneização genética, pela presença de híbridos e pela transposição para montante de P. reticulatum através do Canal da Piracema em Itaipu. Os dados apresentados constituem um avanço na compreensão do parentesco entre P. corruscans e P. reticulatum e representam ferramentas em potencial para programas de conservação biológica, incluindo o monitoramento de introgressão e de integridade genética das populações.
