Abstract Centromochlinae is a widely diverse subfamily with more than 50 species and several taxonomic conflicts due to morphological similarity between Tatia and Centromochlus species. However, cytogenetic studies on this group have been limited to only four species so far. Therefore, here we present the karyotype of Centromochlus schultzi from the Xingu River in Brazil using classic cytogenetic techniques, physical mapping of the 5S and 18S rDNAs, and telomeric sequences (TTAGGG)n. The species had 58 chromosomes, simple NORs and 18S rDNA sites. Heterochromatic regions were detected on the terminal position of most chromosomes, including pericentromeric and centromeric blocks that correspond to interstitial telomeric sites. The 5S rDNA had multiple sites, including a synteny with the 18S rDNA in the pair 24st, which is an ancestral feature for Doradidae, sister group of Auchenipteridae, but appears to be a homoplastic trait in this species. So far, C. schultzi is only the second species within Centromochlus to be karyotyped, but it has already presented characteristics with great potential to assist in future discussions on taxonomic issues in the subfamily Centromochlinae, including the first synteny between rDNAs in Auchenipteridae and also the presence of heterochromatic ITSs that could represent remnants of ancient chromosomal fusions. 5 However far Therefore techniques S TTAGGGn. TTAGGGn TTAGGG n. n (TTAGGG)n chromosomes sites 24st st Doradidae C karyotyped fusions

Abstract The tribe Serrasalmini is a diverse group with paraphyletic genera and taxonomic uncertainties. Several studies have been carried out in this group of fish in order to understand this problem, including the cytogenetic approach. In this study, three species of a clade of Serrasalmini were characterized cytogenetically - Pristobrycon striolatus, Catoprion absconditus and Pygopristis denticulatus. The three species presented diploid number (2n) equal to 62 chromosomes, of one and two arms, with karyotypic formulas and species-specific fundamental numbers. Heterochromatin is centromeric and terminal (bi-telomeric) in most chromosomes, with a conspicuous interstitial block at pair 1 (m) in all three species. The nucleolar organizer regions were multiple and C-band positive, and their location was confirmed via 18S ribosomal DNA mapping; however, with additional sites. The 5S rDNA was located in interstitial region of long arm of pair 1 (m), in the three species (homeologous). Moreover, we observed synteny between 18S and 5S in the species C. absconditus and P. denticulatus, which, according to fiber-FISH, are interspersed. Thus, the maintenance of 2n (62) evidences the diversification of chromosomal formulas within the clade by non-Robertsonian rearrangements and reflects the paraphyly of the related species. uncertainties problem approach study striolatus denticulatus n (2n 6 chromosomes arms speciesspecific specific numbers bitelomeric bi telomeric (bi-telomeric m (m Cband C band positive S mapping however sites m, , homeologous. homeologous . (homeologous) Moreover P which fiberFISH, fiberFISH fiber FISH, FISH fiber-FISH interspersed Thus (62 nonRobertsonian non Robertsonian (homeologous (6 (

Abstract The transposable elements are known by their ability to move and integrate into the genome of the host organism. They are classified in retrotransposons class I, which has, as intermediate of the transposition, the RNA and retrotransposons class II, which is composed of the DNA transposons, whose DNA migrates directly or is copied and inserted into the genome. The retrotransposable element Rex1 is a non-LTR retrotransposon found in several types of organisms. Many studies indicate that the Rex retrotransposons have the capacity to respond to environmental stress. The results found in this study corroborate the hypothesis that this retrotransposon possesses a response to environmental stress, since the chromosomal mapping, obtained through FISH, showed a higher number of markings on animals that were submitted to stress in 48h by the copper sulfate action. Furthermore, we can identify an increase in heterochromatic regions in the chromosomes. In the absolute quantification by Real-Time PCR, we found the largest numbers of Rex1 copies in the samples that were submitted to environmental stress by copper sulfate. organism I has transposition II transposons nonLTR non LTR organisms mapping FISH h action Furthermore chromosomes RealTime Real Time PCR

Abstract Fishes of the Curimatidae family represent one of the most important freshwater ichthyofauna groups of Central and South America, with 117 recognized species distributed in eight genera. In this study, six species - Curimata inornata, Curimatella dorsalis, and Psectrogaster falcata collected from the Lower Araguaia River, Pará, Brazil; Curimata vittata, Curimatella meyeri, and Psectrogaster rutiloides collected from the Catalão Lake, Amazonas, Brazil - were cytogenetically analyzed, investigate the occurrence and distribution of repetitive DNA classes in the karyotypes. All species had 2n=54 metacentric/submetacentric chromosomes. Despite the conservative diploid number, we observed variations in the karyotypic structure among species. Ribosomal DNA (rDNA) 18S and 5S were found in single or multiple sites, with the first report of synteny in Curimatella dorsalis, and the occurrence of several interstitial telomeric sequences (ITSs) in species of the genera Curimatella and Psectrogaster. Interspecific karyotypic diversity both concerning structure and location/position of the nucleolar organizer regions (NOR) and ribosomal DNA, suggesting the occurrence of several non-Robertsonian rearrangements driving the evolution of this family.

RESUMO O DNA barcoding propõe que um fragmento de DNA possa servir para identificar espécies. Em peixes, um fragmento do gene COI tem se mostrado eficaz em muitos estudos com focos diferentes. Nós usamos essa ferramenta molecular para fornecer novas informações sobre a diversidade críptica encontrada no complexo de espécies Hoplias malabaricus. Popularmente conhecida como traíra, H. malabaricus tem uma ampla distribuição na América do Sul. Esse clado mostra diversidade molecular e citogenética, e vários estudos dão suporte à ocorrência de um complexo de espécies. Realizamos análises molecular e cariotípica em indivíduos de H. malabaricus de oito localidades amazônicas, para acessar a diversidade no taxon nominal e elucidar as relações nesse grupo. Usamos 12 amostras em análises citogenéticas e encontramos dois cariomorfos: 2n = 40 (20m + 20sm) (cariomorfo C) e 2n = 42 (22m + 20sm) (cariomorfo A). Usamos 19 amostras em análise molecular, utilizando COI como marcador molecular, análises de máxima verossimilhança e o modelo evolutivo de Kimura-2-parâmetros com estimativa de bootstrap. Encontramos diferenciação relacionada aos cariomorfos com bootstrap de 100%. No entanto, encontramos alta diversidade molecular no cariomorfo C. O padrão observado nos permitiu inferir a presença de diversidade oculta, reforçando a existência de um complexo de espécies.

ABSTRACT DNA barcoding proposes that a fragment of DNA can be used to identify species. In fish, a fragment of cytochrome oxidase subunit I (COI) has been effective in many studies with different foci. Here we use this molecular tool to provide new insights into the cryptic diversity found in the Hoplias malabaricus species complex. Popularly known as trahira, H. malabaricus is widely distributed in South America. The clade shows molecular and cytogenetic diversity, and several studies have supported the occurrence of a complex of species. We performed molecular and karyotypic analysis of H. malabaricus individuals from eight Amazonian localities to assess the diversity present in the nominal taxon, and to clarify relationships within this group. We used 12 samples in cytogenetic analyses and found two karyomorphs: 2n = 40 (20m + 20sm) (karyomorph C) and 2n = 42 (22m + 20sm) (karyomorph A). We used 19 samples in molecular analyses with COI as a molecular marker, maximum likelihood analyses, and the Kimura-2-parameter evolutionary model with bootstrap support. We found karyomorph-related differentiation with bootstrap of 100%. However, we found high molecular diversity within karyomorph C. The observed pattern allowed us to infer the presence of cryptic diversity, reinforcing the existence of a species complex.

Abstract Doradinae (Siluriformes: Doradidae) is the most species-rich subfamily among thorny catfishes, encompassing over 77 valid species, found mainly in Amazon and Platina hydrographic basins. Here, we analyzed seven Doradinae species using combined methods (e.g., cytogenetic tools and Mesquite ancestral reconstruction software) in order to scrutinize the processes that mediated the karyotype diversification in this subfamily. Our ancestral reconstruction recovered that 2n=58 chromosomes and simple nucleolar organizer regions (NOR) are ancestral features only for Wertheimerinae and the most clades of Doradinae. Some exceptions were found in Trachydoras paraguayensis (2n=56), Trachydoras steindachneri (2n=60), Ossancora punctata (2n=66) and Platydoras hancockii whose karyotypes showed a multiple NOR system. The large thorny catfishes, such as Pterodoras granulosus, Oxydoras niger and Centrodoras brachiatus share several karyotype features, with subtle variations only regarding their heterochromatin distribution. On the other hand, a remarkable karyotypic variability has been reported in the fimbriate barbells thorny catfishes. These two contrasting karyoevolution trajectories emerged from a complex interaction between chromosome rearrangements (e.g., inversions and Robertsonian translocations) and mechanisms of heterochromatin dispersion. Moreover, we believe that biological features, such as microhabitats preferences, populational size, low vagility and migratory behavior played a key role during the origin and maintenance of chromosome diversity in Doradinae subfamily.

Abstract In recent decades the diploid numbers recorded in the New World marsupials have been widely discussed in the context of the processes of karyotype evolution in these mammals. While Interstitial Telomeric Sequences (ITS) have long been interpreted as remnants of chromosomal fusion, the biological role of these features, together with their intraspecific variation, has raised a number of questions. In the present study, we analyzed the karyotype of 11 species of Amazonian didelphids, comparing the distribution of the heterochromatin with that of the telomeric signals, and found that, in six species, the ITS coincided with the blocks of heterochromatin. While ITS were found in the X chromosomes of all Marmosa murina individuals, they were variable in all the other species, representing a specific character of each lineage. Our results support the conclusion that ITS may not always be a consequence of chromosomal rearrangements, and that the mechanisms that produce them are still unclear.

Abstract We present new karyotype records for six Proechimys species from the Brazilian Amazon. P. echinothrix from the region of Purus River had 2n = 32 chromosomes and a FN = 58, while P. cuvieri from the region of the Japurá River presented 2n = 28 and FN = 46. All individuals presented hybridization with an 18S rDNA probe in a single chromosome pair, with the exception of P. cuvieri from the Japurá region, which presented a third signal in one of the homologs of pair 1. No ITS were found in any of the individuals. Our data supports the hypothesis that the P. cuvieri population from the Japurá Basin and P. echinothrix from the lower Purus are new taxonomic entities. Our data expand the geographic distribution of the cytotype (2n = 40, FN = 54) described for P. gardneri from the Madeira River, and the cytotype (2n = 46, FN = 50), described for P. guyannensis, as well as the recently-described cytotype of P. goeldii (2n = 16, FN = 14). No clear pattern of chromosomal evolution has yet been defined in Proechimys, despite the considerable karyotypic diversity of the genus.

Resumo Os peixes gigantes da América do Sul do gêneroArapaima, comumente conhecidos como pirarucus, são um dos mais icônicos de Osteoglossiformes. Estudos citogenéticos prévios identificaram suas características cariotípicas, entretanto a caracterização da diferenciação citotaxonômica através de suas distribuições geográficas ainda são desconhecidas. Nesse estudo, nós comparamos características cromossômicas utilizando técnicas de citogenética clássica e molecular em populações das bacias dos rios Amazonas e Tocantins-Araguaia, a fim de verificar se há alguma diferenciação entre representantes desse gênero. Nossos dados revelaram que indivíduos de todas as populações apresentam número diploide de 2n=56 cromossomos e que seus cariótipos são compostos de 14 pares de cromossomos meta- e submetacêntricos e 14 pares de subtelo- e acrocêntricos. Os sítios maiores e menores de rDNA estão localizados em pares cromossômicos separados, onde os sítios maiores de rDNA correspondem a grandes blocos heterocromáticos. Hibridizações genômicas comparativas (CGH) mostraram que o genoma dos espécimes dessas populações é amplamente compartilhado, devido à falta de sinais substanciais específicos. Nossos dados de citogenética comparativa do pirarucu sugerem que embora diferenças genéticas significativas ocorram entre populações, os padrões cariotípicos gerais se mantêm conservados.

Abstract The South American giant fishes of the genus Arapaima, commonly known as pirarucu, are one of the most iconic among Osteoglossiformes. Previously cytogenetic studies have identified their karyotype characteristics; however, characterization of cytotaxonomic differentiation across their distribution range remains unknown. In this study, we compared chromosomal characteristics using conventional and molecular cytogenetic protocols in pirarucu populations from the Amazon and Tocantins-Araguaia river basins to verify if there is differentiation among representatives of this genus. Our data revealed that individuals from all populations present the same diploid chromosome number 2n=56 and karyotype composed of 14 pairs of meta- to submetacentric and 14 pairs of subtelo- to acrocentric chromosomes. The minor and major rDNA sites are in separate chromosomal pairs, in which major rDNA sites corresponds to large heterochromatic blocks. Comparative genomic hybridizations (CGH) showed that the genome of these populations shared a great portion of repetitive elements, due to a lack of substantial specific signals. Our comparative cytogenetic data analysis of pirarucu suggested that, although significant genetic differences occur among populations, their general karyotype patterns remain conserved.

RESUMO Historicamente, há divergências na alocação de espécies entre Centromochlus e Tatia. Este estudo teve como objetivo gerar os primeiros dados citogenéticos para Centromochlus e, através da análise de uma população de Centromochlus heckelii da bacia do rio Amazonas, contribuir como evidência para o dilema histórico taxonômico. Foi encontrado o número diploide de 46 cromossomos e um par heteromórfico nos cariótipos das fêmeas, o que caracteriza um sistema sexual ZZ/ZW. Blocos pálidos de heterocromatina foram localizados na região centromérica de alguns cromossomos; no entanto, o cromossomo exclusivo das fêmeas (W) se apresenta quase todo heterocromático. As AgRONs foram detectadas na posição terminal do braço curto de um par acrocêntrico nos machos e em dois pares cromossômicos nas fêmeas, um par de cromossomos acrocêntricos e o par sexual. Notáveis diferenças entre os dados cromossômicos de Centromochlus heckelii e os dados anteriores das espécies de Tatia são: menor número diploide, presença de sistema de cromossomos sexuais e AgRONs múltiplas em Centromochlus, enquanto espécies de Tatia apresentam AgRON simples e ausência de cromossomos acrocêntricos. Resultados deste estudo mostram que marcadores cromossômicos podem contribuir como evidência para estudos de delimitação taxonômica.

ABSTRACT Historically, there are divergences in the species allocation between Centromochlus and Tatia. This study aimed to generate the first cytogenetic data about Centromochlus and, by analyzing a population of Centromochlus heckelii from the Amazon River basin, to contribute as evidence to a historical taxonomic dilemma. Diploid number of 46 chromosomes and a heteromorphic pair was found in the female karyotypes, thus characterizing a ZZ/ZW sex chromosome system. Pale blocks of heterochromatin were located in centromeric regions of some chromosomes; however, the exclusive female chromosome (W) is almost entirely heterochromatic. AgNORs were detected in terminal position on the short arms of one acrocentric pair in males and two chromosome pairs in females, the acrocentric plus the sex chromosome pair. Notable differences between Centromochlus heckelii and previous data about species of Tatia are: lower diploid number, presence of a sex chromosome system and multiple AgNORs in Centromochlus, while species of Tatia have simple AgNORs and the absence of acrocentric chromosomes. Results in this study show that chromosomal markers could contribute as evidence to taxonomic delimitation studies.

Abstract Cytogenetic studies in the subfamily Potamotrygoninae have provided valuable insights into the understanding of the evolution and diversification of its species. In the present study, the chromosomal features of seven nominal potamotrygonin species are provided: Plesiotrygon iwamae (2n=74, FN=120), Potamotrygon amazona (2n=66, FN=107), P. constellata (2n=66, FN=110), P. leopoldi (2n=64, FN=102), P. motoro (2n=66, FN=106) from four different localities, and P. orbignyi (2n=66, FN=106), P. scobina (2n=66, FN=104), from Central Amazon. Additionally, we found a new karyomorph in P. wallacei. We considered the localization of Nucleolus Organizer Regions (NORs), as well as the pattern of constitutive heterochromatin, as species-specific characters. We found an XX/XY sex chromosome system in P. orbignyi, and we suggest that P. scobina and P. amazona also possess the same sex chromosome system. Overall, the chromosomal evolution in this group appears to have progressed towards a reduction in diploid number, with a concomitant increase in the number of bi-armed and nucleolar chromosomes.

RESUMO Farlowella é um dos gêneros mais diversos de Loricariinae, restrito aos rios da América do Sul. As relações taxonômicas e filogenéticas entre suas espécies são contenciosas e, enquanto os estudos genéticos contribuem para a compreensão dessas relações, o único dado disponível refere-se à descrição cariotípica de apenas uma espécie. No presente estudo, foram analizadas duas espécies amazônicas Farlowella cf. amazonum e F. schreitmuelleri, empregando procedimentos citogenéticos convencionais e moleculares. Ambas as espécies apresentaram número diploide igual a 58 cromossomos, mas com números fundamentais diferentes (NF) de 116 e 112, respectivamente, indicando rearranjos cromossômicos. Bandas C são poucas, especialmente em F. cf. amazonum, e ocorre predominantemente nas regiões centroméricas e em algumas regiões teloméricas, embora F. schreitmuelleri apresenta uma quantidade de heterocromatina muito maior que F. cf. amazonum. Os cromossomos carreadores dos sítios da NOR foram provavelmente os mesmos para ambas as espécies, correspondendo ao primeiro par metacêntrico em F. cf. amazonum e ao 28º acrocêntrico em F. schreitmuelleri. A localização do DNAr 5S foi espécie-específico. Este estudo expandiu os dados citogenéticos disponíveis para espécies de Farlowella e apontou uma remarcável diversidade cromossômica entre espécies/populações, indicando um possível complexo de espécies neste gênero.

ABSTRACT Farlowella is one of the most diverse genera of the Loricariinae, restricted to South America rivers. The taxonomic and phylogenetic relationships among its species are contentious and, while genetic studies would contribute to the understanding of their relationships, the only available datum refer to the karyotype description of only one species. In the present study two Amazonian species, Farlowella cf. amazonum and F. schreitmuelleri, were analyzed using conventional and molecular cytogenetic procedures. Both species had diploid chromosome number 58, but different fundamental numbers (NF) 116 and 112, respectively, indicative of chromosomal rearrangements. C-banding is almost poor, especially in F. cf. amazonum, and occurs predominantly in the centromeric and in some telomeric regions, although genome of F. schreitmuelleri possessed a much larger heterochromatin amount then those of F. cf. amazonum. The chromosomes bearing the NOR sites were likely the same for both species, corresponding to the 1st metacentric pair in F. cf. amazonum and to the 28th acrocentric in F. schreitmuelleri. The location of the 5S rDNA was species-specific marker. This study expanded the available cytogenetic data for Farlowella species and pointed the remarkable karyotype diversity among species/populations, indicating a possible species complex within genus.

Abstract Hoplias malabaricus is a common fish species occurring in white, black and clear water rivers of the Amazon basin. Its large distribution across distinct aquatic environments can pose stressful conditions for dispersal and creates possibilities for the emergence of local adaptive profiles. We investigated the chromosomal localization of repetitive DNA markers (constitutive heterochromatin, rDNA and the transposable element REX-3) in populations from the Amazonas river (white water), the Negro river (black water) and the Tapajós river (clear water), in order to address the variation/association of cytogenomic features and environmental conditions. We found a conserved karyotypic macrostructure with a diploid number of 40 chromosomes (20 metacentrics + 20 submetacentrics) in all the samples. Heteromorphism in pair 14 was detected as evidence for the initial differentiation of an XX/XY system. Minor differences detected in the amount of repetitive DNA markers are interpreted as possible signatures of local adaptations to distinct aquatic environments.

RESUMO Baryancistrus xanthellus é uma espécie da tribo Ancistrini conhecida popularmente como "amarelinho" ou "cascudo pepita de ouro". É um dos peixes ornamentais mais populares e valorizados, devido aos padrões de cor. Também é uma espécie endêmica do rio Xingu, ocorrendo a partir da Volta Grande do Xingu, região onde a Usina Hidrelétrica de Belo Monte está sendo construída, até São Félix do Xingu. O presente estudo teve como objetivo caracterizar citogeneticamente B. xanthellus . Os resultados apontam para a manutenção do 2n=52, considerado a condição mais comum para a tribo, e região organizadora de nucléolo (RON) simples. O mapeamento do DNAr 18S confirmou a marcação da RON e o DNAr 5S foi localizado na posição intersticial de apenas um par cromossômico. A localização dos DNAr 18S e 5S em diferentes pares configura uma apomorfia em Loricariidae. Grandes blocos de heterocromatina estão presentes nos pares 1 e 10 e nas regiões equivalentes à RON e ao DNAr 5S. Os dados obtidos neste estudo corroboram a hipótese filogenética atualmente mais aceita para Ancistrini e demonstram evidências que o gênero Baryancistrus ocupa uma posição basal na tribo.

ABSTRACT Baryancistrus xanthellus is a species from the Ancistrini tribe known commonly as "amarelinho " or "golden nugget pleco". It is one of the most popular and valued ornamental fishes due to its color pattern. Also, it is an endemic species from the Xingu River occurring from Volta Grande do Xingu, region where the Belo Monte Hydropower Dam is being built, to São Félix do Xingu. The current study aimed to cytogenetically characterize B. xanthellus . Results point to the maintenance of 2n=52, which is considered the most common condition for the tribe, and a single nucleolus organizer region (NOR). Mapping of the 18S rDNA confirmed the NOR sites, and the 5S rDNA was mapped in the interstitial position of a single chromosome pair. The 18S and 5S rDNA located in different pairs constitute an apomorphy in Loricariidae. Large blocks of heterochromatin are present in pairs 1 and 10 and in the regions equivalent to NOR and the 5S rDNA. Data obtained in this study corroborated with the currently accepted phylogenetic hypothesis for the Ancistrini and demonstrate evidence that the genus Baryancistrus occupies a basal position in the tribe.

The Serrasalmidae family is composed of a number of commercially interesting species, mainly in the Amazon region where most of these fishes occur. In the present study, we investigated the genomic organization of the 18S and 5S rDNA and telomeric sequences in mitotic chromosomes of four species from the basal clade of the Serrasalmidae family: Colossoma macropomum, Mylossoma aureum, M. duriventre, and Piaractus mesopotamicus, in order to understand the chromosomal evolution in the family. All the species studied had diploid numbers 2n = 54 and exclusively biarmed chromosomes, but variations of the karyotypic formulas were observed. C-banding resulted in similar patterns among the analyzed species, with heterochromatic blocks mainly present in centromeric regions. The 18S rDNA mapping of C. macropomum and P. mesopotamicus revealed multiple sites of this gene; 5S rDNA sites were detected in two chromosome pairs in all species, although not all of them were homeologs. Hybridization with a telomeric probe revealed signals in the terminal portions of chromosomes in all the species and an interstitial signal was observed in one pair of C. macropomum.