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

Resumo O gênero Tetranematichthys possui apenas três espécies, e nenhuma delas tinha sido submetida a análises citogenéticas. Assim, este estudo traz pela primeira vez a análise de Tetranematichthyswallacei, coletado no Igarapé Apaú, bacia do rio Guamá, cidade de Castanhal, estado do Pará, Brasil. O número diploide de 52 cromossomos (32m+18sm+2st, NF = 104) foi encontrado em ambos os sexos, com heterocromatina predominantemente terminal e algumas intersticiais. A sequência telomérica foi observada exclusivamente em regiões terminais. Os sítios de 18S rDNA foram encontrados no par 17sm de todos os exemplares e em apenas um dos homólogos do par 7 em três exemplares. Os sítios de 5S rDNA foram encontrados nos pares 8m e 10m. Tetranematichthyswallacei possui algumas características que dignas de quanto à sua posição filogenética atual, incluindo uma provável redução no número diploide. Além disso, T.wallacei compartilha com Tympanopleuraatronasus o 18S rDNA alocado no braço longo de um grande cromossomo sm (primeiro par), mas não compartilha com Ageneiosus o primeiro par m grande com evidências de fusão, como observado em Ageneiosus inermis. Os dados cromossômicos gerados para T. wallacei, juntamente com os dados dos outros dois táxons de Ageneiosini estudados anteriormente, reforçam propostas de estudos baseados em morfologia que sugerem que a tribo representa o clado mais diferenciado dentro da família. espécies citogenéticas Assim Apaú Guamá Castanhal Pará Brasil 5 32m+18sm+2st, 32m18sm2st msmst 32m 18sm 2st st (32m+18sm+2st 104 sexos intersticiais terminais S 10m atual disso Twallacei T wallacei par, , par) fusão inermis anteriormente família 32m+18sm+2st 10 1

Abstract The genus Tetranematichthys has only three species, and none of them have undergone cytogenetic analyses. Therefore, this study brings for the first time the analysis of Tetranematichthys wallacei, collected from the Igarapé Apaú, Guamá River basin, municipality of Castanhal, Pará State, Brazil. The diploid number found was 52 chromosomes (32m+18sm+2st, NF = 104), in both sexes, with predominantly terminal and some interstitial heterochromatin. Telomeric sequences were observed exclusively in terminal regions. The 18S rDNA sites were found on pair 17sm of all specimens and in only one of the homologous of pair 7 in three specimens. The 5S rDNA sites were found in pairs 8m and 10m. Tetranematichthys wallacei exhibits characteristics worthy of attention regarding its current phylogenetic position, including a probable diploid number reduction. Additionally, it shares with Tympanopleura atronasus the 18S rDNA allocated in the long arm of a large sm chromosome (first pair) but does not share with Ageneiosus the large first m pair with evidence of fusion, as observed in Ageneiosus inermis. The chromosomal data generated for T. wallacei, along with the data from the other two previously studied Ageneiosini taxa, reinforces proposals from morphology-based studies suggesting that the tribe represents the most distinct clade within the family. species analyses Therefore Apaú basin Castanhal State Brazil 5 32m+18sm+2st, 32m18sm2st msmst 32m 18sm 2st st (32m+18sm+2st 104, 104 , 104) sexes heterochromatin regions S 10m position reduction Additionally fusion inermis T taxa morphologybased morphology based family 32m+18sm+2st 10 1

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.

RESUMO Auchenipteridae é dividida nas subfamílias Centromochlinae e Auchenipterinae. Parauchenipterus encontra-se incluído na última e tem sido alvo de discussões relacionadas com a problemática taxonômica de validação ou sinonimização com Trachelyopterus. Foram analisadas citogeneticamente três espécies de duas bacias hidrográficas: Parauchenipterus striatulus do rio Doce, P. galeatus e Trachelyopterus coriaceus, simpátricas do rio Araguaia. Todas as espécies analisadas apresentaram número diploide de 58 cromossomos, com diferença na fórmula cariotípica de P. striatulus. A heterocromatina foi localizada nas regiões terminais de quase todos os cromossomos e na região pericentromérica nos cromossomos acrocêntricos das três espécies. AgNORs e DNAr 18S detectaram RONs simples em um par de cromossomos subtelocêntricos nas três espécies. DNAr 5S foi detectado em três pares de cromossomos submetacêntricos em P. striatulus; em um par de cromossomos metacêntricos e um par submetacêntrico em T. coriaceus; e em apenas um par de cromossomos metacêntricos em P. galeatus. As semelhanças encontradas nos cariótipos das três espécies analisadas indicam a existência de somente Trachelyopterus (não validação de Parauchenipterus) e a diferença encontrada na distribuição de DNAr 5S de P. galeatus em relação às outras populações já estudadas sugere a existência de uma nova unidade taxonômica, portanto P. galeatus compreende um complexo de espécies.

ABSTRACT Auchenipteridae is divided into subfamilies Centromochlinae and Auchenipterinae. Parauchenipterus is included in the latter and is subject of taxonomic discussions concerning its validation or synonymization with Trachelyopterus. Herein, three species from two hydrographic basins were cytogenetically analyzed: Parauchenipterus striatulus from Doce River and two sympatric species, P. galeatus and Trachelyopterus coriaceus, from the Araguaia River. Diploid number of 58 chromosomes was verified for all species, but P. striatulus has different karyotype formula from the others. The three species have heterochromatin located in terminal regions of almost all chromosomes and in pericentromeric region on acrocentric chromosomes. Simple NORs was verified on a subtelocentric chromosome for all species. 5S rDNA sites were detected in three submetacentric chromosome pairs in P. striatulus; in a metacentric chromosome pair and submetacentric pair in T. coriaceus; and in one metacentric chromosome pair in P. galeatus. The similarities found in the karyotypes of the three species suggest the existence of only one genus, Trachelyopterus; therefore, our data refutes the validation of Parauchenipterus. Moreover, the differences in 5S rDNA distribution in P. galeatus in comparison with other populations already studied, indicate the existence of a new taxonomic unit, which suggests a species complex in P. galeatus.

Abstract The aim of this paper was to present the second case of B chromosomes in Auchenipteridae (Trachelyopterus sp.), and to test the hypothesis that the B chromosomes of this species and Parauchenipterus galeatus might have a common origin, since these two species have phylogenetic proximity. Both species have 58 chromosomes in the A complement, heterochromatin preferentially located at terminal region of the most of chromosomes, simple Ag-NORs located at the short arm of a subtelocentric pair, which was confirmed by hybridization with 18S rDNA, two submetacentric pairs carrying 5S rDNA sites, and presence of B chromosomes. The B chromosomes of the two species are small, metacentric, and almost totally heterochromatic, with variation of number intra and interindividual. In addition, for the first time in fish, the telomeric sequence [TTAGGG]n was dispersed along the B chromosomes (both species). The [GATA]n microsatellite were scattered in all chromosomes of the A complement and absent in the B chromosomes, in both species. These aspects confirm the phylogenetic proximity between the genus Parauchenipterus and Trachelyopterus, and they suggest the hypothesis that the B chromosomes of the two species might have common origin, previous to the diversification of these genera.

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.

Ageneiosus é o gênero da família Auchenipteridae mais amplamente distribuído em bacias da América do Sul. Apesar dos estudos cromossômicos nesta família serem escassos, este gênero tem o maior número de espécies analisadas, com número diploide variando de 54 a 56 cromossomos, o que difere do restante da família (2n = 58). Este estudo objetivou analisar Ageneiosus inermisda bacia do rio Araguaia. O número diploide encontrado foi de 56 cromossomos. A heterocromatina se mostrou localizada na região terminal da maioria dos cromossomos, além de um bloco heterocromático pericentromérico no par 1, um par facilmente distinguível no cariótipo pelo seu maior tamanho quando comparado aos outros pares do complemento. AgRONs foram detectadas em somente um par de cromossomos submetacêntricos, que foi confirmado pela FISH. 5S rDNA se mostrou presente em somente um par de cromossomos metacêntricos. A hibridização com a sequência [TTAGGG]n marcou os telômeros de todos os cromossomos, além de um ITS (sequência telomérica intersticial) na região proximal do braço curto do par 1. A sequência repetitiva [GATA]n se mostrou dispersa, com localização preferencial na região terminal dos cromossomos. Ageneiosus apresenta uma organização genômica um pouco diferente quando comparada a outras espécies de Auchenipteridae. As evidências indicam que uma fusão cromossômica originou o primeiro par de cromossomos metacêntricos de A. inermis, rearranjo que parece ser um evento basal para o gênero.

Ageneiosus is the most widely distributed genus of the family Auchenipteridae among South American river basins. Although chromosome studies in the family are scarce, this genus has the largest number of analyzed species, with 2n = 54 to 56 chromosomes, differing from the rest of the family (2n = 58). This study aimed to analyze Ageneiosus inermis from the Araguaia River basin. The diploid number found was of 56 chromosomes. Heterochromatin was allocated in terminal region of most chromosomes, plus a pericentromeric heterochromatic block in pair 1, a pair distinguished by size in relation to other chromosomes pairs. AgNORs were detected in only one submetacentric chromosome pair, which was confirmed by FISH. 5S rDNA was present in only one metacentric chromosome pair. Hybridization with [TTAGGG]n sequence marked the telomeres of all chromosomes, in addition to an ITS in the proximal region of the short arm of pair 1. The repetitive [GATA]n sequence was dispersed, with preferential location in terminal region of the chromosomes. Ageneiosus has a genomic organization somewhat different when compared to other Auchenipteridae species. Evidences indicate that a chromosomal fusion originated the first metacentric chromosome pair in A. inermis, rearrangement which may be a basal event for the genus

Três populações do grupo Hoplias malabaricus das bacias hidrográficas dos rios São Francisco, Araguaia/Tocantins e Xingu foram analisadas citogeneticamente utilizando-se métodos clássicos (coloração com Giemsa, bandamento-C e Ag-RONs) e moleculares (hibridização in situ fluorescente com sondas de rDNA 18S, rDNA 5S e DNA satélite 5SHindIII). Os marcadores cromossômicos foram fundamentais para a caracterização destas populações como pertencentes ao cariomorfo A e para detecção de claras divergências interpopulacionais. Estas diferenças são provavelmente oriundas de diferentes histórias evolutivas do isolamento geográfico entre as populações associado ao modo dispersivo destes organismos, reiterando a diversidade genética do grupo Hoplias malabaricus.

Three populations of the group Hoplias malabaricus from the hydrographic basins of the São Francisco, Araguaia/Tocantins and Xingu Rivers in Brazil were analyzed using classic cytogenetic methods (Giemsa staining, C-banding and Ag-NORs) and molecular methods (fluorescent in situ hybridization with 18S rDNA, 5S rDNA and 5SHindIII satellite DNA probes). The chromosome markers allowed the characterization of these populations as belonging to karyomorph A and the detection of inter-population divergences. These differences likely stem from different evolutionary histories resulting from geographic isolation between populations associated to the dispersive mode of these organisms, reinforcing genetic diversity in the group Hoplias malabaricus.