Chromosomal diversity in three species of Lycosa Latreille, 1804 (Araneae, Lycosidae): Inferences on diversification of diploid number and sexual chromosome systems in Lycosinae.

Lycosa is one of the most speciose genera in Lycosidae, including species with different sexual chromosome systems (SCS). We carried out cytogenetic analyses in three species of Lycosa, revealing that L. erythrognatha and L. sericovittata share 2n ♂ = 22 and SCS X1X20 while L. gr. nordenskjoldi presents 2n ♂ = 19 and SCS XO, composed only of acrocentric chromosomes. All species shared pericentromeric heterochromatin. Nonetheless, one specimen of L. sericovittata carried two chromosomes with terminal heterochromatin and L. gr. nordenskjoldi showed four chromosomes with interstitial heterochromatin plus another chromosome with terminal C-bands. The pericentromeric heterochromatin of all species as well as the terminal heterochromatic blocks in L. sericovittata were CMA3+. The 18S rDNA sites varied in number and type of bearing chromosomes both at inter and intrapopulational levels, with the highest variation in L. gr. nordenskjoldi. These differences may be related to gene dispersal due to the influence of transposition elements and translocation events. Despite these variations, all species shared ribosomal sites in pair 5. This study demonstrated intra and interspecific chromosomal variability of Lycosa, suggesting that chromosomal rearrangements are related to the diversification of diploid number and SCS in this group of spiders.

Chromosomal diversity in three species of Lycosa Latreille, 1804 (Araneae, Lycosidae): inferences on diversification of diploid number and sexual chromosome systems in Lycosinae

Lycosa is one of the most speciose genera in Lycosidae, including species with different sexual chromosome systems (SCS). We carried out cytogenetic analyses in three species of Lycosa, revealing that L. erythrognatha and L. sericovittata share 2n ♂ = 22 and SCS X1X20 while L. gr. nordenskjoldi presents 2n ♂ = 19 and SCS XO, composed only of acrocentric chromosomes. All species shared pericentromeric heterochromatin. Nonetheless, one specimen of L. sericovittata carried two chromosomes with terminal heterochromatin and L. gr. nordenskjoldi showed four chromosomes with interstitial heterochromatin plus another chromosome with terminal C-bands. The pericentromeric heterochromatin of all species as well as the terminal heterochromatic blocks in L. sericovittata were CMA3+. The 18S rDNA sites varied in number and type of bearing chromosomes both at inter and intrapopulational levels, with the highest variation in L. gr. nordenskjoldi. These differences may be related to gene dispersal due to the influence of transposition elements and translocation events. Despite these variations, all species shared ribosomal sites in pair 5. This study demonstrated intra and interspecific chromosomal variability of Lycosa, suggesting that chromosomal rearrangements are related to the diversification of diploid number and SCS in this group of spiders.

First chromosomal analysis in Deinopidae (Araneae) reveals Sex Chromosome System X1X2X3X4, B chromosomes and polymorphism for centric fusion.

Here we present the first cytogentic study concerning Deinopidae and their controversial phylogenetic position. This study karyologically analyzed one population of Deinopis biaculeata Simon, 1906 and five populations of Deinopis plurituberculata Mello-Leitão, 1925. The majority of specimens of D. plurituberculata exhibited 2n♂ = 40 and 2n♀ = 44 telocentric chromosomes (however some of them showed B chromosomes, belongs to Aquidauana and Botucatu population). The Deinopis biaculeata and D. plurituberculata meiosis of males showed 18 autosomal bivalents + X1X2X3X4, n = 22 and n = 18, a rare sex chromosome system (SCS) in spiders. Some individuals of D. plurituberculata from the Campo Grande population exhibited 2n♂ = 39 and 2n♀ = 43, with a metacentric chromosome (heterozygotes for centric fusion). The D. plurituberculata males with the rearrangement exhibit diplotenes with 16 autosomal bivalents + 1 autosomal trivalent + X1X2X3X4 and metaphases II with n = 22 (18 telocentric autosomes + X1X2X3X4), n = 21 (16 telocentric autosomes + a metacentric autosome + X1X2X3X4), n = 18 (18 telocentric autosomes) and n = 17 (16 telocentric autosomes + a metacentric autosome). The Ag-NORs (silver impregnation) are terminally located in a pair, coinciding with secondary constriction, which is the most common configuration for Araneae. The relatively high diploid number in Deinopis corroborates phylogenies that place it in a basal position among Entelegynes, in the UDOH grade (Uloboridae, Deinopidae, Oecobiidae and Hersiliidae). Centric fusion in only one population of D. plurituberculata suggests low dispersion capacity of this species and an absence of homozygotes for fusion suggests their low viability or a need to increase the population sampling of D. plurituberculata exhibiting the rearrangement. B chromosomes were detected in D. plurituberculata, with interpopulacional, intrapopulacional and intraindividual numerical variation, with cells presenting 0 - 3 and 0 - 6 B chromosomes in populations of Aquidauana and Botucatu, respectively.

First chromosomal analysis in Deinopidae (Araneae) reveals sex Chromosome system X1X2X3X4, B chromosomes and polymorphism for centric fusion

Here we present the first cytogentic study concerning Deinopidae and their controversial phylogenetic position. This study karyologically analyzed one population of Deinopis biaculeata Simon, 1906 and five populations of Deinopis plurituberculata Mello-Leitão, 1925. The majority of specimens of D. plurituberculata exhibited 2n♂ = 40 and 2n♀ = 44 telocentric chromosomes (however some of them showed B chromosomes, belongs to Aquidauana and Botucatu population). The Deinopis biaculeata and D. plurituberculata meiosis of males showed 18 autosomal bivalents + X1X2X3X4, n = 22 and n = 18, a rare sex chromosome system (SCS) in spiders. Some individuals of D. plurituberculata from the Campo Grande population exhibited 2n♂ = 39 and 2n♀ = 43, with a metacentric chromosome (heterozygotes for centric fusion). The D. plurituberculata males with the rearrangement exhibit diplotenes with 16 autosomal bivalents + 1 autosomal trivalent + X1X2X3X4 and metaphases II with n = 22 (18 telocentric autosomes + X1X2X3X4), n = 21 (16 telocentric autosomes + a metacentric autosome + X1X2X3X4), n = 18 (18 telocentric autosomes) and n = 17 (16 telocentric autosomes + a metacentric autosome). The Ag-NORs (silver impregnation) are terminally located in a pair, coinciding with secondary constriction, which is the most common configuration for Araneae. The relatively high diploid number in Deinopis corroborates phylogenies that place it in a basal position among Entelegynes, in the UDOH grade (Uloboridae, Deinopidae, Oecobiidae and Hersiliidae). Centric fusion in only one population of D. plurituberculata suggests low dispersion capacity of this species and an absence of homozygotes for fusion suggests their low viability or a need to increase the population sampling of D. plurituberculata exhibiting the rearrangement. B chromosomes were detected in D. plurituberculata, with interpopulacional, intrapopulacional and intraindividual numerical variation, with cells presenting 0 - 3 and 0 - 6 B chromosomes in populations of Aquidauana and Botucatu, respectively.

Chromosome comparison among five species of Neotropical cichlids of Cichlasoma and Gymnogeophagus (Perciformes).

The genera Cichlasoma and Gymnogeophagus belong to the subfamily Cichlinae, the only one in Neotropical cichlids. Cichlasoma dimerus, C. paranaense, C. portalegrense, Gymnogeophagus rhabdotus, and G. lacustris were collected at different points in the Paranapanema and Paraguay basins and the Lagoon of Patos hydrographic system. In addition to conventional analysis, CMA3 fluorochrome staining, and FISH with 18S rDNA probe were performed. All species had a diploid number equal to 48, with interand intraspecific differences in karyotype formulae. All species presented a single AgNOR site, except G. rhabdotus and the C. paranaense population of the Paranapanema River, which revealed more than one pair of nucleolar chromosomes. AgNORs were coincident to 18S rDNA and CMA3. Heterochromatin was distributed in the pericentromeric chromosomal regions and coincident with NORs. For the first time, this work shows cytogenetic data for C. portalegrense, G. lacustris, and G. rhabdotus. Although some results reinforce the idea of conservative chromosome evolution of 2n in Cichlinae, interspecific and populational variations observed confirm that chromosomal rearrangements affect the microstructural karyotype diversification in this group of fish.

Insights on repetitive DNA behavior in two species of Ctenus Walckenaer, 1805 and Guasuctenus Polotow and Brescovit, 2019 (Araneae, Ctenidae): Evolutionary profile of H3 histone, 18S rRNA genes and heterochromatin distribution.

Ctenidae represents one of the most representative spider families in the tropical forests of Brazil. Its largest genus, Ctenus, has approximately 220 species out of the more than 520 Ctenidae species described, and several authors consider it polyphyletic. Chromosomal data are only available for four species of Ctenus, representing a large gap in the cytogenetic knowledge about the group. This study provided cytogenetic data on two Ctenus species and one Guasuctenus (previously described as Ctenus). All showed 2n♂ = 28 (26+X1X20). Guasuctenus longipes presented two chromosome pairs containing 18S rDNA genes and C. medius, however C. ornatus showed only one chromosome pair with the 18S rDNA gene. Hybridization data using histone H3 probe indicated specific profiles: histone H3 genes were found in one chromosome pair in G. longipes, in three pairs in C. medius, and in four pairs in C. ornatus. Furthermore, supernumerary chromosomes were identified in C. ornatus presenting a meiotic behavior similar to that of sex chromosomes; and a trivalent was found in C. medius, formed by the association of one sex chromosome and an autosomal bivalent, indicating the importance of these events for the diversification of sex chromosomes in spiders. The C-banding pattern was similar between C. medius and C. ornatus with regard to the number and locations of heterochromatic bands, suggesting that heterochromatin amplification and dispersion, affect karyotypic evolution in the genus. Cytogenetic data showed similarity between C. medius and C. ornatus, and differentiation of G. longipes congruent with morphological data. Moreover, although more comparative analyses are needed to specify composition of the dispersed heterochromatin in Ctenus, the mapping of heterochromatic bands provided insights about the evolution of the karyotypes in this genus.

Insights on repetitive DNA behavior in two species of Ctenus Walckenaer, 1805 and Guasuctenus Polotow and Brescovit, 2019 (Araneae, Ctenidae): evolutionary profile of H3 histone, 18S rRNA genes and heterochromatin distribution

Ctenidae represents one of the most representative spider families in the tropical forests of Brazil. Its largest genus, Ctenus, has approximately 220 species out of the more than 520 Ctenidae species described, and several authors consider it polyphyletic. Chromosomal data are only available for four species of Ctenus, representing a large gap in the cytogenetic knowledge about the group. This study provided cytogenetic data on two Ctenus species and one Guasuctenus (previously described as Ctenus). All showed 2n? = 28 (26+X1X20). Guasuctenus longipes presented two chromosome pairs containing 18S rDNA genes and C. medius, however C. ornatus showed only one chromosome pair with the 18S rDNA gene. Hybridization data using histone H3 probe indicated specific profiles: histone H3 genes were found in one chromosome pair in G. longipes, in three pairs in C. medius, and in four pairs in C. ornatus. Furthermore, supernumerary chromosomes were identified in C. ornatus presenting a meiotic behavior similar to that of sex chromosomes; and a trivalent was found in C. medius, formed by the association of one sex chromosome and an autosomal bivalent, indicating the importance of these events for the diversification of sex chromosomes in spiders. The C-banding pattern was similar between C. medius and C. ornatus with regard to the number and locations of heterochromatic bands, suggesting that heterochromatin amplification and dispersion, affect karyotypic evolution in the genus. Cytogenetic data showed similarity between C. medius and C. ornatus, and differentiation of G. longipes congruent with morphological data. Moreover, although more comparative analyses are needed to specify composition of the dispersed heterochromatin in Ctenus, the mapping of heterochromatic bands provided insights about the evolution of the karyotypes in this genus

Insights on repetitive DNA behavior in two species of Ctenus Walckenaer, 1805 and Guasuctenus Polotow and Brescovit, 2019 (Araneae, Ctenidae): evolutionary profile of H3 histone, 18S rRNA genes and heterochromatin distribution

Ctenidae represents one of the most representative spider families in the tropical forests of Brazil. Its largest genus, Ctenus, has approximately 220 species out of the more than 520 Ctenidae species described, and several authors consider it polyphyletic. Chromosomal data are only available for four species of Ctenus, representing a large gap in the cytogenetic knowledge about the group. This study provided cytogenetic data on two Ctenus species and one Guasuctenus (previously described as Ctenus). All showed 2n? = 28 (26+X1X20). Guasuctenus longipes presented two chromosome pairs containing 18S rDNA genes and C. medius, however C. ornatus showed only one chromosome pair with the 18S rDNA gene. Hybridization data using histone H3 probe indicated specific profiles: histone H3 genes were found in one chromosome pair in G. longipes, in three pairs in C. medius, and in four pairs in C. ornatus. Furthermore, supernumerary chromosomes were identified in C. ornatus presenting a meiotic behavior similar to that of sex chromosomes; and a trivalent was found in C. medius, formed by the association of one sex chromosome and an autosomal bivalent, indicating the importance of these events for the diversification of sex chromosomes in spiders. The C-banding pattern was similar between C. medius and C. ornatus with regard to the number and locations of heterochromatic bands, suggesting that heterochromatin amplification and dispersion, affect karyotypic evolution in the genus. Cytogenetic data showed similarity between C. medius and C. ornatus, and differentiation of G. longipes congruent with morphological data. Moreover, although more comparative analyses are needed to specify composition of the dispersed heterochromatin in Ctenus, the mapping of heterochromatic bands provided insights about the evolution of the karyotypes in this genus

The Karyotypic Diversification of Calophysines and the Exallodontus-Propimelodus Clade (Pimelodidae, Siluriformes): A Cytotaxonomic and Evolutionary Approach in Pimelodidae Based on Ancestral State Reconstruction.

Pimelodidae family is one of the most diverse and widely distributed fish groups in South America. Phylogenetic analysis in the family have recently indicated the existence of two main clades: "sorubiminae" and the OCP clade, including Pimelodus ornatus, "calophysines" and "pimelodines." The aim of this study was to investigate the karyotype of three Amazonian Pimelodidae species: Calophysus macropterus, Propimelodus eigenmanni, and Exallodontus aguanai associating them to the literature, seeking to reconstruct probable ancestral characters. C. macropterus has 2n = 50, 20m+20sm+10a (fundamental number [FN] = 90), simple interstitial nucleolar organizing regions (NORs), and four 5S rDNA sites terminals, two in synteny with the 18S rDNA. P. eigenmanni has 2n = 56, 28m+20sm+2st+6a (FN = 106), simple NORs, and two 5S rDNA sites terminals. E. aguanai has 2n = 56, 36m+12sm+2st+6a (FN = 106) and 18S and 5S rDNA sites interstitial syntenic in the chromosome 1. All species exhibited a higher amount of heterochromatin, differing from the pattern of the family, and strong marking associated with NORs. The integration between molecular phylogenetic data and karyotype data indicated a high probability that 2n = 56 and simple terminals NORs in the short arm are ancestral characters in Pimelodidae, evidenced in "sorubiminae." In the OCP clade derived traits were observed resulting from chromosomal changes that played a critical role in the karyotype evolution of the group.

Karyotype diversity between species of Crenicichla (Perciformes, Cichlidae) from different Brazilian hydrographic basins.

Crenicichla is the largest genus in the Cichlidae family in South America. The genus includes 100 valid species that are popularly known in Brazil as jacundás or joaninhas and are widely distributed in rivers east of the Andes. Cytogenetic analyses were carried out on seven species in this genus. All species showed a diploid number of 48 with interspecific differences in karyotype formulas and AgNORs located in interstitial position on the short arm of the largest metacentric pair, except for the two populations from C. britskii. Population A showed terminal markings on the long arm of the fifth pair of the complement, and population B showed up to two marked chromosome pairs. FISH with an 18S rDNA probe was coincident with AgNORs and CMA3, except for pair 6 from population B of C. britskii that did not presented positive CMA3 sites. This work presents first cytogenetic data for C. haroldoi, C. maculata, and C. punctata, and the results show karyotypic patterns similar to those in the literature. However, the diversity found in populations of C. britskii represents new information about the evolution of the karyotype of the Cichlidae family, which has been conservative. Furthermore, the data could assist in phylogenetic studies of Crenicichla.

Genetic and Chromosomal Differentiation of Rhamdia quelen (Siluriformes, Heptapteridae) Revealed by Repetitive Molecular Markers and DNA Barcoding.

Rhamdia quelen, a species of Heptapteridae, is considered to be a complex because of taxonomic and phylogenetic inconsistencies. Determining the physical location of repetitive DNA sequences on the chromosomes and the DNA barcode might increase our understanding of these inconsistencies within different groups of fish. To this end, we analyzed R. quelen populations from two river basins in Brazil, Paraguay and Parana, using DNA barcoding and different chromosomal markers, including U2 snDNA, which has never been analyzed for any Rhamdia species. Cytochrome c oxidase I gene sequence analysis revealed a significant differentiation among populations from the Miranda and Quexada rivers, with genetic distances compatible to those found among different species in neotropical fishes. Our results, in general, revealed a conservative chromosomal evolution in R. quelen and a differential distribution of some markers, such as 5S rDNA and U2 snDNA, in different populations. We suggest that R. quelen must undergo a major revision in its morphological, genetic, and cytogenetic molecular and taxonomic structure to elucidate possible operational taxonomic units.

Cytogenetic Trends in Two Families of the Neotropical Catfishes: Heptapteridae and Pseudopimelodidae (Siluriformes).

Several neotropical Siluriformes groups suffered important taxonomic revisions based on the evaluation of morphological and molecular characteristics that allow the construction of new phylogenetic hypothesis. In the present study were cytogenetically analyzed six species belonging to Heptapteridae (Cetopsorhamdia iheringi, Phenacorhamdia tenebrosa, Rhamdella eriarcha, Pimelodella meeki, Pimelodella australis, Heptapterus mustelinus) and two to Pseudopimelodidae families (Microglanis cottoides and Microglanis cibelae) by means of differential staining techniques to describe more precisely cytogenetic similarities and differences. The diploid number of R. eriarcha with 2n = 58 and M. cibelae with 2n = 56 were reported for the first time. Also, the lowest chromosome number (2n = 48) for P. tenebrosa was described. The chromosome-banding techniques for to put in evidence nucleolar organizers impregnated by silver nitrate ([AgNORs], chromomycin A3 [CMA3], and rDNA 18S) showed for all studied species conserved patterns, characteristic for each family. The rDNA 5S showed high variability among species or populations of both families, these regions could be simple or multiple, syntenic, or not with rDNA18S. The chromosome markers showed that both families are related not only from a morphologic point of view but also by their karyotypic characteristics, however, some of the present cytogenetic results evidence the importance of new morphologic, molecular, and phylogenetic studies to improve the knowledge of these fish groups.

First chromosome data on Steindachneridion doceanum (Siluriformes: Pimelodidae): a critically endangered catfish endemic of the Doce River basin, Brazil

The present report represents the first cytogenetic description of Steindachneridion doceanum, great catfish which is currently at high extinction risk and it is listed as threatened on the red list of the Brazilian Ministry of the Environment, also are suggested karyotype relationships with other species of the same genus endemic from other river basins. The results revealed a diploid number of 2n = 56 and the karyotype composed of 18 metacentric, 20 submetacentric, 10 subtelocentric and 8 acrocentric chromosomes (NF = 104). The AgNORs and CMA3 signals were coincident in location occupying the short arm of an acrocentric chromosome pair (25th), in a secondary constriction. The 5S rDNA genes were localized on the short arms of one subtelocentric pair. C-banding revealed terminal blocks on the short arms on many chromosomes as well as terminal positive bands at the both ends of a submetacentric pair. C banding also revealed a large heterochromatic block in the secondary constriction (25th) region that was coincident with the AgNORs sites and CMA3+ bright bands. In spite S. doceanum represent an endemic taxon, in spite their geographic isolation their cytogenetic characteristics show similarities with other species of the genus.(AU)

O presente trabalho apresenta a primeira descrição citogenética de Steindachneridion doceanum, grande bagre que se encontra atualmente em alto risco de extinção e listado como ameaçado na lista vermelha do Ministério do Meio Ambiente, também sugere relações cariotípicas com outras espécies do mesmo gênero, endêmicas de outras bacias hidrográficas. Os resultados revelaram um número diplóide de 56 cromossomos e o cariótipo composto por 18 elementos metacêntricos, 20 submetacêntricos, 10 subtelocêntricos e 8 acrocêntricos (NF = 104). As marcações AgNORs e CMA3 foram coincidentes ocupando o braço curto de um par de cromossomos acrocêntricos (par 25), em uma constrição secundária. Os genes 5S rDNA foram detectados nos braços curtos de um par subtelocêntrico. A banda C revelou blocos terminais nos braços curtos em vários cromossomos, bem como blocos terminais nas duas extremidades de um par submetacêntrico. A banda C também evidenciou um grande bloco heterocromático na constrição secundária (par 25) coincidente com os sítios AgNORs e as bandas CMA3 positivas. Apesar de S. doceanum representar um táxon endêmico, suas características citogenéticas mostram semelhanças com outras espécies do gênero das quais se encontra geograficamente isolado.(AU)

Cytogenetic analysis of five Ctenidae species (Araneae): detection of heterochromatin and 18S rDNA sites.

The present study aimed to cytogenetically analyse five Ctenidae species Ctenus ornatus (Keyserling, 1877), Ctenus medius (Keyserling, 1891), Phoneutria nigriventer (Keyserling, 1891), Viracucha andicola (Simon, 1906), and Enoploctenus cyclothorax (Philip Bertkau, 1880), from Brazil. All species presented a 2n♂ = 28 except for V. andicola, which showed 2n♂ = 29. Analysis of segregation and behavior of sex chromosomes during male meiosis showed a sex chromosome system of the type X1X20 in species with 28 chromosomes and X1X2X30 in V. andicola. C banding stained with fluorochromes CMA3 and DAPI revealed two distributions patterns of GC-rich heterochromatin: (i) in terminal regions of most chromosomes, as presented in C. medius, P. nigriventer, E. cyclothorax and V. andicola and (ii) in interstitial regions of most chromosomes, in addition to terminal regions, as observed for C. ornatus. The population of Ubatuba (São Paulo State) of this same species displayed an additional accumulation of GC-rich heterochromatin in one bivalent. Fluorescent in situ hybridization revealed that this bivalent corresponded to the NOR-bearing chromosome pair. All analyzed species have one bivalent with 18S rDNA site, except P. nigriventer, which has three bivalents with 18S rDNA site. Karyotypes of two species, C. medius and E. cyclothorax, are described for the first time. The latter species is the first karyotyped representative of the subfamily Acantheinae. Finally, 18S rDNA probe is used for the first time in Ctenidae at the present study.

First report of B chromosomes in three neotropical thorny catfishes (Siluriformes, Doradidae).

The family Doradidae (Siluriformes) is an important group of fishes endemic to freshwater ecosystems in South America. Some cytogenetic studies have been conducted focused on the group; however, there are no reports on the occurrence of B chromosomes for the family. In this paper the chromosomal characteristics of Platydoras armatulus (Valenciennes, 1840), Pterodoras granulosus (Valenciennes, 1821) and Ossancora punctata (Kner, 1855) were investigated through classical cytogenetics approaches. The conventional staining reveals 2n=58 in Platydoras armatulus and Pterodoras granulosus, however with distinct karyotypic formulae, possibly originated by pericentric inversions. In Ossancora punctata a derivate karyotype was described with 2n=66 and predominance of acrocentric chromosomes. The C banding pattern was resolutive in discriminating the three species, being considered an important cytotaxonomic marker. All species showed B chromosomes totally heterochromatic with non-Mendelian segregation during meiosis and low frequencies in mitotic cells. The probably origin of these additional elements was through fragmentations of chromosomes of the standard complement, which occurred recently and independently in these three species. The diploid number observed in Ossancora punctata is an evidence of centric fusions and up to the moment it is the highest diploid number reported for Doradidae.

Cytogenetic studies in the redtail catfish, Phractocephalus hemioliopterus (Bloch & Schneider, 1801) (Siluriformes, Pimelodidae) a giant fish from Amazon basin.

The objective of this study was to cytogenetically analyze Phractocephalus hemioliopterus comparing the findings with other data to infer relationships among Pimelodidae species. The results revealed a diploid number of 2n = 56 and the karyotype composed of 16 metacentric, 20 submetacentric, 6 subtelocentric and 14 acrocentric chromosomes (FN = 98). The Ag-NORs, 18S rDNA and CMA3 signals were coincident in location occupying the short arm of an acrocentric chromosome pair (23th), in a secondary constriction. The 5S rDNA genes were localized near the centromere on the short arms of one submetacentric chromosome pair. C-bands were localized predominantly in the terminal regions of chromosomes, including the AgNORs and a small metacentric pair with a conspicuous positive band on interstitial region. This chromosome pair could be considered a species-specific cytogenetic marker.

Repetitive DNA in the Catfish Genome: rDNA, Microsatellites, and Tc1-Mariner Transposon Sequences in Imparfinis Species (Siluriformes, Heptapteridae).

Physical mapping of repetitive DNA families in the karyotypes of fish is important to understand the organization and evolution of different orders, families, genera, or species. Fish in the genus Imparfinis show diverse karyotypes with various diploid numbers and ribosomal DNA (rDNA) locations. Here we isolated and characterized Tc1-mariner nucleotide sequences from Imparfinis schubarti, and mapped their locations together with 18S rDNA, 5S rDNA, and microsatellite probes in Imparfinis borodini and I. schubarti chromosomes. The physical mapping of Tc1/Mariner on chromosomes revealed dispersed signals in heterochromatin blocks with small accumulations in the terminal and interstitial regions of I. borodini and I. schubarti. Tc1/Mariner was coincident with rDNA chromosomes sites in both species, suggesting that this transposable element may have participated in the dispersion and evolution of these sequences in the fish genome. Our analysis suggests that different transposons and microsatellites have accumulated in the I. borodini and I. schubarti genomes and that the distribution patterns of these elements may be related to karyotype evolution within Imparfinis.

Chromosomal Mapping of Repetitive DNA Sequences in the Genus Bryconamericus (Characidae) and DNA Barcoding to Differentiate Populations.

The mapping of repetitive DNA sites by fluorescence in situ hybridization has been widely used for karyotype studies in different species of fish, especially when dealing with related species or even genera presenting high chromosome variability. This study analyzed three populations of Bryconamericus, with diploid number preserved, but with different karyotype formulae. Bryconamericus ecai, from the Forquetinha river/RS, presented three new cytotypes, increasing the number of karyotype forms to seven in this population. Other two populations of Bryconamericus sp. from the Vermelho stream/PR and Cambuta river/PR exhibited interpopulation variation. The chromosome mapping of rDNA sites revealed unique markings among the three populations, showing inter- and intrapopulation variability located in the terminal region. The molecular analysis using DNA barcoding complementing the cytogenetic analysis also showed differentiation among the three populations. The U2 small nuclear DNA repetitive sequence exhibited conserved features, being located in the interstitial region of a single chromosome pair. This is the first report on its occurrence in the genus Bryconamericus. Data obtained revealed a karyotype variability already assigned to the genus, along with polymorphism of ribosomal sites, demonstrating that this group of fish can be undergoing a divergent evolutionary process, constituting a substantive model for studies of chromosomal evolution.

Cytogenetic analysis of five Ctenidae species (Araneae): detection of heterochromatin and 18S rDNA sites

The present study aimed to cytogenetically analyse five Ctenidae species Ctenus ornatus (Keyserling, 1877), Ctenus medius (Keyserling, 1891), Phoneutria nigriventer (Keyserling, 1891), Viracucha andicola (Simon, 1906), and Enoploctenus cyclothorax (Philip Bertkau, 1880), from Brazil. All species presented a 2n(SIC) = 28 except for V. andicola, which showed 2n(SIC) = 29. Analysis of segregation and behavior of sex chromosomes during male meiosis showed a sex chromosome system of the type X(1)X(2)0 in species with 28 chromosomes and X(1)X(2)X(3)0 in V. andicola. C banding stained with fluorochromes CMA(3) and DAPI revealed two distributions patterns of GC-rich heterochromatin: (i) in terminal regions of most chromosomes, as presented in C. medius, P. nigriventer, E. cyclothorax and V. andicola and (ii) in interstitial regions of most chromosomes, in addition to terminal regions, as observed for C. ornatus. The population of Ubatuba (Sao Paulo State) of this same species displayed an additional accumulation of GC-rich heterochromatin in one bivalent. Fluorescent in situ hybridization revealed that this bivalent corresponded to the NOR-bearing chromosome pair. All analyzed species have one bivalent with 18S rDNA site, except P. nigriventer, which has three bivalents with 18S rDNA site. Karyotypes of two species, C. medius and E. cyclothorax, are described for the first time. The latter species is the first karyotyped representative of the subfamily Acantheinae. Finally, 18S rDNA probe is used for the first time in Ctenidae at the present study.

Isolation and characterization of 5S rDNA sequences in catfishes genome (Heptapteridae and Pseudopimelodidae): perspectives for rDNA studies in fish by C0t method.

Sequences of 5S ribosomal RNA (rRNA) are extensively used in fish cytogenomic studies, once they have a flexible organization at the chromosomal level, showing inter- and intra-specific variation in number and position in karyotypes. Sequences from the genome of Imparfinis schubarti (Heptapteridae) were isolated, aiming to understand the organization of 5S rDNA families in the fish genome. The isolation of 5S rDNA from the genome of I. schubarti was carried out by reassociation kinetics (C0t) and PCR amplification. The obtained sequences were cloned for the construction of a micro-library. The obtained clones were sequenced and hybridized in I. schubarti and Microglanis cottoides (Pseudopimelodidae) for chromosome mapping. An analysis of the sequence alignments with other fish groups was accomplished. Both methods were effective when using 5S rDNA for hybridization in I. schubarti genome. However, the C0t method enabled the use of a complete 5S rRNA gene, which was also successful in the hybridization of M. cottoides. Nevertheless, this gene was obtained only partially by PCR. The hybridization results and sequence analyses showed that intact 5S regions are more appropriate for the probe operation, due to conserved structure and motifs. This study contributes to a better understanding of the organization of multigene families in catfish's genomes.