Karen Ventura

The taxonomic status of the endangered thin-spined porcupine, Chaetomys subspinosus (Olfers, 1818), based on molecular and karyologic data

BackgroundThe thin-spined porcupine, also known as the bristle-spined rat, Chaetomys subspinosus (Olfers, 1818), the only member of its genus, figures among Brazilian endangered species. In addition to being threatened, it is poorly known, and even its taxonomic status at the family level has long been controversial. The genus Chaetomys was originally regarded as a porcupine in the family Erethizontidae, but some authors classified it as a spiny-rat in the family Echimyidae. Although the dispute seems to be settled in favor of the erethizontid advocates, further discussion of its affinities should be based on a phylogenetic framework. In the present study, we used nucleotide-sequence data from the complete mitochondrial cytochrome b gene and karyotypic information to address this issue. Our molecular analyses included one individual of Chaetomys subspinosus from the state of Bahia in northeastern Brazil, and other hystricognaths.ResultsAll topologies recovered in our molecular phylogenetic analyses strongly supported Chaetomys subspinosus as a sister clade of the erethizontids. Cytogenetically, Chaetomys subspinosus showed 2n = 52 and FN = 76. Although the sexual pair could not be identified, we assumed that the X chromosome is biarmed. The karyotype included 13 large to medium metacentric and submetacentric chromosome pairs, one small subtelocentric pair, and 12 small acrocentric pairs. The subtelocentric pair 14 had a terminal secondary constriction in the short arm, corresponding to the nucleolar organizer region (Ag-NOR), similar to the erethizontid Sphiggurus villosus, 2n = 42 and FN = 76, and different from the echimyids, in which the secondary constriction is interstitial.ConclusionBoth molecular phylogenies and karyotypical evidence indicated that Chaetomys is closely related to the Erethizontidae rather than to the Echimyidae, although in a basal position relative to the rest of the Erethizontidae. The high levels of molecular and morphological divergence suggest that Chaetomys belongs to an early radiation of the Erethizontidae that may have occurred in the Early Miocene, and should be assigned to its own subfamily, the Chaetomyinae.

Non-telomeric sites as evidence of chromosomal rearrangement and repetitive (TTAGGG)n arrays in heterochromatic and euchromatic regions in four species of Akodon (Rodentia, Muridae).

Comparative studies among four species – Akodonazarae (2n = 38), A. lindberghi (2n = 42), A. paranaensis (2n = 44) and A. serrensis (2n = 46) – employing classic cytogenetics (C- and G-bands) and fluorescence in situ hybridization with telomeric (TTAGGG)n sequencesare reported here. Non-telomeric signals in addition to the regular telomeric sites were detected in three species:A. azarae, A. lindberghi and A. serrensis. One interstitial telomeric site (ITS) was observed proximally at the long arm of chromosome 1 of A. azarae. The comparison of G-banding patterns among the species indicated that the ITS was due to a tandem fusion/fission rearrangement. Non-telomeric signals of A. lindberghi and A. serrensis were not related to chromosomal rearrangements; instead, the sequences co-localized with (i) heterochromatic regions of all chromosomes in A. serrensis; (ii) some heterochromatic regions in A. lindberghi, and (iii) both euchromatic and heterochromatic regions in the metacentric pair of A. lindberghi. These exceptional findings revealed that ITS in Akodon can be related to chromosomal rearrangements and repetitive sequences in the constitutive heterochromatin and that the richness of TTAGGG-like sequences in the euchromatin could be hypothesized to be a result of amplification of the referred sequence along the chromosome arms.

Phylogeographic Structure and Karyotypic Diversity of the Brazilian Shrew Mouse (Blarinomys breviceps, Sigmodontinae) in the Atlantic Forest

Blarinomys breviceps possesses cryptic and burrowing habits with poorly documented genetics and life history traits. Due to its rarity, only a few specimens and DNA sequences have been deposited in collections worldwide. Here, we present the most comprehensive cytogenetic and molecular characterization of this rare genus. Phylogenetic analyses based on partial cytochrome b sequences were performed, attempting to establish the relationships among individuals with distinct karyotypes along the geographic distribution of the genus in the Atlantic Forest. Classical and molecular cytogenetics, using banding patterns and FISH of telomeric and whole chromosome X-specific painting probes (obtained from the Akodontini Akodon cursor) were used to characterize and compare the chromosomal complements. Molecular phylogenetic analyses recovered 2 main geographically structured clades, northeastern and southeastern with pairwise sequence divergences among specimens varying between 4.9 and 8.4%. Eight distinct karyomorphs are described: (A) 2n = 52 (50A, XX), (B) 2n = 52 (48A, XY+2Bs), (C) 2n = 45 (42A, XY+1B), (D) 2n = 43 (37A, XX+4Bs), (E) 2n = 37 (34A, XY+1B), (F) 2n = 34 (32A, XX), (G) 2n = 31 (27A, XX+2Bs), (H) 2n = 28 (26A, XY), all with the same number of autosomal arms (FNA = 50). Variation of 0–4 supernumerary chromosomes (Bs) presenting heterogeneity in morphology and distribution of interstitial telomeric sequences (ITSs) is reported. ITSs are also found in some metacentric autosomes. The phylogeographic separation between 2 major lineages with high levels of genetic divergence, and the wide karyotypic diversity indicate that B. breviceps is a diverse group that warrants taxonomic re-evaluation.

An undescribed karyotype for Thaptomys (2n = 50) and the mechanism of differentiation from Thaptomys nigrita (2n = 52) evidenced by FISH and Ag-NORs

Abstract Thaptomys nigrita is a monotypic species with 2n = 52 from Akodontini tribe. The karyotype is composed by 25 pairs of autosome being 24 acrocentric decreasing in size and a small metacentric pair. X and Y are respectively a medium size acrocentric and a small submetacentric. In this paper we report for the first time a karyotype with 2n = 50 for an undescribed species of genus Thaptomys. This new karyotype is encompasses by 24 pairs of acrocentric autosomes decreasing in size; X and Y chromosomes are respectively a large acrocentric and a small submetacentric; heterochromatic blocks are observed in the pericentromeric regions of all autosomes and of the X, whereas the long arm of the Y is entirely heterochromatic. Multiple Ag-NORs are located at the telomeric regions of the long arm of the autosomes, and a single chromosome pair (24) presents Ag-NORs in both telomeric regions, which is similar to the pattern observed in the metacentric autosome pair 25 of Thaptomys nigrita with 2n = 52. It can be suggested that this pair 24 has undergone a pericentric inversion and originated the acrocentric pair in Thaptomys sp. with 2n = 50. G-banding pattern and interstitial telomeric signal (ITS) by FISH suggest that the karyotype differentiation between both karyomorphs with 2n = 52 in Thaptomys nigrita and 2n = 50 of Thaptomys sp. was due to a tandem fusion involving respectively pairs 2 and 24 from the former resulting in pair 2 of the latter. We propose that this new karyotype with 2n = 50 belongs to a new and cryptic species for the genus Thaptomys, since these two entities seem to be morphologically indistinguishable and the geographic localization plus the chromosome rearrangements can represent a reproductive barrier between these two forms.

The phylogenetic position of the enigmatic Atlantic forest-endemic spiny mouse Abrawayaomys (Rodentia: Sigmodontinae)

BackgroundThe phylogenetic position of the sigmodontine genus Abrawayaomys, historically assigned to the tribe Thomasomyini or considered a sigmodontine incertae sedis, was assessed on the basis of nuclear and mitochondrial DNA sequences obtained from four individuals from different localities in the Atlantic forest of Brazil. Sequences of Abrawayaomys were analyzed in the context of broad taxonomic matrices by means of maximum-likelihood (ML) and Bayesian analyses (BA).ResultsThe phylogenetic position of Abrawayaomys differed depending on the gene analyzed and the analysis performed (interphotoreceptor retinoid-binding protein (IRBP) ML: sister to Thomasomyini; IRBP BA: sister to Akodontini; cytochrome (Cyt) b ML: sister to Neotomys; and Cyt b BA: sister to Reithrodontini). With the sole exception of the BA based on Cyt b sequences, where the Abrawayaomys-Reithrodon clade had strong support, all sister-group relationships involving Abrawayaomys lacked any significant support.ConclusionsAs such, Abrawayaomys constitutes the only representative so far known of one of the main lineages of the sigmodontine radiation, differing from all other Atlantic forest sigmodontine rodents by having a unique combination of morphological character states. Therefore, in formal classifications, it should be regarded as a Sigmodontinae incertae sedis.

Thaptomys Thomas 1915 (Rodentia, Sigmodontinae, Akodontini) with karyotypes 2n = 50, FN = 48, and 2n = 52, FN = 52: Two monophyletic lineages recovered by molecular phylogeny

A novel karyotype with 2n = 50, FN = 48, was described for specimens of Thaptomys collected at Una, State of Bahia, Brazil, which are morphologically indistinguishable from Thaptomys nigrita, 2n = 52, FN = 52, found in other localities. It was hence proposed that the 2n = 50 karyotype could belong to a distinct species, cryptic of Thaptomys nigrita, once chromosomal rearrangements observed, along with the geographic distance, might represent a reproductive barrier between both forms. Phylogenetic analyses using maximum parsimony and maximum likelihood based on partial cytochrome b sequences with 1077 bp were performed, attempting to establish the relationships among the individuals with distinct karyotypes along the geographic distribution of the genus; the sample comprised 18 karyotyped specimens of Thaptomys, encompassing 15 haplotypes, from eight different localities of the Atlantic Rainforest. The intra-generic relationships corroborated the distinct diploid numbers, once both phylogenetic reconstructions recovered two monophyletic lineages, a northeastern clade grouping the 2n = 50 and a southeastern clade with three subclades, grouping the 2n = 52 karyotype. The sequence divergence observed between their individuals ranged from 1.9% to 3.5%.

Karyotype characterization and nucleolar organizer regions of marsupial species (Didelphidae) from areas of Cerrado and Atlantic Forest in Brazil

The karyotypes of 23 specimens belonging to 16 species from nine genera of Brazilian marsupials (family Didelphidae) were studied. The animals were collected in eight localities of Cerrado or Atlantic Forest biomes in the states of Goias, Tocantins and Sao Paulo. The karyotypes were analyzed after conventional Giemsa staining and silver staining of the nucleolus organizer regions (Ag-NORs). New karyotypic data were obtained for Gracilinanus microtarsus (2n = 14, FN = 24),Marmosops paulensis (2n = 14, FN = 24), Micoreus paraguayanus (2n = 14, FN = 20) and Monodelphis rubida (2n = 18, FN = 32) and are discussed in detail. The karyotypes of G. microtarsus, M. paulensis and M. paraguayanus include three large pairs of submetacentrics (pairs 1, 2 and 3) and a medium-sized metacentric or submetacentric pair 4. Pairs 5 and 6 are small submetacentrics in G. microtarsus and M. paulensis and acrocentrics in M. paraguayanus. M. paulensis presented a single Ag-NOR in pair 6 (6p6p), while M. paraguayanus exhibited multiple Ag-NORs in pairs 5 and 6 (5pq5pq6p6p). There was variation in size and morphology of the sex chromosomes among these species. Monodelphis rubida presented a karyotype with 2n = 18 and FN = 32 composed of a large submetacentric pair 1, a medium-sized metacentric pair 2 and six pairs of submetacentrics (pairs 3 through 8). The X was a small acrocentric and the Y was dot-like. A single Ag-NOR bearing pair (5p5p) characterized M. rubida. Relevant karyotypic information was obtained for 19 specimens belonging to 12 species collected in areas sampled for the first time [Caluromys lanatus and C. philander (2n = 14, FN = 20), Gracilinanus emiliae (2n = 14, FN = 24), Marmosa murina, Metachirus nudicaudatus and Micoureus demerarae (2n = 14, FN = 20), Monodelphis americana (2n = 18, FN = 32) and M. domestica (2n = 18, FN = 20), and Didelphis marsupialis, Philander frenata, P. opossum and P. sp (2n = 22, FN = 20)]. Although the karyotypes were relatively conserved with respect to the morphology of the autosomes among species with the same diploid number, some differences regarding FN, sex chromosomes morphology and Ag-NORs patterns were detected.

A New Allopatric Lineage of the Rodent Deltamys (Rodentia: Sigmodontinae) and the Chromosomal Evolution in Deltamys kempi and Deltamys sp.

Deltamys Thomas 1917 is a poorly studied and rarely collected taxon of Akodontini (Sigmodontinae). The single described species, Deltamys kempi (DKE), has a basic karyotype with a diploid number of 2n = 37 in males and 2n = 38 in females, a fundamental number FN = 38 for both sexes, and an X1X1X2X2/X1X2Y sex determination system. Herein, a new allopatric form, Deltamys sp. (DSP), is reported, based on specimens from southern Brazil, with 2n = 40, FN = 40 and XX/XY sex chromosomes. We describe the karyotype and mechanism of chromosomal differentiation between both Deltamys complements. Phylogenetic analyses, based on the complete sequence (1,140 bp) of the mitochondrial cytochrome b gene, grouped Deltamys sp. as sister species to D. kempi, with up to 12% genetic divergence between them. The GTG-banding patterns show complete autosomal correspondence between D. kempi and Deltamys sp. and identify a tandem rearrangement involving DSP7, DSP19 and DKE4 that is responsible for the differences in 2n and FN. Chromosome painting with Akodon paranaensis chromosome 21 (a small metacentric akodont marker) paint revealed total homology with the smallest acrocentric Deltamys sp. chromosome, DSP19. This suggests the occurrence of a pericentric inversion or centromeric shift when compared to other akodontines, with a posterior tandem rearrangement giving rise to DKE4. In DKE, large blocks of pericentromeric constitutive heterochromatin are present on the autosomes and the X, and the Y/autosome has an entirely heterochromatic short arm. In DSP, small heterochromatic blocks are observed on autosomes and X, and the Y is a very small, mostly heterochromatic acrocentric. The cytogenetic analyses suggest that the Deltamys sp. karyotype is ancestral, with the derived condition resulting from a tandem fusion (DSP7 + DSP19) and the Y/autosome translocation giving rise to the multiple sex chromosome system. The autosomal rearrangements, the differences in CBG-banding patterns and Ag-NOR localization, as well as the presence of X1X1X2X2/X1X2Y and XX/XY sex determination mechanisms, possibly acting as a reproductive barrier, and the phylogenetic position within the Deltamys genus, with high genetic divergence, call for a taxonomic review of the genus.

Karyotypic analyses and morphological comments on the endemic and endangered Brazilian painted tree rat Callistomys pictus (Rodentia, Echimyidae)

The genus Callistomys belongs to the rodent family Echimyidae, subfamily Echimyinae, and its only living representative is Callistomys pictus, a rare and vulnerable endemic species of the state of Bahia, Brazil. Callistomys has been previously classified as Nelomys, Loncheres, Isothrix and Echimys. In this paper we present the karyotype of Callistomys pictus, including CBG and GTG-banding patterns and silver staining of the nucleolus organizer regions (Ag-NORs). Comments on Callistomys pictus morphological traits and a compilation of Echimyinae chromosomal data are also included. Our analyses revealed that Callistomys can be recognized both by its distintinctive morphology and by its karyotype.

The lowest diploid number in Testudines: Banding patterns, telomeric and 45S rDNA FISH in Peltocephalus dumerilianus, 2n = 26 and FN = 52 (Pleurodira, Podocnemididae)

The karyotype of the big-headed Amazon River turtle, Peltocephalus dumerilianus, is characterized based on a sample of seven juveniles from Reserva Biológica do Rio Trombetas, Pará State, Brazil (1°30′ S, 56°34′ W). Here we present the first results on GTG and CBG-banding patterns, Ag-NOR staining and FISH, with telomeric and 45S rDNA sequences as probes. A cytogenetic comparison with related Podocnemidae is also provided.