Cytogenetic analysis of Phyllomedusa distincta Lutz, 1950 (2n = 2x = 26), P. tetraploidea Pombal and Haddad, 1992 (2n = 4x = 52), and their natural triploid hybrids (2n = 3x = 39) (Anura, Hylidae, Phyllomedusinae)

Background: Natural polyploidy has played an important role during the speciation and evolution of vertebrates, including anurans, with more than 55 described cases. The species of the Phyllomedusa burmeisteri group are mostly characterized by having 26 chromosomes, but a karyotype with 52 chromosomes was described in P. tetraploidea. This species was found in sintopy with P. distincta in two localities of São Paulo State (Brazil), where triploid animals also occur, as consequence of natural hybridisation. We analyse the chromosomes of P. distincta, P. tetraploidea, and their triploid hybrids, to enlighten the origin of polyploidy and to obtain some evidence on diploidisation of tetraploid karyotype.Results: Phyllomedusa distincta was 2n = 2x = 26, whereas P. tetraploidea was 2n = 4x = 52, and the hybrid individuals was 2n = 3x = 39. In meiotic phases, bivalents were observed in the diploid males, whereas both bivalents and tetravalents were observed in the tetraploid males. Univalents, bivalents or trivalents; metaphase II cells carrying variable number of chromosomes; and spermatids were detected in the testis preparations of the triploid males, indicating that the triploids were not completely sterile. In natural and experimental conditions, the triploids cross with the parental species, producing abnormal egg clutches and tadpoles with malformations. The embryos and tadpoles exhibited intraindividual karyotype variability and all of the metaphases contained abnormal constitutions. Multiple NORs, detected by Ag-impregnation and FISH with an rDNA probe, were observed on chromosome 1 in the three karyotypic forms; and, additionally, on chromosome 9 in the diploids, mostly on chromosome 8 in the tetraploids, and on both chromosome 8 and 9 in the triploids. Nevertheless, NOR-bearing chromosome 9 was detected in the tetraploids, and chromosome 9 carried active or inactive NORs in the triploids. C-banding, base-specific fluorochrome stainings with CMA3 and DAPI, FISH with a telomeric probe, and BrdU incorporation in DNA showed nearly equivalent patterns in the karyotypes of P. distincta, P. tetraploidea, and the triploid hybrids.Conclusions: All the used cytogenetic techniques have provided strong evidence that the process of diploidisation, an essential step for stabilising the selective advantages produced by polyploidisation, is under way in distinct quartets of the tetraploid karyotype. © 2013 Gruber et al.; licensee BioMed Central Ltd.

Diferenciação gonadal em Piaractus mesopotamicus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Desempenho zootécnico e caracterização da linhagem germinativa de peixes diploides e triploides de lambari (Astyanax altiparanae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Cytogenetics of three Brazilian Goniosoma species: A new record for diploid number in Laniatores (Opiliones, Gonyleptidae, Goniosomatinae)

Currently, 60 species of harvestmen have been karyotyped and all of these are from the Nearctic and Palearctic regions. This work is the first cytogenetic report of three gonyleptid species of the suborder Laniatores: Goniosoina aff. badiuln, G. proxiinuni and G. spelaeum of the Neotropical region, from the southeastern region of Brazil. Conventional Giemsa stain chromosome preparations were obtained from embryonic cells and adult male testes. Embryo mitotic plates of G. aff. badium and G. proximum indicated 88 chromosomes, and mitotic spermatogonial plates of G. spelaeum males revealed intra- and interindividual variation of chromosome number, ranging from 92-109 chromosomes. In the three analyzed species, the mitotic chromosomes were meta- or subinetacentric with no obvious sex chromosomes being identified during mitosis. Prophase I spermatocytes of G. spelaeum also revealed intra- and interindividual bivalent number variation and furthermore indicated the presence of multivalence. The karyotypes of these three Goniosoina species exhibited the largest chromosome pair with a negative heteropycnosis in the distal region of the shortest arrn chromosomes of G. spelaeum submitted to silver impregnation evidenced this negative heteropycnotic region as nucleolus organizer region (NOR). These results, when compared with cytogenetic data of other Laniatores species from the Palearctic region, indicated that a new record for diploid chromosome number probably characterize the genus Goniosoma in the Neotropical region.

Cytogenetic analysis of the neotropical spider Nephilengys cruentata (Araneomorphae, tetragnathidae): standard staining, NORs, C-bands and base-specific fluorochromes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Female pintado catfish choose well-fed males

This study tested the preference of pintado catfish females (Pseudoplatystoma coruscans) for associating with well-fed or food-deprived males in simultaneous choice tests. Females were tested under three different treatments in which: (1) females could choose on the basis of multiple cues from the well-fed or food-deprived males (visual plus chemical); (2) only chemical cues were presented; (3) only visual cues were presented. Females chose well-fed males when chemical cues were presented (either visual + chemical or only chemical cues). When only visual cues were presented, females spent an equal amount of time in the choice apparatus compartments. Chemical signals, either directly through control of pheromone production, or indirectly by excreted metabolites, are likely to be an important source of information about nutritional condition in mating choice.

Genetic relationships among Arachis hypogaea L. (AABB) and diploid Arachis species with AA and BB genomes

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid, with two types of genomes, classified as AA and BB, according to cytogenetic characters. Similar genomes to those of A. hypogaea are found in the wild diploid species of section Arachis, which is one of the nine Arachis sections. The wild species have resistances to pests and diseases that affect the cultivated peanut and are a potential source of genes to increase the resistance levels in peanut. The aim of this study was to analyze the genetic variability within AA and BB genome species and to evaluate how they are related to each other and to A. hypogaea, using RAPD markers. Eighty-seven polymorphic bands amplified by ten 10-mer primers were analyzed. The species were divided into two major groups, and the AA and the BB genome species were, in general, separated from each other. The results showed that high variation is available within species that have genomes similar to the AA and the BB genomes of A. hypogaea.

The karyotype of Cabassous unicinctus (Dasypodidae, Xenarthra)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Localization of 5-methylcytosine in metaphase chromosomes of diploid and triploid pacu fish, Piaractus mesopotamicus (Pisces, Characiformes)

The distribution of 5-methylcytosine (5-MeC) was investigated in fish chromosomes by indirect immunofluorescence using a highly specific 5-MeC monoclonal antibody. Diploid and artificially produced triploid specimens of the pacu fish, Piaractus mesopotamicus, were analyzed. The strong immunofluorescent signals were coincident with the heterochromatic regions of both diploids and triploids in a pattern that matched the C-banding pattern. In the euchromatin, heterogeneous labeling was observed along the chromatids. The weakness of this labeling hindered comparison of the fluorescence labeling of homologous chromosomes from diploid and triploid individuals. However, no striking differences were observed. The possibility that the euchromatin labeling by the 5-MeC antibody is related to the occurrence of mildly repetitive sequences in the genome of Piaractus is discussed.

Cytogenetic studies in three species of Lutjanus (Perciformes: Lutjanidae: Lutjaninae) from the Isla Margarita, Venezuela

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Karyotypic conservatism in samples of Characidium cf. zebra (Teleostei, Characiformes, Crenuchidae): physical mapping of ribosomal genes and natural triploidy

Basic and molecular cytogenetic analyses were performed in specimens of Characidium cf. zebra from five collection sites located throughout the Tietê, Paranapanema and Paraguay river basins. The diploid number in specimens from all samples was 2n = 50 with a karyotype composed of 32 metacentric and 18 submetacentric chromosomes in both males and females. Constitutive heterochromatin was present at the centromeric regions of all chromosomes and pair 23, had additional interstitial heterochromatic blocks on its long arms. The nucleolar organizer regions (NORs) were located on the long arms of pair 23, while the 5S rDNA sites were detected in different chromosomes among the studied samples. One specimen from the Alambari river was a natural triploid and had two extra chromosomes, resulting in 2n = 77. The remarkable karyotypic similarity among the specimens of C. cf. zebra suggests a close evolutionary relationship. on the other hand, the distinct patterns of 5S rDNA distribution may be the result of gene flow constraints during their evolutionary history.

Chromosomal diversification in populations of Characidium cf. gomesi (Teleostei, Crenuchidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Occurrence of ZZ/ZW sex chromosomes in Thoracocharax stellatus fish (Characiformes, Gasteropelecidae) from the Araguaia River, south America

The karyotypic and chromosomal characteristics of the hatchetfish Thoracocharax stellatus from the Araguaia River, Brazil (Araguaia-Tocantins basin) were analyzed using Giemsa, AgNO3, and CMA(3) fluorescent staining, and C-banding. The diploid chromosome number was 54 and the karyotypes of females and males were composed of six metacentrics, six submetacentrics, six subtelocentrics and 36 acrocentrics. Two impaired acrocentric chromosomes were detected in the female karyotype. C-banding showed heterochromatic blocks at several chromosomes and an entirely heterochromatic acrocentric chromosome in females that was lacking in the male karyotype. This discovery indicated a heteromorphic sex chromosome system of the ZZ/ZW type. Ag-staining and CMA(3) fluorescence revealed one major chromosome pair bearing the NORs with the presence of additional signals in some metaphases. Both heterochromatic segments associated with Ag-NORs and the W chromosome were positively stained by CMA(3). Considering the present data and previous findings it is hypothesized that the occurrence of ZW sex chromosome system is widespread in the genus Thoracocharax.

Cytogenetic characterization of Eurysternus caribaeus (Coleoptera: Scarabaeidae): evidence of sex-autosome fusion and diploid number reduction prior to species dispersion

Mitotic and meiotic chromosomes of several populations of Eurysternus caribaeus (Coleoptera: Scarabaeidae) were analysed through conventional staining, C-banding, base-specific fluorochromes, silver nitrate staining and fluorescent in situ hybridization (FISH). All specimens showed 2n = 8 in their karyotypes, with a neo-XY sex system (Y is a submetacentric and X a metacentric) and three pairs of submetacentric autosomes. The analysis of constitutive heterochromatin (CH) revealed small blocks located in the centromeric region of all chromosomes which do not present positive staining under the fluorochromes CMA3 and DAPI. Silver nitrate staining revealed that the nucleolar organizer region (NORs) is associated with the sex chromosomes. The FISH technique revealed that rDNA sites in the X and Y are different in size. Data from different populations indicate that the diploid number reduction (2n = 8) observed in E. caribaeus is established and presumably has preceded the dispersion of this species. Moreover, this reduction occasioned the translocation of rDNA sites to the sex chromosomes, X and Y, an uncommon pattern in Scarabaeidae that was observed for the first time by the FISH in this work.