Cytogenetics of four foam-nesting frog species of the Physalaemus gracilis group (Anura, Leptodactylidae).

Intending to increase the knowledge about cytogenetics of Physalaemus and the sparsely studied P. gracilis group, we analyzed the karyotypes of P. carrizorum, P. gracilis, P. lisei, and P. sp. aff. gracilis. We studied chromosome morphology, heterochromatin patterns, Ag-NORs location and mapped the repetitive DNA sequence PcP190. All species showed diploid karyotypes composed of 22 bi-armed chromosomes and similar C- bands and Ag-NOR patterns. C-bands were mainly centromeric and pericentromeric; non-centromeric C-bands were detected on the telomeres of pair 1 in P. lisei, although polymorphic, and interstitially on pair 10 of P. gracilis. This last character is useful to distinguish P. gracilis from its sibling species P. sp. aff. gracilis. The Ag-NOR sites were detected on the long arms of chromosome pair 8 but with a variable position among species. Clusters of PcP190 showed centromeric and pericentromeric positions coincident with conspicuous C-bands, on pairs 2 and 9 in P. gracilis and P. sp. aff. gracilis, pair 3 in P. carrizorum, and pair 7 in P. lisei. These results significantly increase the knowledge about Physalaemus cytogenetics and encourage further studies on the satellite PcP190 in other genera of Leiuperinae to better understand its taxonomic distribution and the evolutionary dynamics.

Phylogeny and chromosomal diversification in the Dichroplus elongatus species group (Orthoptera, Melanoplinae).

In an attempt to track the chromosomal differentiation in the Dichroplus elongatus species group, we analyzed the karyotypes of four species with classical cytogenetic and mapping several multigene families through fluorescent in situ hybridization (FISH). We improved the taxon sampling of the D. elongatus species group adding new molecular data to infer the phylogeny of the genus and reconstruct the karyotype evolution. Our molecular analyses recovered a fully resolved tree with no evidence for the monophyly of Dichroplus. However, we recovered several stable clades within the genus, including the D. elongatus species group, under the different strategies of tree analyses (Maximum Parsimony and Maximum Likelihood). The chromosomal data revealed minor variation in the D. elongatus species group's karyotypes caused by chromosome rearrangements compared to the phylogenetically related D. maculipennis species group. The karyotypes of D. intermedius and D. exilis described herein showed the standard characteristics found in most Dichroplini, 2n = 23/24, X0♂ XX♀, Fundamental number (FN) = 23/24. However, we noticed two established pericentric inversions in D. intermedius karyotype, raising the FN to 27♂/28♀. A strong variation in the heterochromatic blocks distribution was evidenced at interespecific level. The multigene families' mapping revealed significant variation, mainly in rDNA clusters. These variations are probably caused by micro chromosomal changes, such as movement of transposable elements (TEs) and ectopic recombination. These observations suggest a high genomic dynamism for these repetitive DNA sequences in related species. The reconstruction of the chromosome character "variation in the FN" posits the FN = 23/24 as the ancestral state, and it is hypothesized that variations due to pericentric inversions has arisen independently three times in the evolutionary history of Dichroplus. One of these independent events occurred in the D. elongatus species group, where D. intermedius is the unique case with the highest FN described in the tribe Dichroplini.

Neo-sex Chromosomes in the Maculipennis Species Group (Dichroplus: Acrididae, Melanoplinae): The Cases of D. maculipennis and D. vittigerum.

South American melanopline grasshoppers display a disproportionate number of derived karyotypes, including many cases of neo-sex chromosome systems. This is especially true of the genus Dichroplus and its Maculipennis species group. We analyzed the karyotype and neo-sex chromosomes in mitosis and meiosis of Dichroplus maculipennis and D. vittigerum from Argentina using conventional and fluorescent cytogenetic protocols in order to elucidate the behavior and origin of these neo-XY systems in relation to the current phylogeny of this group. Our results showed that D. maculipennis (2n = 22♂/22♀; neoXY/neoXX) and D. vittigerum, whose karyotype is described here for the first time (2n = 18♂/18♀; neoXY/neoXX), show highly evolved neo-XY systems, although with significant differences between them. Furthermore, both species differ for two autosomal fixed Robertsonian fusions present in D. vittigerum. Analysis of karyotypic character state optimization strongly suggests the independent origin and evolution of neo-sex systems within this species group.

Cytogenetic characterization and B chromosome diversity in direct-developing frogs of the genus Oreobates (Brachycephaloidea, Craugastoridae).

Oreobates Jiménez de la Espada, 1872 is a large group of South American frogs with terrestrial reproduction and direct development, located in the superfamily Brachycephaloidea. About 260 brachycephaloidean species have been cytogenetically studied so far, at least with standard techniques. However, this information represents fewer than 17% species of the family Craugastoridae Hedges, Duellman & Heinicke, 2008, where the genus Oreobates is included. In the present work, using a diversity of standard and molecular techniques, we describe the karyotype of Oreobates barituensis Vaira & Ferrari, 2008, Oreobates berdemenos Pereyra, Cardozo, Baldo & Baldo, 2014 and Oreobates discoidalis (Peracca, 1895), from northwestern Argentina. The three species analyzed showed a diploid karyotype with 2n = 22 biarmed chromosomes, fundamental number (FN) = 44, nucleolus organizer regions (NORs) located pericentromerically on pair 7, and a centromeric and pericentromeric C-banding pattern. We observed variations in the chromosome number in Oreobates barituensis due the presence of two morphs of B chromosomes, one medium-sized telocentric (BT) and another subtelocentric and smaller (Bst). Both B chromosomes are mitotically stable and were recorded in all somatic and germinal cells analyzed. The BT chromosome occurred at a maximum of one per individual (2n = 22+BT), and the other one was observed single (2n = 22 + Bst) or as a pair in two doses (2n = 22 + 2BT). We additionally observed other supernumerary chromosomes in the three species analyzed, all of them euchromatic, small, dot-shaped and with instability during mitoses, showing a frequency of occurrence below 50% in studied specimens. The occurrence of polymorphic and spontaneous chromosomal rearrangements and supernumerary chromosomes is a recurrent feature reported in frogs with terrestrial habits (Brachycephaloidea and Hemiphractidae Peters, 1862), which suggests that Brachycephaloidea may be a promising group for studying the origin and maintenance of B chromosomes in anurans.