The karyotype of Holoaden luederwaldti (Anura, Strabomantidae), with report of natural triploidy.

The genus Holoaden includes three species described so far, but the only published cytogenetic data is from Holoaden bradei, with the karyotype 2n = 18, based on conventional staining. In the present paper we report, for the first time, data on chromosomes of H. luederwaldti, which presented 2n = 18 and a case of natural triploidy, with 2n = 3x = 27. In this sample, another karyotypic variation was observed due to the occurrence of two types of chromosome 8, which present submetacentric or subtelocentric morphologies. Homomorphic subtelocentric or heteromorphic condition was observed among the diploid specimens, whereas the triploid had one submetacentric and two subtelocentric chromosomes 8. In all specimens, Ag-NOR was located in the long arms of chromosomes 8, at the interstitial region when subtelocentric, or in the proximal region when submetacentric, confirmed by fluorescent in situ hybridization with the HM123 probe. The C bands showed centromeric distribution and distribution at Ag-NOR site. The centromeric heterochromatin was fluorescent with DAPI staining, whereas the Ag- NOR displayed bright fluorescence with CMA3. Fluorescent in situ hybridization using a telomeric probe labelled exclusively the telomere regions. Although the same 2n = 18 chromosome numbers have been observed in H. luederwaldti and H. bradei, some differences in both karyotypes can be visualized, mainly with regard to the morphology of the last chromosome pairs.

Karyotypic similarity among Barycholos ternetzi and five species of the genus Eleutherodactylus from southeastern Brazil (Anura, Brachycephalidae).

Comparative cytogenetic analyses were carried out in six species of Brachycephalidae from southeastern Brazil. Barycholos ternetzi, Eleutherodactylus binotatus, Eleutherodactylus guentheri, Eleutherodactylus juipoca, Eleutherodactylus parvus and Eleutherodactylus sp. have 2n=22 karyotypes with a marked variation in the morphology of chromosome pairs 8, 10 and 11, which are of telocentric or metacentric types, resulting in FN=38, 40 and 44. Eleutherodactylus have a single chromosome pair bearing Ag-NOR, i.e. pair 1 in E. binotatus, pair 6 in E. guentheri and E. parvus, and pair 11 in E. juipoca and Eleutherodactylus sp. In contrast, B. ternetzi showed Ag-positive sites in the chromosome pairs 1, 4, 5, 9 and 11, and only one to three labelings per metaphase in each individual. Nevertheless, the main chromosome pair with Ag-NOR in the species seems to be the 11th, like in E. juipoca and Eleutherodactylus sp. The NOR site was confirmed by fluorescence in situ hybridization (FISH) technique in E. binotatus and in B. ternetzi, bearing 1p1p and 9p11p11p Ag-NOR pattern, respectively. All the species exhibited predominantly centromeric C-banding pattern, but interstitial bands have also been observed in some cases. In E. binotatus, there is an indication of geographical difference in the distribution of the interstitial C-bands. The fluorochromes GC-specific chromomycin A(3) (CMA(3)) and AT-specific 4',6-diamidino-2-phenylindole (DAPI), with distamycin A (DA) counterstaining, provided the molecular content of some repetitive regions in the karyotypes of the species. One male of E. binotatus presented an extensive heteromorphism, involving at least five different pairs, probably as a consequence of multiple reciprocal translocations. Such rearrangements might be responsible for the multivalent chain seen in the meiosis of this specimen, as well as in another male, although not exhibiting chromosome heteromorphism. The remaining males and those belonging to the other species have always shown 11 bivalents in diplotene and metaphase I cells. In all male specimens, metaphases II presented 11 chromosomes. Despite the observed discrepancies, the five species of Eleutherodactylus have a great uniformity in the 2n=22 karyotypes, suggesting an assemblage of species from southeastern and southern Brazil, in contrast to northern and northeastern assemblage which is characterized by higher diploid numbers. Undoubtedly, B. ternetzi could be included in that proposed assemblage, due to its karyotypic similarity with the Eleutherodactylus species, as evidenced in the present study. This fact strongly supports the close relationships of both genera, previously inferred on the basis of several characters shared by their species.