Molecular cytogenetics and phylogenetic analysis of Brazilian leaf frog species of the genera Phyllomedusa and Phasmahyla (Hylidae: Phyllomedusinae)

In this study we describe the karyotypes and molecular phylogenetic relationships of the leaf frogs Phyllomedusa bahiana Lutz, 1925, Phyllomedusa burmeisteri Boulenger, 1882, Phyllomedusa nordestina Caramaschi, 2006, Phyllomedusa rohdei Mertens, 1926, Phyllomedusa hypochondrialis (Daudin, 1800), and Phasmahyla spectabilis Cruz, Feio and Nascimento, 2008. We analyzed the karyotypes using C-banding, fluorochrome staining, and fluorescence in situ hybridization of telomeric probe, and inferred phylogeny using nuclear tyrosinase and mitochondrial 16S rDNA sequences. Heterochromatin distribution in P. nordestina diverged from the other species, and P. bahiana and P. rohdei showed evident interstitial telomere sequences. Molecular analyzes confirmed the current taxonomic classification, grouping mitochondrial DNA sequences of each species, and usually without haplotype sharing in the nuclear gene. We also extended the distribution of P. burmeisteri to northern Bahia state and restricted P. bahiana to southern Bahia state, with a discrete extension of the range of P. bahiana to the north and northeast of Minas Gerais state. The tyrosinase haplotype network showed a haplotype shared between P. bahiana and P. burmeisteri from Porto Seguro, Bahia. Based on this, we suggest a new distribution in the overlapping area between these two species, which according to our data ranges between Porto Seguro and Camacan, in Bahia state.