A new species ofLeptodactylusfrog (Anura: Leptodactylidae) from the South American Gran Chaco, morphologically similar and previously confused with the widespreadLeptodactylus mystacinus, is described through the use of multiple sources of evidence (molecular, external morphology, coloration, osteology, bioacoustics, and behavior). The phylogenetic analysis with partial sequences of mitochondrial rDNA genes (12S and 16S) recovered the new species within theL. fuscusgroup, being highly divergent (>3% genetic distance in 16S). The new species was recovered as sister taxa ofL. mystacinus, from which it is distinguished by tympanum coloration, cephalic index, dorsum and legs coloration, and some osteological differences in nasals and prevomers. This new frog is characterized by a moderate body size (SVL 46.80–66.21 mm), distinctive color pattern (reddish dorsal surfaces of body with noticeable black stripes in the dorsolateral folds), a circular and dark tympanum with dark tympanic annuli, and behavior of males that call on top of fallen logs and tree branches close to the ground.

Dendropsophini is the most species-rich tribe within Hylidae with 234 described species. Although cytogenetic information is sparse, chromosome numbers and morphology have been considered as an important character system for systematic inferences in this group. Using a diversity of standard and molecular techniques, we describe the previously unknown karyotypes of the genera Xenohyla, Scarthyla and Sphaenorhynchus and provide new information on Dendropsophus and Lysapsus. Our results reveal significant karyotype diversity among Dendropsophini, with diploid chromosome numbers ranging from 2n = 22 in S. goinorum, 2n = 24 in Lysapsus, Scinax, Xenohyla, and almost all species of Sphaenorhynchus and Pseudis, 2n = 26 in S.carneus, 2n = 28 in P.cardosoi, to 2n = 30 in all known Dendropsophus species. Although nucleolar organizer regions (NORs) and C-banding patterns show a high degree of variability, NOR positions in 2n = 22, 24 and 28 karyotypes and C-banding patterns in Lysapsus and Pseudis are informative cytological markers. Interstitial telomeric sequences reveal a diploid number reduction from 24 to 22 in Scarthyla by a chromosome fusion event. The diploid number of X.truncata corroborates the character state of 2n = 30 as a synapomorphy of Dendropsophus.

We studied the external and oral cavity morphology of the tadpoles of eight species of Hypsiboas in the H. albopunctatus, H. faber, H. punctatus and H. pulchellus species groups. After a review of the available information about larval external and oral cavity morphology, no character state seems to be synapomorphic for Hypsiboas. The presence of a fleshy projection in the inner margin of the nostrils and rounded vacuities of the anteromedial surface of the choanae (pending the confirmation of the latter in Hyloscirtus and Myersiohyla) seems to be synapomorphic for the tribe Cophomantini, as previously noticed by other authors. Some putative synapomorphies are suggested for some species groups of Hypsiboas, but a denser sampling is needed to study the taxonomic distribution of these character states, in order to determine which clades they may support. The presence of lateral flaps with labial teeth in the oral disc is a variable feature of many species in the H. faber and H. pulchellus groups. A spiracular tube free from the body wall is present in some species, mostly in the H. albopunctatus group, but also in the H. rufitelus, H. faber and H. pulchellus groups. Unique ventrolateral cumules of neuromasts are present in H. faber, and also in some species of other groups of Hypsiboas and of the sister genus Aplastodiscus. Our results highlight the importance of studying the taxonomic distribution of many character states that were sometimes overlooked in tadpole descriptions but seem relevant to test phylogenetic hypothesis.

The hylid tribe Cophomantini is a diverse clade of Neotropical treefrogs composed of the genera Aplastodiscus, Boana, Bokermannohyla, Hyloscirtus, and Myersiohyla. The phylogenetic relationships of Cophomantini have been comprehensively reviewed in the literature, providing a suitable framework for the study of chromosome evolution. Employing different banding techniques, we studied the chromosomes of 25 species of Boana and 3 of Hyloscirtus; thus providing, for the first time, data for Hyloscirtus and for 15 species of Boana. Most species showed karyotypes with 2n = 2x = 24 chromosomes; some species of the B. albopunctata group have 2n = 2x = 22, and H. alytolylax has 2n = 2x = 20. Karyotypes are all bi-armed in most species presented, with the exception of H. larinopygion (FN = 46) and H. alytolylax (FN = 38), with karyotypes that have a single pair of small telocentric chromosomes. In most species of Boana, NORs are observed in a single pair of chromosomes, mostly in the small chromosomes, although in some species of the B. albopunctata, B. pulchella, and B. semilineata groups, this marker occurs on the larger pairs 8, 1, and 7, respectively. In Hyloscirtus, NOR position differs in the three studied species: H. alytolylax (4p), H. palmeri (4q), and H. larinopygion (1p). Heterochromatin is a variable marker that could provide valuable evidence, but it would be necesserary to understand the molecular composition of the C-bands that are observed in different species in order to test its putative homology. In H. alytolylax, a centromeric DAPI+ band was observed on one homologue of chromosome pair 2. The band was present in males but absent in females, providing evidence for an XX/XY sex determining system in this species. We review and discuss the importance of the different chromosome markers (NOR position, C-bands, and DAPI/CMA₃ patterns) for their impact on the taxonomy and karyotype evolution in Cophomantini.