Description and evolution of the larynx of the Physalaemus olfersii species group, with remarks on the laryngeal anatomy of the P. cuvieri clade (Amphibia: Anura: Leiuperinae).

The anuran larynx is an organ of great evolutionary interest because it impacts male reproductive success in courtships. However, little is known about the diversity of the larynx's anatomy, evolutionary history and systematics importance. Here, we describe and compare the anatomy of the larynx of 10 Physalaemus species of the P. cuvieri clade, focusing on the P. olfersii species group. We also reconstructed the ancestral states and tested the phylogenetic signal for the anatomical features. In all the species, the larynx has a general globular shape with the arytenoid cartilages covering almost its entire dorsal surface, while the anterior process of the cricoid cartilages covers most of the ventral surface. The size of the secondary fibrous mass, the thickness of the vocal membrane, and the attachment position of the vocal membrane's free edge considerably differ among the species. Moreover, only four species of a single clade in the P. olfersii species group have the primary fibrous mass well-developed with a suspended region in the dorsolateral passage. We found a significant phylogenetic signal for all these characters. Ancestral reconstructions pointed to reduction tendencies in the thickness of the vocal membrane and the size of the secondary fibrous mass, and a shift of the ventral attachment of the vocal membrane, increasing the angle of its free edge along the phylogeny. This latter trait can diagnose the entire Physalaemus olfersii group, which has the ventral ends of the arytenoids positioned posteriorly, giving this group the steepest angles for the vocal membrane's free edge in relation to the frontal plane. Based on our results, the larynges can contribute to the Physalaemus olfersii species group's systematics and could be elucidative to understand the evolution of the genus. High levels of anatomical and bioacoustical complexity and diversity observed in the group support the expected correlation between vocal anatomy and bioacoustical signal.