Systematics of the Boana semilineata species group (Anura: Hylidae), with a description of two new species from Amazonian Ecuador

The combination of genetic and phenotypic characters for species delimitation has allowed the discovery of many undescribed species of Neotropical amphibians. In this study, we used DNA sequences (genes 12S, 16S, ND1 and COI) and morphologic, bioacoustic and environmental characters of the Boana semilineata group to evaluate their phylogenetic relationships and assess their species limits. In addition, we included DNA sequences of several species of Boana to explore cryptic diversity in other groups. We found three Confirmed Candidate Species (CCS) within the B. semilineata group. Holotype examination of Hyla appendiculata shows that it is a valid species that corresponds to one of the CCS, which is here transferred to Boana. We describe the two remaining CCS. Our phylogeny highlights a number of secondary but meaningful observations that deserve further investigation: (1) populations of B. pellucens from northern Ecuador are more closely related to B. rufitela from Panama than to other Ecuadorian populations of B. pellucens; (2) we report, for the first time, the phylogenetic relationships of B. rubracyla showing that it is closely related to B. rufitela and B. pellucens; and (3) B. cinerascens and B. punctata form two species complexes consisting of several unnamed highly divergent lineages. Each of these lineages likely represents an undescribed species.