Köhler, J., Koscinski, D., Padial, J. M., Chaparro, J. C., Handford, P., Lougheed, S. C. & De la Riva, I. (2010). Systematics of Andean gladiator frogs of the Hypsiboas pulchellus species group (Anura, Hylidae). -Zoologica Scripta, 39, 572-590.We revisit the taxonomic status of Andean species and populations of frogs of the Hypsiboas pulchellus group using multiple lines of evidence potentially indicative of evolutionary lineage divergence in anurans: differences in qualitative morphological or bioacoustic character states, no overlap in quantitative characters of advertisement calls, and monophyly of gene genealogies. We found qualitative and quantitative morphological characters to be extremely variable among species and populations of the group and thus of very limited use in assessing lineage divergence. In contrast, phylogenetic analyses based on 16S rRNA and cytochrome b sequences resolved highly supported clades that are in concordance with bioacoustic differences. The results support the specific distinctness of most nominal species recognized in the group, including the Bolivian Hypsiboas balzani and Hypsiboas callipleura, two species that were considered to be synonymous, and revealed the presence of an undescribed species from southern Peru, which is here described as Hypsiboas gladiator sp. n. In contrast, Hypsiboas andinus and Hypsiboas riojanus were mutually paraphyletic, and showed no differences in morphology and acoustic characters. Consequently, we regard the former as a junior synonym of the latter. However, we discovered that populations of H. riojanus from central Bolivia exhibit some degree of genetic differentiation and advertisement call differences with respect to Argentine populations, but sampling of these Bolivian populations is too sparse to draw taxonomic conclusions. Our phylogenetic results support the hypothesis that ancestral lineages of the Andean H. pulchellus group experienced successive splitting events along a latitudinal gradient from north to south. © 2010 The Authors. Zoologica Scripta © 2010 The Norwegian Academy of Science and Letters.