Museomics resolves 200 years of taxonomic uncertainty: Strix pulsatrix (Strigiformes, Strigidae) is a valid species

The identification of valid species is central to biology, and genetic data have been essential in uncovering new taxonomic units across groups. For polytypic taxa, genetics helps distinguish valid units from natural variation. Pulsatrix perspicillata (Spectacled Owl) is a widespread polytypic species with six recognized subspecies. We used genetic data and museomics techniques to: (i) test the validity of Strix pulsatrixWied, 1820 (=Pulsatrix perspicillata pulsatrix) as a species distinct from Pulsatrix perspicillata; and (ii) estimate the phylogenetic relationships and divergence times within the genus Pulsatrix Kaup, 1848. We conducted population structure and phylogenetic analyses using SNPs matrices and alignments of Ultraconserved Elements (UCEs) from 16 individuals of Pulsatrix spp., including four historical samples of P. p. pulsatrix. Additionally, we sequenced the mtDNA-ND2 gene from 38 samples representing all known Pulsatrix species to further elucidate phylogenetic relationships and estimate divergence times. Our results indicate that Pulsatrix perspicillata pulsatrix represents an independent lineage from P. perspicillata, as supported by its reciprocal monophyly and distinct population structure. UCE analyses also grouped P. koeniswaldiana (Tawny-browed Owl) and P. melanota (Band-bellied Owl) into a distinct cluster. Phylogenetic analyses based on both UCEs and mtDNA-ND2 sequences support the monophyly of the genus Pulsatrix. Pulsatrix koeniswaldiana and P. melanota form a monophyletic group that is sister to P. perspicillata. Pulsatrix diverged in the Late Miocene and diversified in the Pliocene-Quaternary. The evolutionary history of Pulsatrix appears to have been influenced by (a) the final phase of the Andean uplift and (b) climatic oscillations during the Pleistocene. Pulsatrix p. pulsatrix shows genetic divergence consistent with a species-level split from P. perspicillata, and we henceforth recognize it as Pulsatrix pulsatrix (Wied, 1820). We recommend future studies to assess its current distribution and inform the development of conservation strategies.