DNA barcoding as a tool for elucidating species delineation in wide-ranging species as illustrated by owls (Tytonidae and Strigidae).

The mitochondrial cytochrome c-oxidase subunit I (cox1) can serve as a fast and accurate marker for the identification of animal species, and for the discovery of new species across the tree of life. Distinguishing species using this universal molecular marker, a technique known as DNA barcoding, relies on the identifying the gap between intra- and interspecific divergence. One of the difficulties could be wide-ranging, cosmopolitan species that show large amounts of morphological variation. The barn owl Tyto alba is a case in point. It occurs worldwide and varies morphologically, leading to the recognition of many subspecies or, more recently, species. We analysed data from the cox1 gene for 31 individuals of seven subspecies, and compared this with 214 sequences from 29 other owl species. Phylogenetic analysis of the T. alba samples gives very strong support for an Old World alba-clade (three subspecies) and a New World furcata-clade (four subspecies) that are genetically equidistant. The amount of intraspecific variation within each of these clades ranges from 0.66-0.99%, but variation among these clades ranges from 5.33-6.20%. Combined these data suggest that barn owl of the Old World is indeed best considered a separate species different from that of the New World. For combined dataset, sample size of owl species (n between 1 and 21 sequences) increased with geographic range size but we did not find significant relationships between interspecific divergence and sample size or between interspecific divergence and geographic range. For 21/24 species of owls with sample sizes of n ≥4 the maximum interspecific divergences was ≤ 3.00%. However, similar to those found in barn owls, the largest amount of divergence (3.23-4.09%) was present in two other wide-ranging species (Strix nebulosa and Aegolius funereus) raising the possibility of multiple species in other wide-ranging owls as well.