Females do count: Documenting Chironomidae (Diptera) species diversity using DNA barcoding

Because the family Chironomidae, or non-biting midges, is one of the most species-rich groups of macroinvertebrates in freshwater habitats, species-level identifications of chironomids are important for biodiversity assessments in these ecosystems. Morphology-based species identifications from adult female chironomids usually are considerably more difficult than from adult males, or even impossible; thus, the females are often neglected in community assessments. We used DNA barcoding to investigate how inclusion of the females influenced the species count from springs and spring brooks at Sølendet Nature Reserve in Central Norway. By means of the barcodes we were able to identify 77.6% of the females to species by associating them with males from the study site or from other regions, whereas the remaining, unassociated females could be identified to genus level only. The number of recorded species increased by 27% when females were included. We also found that DNA barcoding is effective for the detection of taxonomically challenging species and species groups. Using DNA barcoding in combination with traditional taxonomy, we recognised at least five species new to science and three species and one genus new to Norway.