Performance of cytochrome c oxidase subunit I (COI), ribosomal DNA Large Subunit (LSU) and Internal Transcribed Spacer 2 (ITS2) in DNA barcoding of Collembola

• All tested markers proved efficient in discriminating species of Collembola.
• Genetic divergence was highest for COI, followed by ITS2 and LSU.
• ITS2 has a potential to be used also for metabarcoding analyses.

Studies in soil meiofauna with traditional methods are highly laborious and identification exclusively by means of morphological traits usually underestimates species richness. Although the mitochondrial DNA Cytochrome c Oxidase subunit I gene (COI, CO1, cox1, coxI) has been proposed as a standard DNA barcode for animals, nuclear ribosomal DNA markers such as Large Subunit (LSU, 28S) and Internal Transcribed Spacer (ITS) have proven efficient to distinguish among species of animals, eukaryotic microbes and plants. The COI and LSU markers discriminate among closely related species of Collembola, but the efficiency of ITS2 has never been tested. We evaluated the relative performance of these three markers for DNA barcoding of Collembola. All three markers proved highly efficient in discriminating species, but LSU exhibited very low differences among certain closely related species. We conclude that ITS2 may serve as an alternative barcode to identify Collembola and has a potential to be used for metabarcoding analyses.