View article

Wild bees are crucial organisms for terrestrial environments. Their ongoing decline could cause irreparable damage to ecosystem services vital to plant reproduction and human food production. The importance of taking swift action to prevent further declines is widely acknowledged, but the current deficit of reliable taxonomic information complicates the development of efficient conservation strategies targeting wild bees. DNA metabarcoding can help to improve this situation by providing rapid and standardized mass identification. This technique allows the analysis of large numbers of specimens without the need for specialized taxonomic knowledge by matching high-throughput sequencing reads against public DNA barcode reference libraries. However, the validation of this approach for wild bees requires the evaluation of potential error sources on a regional scale. Here we analyzed the effects of three potential error sources on a metabarcoding pipeline customized for the wild bee fauna of Luxembourg. In an in silico study, we checked the completeness of the BOLD reference library for 349 species found in the country, the correspondence between molecular and morphological species delimitation for these taxa, and the amplification efficiency of three commonly used metabarcoding primer pairs (mlCOlintF/HCO2198, LepF1/MLepF1-Rev and BF2/BR2). The detection power of the pipeline was evaluated based on the species recovery rates from mock communities of known composition under variable DNA concentration treatments. The reference barcode library evaluation results show that 97% of the species have at least a single barcode …