A Complete Reference DNA Barcode Library for Austrian Bumblebees

Bumblebees (Bombus spp.) are essential pollinators in natural and agricultural ecosystems but face increasing threats across Europe from habitat loss, climate change, and intensive land use. Austria hosts 42 recognized bumblebee species, yet comprehensive molecular data have been lacking. Here, we present the first complete DNA barcode reference library for the Austrian bumblebee fauna, generated as part of the Austrian Barcode of Life initiative. This reference library includes 586 partial mitochondrial COI sequences. DNA barcoding successfully identified all species, with distinct Barcode Index Numbers (BINs) and no BIN sharing observed, demonstrating its reliability as a complementary method to traditional morphology-based identification. Intraspecific genetic diversity was generally low, though B. jonellus exhibited notable mitochondrial structure with a complex biogeographic pattern. Our results underscore the value of DNA barcoding as a straightforward tool for accurate species identification and biodiversity monitoring, even for non-experts, while also highlighting cryptic genetic variation within widely distributed species. This reference library provides a robust framework for taxonomic, ecological, and conservation research, and supports future metabarcoding-based monitoring efforts in Austria and beyond.