A custom regional DNA barcode reference library for lichen-forming fungi of the Intermountain West, USA, increases successful specimen identification

DNA barcoding approaches provide powerful tools for characterizing fungal diversity. However, DNA barcoding is limited by poor representation of species-level diversity in fungal sequence databases. Can the development of custom, regionally focused DNA refer-ence libraries improve species-level identification rates for lichen-forming fungi? To explore this question, we created a regional ITS database for lichen-forming fungi (LFF) in the In-termountain Region of the United States. The custom database comprised over 4800 se-quences and represented over 600 formal and provisional species. Lichen communities were sampled at 11 sites throughout the Intermountain Region, and LFF diversity was character-ized using high-throughput ITS2 amplicon sequencing. We compared the species-level iden-tification success rates from our bulk community samples using our regional ITS database and the widely used UNITE database. The custom regional database resulted in signifi-cantly higher species-level assignments of operational taxonomic units (OTUs) than the UNITE database . These results highlight that developing regional databases may accelerate a wide range of LFF research by improving our ability to characterize species-level diversity using DNA barcoding.