Sample culturing and processing
Swab samples were obtained from population of free-living easter water dragons (Intellagama lesueurii ) with a history of infection withN. barbatae in Brisbane city’s Roma Street Parkland (-27°27’46’S, 153°1’11’E), Queensland, Australia. Skin lesions from animals displaying characteristic signs of infection were sampled with sterile rayon dry transfer swabs (Copan Diagnostics Inc.) and placed on ice before transportation to the laboratory to be stored at -20oC until processed to extract DNA using the Wizard Genomic DNA Purification Kit (Promega). Type cultures were purchased from the UAMH Centre for Global Microfungal Biodiversity in Toronto, Canada. N. barbatae strain USC001 was isolated and sequenced previously (Powell et al., 2021). Fungal cultures were grown on potato dextrose agar at 25oC for up to three weeks to obtain sufficient growth. Plate cultures were scraped, and DNA was purified using the DNeasy Plant Mini kit (Qiagen) with 5 mm Stainless Steel Beads (Qiagen). Purified DNA was then quantified and stored at -20oC. Short-read sequencing was undertaken using the Nextera DNA Flex library kit and run on the Illumina NextSeq platform at Centre for the Analysis of Genome Evolution & Function at the University of Toronto.