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.