DNA metabarcoding has played a pivotal role in advancing our understanding of the diversity and function of soil-inhabiting fungi. The Australian Microbiome Initiative has produced an extensive soil fungal metabarcoding dataset of more than 2000 plots across a breadth of ecosystems in Australia and Antarctica. Sequence data requires rigorous approaches for the integration of species occurrences into biodiversity platforms, addressing biases due to false positives or overinflated diversity estimates, among others. To tackle such biases, we conducted a rigorous analysis of the fungal dataset following best practices in fungal metabarcoding and integrated it with over 100 predictor variables to fast-track data exploration. We carefully validated our methodology based on studies conducted on historical versions of the dataset. Our approach generated robust information on Australian soil fungi that can be leveraged by end-users interested in biodiversity, biogeography, and conservation. This novel resource will unlock new frontiers in soil fungal research within the Southern Hemisphere and beyond.