The preserved Cerrado from Northeastern Brazil presents different physicochemical properties and plant diversity, which can influence the fungal communities. Therefore, we evaluated the fungal diversity in preserved sites, at Sete Ci-dades National Park, across a gradient of vegetation that in-cluded Campo graminoide, Cerrado stricto sensu, Cerradao, and Floresta decidual. Of all of the operational taxonomic units (OTUs) obtained, the Floresta decidual presented the highest richness. Ascomycota were the most abundant phy-lum (45%), followed by Basidiomycota (32%). Basal fungi and other phyla accounted for 23% of the total dataset. Agaricomycetes, Eurotiomycetes, Lecanoromycetes, Basi-diobolus, Dothideomycetes, and Taphrinomycetes were the most abundant classes of fungi found across the gradient of Cerrado vegetation. In conclusion, our study suggests that the Brazilian Cerrado from Sete Cidades National Park pre-sents a high fungal diversity and includes sources of new fungal species for biotechnological purposes.