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Data include soil and litter measurements for moisture, pH, and carbon-to-nitrogen ratio. Samples were collected from 8 different ecoregions, as determined by NEON, at various NEON/LTER and/or other experimental sites. Soil cores and litter samples were taken in the spring and fall of 2022. 

The Ordway-Swisher Biological Station (OSBS) is a 38-km2 reserve owned by the University of Florida and is part of the National Ecological Observatory Network (NEON). The reserve contains several iconic Florida habitats, such as sandhill, mesic hammock, and scrubby flatwoods. While plants and animals have been extensively studied at OSBS, the fungi remain poorly known. Fungal inventories are critical to increase knowledge of both fungal diversity and species ranges, and thus to provide foundational data for a wide array of applications in ecology and resource management. Here, we present the results of a nine-year effort to collect, preserve, and DNA barcode the macrofungi at OSBS. This effort generated >1200 vouchered specimens and 984 ITS rDNA sequences, representing more than 546 species. Our sampling was dominated by Basidiomycota and revealed a high diversity of symbiotic ectomycorrhizal fungi, particularly species of Amanita, Cortinarius, and Russula. Sampling curves and both Chao1 and Jacknife1 richness estimators suggest that our DNA barcoding efforts captured only about half of the macrofungi species and that a more complete inventory would detect 897–1177 macrofungi species at OSBS. Our sampling found more species of macrofungi at OSBS than the known number of vertebrate animal species at the reserve and our estimates also suggest that there are likely more macrofungi species than plant species at OSBS. This study is the first comprehensive macrofungi inventory within a NEON site and highlights the importance of long-term monitoring to provide novel data on fungal diversity, community structure, conservation, biogeography, and taxonomy.