Whilst the importance of fungal primary biological aerosol particles (PBAPs) has been recognised, few studies have empirically assessed how land-use patterns influence them. Here, we show the impacts of different land-use patterns on fungal PBAPs within the Brazilian Atlantic Forest biodiversity hotspot. Spanning a distance of ca. 600 km within this biome, we collected fungal bioaerosols in the following land-use patterns: a 25-year-old coffee plantation, a 2.5-year-old Eucalyptus plantation, a 3-months-old maize crop, a 2-year-old and another 7-year-old native forest reforestation area, and a native forest fragment. Using the portable Burkard air sampler, a total of 14 morphotype-species were found. Cladosporium sp. comprised as much as about 95% of all fungal spores collected, being present in all samples (frequency of 100%). Forest systems had as much as 5-to-19-fold more fungal spores in the air than maize crops. Sampling height and time (morning vs. afternoon) did not influence fungal concentration and number of species. In addition, using data derived from an eddy covariance tower in the Eucalyptus site, we estimated the emission rate to be 6649 spores m-2 s-1. Our study confirms that land-use patterns affect fungal PBAPs, and that replacing large areas of native Atlantic Forest by monospecific stands, a homogenisation of airborne fungi is occurring, with unknown consequences for climate regulation.