Mulching (cutting of vegetation without removal of clippings) is used as a low-cost method for maintaining remote or abandoned grasslands in Slovakia. The likely consequence of mulching is seasonal nutrient enrichment resulting from decomposition of plant litter by saprotrophic organisms. The potential changes in biodiversity of the ecosystem caused by long-term application of mulching are to date only very poorly understood. In order to examine the impact of mulching on soil mycobiota, we compared six different grassland management regimes applied over nine years on a sub-montane oligotrophic Nardus pasture in the Central Slovakia. The diversity of soil fungi was assessed using DNA metabarcoding of the ITS2 regions of the nrRNA locus performed by Illumina MiSeq. We focused on a particular group of macrofungi which is characteristic of traditionally managed and undisturbed European grasslands, and which are often the dominant soil fungi in these habitats. These are collectively known as CHEGD fungi (the acronym of the constituent taxa: Clavariaceae, Hygrophoraceae, Entolomataceae, Geoglossaceae and Dermoloma). We compared the relative abundance and diversity of CHEGD fungi with the total fungal and plant diversity. CHEGD fungi were dominant across all management treatments. Although there were no statistical effects of treatments on total fungal richness and diversity, CHEGD fungi and vascular plants diversity and richness were lower on plots where mulching or no management were imposed, suggesting that such management regimes would have a negative impact on grassland fungi. However, no single treatment covered the total CHEGD diversity of the study, indicating that the localized use of mulching in addition to traditional managements can enhance overall diversity of grasslands in the area. Our results also suggest that the impact of mulching depends on the season when the grassland is mulched and it might be reduced by combination with other management treatments. The high relative abundance and sensitivity of CHEDG fungi in oligotrophic grasslands to management treatments makes them excellent indicators of grassland natural quality and is consistent with the ecological importance of this fungal group.