Agricultural practices can affect the diversity and community structure of soil fungi. This study investigates the impact of long-term agricultural practices on soil fungal diversity in the Loess Plateau of northwestern China. Different tillage practices have been implemented for 13 years, and their impact on soil fungi is assessed using high-throughput Illumina Sequencing. This study found a total of 2071 fungal Amplicon Sequence Variants (ASVs), and these were assigned to 25 different phyla, 372 families, and 496 genera. The fungal communities were dominated by Ascomycota (52.1%), followed by Zygomycota (14.3%) and Basidiomycota (9.0%). In general, the soil exhibited higher fungal community abundance, richness, and diversity in winter than in summer. Notably, no-tillage or stubble retention resulted in greater diversity than conventional tillage, with no-tillage combined with stubble retention resulting in the highest fungal richness, diversity, and network complexity in both summer and winter. These findings indicate that no-tillage with stubble retention is beneficial for biological soil components, which favors the establishment of abundant and diverse soil fungal communities in the Loess Plateau of China. The present study expands the knowledge of fungal communities in agro-ecosystems and the long-term ecosystem benefits of tillage practices.