Fungi are one of the most diverse groups of organisms and considered as one of the least-explored biodiversity resources. Soil fungal community was investigated in two agricultural sites in Khartoum state, Sudan, during two seasons. A total of 42 soil samples were collected, their physicochemical properties were determined, then subjected to metabarcoding and metagenomic analyses. fungal community composition, diversity and microbial trophic modes were determined utilizing R software packages. From both sites, a total of 15 different phyla were detected, out of them, 11 were the most abundant and frequent. Ascomycota was the dominant phylum (86.54% total abundance), followed by the Basidiomycota (8.29%). The dominant class was Sordariomycetes (41.02%), followed by Dothideomycetes (19.80%). Aspergillus (6.2%), Curvularia (6.0%), Neurospora (5.8%) and Fusarium (4.9%) were the most abundant genera. Deniquelata for the first time being recorded in Sudan. Apha diversity measures revealed sample richness ranging from 71 to 361 ASVs, and Shannon index ranging from 2.794 to 5.087. The two sites had significantly different alpha diversity. Land-use types were also significantly different in their diversity regardless of site. Season had no effect on alpha diversity of soil fungal communities. Beta diversity analysis indicated significant differences between the two sites and the different land-use types. No significant differences in the community structure recorded between the two seasons. The dominant trophic mode among the assigned ASVs in soil mycobiome was saprotroph mode (22.11%). Results of this study reveals that fungal community structure is affected by site and land-use type. It gives a comprehensive database for the mycobiome of the agricultural soil in Khartoum state.