This research was throughout 2018-2020, filamentous fungi of agricultural importance were isolated from the Cerro Las Tetillas, located in the municipality of Salvatierra, Guanajuato. The hill was divided into three strata: upper, middle and lower. Two hundred soil samples were taken at depths of 0-15 and 15-30 cm. These were processed in the biology laboratory, belonging to the Higher Technological Institute of Salvatierra. Two isolation techniques were used: direct seeding and serial dilutions, over Petri dishes with potato dextrose agar culture medium and 100 mg of streptomycin to control bacteria. One hundred and thirteen axenic strains corresponding to 16 genera identified from “Cerro Tetillas” were obtained. At the upper stratum, 13 genera were identified, while in the middle and lower strata, nine genera were identified in each one of them. The diversity of soil fungi found in the different strata of the hill may be due to physicochemical characteristics of soil, native vegetation, interaction and abundance of soil fungi species, human disturbance and rain runoff.