Irish potato and tomato are among the most widely cultivated crops in Uganda. In 2020, samples of affected potato tubers and tomato fruits were collected from farms across four regions in Uganda for analysis. A total of 22 strains of Fusarium spp. were isolated from potato tubers and seven strains were isolated from tomato fruits. Identification of the fungal species was accomplished using cultural and morphological characteristics, as well as DNA sequencing targeting specific regions: ITS1–5.8S–ITS2, parts of the elongation factor 1 (tef 1) gene, and beta-tubulin (β-tub) gene. The analysis of the isolated strains from potato tubers revealed the presence of Fusarium incarnatum-equisety species complex, F. sambucinum species complex, F. oxysporum species complex, F. solani species complex. Additionally, F. incarnatum-equiseti species complex was detected in tomato fruits. All the investigated strains exhibited the ability to successfully infect both injured tomato fruits and potato tubers. Tested strains were susceptible to difenoconazole (ЕС50 = 0.08–8.5 mg/L) and thiabendazole (EC50 = 0.67–5.1 mg/L).