A high frequency of plants showing symptoms of wilting and root rot has been observed in tomato‐producing areas in Santarém and Belterra, Pará, Brazil. These plants showed symptoms ranging from yellowing to wilting, with roots displaying reduced development and darkening of the xylem. Four fungal isolates, derived from a collection of 12 obtained from symptomatic plants, were chosen for identification using both morphological and phylogenetic species concepts. To determine the identity of the fungi, DNA was extracted and used in PCR reactions. PCR products were sequenced from the TEF‐1α and RBP2 regions and compared with sequences in GenBank using BLASTn. Based on the phylogenetic analyses, the isolates were identified as Fusarium falciforme and Fusarium suttonianum, belonging to the species complex Fusarium solani – (FSSC), and Fusarium triseptatum, belonging to the species complex Fusarium oxysporum – (FOSC). The fourth isolate, COUFPI 295, grouped within a clade of F. kalimantanense and F. sangayamense from FOSC. The fulfilment of Koch's postulates confirmed that all isolates induced symptoms of root rot and concomitant reduction in root mass, leading to observable wilting symptoms. This is the first report of F. suttonianum, F. falciforme and F. triseptatum causing root rot and wilt in tomato plants in Brazil.