This work was carried out with the objective of performing a molecular and aggressiveness characterization of F. solani and F. oxysporum f.sp. passiflorae collected in the Pantanal, Cerrado and Amazon biomes. We selected the most aggressive isolates for use in breeding programs aiming resistance to Collar Rot and Fusariosis. For inoculation of the isolates of F. oxysporum f.sp. passiflorae the washed root method was used. The molecular characterization of the isolates was carried out by partial sequencing of the Transcribed Internal Spacer of the rDNA region. The isolates of F. solani formed two distinct groups in relation to aggressiveness. Among all isolates, FSUNEMAT 40 and FSUNEMAT 46 were the most aggressive. The model with K=2 was taken as the best model to explain the genetic structure of the F. solani populations, with clear combinations of genes from both gene pools. There were three groups with respect to the aggressiveness of the isolates of F. oxysporum f.sp. passiflorae, with the isolated FOUNEMAT 22 being the most aggressive. In view of these results, the isolates of F. solani collected in P. edulis in the state of Mato Grosso presented a high molecular variability independent from the biome of origin, and this was also observed in the tests of aggressiveness. The results indicate the need to consider the molecular variability and the aggressiveness of the pathogens in the evaluation of genotypes of sour passion fruit in programs of selection of resistant cultivars.