ABSTRACT Macrophomina (Botryosphaeriaceae) is one of the main genera of soilborne phytopathogenic fungi, which causes root and seed rot in more than 800 host plants worldwide. Recent phylogenetic studies identified the species M. phaseolina and M. pseudophaseolina in Trianthema portulacastrum and Boerhavia diffusa in melon and watermelon production areas in northeastern Brazil. Therefore, the objective of this study was: i) to verify the effect of temperature and salinity on the mycelial growth of M. phaseolina, M. pseudophaseolina and M. euphorbiicola, ii) to assess their pathogenicity on melon and watermelon seedlings, and iii) to determine their sensitivity to the fungicide carbendazim. The optimal temperature for mycelial growth rate (MGR) for Macrophomina spp. ranged from 27.18 ºC (CMM4771 – M. pseudophaseolina) to 31.80 ºC (CMM4763 – M. phaseolina). For the effect of salinity on mycelial growth of Macrophomina isolates, the EC50 ranged from 103.76 (CMM4868 – M. euphorbiicola) to 315.25 mM (CMM4801 – M. pseudophaseolina). The pathogenicity test demonstrated that M. phaseolina, M. pseudophaseolina and M. euphorbiicola are pathogenic on melon with M. phaseolina exhibiting a higher level of virulence. Macrophomina euphorbiicola isolates did not cause disease in watermelon. The most sensitive isolates to the fungicide carbendazim were CMM4868, CMM4867 (M. euphorbiicola) and CMM1531 (M. phaseolina) with EC50 of 0.003, 0.012 and 0.012 mg.L-1 a.i., respectively. All Macrophomina spp. used in these experiments were pathogenic to the tested melon and watermelon cultivars with the exception of the M. euphorbiicola isolate that did not cause damage to watermelon.