Bacterial antagonists of Helminthosporium solani were submitted to different tests in order to determine the role of antibiosis in their antagonistic interaction. Among the bacterial strains tested, seven (Alcaligenes piechaudii, Aquaspirillum autotrophicum, Cellulomonas fimi, Pseudomonas chlororaphis, Pseudomonas putida (strains 94-19 and E-30) and Streptomyces griseus) were shown to produce agar diffusible metabolites inhibiting H. solani mycelial growth and/or conidial germination. Differential activity was revealed when diffusible metabolites were extracted from either pure cultures of these antagonists or dual culture in the presence of H. solani. The results presented indicate that the methodology employed could be a decisive factor in whether or not antibiosis can be identified as a mode of action of biocontrol agents.