Colletotrichum falcatum is the major fungal pathogen causing sugarcane red rot. Four antagonistic bacterial
strains exhibiting biocontrol activity against this pathogen in greenhouse conditions were characterized for
production of different antifungal metabolites and biocontrol determinants to elucidate the mechanism of action
involved in their antagonistic activity. The strains were also evaluated under field conditions to assess their biocontrol
potential. All the strains produced hydrogen cyanide (HCN), and volatile and diffusible antibiotics. In
addition, the Ochrobactrum intermedium strain NH-5 produced siderophores and the broad spectrum antibiotic
2, 4-diacetylphloroglucinol (2,4-DAPG); Pseudomonas sp. NH-203 produced siderophores, and Pseudomonas sp.
NH-276 produced protease. Two strains, Ochrobactrum intermedium NH-5 and Stenotrophomonas maltophilia NH-
300, exhibited good biocontrol activity, suppressing red rot by 44–52% on two sugarcane varieties, SPF-234 and
Co-1148, in field experiments. The strains gave consistent results in three consecutive years and showed potential
to be used as biopesticides.