Actinobacteria of the genus Streptomyces are part of the soil microbiota from rice-growing areas and along with other microorganisms, such as Trichoderma spp., combine to act as natural enemies against the destructive soil-borne pathogen Rhizoctonia solani, causal agent of rice sheath blight disease. In this study, seven actinobacteria of the genus Streptomyces and three fungi of the genera Trichoderma and Purpureocillium were isolated from soils cultivated with rice using the serial dilution method. Streptomyces spp. M2A2 was selected for its ability to significantly reduce the in vitro growth of R. solani by 52% after 96 h by antibiosis in dual culture, while in the control treatment the mycelial growth was 100%. Furthermore, biocontrol efficacy of treated plants of the susceptible cultivar Fedearroz 68 with actinobacteria was confirmed and the onset of symptoms were delayed up to 14 days, compared to the control treatment. Rice plants treated with Streptomyces spp. M2A2 showed lesions of R. solani reaching 0.7% of the plant height, the effectiveness of this treatment was similar to the difenoconazole treatment, whereas in the control treatment, the lesions covered 34% of the plant height. When compared to the antagonist fungus Trichoderma spp. M2H1, Streptomyces spp. M2A2 presented a better performance of biological control. The results clearly demonstrated that Streptomyces spp. M2A2 isolate from soils of rice growing areas has biocontrol efficiency against R. solani and therefore can be a promising biocontrol agent.