<p>Purpose:&#xa0;<i>Colletotrichum graminicola</i> is a significant threat to maize production at global scale. Conventional chemical-based plant disease management strategies pose multiple risks to human health and contribute towards environmental degradation. Biochar offers a sustainable and eco-friendly alternative for the plant disease management. Methods: This study evaluated the combined effects of maize biochar and <i>Bacillus subtilis</i> as a bio-control agent, on the suppression of maize anthracnose. A mixture of soil with 3 and 6% concentrations of biochar separately and in combination with or without <i>B. subtilis</i> was used against <i>C. graminicola</i> in maize. Additionally, in vitro dual culture assays were performed to study the antagonistic activity of <i>B. subtilis</i> against the pathogen in the presence of biochar. Results: The highest mycelial growth inhibition (57%) was observed with <i>B. subtilis</i> alone, as compared to the control treatments. Soil amendment with 6% biochar resulted into disease severity index of 20% in the presence of <i>B. subtilis</i>, with the reduction in disease incidence to 10%. Higher dose of biochar in soil not only improved the maize growth parameters but also the activities of key defense related enzymes, including peroxidase and catalase, particularly in plants inoculated with <i>C. graminicola</i>. The highest chlorophyll content was recorded in the treatment with 6% biochar in the absence of the pathogen. Conclusion: Overall, the application of 6% maize biochar, especially in combination with <i>B. subtilis</i>, significantly reduced anthracnose symptoms in maize and improved physiological and biochemical plant response towards pathogen as compared to the 3% biochar. To the best of our knowledge, this study is the first to report the synergistic efficacy of biochar and <i>B. subtilis</i> in managing maize anthracnose.</p>