Abstract
Black shank disease in tobacco, caused by Phytophthora nicotianae, can lead to yield losses of 30%-50% upon outbreak. Recently, biochar derived from agricultural waste has shown significant potential in controlling soil-borne diseases, though its mechanisms remain unclear. Over a 3-year observation period, we found that the incidence of black shank was significantly lower in plots amended with biochar compared with normal cultivation plots. To investigate the underlying mechanisms, we studied both the direct and indirect effects of biochar on black shank. Direct antifungal assays indicated that biochar reduced the total number of sporangia by 53.91%. Further pot experiments revealed a 62.34% reduction in the P. nicotianae population in the soil following biochar application. Additionally, biochar application led to notable changes in soil physicochemical properties and microbial community composition. Microbial species analysis showed that biochar promoted the aggregation of beneficial microbes such as Sphingomonas, Flavisolibacter, and Mucoromycota. Functional predictions using the PICRUSt 2 software revealed that biochar enhances bacterial functions related to antimicrobial substance synthesis (Tetracycline biosynthesis), detoxification metabolism (D-arginine and D-ornithine metabolism, arginine and proline metabolism), and lipid and fatty acid metabolism (Lipopolysaccharide biosynthesis, fatty acid biosynthesis), while fungal functions showed no significant changes. This suggests that rhizosphere bacteria play a more prominent role in the suppression of black shank by biochar, a finding supported by partial least squares path modeling analysis. Therefore, we hypothesize that biochar not only directly inhibits P. nicotianae growth but also regulates the composition of the rhizosphere microbial community, inducing the production of antimicrobial substances by rhizosphere bacteria, effectively preventing P. nicotianae invasion.IMPORTANCEBlack shank, a global soil-borne fungal disease in tobacco, currently lacks effective control methods. Notably, biochar derived from agricultural waste has shown significant potential in controlling soil-borne diseases. Over a 3-year observation period, we found that plots amended with biochar had a significantly lower incidence of black shank compared with normal cultivation plots. However, the mechanisms of disease suppression remained unclear. Through in vitro antifungal assays and pot experiments, we discovered that tobacco-derived biochar can directly inhibit the growth of the pathogen. Additionally, biochar regulates the composition of the rhizosphere microbial community, inducing rhizosphere bacteria to produce antimicrobial substances, effectively preventing pathogen invasion. This discovery reveals both the direct and indirect mechanisms by which biochar suppresses black shank in tobacco. It provides a scientific basis for developing green control technologies for black shank and offers theoretical support for the application of biochar in managing soil-borne diseases in tobacco cultivation areas.