Enhancement of Mycelial Growth and Antifungal Activity by Combining Fermentation Optimization and Genetic Engineering in S10.

The biocontrol strain S10 was isolated from tomato leaf mold. The fermentation broth of strain S10 can effectively control Fusarium head blight (FHB), caused by . Enhancing antifungal activity is essential in advancing its commercialization. In this study, we aimed to improve the antifungal activity of S10 by integrating fermentation optimization and genetic engineering. Single-factor experiments revealed that seven parameters, namely corn flour, yeast extract, NaNO, CaCO, KHPO, KCl, ZnSO·7HO, and MnCl·4HO, were identified as significant components. A Plackett-Burman design (PDB) indicated that corn flour, yeast extract, and ZnSO·7HO were the most critical variables affecting its inhibitory activity and mycelial biomass. The fermentation medium was further determined based on the steepest climbing experiment and a Box-Behnken design (BBD), and the mycelial dry weight of S10 was improved from 2.13 g/L in Gauze's synthetic No. 1 medium to 8.12 g/L in the optimized medium, closely aligning with the predicted value of 7.98 g/L. Under the optimized medium, the antifungal rate of increased from 67.36 to 82.2%. The spore suspension of strain S10 cultured in the optimized medium substantially improved its biocontrol efficacy against FHB. Moreover, disruption of the key gene led to increased antifungal activity of strain S10 against . Importantly, the antifungal activity of Δ was greatly increased under the optimized fermentation medium. This study suggests that the gene negatively regulates bioactive compound biosynthesis, and the optimized medium provides favorable conditions for the growth of S10. These observations establish an extended basis for the large-scale bioactive metabolite secretion of S10, providing a strong foundation for sustainable FHB management in agriculture.