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The aim of this study was to develop a protocol for the production of fungal bio-pesticides with high efficiency, low cost, and non-polluting fermentation, while also increasing their survival rate under field conditions. This is the first study to develop biocontrol Trichoderma harzianum transformants TS1 that are resistant to benzimidazole fungicides. Agricultural corn stover and wheat bran waste were used as a medium and inducing carbon source for solid fermentation. Spore production was observed, and the method was optimized using single-factor tests with 4 factors at 3 levels in an orthogonal experimental design to determine the optimal culture conditions for T. harzianum TS1. In this step, we determined the best conditions for fermenting the biocontrol fungi. The optimal culture conditions for T. harzianum TS1 were cultivated for 8 days, a ratio of straw to wheat bran of 1:3, ammonium persulfate as the nitrogen source, and a water content of 30 mL. Under optimal culture conditions, the sporulation of T. harzianum TS1 reached 1.49 x 10(10) CFU/g, which was 1.46-fold higher than that achieved before optimization. Increased sporulation of T. harzianum TS1 results in better utilization of space and nutrients to achieve control of plant pathogens. This method allows for the recycling of agricultural waste straw.

Optimization of solid-state fermentation conditions for Trichoderma harzianum using an orthogonal test