Bioactive proteins from Trichoderma for the control of Fusarium: Evaluation of antagonist efficacy and potential for agronomic application

Abstract

The identification of Trichoderma proteins has significantly advanced in recent years, highlighting their potential as alternatives to chemical pesticides. This study investigated the in vitro antagonistic effects of Trichoderma harzianum (Pista-9) against Fusarium oxysporum, Fusarium graminearum, and Fusarium verticillioides. Additionally, the differential expression of proteins in response to co-cultivating Pista-9 and F. oxysporum in a low-carbon medium to promote fungal competition was evaluated. A liquid culture was prepared by adding 100μL of 10⁷ conidia/mL. As a test, Pista-9 was grown in the presence of the F. oxysporum live and heat-inactivated. The cultures were incubated at 28 °C for 7 days at 180 rpm. Then the solutions were filtered, and ammonium sulfate 0–85 % saturation was added. After the precipitation process, the protein extract was analyzed by one-dimensional electrophoresis and LC-MS/MS. The effect of protein extracts (300 μg/mL) on the growth of F. oxysporum was evaluated in vitro. In vitro antagonism tests demonstrated that Pista-9 is an effective biocontrol agent. Electrophoresis revealed that extracts presented majority bands ranging from 10 to 75 kDa and proteins such as glutaminase, metalloproteinase, chitinase, and peptide hydrolase were identified. The extract from Pista-9 was able to inhibit 71 % of F. oxysporum growth. Furthermore, the growth-promoting potential of the extract was evaluated through its application on tomato leaves. The results showed that the extract promoted growth and increased biomass. Proteins from Pista-9 hold significant potential as bioactive molecules for controlling phytopathogens and promoting plant growth, representing a valuable resource for biotechnological applications aimed at developing sustainable agricultural solutions.