Involvement of lytic enzymes and secondary metabolites produced by Trichoderma spp. in the biological control of Pythium myriotylum.

Abstract

This investigation was aimed to evaluate the antimicrobial activities and involvement of extracellular lytic enzymes produced by four strains of Trichoderma in the inhibition of Pythium myriotylum. Antagonistic effects were tested by dual culture. Activities of lytic enzymes were evaluated from the filtrate of each strain after cultivation in selected media. Organic extracts were obtained from liquid media subsequent to the cultivation of Trichoderma in potato dextrose broth (PDB). Non-volatile organic compounds such as polyphenols and flavonoids were evaluated spectrophotometrically while volatile organic compounds (VOCs) were analyzed by gas chromatography coupled with mass spectrometry (GC-MS). The antimicrobial activity of the organic extracts was determined using the poisoning method. Results have shown that all the strains were antagonists against P. myriotylum. T. erinaceum (IT-58), T. gamsii (IT-62), T. afroharzianum (P8), and T. harzianum (P11) that were found to produce cellulase, protease, and xylanase. Over 20 compounds were identified in each extract, including esters, lactones, and organic acids. The organic extracts also contained high amounts of polyphenolic compounds and flavonoids and significantly inhibited the mycelial growth of P. myriotylum. The minimal inhibition concentrations were 80μg/μL, 40μg/μL, 20μg/μL, and 10μg/μL, for extracts obtained from T. erinaceum (IT-58), T. gamsii (IT-62), T. afroharzianum (P8), and T. harzianum (P11), respectively. There was significant correlation between the production of total polyphenol and flavonoid content and the antagonistic effects of the tested strains.