ANTIFUNGAL, SYNERGISTIC INTERACTION BETWEEN CHITINOLYTIC ENZYMES FROM TRICHODERMA-HARZIANUM AND ENTEROBACTER-CLOACAE

Biocontrol strains from the genera Enterobacter and Pseudomonas and two chitinolytic enzymes from Trichoderma harzianum isolate P1 were combined and tested for antifungal activity in bioassays. Inhibitory effects on spore germination and germ tube elongation of Botrytis cinerea, Fusarium solani, and Uncinula necator were synergistically increased by mixing fungal enzymes and cells of Enterobacter cloacae but not of Pseudomonas spp. Culture filtrate of E. cloacae contained antifungal compounds and produced moderate levels of inhibition, either in plate assays or in bioassays conducted in potato-dextrose broth. However, the combination of bacterial culture filtrate with fungal chitinolytic enzymes generated only an additive response, indicating that the presence of bacterial cells was required for a synergistic effect. Chitinolytic enzyme activity in the presence of chitinous substrates enhanced the growth of E. cloacae and readily restored the ability of bacterial cells to bind to hyphae of the pathogens despite high concentrations of D-glucose or sucrose in the medium. The results of this study suggest that transgenic bacteria, capable of binding to fungal cell walls and expressing fungal genes encoding cell wall-degrading enzymes, may be powerful biocontrol agents.