Enhancement of Lytic Enzymes Activity and Antagonistic Traits of Trichoderma harzianum Using γ-Radiation Induced Mutation

Trichoderma species are known as effective agents used for biological control of plant pathogenic fungi. The Trichoderma harzianum and its mutant isolates were cultured and their traits including, mycelial growth, antagonistic activity and extracellular proteins and enzymes production (Chitinase and Cellulase) were investigated to select the most effective mutant isolates against plant pathogenic fungus Rhizoctonia solani. Also, the purity and composition of enzyme-rich protein samples were evaluated under denaturing gel electrophoresis. This study clearly showed the possibility of improving mycelia growth rate (from 1.18 to 1.33 cm d-1), the antagonistic capability of Trichoderma (from 54.9% growth inhibition of R. solani to 66%), extracellular proteins and enzymes production for biological control of plant diseases through mutation with γ-radiation. Also, compared to wild type strain, protein production in the mutant isolates increased. Moreover, the highest specific chitinase enzyme activities were observed in mutant isolates T. h M8 (42.48 U mg-1) and T. h M15 (38.25 U mg-1). Trichoderma mutant of T. h M8 maintained higher mycelia growth rate and higher ability to inhibit growth of R. solani. The SDSPAGE profiles had several enzyme protein bands such as CelloBioHydrolases (CBHs), EndoGlucanases (EGs), β-Glucosidases (BGLs), endochitinases, and β-(1, 4)-N-acetyl glucoaminidases. SDS-PAGE analysis indicated the presence of different protein bands in the range of 10.5 to 245 KDa. Interestingly, expression of chitinase in 95 percent of mutants was higher than wild type of T. harzianum. The results showed that gamma mutation could increase the efficiency and amount of enzymes in T. harzianum, while these enzymes are involved in antagonistic properties of T. harzianum.