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The present study was carried out in Ain Shams Center of Genetic engineering and biotechnology faculty of agriculture (ACGEB) Trichoderma strains are known to produce extracellular lytic enzymes, including chitinase, cellulase, and protease which have a wide range of applications in the biological controlled, agriculture and other industries. Because of the importance of these products, the present study aimed to genetically improve extracellular lytic enzyme. This study aimed to carry out genetic improvement for some strains of Trichoderma spp. to increase its productivity of enzymes through mutation induction UV (ultra violet irradiation) as one of physical mutants.
Isolation and characterization of Trichoderma strains
1.1 four isolates were isolated from soil samples collected from el Behera Peninsula one of them was Trichoderma haizinum(T1) and three isolates were Trichoderma viride (T2, T3 and T4) and all isolates defined according to the morphological specifications of colonies and the microscopy inspection.
1.2 Antagonism experiment carried out between Trichoderma fungi and Fusarium oxysporum , Rhizoctonia solani and Pythium sp pathogenic fungi were plated in petri plates in vitro and shown that the strains T1 and T2 were the most efficient strains in antithesis with one of resistance means of Trichoderma.
1.3 According to the importance of Trichoderma strains in this study and the ability of using them in Biocontrol field. So, it was a successful try to employ those strains on the biochemical and molecular level using five primers in ISSR PCR technique.
1.4 An evaluation of strains ability to produce enzymes carried and showed that (T1) Trichoderma harizinum was the highest in chitinase production (Enzyme activity 160 u/ml /min), while the strain (T2) Trichoderma viride was the highest in β-1,3-glucanase production (Enzyme activity 225 u/ml /min).
2. Genetic Improvement of Trichoderma strains to inc analyzer enzymes production.
2.1 The highest strains of Chitinase and β-1,3-glucanase enzyme production had mutation exposure with a volume (300 ug/100 ml) and it was in the volume of produced enzymes.
2.2 A second mutation phase carried out with the same concentration of mutant material and enzyme measurement once more, the percentage of enzyme production was to the first phase of mutation and the T112 strain was the highest in chitinase production (215 u/ml /min) with a percentage (236%), while the T111 strain was the highest in β-1,3-glucanase production (265 u/ml/min) with a percentage (135%) compared with the original strain.
According to the results obtained, the study indicated to the importance of Trichoderma strains in biotechnology applications field and explained the ability of its usage as biological compost due to its ability on producing the enzymes which analyze the pathogenic plants fungi walls.