Abstract
Trichoderma reesei strains belong to the group of industrially important microorganisms because of their ability to produce cellulase enzymes. A method for strain improvement is protoplast fusion. We tested the potential of electrofusion in protoplast hybridization in the case of two auxotrophic strains (ade − lys − and ade − leu −). After dielectrophoretic cell collection (500 V/cm, 1 MHz) we used rectangular fusion pulses in a range of electric field strengths E = 1–4.5 kV/cm and pulse durations τ = 25–120 μs (number of pulses N = 1–5). An optimal yield of hybrids able to form colonies on minimal/ade-medium was found at E = 4.1 kV/cm, τ = 75 μs, and N = 5.28% of the fusants selected statistically were stable and most of them showed a changed behaviour in catabolite repression. These results indicate new possibilities for strain improvement in T. reesei.
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