• Trichoderma virens
• textile dyes
• moonlight enzyme
• lignin
• laccase
• Coprinus cinereus
• ABTS

The goal of this work was to investigate laccase functions in Trichoderma virens. The strain under study, I10, was known to efficiently antagonize plant pathogens and to be involved in mycoparasitism against sclerotial fungi. For this a laccase activity could be needed to penetrate cell walls of sclerotia rind, the external layer mostly rich in melanin. Six putative laccase genes, likely associated to different biological roles, were previously identified in the T. virens genome. These possible laccase functions could be both interesting for biocontrol and biotechnological applications related to ligninolysis
The analysis performed in this thesis work included the development of a biochemical and molecular screening test for laccase activity and a comparison with the lignocellulolytic basidiomycete Coprinus cinereus in biochemical and molecular assays.
The main issues to be assessed were:
• T. virens production of extracellular laccases in the presence of lignocellulosic biomass;
• the differential expression of the T. virens six putative laccase genes;
• the T. virens laccase production compared to the one of the model organism C. cinereus.
In order to accomplish these tasks, several experimental trials have been set during a hosting period at the laboratories of the Section for Sustainable Biotechnology - Copenhagen Institute of Technology, Aalborg University, Denmark. The general experimental design can be resumed as follows:
• growth of fungal cultures on both a lignocellulosic and non-lignocellulosic media;
• analysis of laccase activity according to the ABTS assay;
• RT-PCR with primers specific for the six T. virens putative laccase genes and C. cinereus lcc1 gene.