Proteomics of the biological control strain Trichoderma atroviride

Trichoderma atroviride has a natural ability to parasitise phytopathogenic fungi such as Rhizoctonia solani and Botrytis cinerea therefore providing an environmentally sound alternative to chemical fungicides in the management of these pathogens. Two-dimensional electrophoresis was used to display cellular protein patterns of T. atroviride (T. harzianum P1) grown on media containing either glucose or R. solani cell walls. Protein profiles were compared to identify T. atroviride proteins up-regulated in the presence of theR. solani cell walls. Twenty four protein spots were identified using matrix assisted laser desorption ionisation mass spectrometry, liquid chromatography mass spectrometry and Nterminal sequencing. Three novel proteases to T. atroviride were up-regulated and identified as vacuolar serine protease, vacuolar protease A and trypsin-like protease. Two of these proteases, vacuolar protease A and vacuolar serine protease have been sequenced and cloned using chromosome walking PCR. Vacuolar protease A has two predicted introns, a predicted signal peptide and contains an aspartic proteinase conserved domain. Vacuolar serine protease has one predicted intron, a predicted signal peptide and contains a peptidase S8 conserved domain.