Tracking the roots of cellulase hyperproduction by the fungus Trichoderma reesei using massively parallel DNA sequencing

• DOI: 10.1073/pnas.0905848106
• Publication Date: 2009-09-03T02:00:35Z
• Authors: Stéphane Le Crom; Wendy Schackwitz; Len Pennacchio; Jon K. Magnuson; David E. Culley; James R. Collett; Joel Martin; Irina S. Druzhinina; Hugues Mathis; Frédéric Monot; Bernhard Seiboth; Barbara Cherry; Michael Rey; Randy Berka; Christian P. Kubicek; Scott E. Baker; Antoine Margeot
• Keywords: Hypocrea; Massive parallel sequencing; Fungal protein

Trichoderma reesei (teleomorph Hypocrea jecorina) is the main industrial source of cellulases and hemicellulases harnessed for hydrolysis biomass to simple sugars, which can then be converted biofuels such as ethanol other chemicals. The highly productive strains in use today were generated by classical mutagenesis. To learn how cellulase production was improved these techniques, we performed massively parallel sequencing identify mutations genomes two hyperproducing (NG14, its direct descendant, RUT C30). We detected a surprisingly high number mutagenic events: 223 single nucleotides variants, 15 small deletions or insertions, 18 larger deletions, leading loss more than 100 kb genomic DNA. From events, report previously undocumented non-synonymous 43 genes that are mainly involved nuclear transport, mRNA stability, transcription, secretion/vacuolar targeting, metabolism. This homogeneity functional categories suggests multiple changes necessary improve not simply few clear-cut events. Phenotype microarrays show some result strong carbon assimilation pattern mutants with respect wild-type strain QM6a. Our analysis provides genome-wide insights into induced mutagenesis filamentous fungus areas generation enhanced T. applications biofuel production.