Project description:Trichoderma reesei is the main industrial producer of cellulases and hemicellulases used to depolymerize biomass in many biotechnical applications. Many production strains in use have been generated by classical mutagenesis. In this study we characterized genomic alterations in hyperproducing mutants of T. reesei by high-resolution comparative genomic hybridisation tiling array. We carried out aCGH analysis of four hyperproducing strains (QM9123, QM9414, NG14 and RutC-30) using QM6a genome as a reference. ArrayCGH analysis identified dozens of mutations in each strain analyzed.
Project description:Trichoderma reesei is the main industrial producer of cellulases and hemicellulases used to depolymerize biomass in many biotechnical applications. Many production strains in use have been generated by classical mutagenesis. In this study we characterized genomic alterations in hyperproducing mutants of T. reesei by high-resolution comparative genomic hybridisation tiling array. We carried out aCGH analysis of four hyperproducing strains (QM9123, QM9414, NG14 and RutC-30) using QM6a genome as a reference. ArrayCGH analysis identified dozens of mutations in each strain analyzed. 2.1 million oligonucleotide probe custom aCGH (HD2 format, RocheNimblegen) was designed according to T. reesei strain QM6a genome v2.0 (http://genome.jgi-psf.org/Trire2/Trire2.home.html). 14 samples are included in this set; 3 replicates of each strain (except two replicates of QM9123) were analyzed (four mutant strains and QM6a control strain for self-hybridization)
Project description:Investigation of whole genome gene expression level changes in response to different light conditions of the T. reesei QM9414 parental strain and the deletion strains delta-phlp1, delta-gnb1 and delta gng1, cultivated on 1 % microcrystalline cellulose. The mutants analyzed in this study are further described in Tisch et al. 2011: Carbohydrate degradation is significantly regulated by light and the phosducin like protein PhLP1 in Trichoderma reesei (Hypocrea jecorina). We used two biological replicates of four T. reesei strains (QM9414, delta-phlp1, delta-gnb1 and delta-gng1), cultivated in constant light (LL, 1800 lux) or constant darkness (DD) on microcrystalline cellulose.
Project description:Investigation of whole genome gene expression level changes in response to different light conditions of the T. reesei QM9414 parental strain and the deletion strains delta-phlp1, delta-gnb1 and delta gng1, cultivated on 1 % microcrystalline cellulose. The mutants analyzed in this study are further described in Tisch et al. 2011: Carbohydrate degradation is significantly regulated by light and the phosducin like protein PhLP1 in Trichoderma reesei (Hypocrea jecorina).
Project description:The ascomycete Trichoderma reesei is one of the most well studied cellulolytic fungi and widely used in the biotechnology industry, as in the production of second-generation bioethanol. Carbon catabolite repression (CCR) mechanism adopted by T. reesei is mediated by the transcription factor CRE1 and consists in the repression of genes related to the production of cellulase when a readily available carbon source is present in the medium. Using RNA sequencing this study aims to contribute to understanding of CCR during growth in cellulose and glucose, by comparing the mutant strain of T. reesei Δcre1 with its parental, QM9414.
Project description:We perform a self hybridisation comprative genomic hybridization (CGH) in order to validate the probe tiling design we done on Trichoderma reesei. This hybridization was done using QM6a wild type strain.
Project description:Investigation of whole genome gene expression level changes in response to different light conditions of the T. reesei QM9414 deletion strains delta-blr1, delta-blr2 and delta env1 cultivated on 1% microcrystalline cellulose. Perception and proper interpretation of environmental signals is crucial for survival in any natural habitat. Although the biotechnological workhorse Trichoderma reesei (Hypocrea jecorina) is predominantly known for its capability of efficient plant cell wall degradation, recent studies show that it has not lost its evolutionary heritage. Transmission of nutrient signals via the heterotrimeric G protein pathway has been shown to be influenced by light. We show that this interconnection is mainly established by the light regulatory protein ENV1 and the phosducin-like protein PhLP1 via mutual transcriptional regulation and influence on GNB1 (G protein beta subunit) function. ENV1 thereby exerts a more severe effect on gene transcription than BLR1 or BLR2. Lack of either one of the photoreceptors or PhLP1, GNB1 or GNG1 leads to a partial shutdown of processes upregulated in light, indicating that heterotrimeric G protein signalling exerts its major function in light and is a target of the light response machinery. Consequently, signals transmitted via the G protein pathway are of different relevance in light and darkness. Investigation of regulation of glycoside hydrolases as one of the major output pathways of this mechanism revealed that 79% of all genes belonging to this group, representing all GH-families available in T. reesei, are potentially responsive to light. We conclude that ENV1 is a key factor in connecting nutrient signalling with light response and establishes a signalling output pathway independent of BLR1 and BLR2. We used two biological replicates of three T. reesei strains (delta-blr1, delta-blr2 and delta-env1), cultivated in constant light (LL, 1800 lux) or constant darkness (DD) on microcrystalline cellulose. The strains used in this study were cultivated, hybridized and analyzed together with strains and samples from GSE27581; the corresponding wild-type strain QM9414 samples have accession numbers GSM683732, GSM683733, GSM683734 and GSM683735.
Project description:We perform a self hybridisation comprative genomic hybridization (CGH) in order to validate the probe tiling design we done on Trichoderma reesei. This hybridization was done using QM6a wild type strain. One biological replicate
Project description:The mechanism of carbon catabolite repression (CCR) mediated by CRE1 in Trichoderma reesei emerged as a way to adapt to the environment in which the fungus is found. In situations where there is the presence of readily available carbon sources such as glucose, the fungus activates this mechanism and inhibits the production of cellulolytic complex enzymes to avoid unnecessary energy expenditure. CCR has been well described for the growth of T. reesei in cellulose and glucose, however, little is known about this process when the carbon source available to the fungus is sophorose, one of the most potent inducer of cellulase production. Thus, we performed high-throughput RNA sequencing using the Illumina/HiSeq-2000 to contribute to the understanding of CCR during cellulase formation in the presence of sophorose, by comparing the mutant Δcre1 with its parental strain, QM9414. Of the 9129 genes present in the genome of T. reesei, 184 were up- and 344 down-regulated in the mutant strain Δcre1 compared to QM9414. Genes belonging to CAZy, transcription factors and transporters are among the gene classes that were repressed by CRE1 in the presence of sophorose, most of which was regulated by CRE1 in an indirect way. Overall, there was a similarity in the profile of repressed genes when compared with another inducing carbon source, cellulose. These results contribute to a better understanding of CRE1-meadiated CCR in T. reesei when glucose comes from a potent inducer as sophorose, which can be very useful in improving the production of cellulases by the biotechnology sector.