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The Hypocrea jecorina (Trichoderma reesei) hypercellulolytic mutant RUT C30 lacks a 85 kb (29 gene-encoding) region
Verena Seidl1, Christian Gamauf, Irina S Druzhinina
1Research Area Gene Technology and Applied Biochemistry, Institute of Chemical Engineering, Vienna University of Technology, Getreidemarkt 9/166-5, A-1060 Wien, Austria. vseidl@mail.zserv.tuwien.ac.at
Background:
The hypercellulolytic mutant Hypocrea jecorina (anamorph Trichoderma reesei) RUT C30 is the H. jecorina strain most frequently used for cellulase fermentations and has also often been employed for basic research on cellulase regulation. This strain has been reported to contain a truncated carbon catabolite repressor gene cre1 and is consequently carbon catabolite derepressed. To date this and an additional frame-shift mutation in the glycoprotein-processing beta-glucosidase II encoding gene are the only known genetic differences in strain RUT C30.
Results:
In the present paper we show that H. jecorina RUT C30 lacks an 85 kb genomic fragment, and consequently misses additional 29 genes comprising transcription factors, enzymes of the primary metabolism and transport proteins. This loss is already present in the ancestor of RUT C30--NG 14--and seems to have occurred in a palindromic AT-rich repeat (PATRR) typically inducing chromosomal translocations, and is not linked to the cre1 locus. The mutation of the cre1 locus has specifically occurred in RUT C30. Some of the genes that are lacking in RUT C30 could be correlated with pronounced alterations in its phenotype, such as poor growth on alpha-linked oligo- and polyglucosides (loss of maltose permease), or disturbance of osmotic homeostasis.
Conclusion:
Our data place a general caveat on the use of H. jecorina RUT C30 for further basic research.
Insights
The widely used Hypocrea jecorina RUT C30 strain has a large genomic deletion, not just the known cre1 mutation. This deletion impacts gene function and cellular processes, questioning its suitability for research.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Hypocrea jecorina (Trichoderma reesei) RUT C30 is a key strain for cellulase production and research.
- Previous studies identified a truncated carbon catabolite repressor gene (cre1) and a frame-shift mutation in beta-glucosidase II as the only known genetic differences.
- Strain RUT C30 is widely used for cellulase fermentations and basic research on cellulase regulation.
Purpose of the Study:
- To investigate the complete genetic makeup of the Hypocrea jecorina RUT C30 strain.
- To identify all genetic differences between strain RUT C30 and its ancestors.
- To assess the implications of identified genetic variations on strain phenotype and research applications.
Main Methods:
- Genomic analysis of H. jecorina RUT C30.
- Comparison of RUT C30 genome with its ancestor NG 14.
- Phenotypic analysis of strain RUT C30.
Main Results:
- H. jecorina RUT C30 is missing an 85 kb genomic fragment containing 29 genes, including transcription factors, metabolic enzymes, and transport proteins.
- This deletion is present in the ancestor strain NG 14 and occurred near a palindromic AT-rich repeat (PATRR), independent of the cre1 locus.
- The cre1 mutation is specific to RUT C30, and the genomic deletion correlates with altered phenotypes like poor growth on specific carbohydrates and osmotic imbalance.
Conclusions:
- The genetic makeup of H. jecorina RUT C30 is more complex than previously understood, with a significant genomic deletion.
- The identified genetic variations, particularly the large deletion, can significantly affect the strain's phenotype and metabolic capabilities.
- Caution is advised when using H. jecorina RUT C30 for basic research due to these uncharacterized genetic differences.
