Deletion of the cpku80 gene in the chestnut blight fungus, Cryphonectria parasitica, enhances gene disruption efficiency.

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TitleDeletion of the cpku80 gene in the chestnut blight fungus, Cryphonectria parasitica, enhances gene disruption efficiency.
Publication TypeJournal Article
Year of Publication2008
AuthorsLan, X, Yao, Z, Zhou, Y, Shang, J, Lin, H, Nuss, DL, Chen, B
JournalCurr Genet
Volume53
Issue1
Pagination59-66
Date Published2008 Jan
ISSN0172-8083
KeywordsAscomycota, Cloning, Molecular, DNA Repair, DNA, Fungal, Fungal Proteins, Gene Expression Regulation, Fungal, Genes, Fungal, Molecular Sequence Data, Plant Diseases, Sequence Analysis, DNA, Virulence
Abstract

The chestnut blight fungus, Cryphonectria parasitica, and associated virulence-attenuating hypoviruses have emerged as an important model system for studying molecular mechanisms underlying fungal-plant pathogenic interactions. As more gene sequence information becomes available as a result of C. parasitica express sequence tags (ESTs) and ongoing whole genome sequencing projects, the development of an efficient gene disruption system has become an urgent need for functional genomics studies of this important forestry pathogen. Here, we report the cloning of the C. parasitica gene cpku80 that encodes a key component of the nonhomologous end joining DNA repair pathway and the construction of a corresponding deletion mutant strain. The cpku80 mutant was indistinguishable from the parental wild-type strain EP155 in colony morphology, ability to support hypovirus replication, conidiation and virulence. As predicted, the Deltacpku80 strain did exhibit an increased sensitivity to the mutagen methyl methanesulfonate. A test with three selected genes resulted in a gene disruption efficiency of about 80% for the Deltacpku80 strain, a significant increase over the 2-5% levels of homologous recombination generally observed for the wild-type strain EP155. This efficient homologous recombination system provides a powerful tool for large-scale analysis of gene functions in C. parasitica.

DOI10.1007/s00294-007-0162-x
Alternate JournalCurr. Genet.
PubMed ID17972079
Grant ListGM55981 / GM / NIGMS NIH HHS / United States