title: Improved gene targeting in Magnaporthe grisea by inactivation of MgKU80 required for non-homologous end joining.
abstract: The ascomycete Magnaporthe grisea is a model species for the study of plant fungal interactions. As in many filamentous fungi, targeted gene replacement occurs at low frequency in M. grisea (average 7%). mus52/KU80 is a gene essential for non-homologous end joining (NHEJ) of DNA double-strand breaks. Its deletion increases the frequency of targeted gene replacement in fungi [Ninomiya, Y., Suzuki, K., Ishii, C., Inoue, H., 2004. Highly efficient gene replacements in Neurospora strains deficient for non-homologous end joining. Proc. Natl. Acad. Sci. USA 101(33), 12248-53]. M. grisea KU80 deletion mutants were constructed and displayed wild-type phenotypes regarding pathogenicity, growth, sporulation and mating. MgADE4 targeted gene replacement frequency was increased in Deltaku80 mutants (80% vs 5%) and high frequencies (>80%) were observed at seven other loci. However, the deletion of MgKU80 did not increase the frequency of ACE1 replacement indicating that this locus has an intrinsic reduced ability for gene replacement. These results open the way to large-scale reverse genetics experiments in M. grisea facilitating the study of the infection process.
authors: Biochemistry, Bayer CropScience, Lyon, France.
authors: François
authors: F
authors: Villalba
authors: Jérôme
authors: J
authors: Collemare
authors: Patricia
authors: P
authors: Landraud
authors: Karine
authors: K
authors: Lambou
authors: Viviane
authors: V
authors: Brozek
authors: Bénédicte
authors: B
authors: Cirer
authors: Damien
authors: D
authors: Morin
authors: Christophe
authors: C
authors: Bruel
authors: Roland
authors: R
authors: Beffa
authors: Marc-Henri
authors: MH
authors: Lebrun
mesh_terms: D004268:DNA-Binding Proteins / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods*; D005800:Genes, Fungal; D020082:Magnaporthe / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D012275:Oryza / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology; D011995:Recombination, Genetic; D014774:Virulence
chemical_list: D004268:DNA-Binding Proteins; D005656:Fungal Proteins