Improved gene targeting in Magnaporthe grisea by inactivation of MgKU80 required for non-homologous end joining.
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.
Villalba
François
F
Biochemistry, Bayer CropScience, Lyon, France.
Collemare
Jérôme
J
Landraud
Patricia
P
Lambou
Karine
K
Brozek
Viviane
V
Cirer
Bénédicte
B
Morin
Damien
D
Bruel
Christophe
C
Beffa
Roland
R
Lebrun
Marc-Henri
MH
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
D004268:DNA-Binding Proteins; D005656:Fungal Proteins