title: Ku70 or Ku80 deficiencies in the fungus Botrytis cinerea facilitate targeting of genes that are hard to knock out in a wild-type context.
abstract: The filamentous ascomycete Botrytis cinerea is one of the most studied models for understanding the necrotrophic behaviour of phytopathogenic fungi. The genomes of two strains of B. cinerea have been sequenced (B05.10 and T4), which may contribute to elucidating the virulence polymorphism in this fungus. In this study, both strains were genetically modified in order to construct recipient strains designed to target genes that are hard to knock out. Deletions of BcKu70 gene in B05.10 strain and BcKu80 gene in T4 strain both affected the nonhomologous end-joining (NHEJ) DNA repair mechanism. NHEJ is responsible for the ectopic integration of gene replacement cassettes during fungal transformation and leads to a lower frequency of homologous recombination (HR). Ku deficiencies in B. cinerea did not disturb in vitro or in planta growth, but clearly improved HR efficiency for the putative sesquiterpene cyclase-encoding gene Cnd15, which was hard to knock out in a wild-type strain.
authors.affiliation: UMR1290 BIOGER-CPP, INRA-AgroParisTech, Versailles, France. mathias.choquer@univ-lyon1.fr
authors.forename: Mathias
authors.initials: M
authors.lastname: Choquer
authors.forename: Guillaume
authors.initials: G
authors.lastname: Robin
authors.forename: Pascal
authors.initials: P
authors.lastname: Le Pêcheur
authors.forename: Corinne
authors.initials: C
authors.lastname: Giraud
authors.forename: Caroline
authors.initials: C
authors.lastname: Levis
authors.forename: Muriel
authors.initials: M
authors.lastname: Viaud
mesh_terms: D020171:Botrytis / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D009154:Mutation; D010802:Phylogeny; D010935:Plant Diseases / Q000382:microbiology; D011995:Recombination, Genetic
chemical_list: D004268:DNA-Binding Proteins; D005656:Fungal Proteins