title: A ku70 null mutant improves gene targeting frequency in the fungal pathogen Verticillium dahliae.
abstract: To overcome the challenges met with gene deletion in the plant pathogen Verticillium dahliae, a mutant strain with impaired non-homologous end joining DNA repair was generated to improve targeted gene replacement frequencies. A V. dahliae 991 ΔVdku70 null mutant strain was generated using Agrobacterium tumefaciens-mediated transformation. Despite having impaired non-homologous end joining DNA repair function, the ΔVdku70 strain exhibited normal growth, reproduction capability, and pathogenicity when compared with the wild-type strain. When the ΔVdku70 strain was used to delete 2-oxoglutarate dehydrogenase E2, ferric reductase transmembrane component 3 precursor, and ferric reductase transmembrane component 6 genes, gene replacement frequencies ranged between 22.8 and 34.7% compared with 0.3 and 0.5 % in the wild-type strain. The ΔVdku70 strain will be a valuable tool to generate deletion strains when studying factors that underlie virulence and pathogenesis in this filamentous fungus.
authors.forename: Xiliang
authors.initials: X
authors.lastname: Qi
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Su
authors.forename: Huiming
authors.initials: H
authors.lastname: Guo
authors.forename: Juncang
authors.initials: J
authors.lastname: Qi
authors.forename: Hongmei
authors.initials: H
authors.lastname: Cheng
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D034961:Antigens, Nuclear / Q000235:genetics*; D059766:DNA End-Joining Repair; D004271:DNA, Fungal / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D005656:Fungal Proteins / Q000378:metabolism; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D018390:Gene Targeting; D059765:Homologous Recombination / Q000235:genetics; D000072200:Ku Autoantigen; D016254:Mutagenesis, Insertional; D009154:Mutation; D010935:Plant Diseases / Q000382:microbiology; D012639:Seeds / Q000382:microbiology; D014026:Nicotiana / Q000382:microbiology; D020172:Verticillium / Q000235:genetics* / Q000254:growth & development; D014774:Virulence / Q000235:genetics*
chemical_list: D034961:Antigens, Nuclear; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D000072200:Ku Autoantigen