A ku70 null mutant improves gene targeting frequency in the fungal pathogen Verticillium dahliae

作者：Qi, Xiliang; Su, Xiaofeng; Guo, Huiming; Qi, Juncang^*; Cheng, Hongmei

来源：World Journal of Microbiology and Biotechnology, 2015, 31(12): 1889-1897.

DOI：10.1007/s11274-015-1907-1

• DNA end-joining repair
• Verticillium dahliae
• Homologous recombination
• Delta Vdku70 strain
• Targeted gene replacement

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 Delta Vdku70 null mutant strain was generated using Agrobacterium tumefaciens-mediated transformation. Despite having impaired non-homologous end joining DNA repair function, the Delta Vdku70 strain exhibited normal growth, reproduction capability, and pathogenicity when compared with the wild-type strain. When the Delta Vdku70 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 Delta Vdku70 strain will be a valuable tool to generate deletion strains when studying factors that underlie virulence and pathogenesis in this filamentous fungus.

• 出版日期2015-12
• 单位中国农业科学院; 石河子大学