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

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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.

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Acknowledgments

This work was supported by a Grant from the Key Project for Breeding Genetic Modified Organisms (2014ZX08005-004). We would like to thank Dr. Latifur Rehman, for reading this manuscript.

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Authors and Affiliations

  1. Agriculture College of Shihezi University, Shihezi, 832000, China

    Xiliang Qi & Juncang Qi

  2. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Xiliang Qi, Xiaofeng Su, Huiming Guo & Hongmei Cheng

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  1. Xiliang Qi
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  2. Xiaofeng Su
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  3. Huiming Guo
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  4. Juncang Qi
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  5. Hongmei Cheng
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Correspondence to Juncang Qi or Hongmei Cheng.

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Qi, X., Su, X., Guo, H. et al. A ku70 null mutant improves gene targeting frequency in the fungal pathogen Verticillium dahliae . World J Microbiol Biotechnol 31, 1889–1897 (2015). https://doi.org/10.1007/s11274-015-1907-1

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  • DOI: https://doi.org/10.1007/s11274-015-1907-1

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