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FreB is involved in the ferric metabolism and multiple pathogenicity-related traits of Verticillium dahliae

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Abstract

Ferric reductases are integral membrane proteins involved in the reduction of environmental ferric iron into the biologically available ferrous iron. In the most overwhelming phytopathogenic fungus, Verticillium dahliae, these ferric reductase are not studied in details. In this study we explored the role of FreB gene (VDAG_06616) in the ferric reduction and virulence of V. dahliae by generating the knockout mutants (ΔFreB) and complementary strains (ΔFreB-C) using protoplast transformation. When cultured on media supplemented with FeSO4, FeCl3 and no iron, ΔFreB exhibited significantly reduced growth and spore production especially on media with no iron. Transmembrane ferric reductase activity of ΔFreB was decreased up to 50% than wild type strains (Vd-wt). The activity was fully restored in ΔFreB-C. Meanwhile, the expression levels of other related genes (Frect-4, Frect-5, Frect-6 and Met) were obviously increased in ΔFreB. Compared with the Vd-wt and ΔFreB-C, ΔFreB-1 and ΔFreB-2 were impaired in colony diameter and spore number on different carbon sources (starch, sucrose, galactose and xylose). ΔFreB-1 and ΔFreB-2 were also highly sensitive to oxidative stress as revealed by the plate diffusion assay when 100 µM H2O2 was applied to the fungal culture. When Nicotiana benthamiana plants were inoculated, ΔFreB exhibited less disease symptoms than Vd-wt and ΔFreB-C. In conclusion, the present findings not only indicate that FreB mediates the ferric metabolism and is required for the full virulence in V. dahliae, but would also accelerate future investigation to uncover the pathogenic mechanism of this fungus.

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Acknowledgements

This work was supported by a grant from the National Natural Science Foundation of China (31772244), the National Nonprofit Industry Research (201503109) and the Agricultural Science and Technology Innovation Program of CAAS.

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

      Latifur Rehman, Xiaofeng Su, Xiaokang Li, Xiliang Qi, Huiming Guo & Hongmei Cheng

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    1. Latifur Rehman
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    Correspondence to Hongmei Cheng.

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    Communicated by M. Kupiec.

    Latifur Rehman and Xiaofeng Su contributed equally.

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    Rehman, L., Su, X., Li, X. et al. FreB is involved in the ferric metabolism and multiple pathogenicity-related traits of Verticillium dahliae . Curr Genet 64, 645–659 (2018). https://doi.org/10.1007/s00294-017-0780-x

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