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C239S Mutation in the β-Tubulin of Phytophthora sojae Confers Resistance to Zoxamide

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https://ir.library.oregonstate.edu/concern/articles/7m01bn33t

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  • Zoxamide is the sole β-tubulin inhibitor registered for the control of oomycete pathogens. The current study investigated the activity of zoxamide against Phytophthora sojae and baseline sensitivity was established with a mean EC₅₀ of 0.048 μg/ml. The data is critical for monitoring changes in zoxamide-sensitivity in the field. Three stable resistant mutants with a high resistance level were obtained by selection on zoxamide amended media. Although the development of resistance occurred at a low frequency, there were no apparent fitness penalty in the acquired mutants in terms of growth rate, sporulation, germination and pathogenicity. Based on the biological profiles and low mutagenesis rate, the resistance risk of P. sojae to zoxamide can be estimated as low to medium. Further investigation revealed all the zoxamide-resistant mutants had a point mutation of C239S in their β-tubulin. Zoxamide also exhibited high activity against most species from the genus Pythium in which only Pythium aphanidermatum was found naturally resistant to zoxamide and harboring the natural point mutation S239 in the β-tubulin. Back-transformation in P. sojae with the mutated allele (S239) confirmed the C239S mutation can induce resistance to zoxamide, and the resistance level was positively related to the expression level of the mutated gene. In contrast, the overexpression of the wild type gene was unable to cause zoxamide resistance. It is the first report on the resistance molecular mechanism of zoxamide in oomycetes. Based on our study, C239 is supposed to be a key target site of zoxamide, which distinguishes zoxamide from benzimidazoles and accounts for its low resistance risk. The result can provide advice on the design of new β-tubulin inhibitors in future.
  • KEYWORDS: beta-tubulin inhibitor, zoxamide, resistance risk, molecular mechanism of resistance, C239S, oomycetes
  • This is the publisher’s final pdf. The published article is copyrighted by the author(s) and published by Frontiers Media. The published article can be found at: http://journal.frontiersin.org/journal/microbiology
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  • Cai, M., Miao, J., Song, X., Lin, D., Bi, Y., Chen, L., ... & Tyler, B. M. (2016). C239S Mutation in the β-Tubulin of Phytophthora sojae Confers Resistance to Zoxamide. Frontiers in Microbiology, 7, 762. doi:10.3389/fmicb.2016.00762
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  • This work was funded by the Special Fund for Agro-scientific Research in the Public Interest (No. 201303023), National Natural Science Foundation of China (NO. 31272061) and Chinese Universities Scientific Fund (No. 15053201).
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file details CaiC239SMutationBetaTubulin.pdf 07/25/2017 Público
file details CaiC239SMutationBetaTubulinDataSheet1.pdf 07/25/2017 Público