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. 2018 Nov 20;8:17069. doi: 10.1038/s41598-018-35223-8

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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In planta inoculation assays for Ppt-associated gene deletion strains highlights the key role of lysine biosynthesis for virulence. (A) Upper panel: All PPT associated disruption strains, ∆ZtPpt-1, ∆ZtPpt-2, ∆ZtPks1-1, ∆ZtPks1-2, ∆ZtNrps1-1, ∆ZtNrps1-2 and ∆ZtAar, and the two wild type strains IPO323 and ∆Ztku70 after 22 days post inoculation (dpi) of the susceptible wheat cv, Riband; lower panel: the Z. tritici lysine auxotrophic PPT associated strains ∆ZtPpt-1, ∆ZtPpt-2 and ∆ZtAar, and the two wildtype strains after 22 (dpi) with lysine added to starting inoculum. (B) The WT ∆Ztku70, and ∆ZtPks1-1 and ∆ZtPks1-2 melanin synthetase mutants viewed under a light microscope after 22 dpi. White arrows mark pycnidia, which in the melanin synthetase mutants are unpigmented. (C) Spores collected from infected leaves after 22 dpi. Left portion of plot: all PPT associated strains and the WTs without lysine added to fungal inoculum; right portion of plot: lysine auxotrophic PPT associated strains and the WTs with lysine added to fungal inoculum. Asterisks represent significant differences in spore counts relative to the WT that constituted the background strain (either IPO323 for ΔZtAar or ΔZtKu70 for the rest) at α < 0.05. Black horizontal bars represent median values, boxes represent second and third quartiles and whiskers represent interquartile range.