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. 2018 Jun 5;8:8615. doi: 10.1038/s41598-018-27000-4

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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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Silencing of NbHAOX8, NbGOX1 and NbGOX4 differently affected resistance to Xoo and Sclerotinia sclerotiorum in N. benthamiana. Disease resistance was evaluated in the newly, fully expanded leaves of VIGS-treated plants at 21 d after agro-inoculation. For each treatment, at least 6 leaves from 6 silenced plants (one leaf from each plant) were examined. Each experiment was conducted three times independently. (A) Symptoms of Xoo-induced HR in the gene-silenced plants. Photographs were taken at 18 hpi. (B) Severity of Xoo-induced HR in the gene-silenced plants at 18 hpi. HR is classified based on the intensity of cell death in inoculated areas: I, HR area smaller than 30% of infiltrated leaf area; II, HR area larger than 30% but smaller than 90% of infiltrated leaf area; III, HR area larger than 90% of the infiltrated leaf area. HR severity is indicated as the percentage of leaves exhibiting HR over total infiltrated leaves. (C) Statistical analysis of Xoo bacterial number in the infiltrated areas at 0 and 24 hpi. Data were analyzed using SPSS. Significant difference is indicated as different lowercase letters (P < 0.05, DMRT). (D) Disease symptoms of the gene-silenced plants after inoculation with Ss. Photographs were taken at 26 hpi. (E) Statistical analysis of lesion diameter. Significant difference of lesion diameter is indicated as lowercase letters (P < 0.05, DMRT).