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. 2017 Oct 18;7:13444. doi: 10.1038/s41598-017-13870-7

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© The Author(s) 2017

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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Symptoms (A) and accumulation of CMV and the different 2b protein mutants (B) in N. benthamiana plants. Infections with Rs-SPS/40-42/APA, Rs-SPS/40-42/DPD were asymptomatic during the monitored period (six weeks). Infection with Rs-SPS/40-42/SPD and Rs-SPS/40-42/DPS induced mild symptoms: mild leaf distortion, mild systemic mosaic, and mild stunting (A). Rs-SPS/40-42/APS, Rs-SPS/40-42/SAS and Rs-SPS/40-42/SPA exhibited severe symptoms, including systemic mosaic, leaf distortion, and chlorosis. However, symptoms induced by these mutants were never as severe as those induced by Rs-CMV. CMV accumulation in systemically infected leaves analysed by northern hybridization at 7 dpi. The viral RNA accumulation was in accordance with the strength of the symptoms. The radiolabelled probe was specific for Rs-CMV RNA 3. Ethidium bromide-stained rRNA from the same volume of the sample is shown below each lane.