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. 2018 Mar 28;9:1268. doi: 10.1038/s41467-018-03658-2

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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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Fig. 8 — Silencing of NbBeclin1 or NbATG8a promotes TuMV infection in N. benthamiana. a GFP fluorescence and viral symptoms in plants pre-inoculated with TRV1 together with TRV2-GUS (control), TRV2-NbBeclin1, or TRV2-NbATG8a for 7 days and then infected by TuMV-GFP. Plants were photographed under UV light at 3 and 6 dpi and under regular light at 6 dpi. b Quantification of TuMV genomic RNA in the above plants. RNA was extracted from TuMV-GFP-inoculated leaves at 3 dpi or systemically infected leaves at 6 dpi and 30 dpi. The values are presented as means of fold change ±SD relative to the control plants (pretreated with TVR1 and TRV2-GUS). Error bars represent SD. Three independent experiments, each consisting of three biological replicates, were carried out. Values from one representative result were used to plot a histogram and were normalized against NbActin transcripts in the same sample. The data were analyzed using Student’s t-test (two-sided, *P < 0.05, **P < 0.01). c Protoplast transfection assay. Protoplasts isolated from N. benthamiana plants pre-inoculated with TRV1 together with TRV2-GUS, TRV2-NbBeclin1, or TRV2-NbATG8a were transfected with TuMV-GFP. RNA was extracted at 16, 24, and 48 hpt and quantified by qRT-PCR to analyze TuMV genomic RNA accumulations. Values are presented as means ± SD, and error bars represent SD (n = 3 biological replicates). The data were analyzed using Student’s t-test and asterisks denote significant differences compared to the TuMV-infected protoplasts from control plants pre-treated with TRV1 and TRV-GUS (two-sided, *P < 0.05, **P < 0.01)