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. 2017 May 15;7:1908. doi: 10.1038/s41598-017-02017-3

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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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Comparison of virus accumulation in protoplasts of Nicotiana tabacum BY2 cells with filtered untreated or heat inactivated virus inocula. Quantitative Reverse transcriptase PCR (qRT-PCR) data from Nicotiana tabacum BY2 cells were used to obtain absolute quantification of viral RNA at T0 and T48 time points with heat-inactivated (purple for T0 and red for T48) or untreated (green for T0, yellow for T48) viral inocula. In Panel A: Quantification graph of PaTV1 confirmed the ability to replicate when the viral purification is untreated, whereas when inactivated before transfection the final RNA amount at T48 (red bar) was lower than at T0. In panels C quantification of the Partitivirus Penicillium aurantiogriseum partitivirus 1 (PaPV1): similarly to PaTV1, PaPV1 can replicate in untreated biological replicates whereas the RNA accumulation level decreased when thermally inactivated suspension was used for transfection. Panels B,D,E and F show data for the other viruses: none of them displays a statistically significant increase in virus RNA load. *P < 0.05 **P < 0.01 and ***P < 0.001.