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. 2017 Apr 3;7:539. doi: 10.1038/s41598-017-00533-w

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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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Asn174 in the Large subunit is essential for a systemic infection. (a) For each mutant and WT virion 15 grams of leaves were analysed. Coomassie blue stained SDS PAGE shows the mutation N174A results in similar levels of the Large (L) and small (S) coat proteins to those found with WT virus being produced in infiltrated leaves. By contrast the N174D mutation resulted in a substantial reduction in the amounts of the two coat proteins. Both mutations inhibit CPMV from causing a systemic infection, with the L and S coat proteins being undetectable in extracts. (b) Coomassie blue and ethidium bromide (EtBr) stained native agarose gels of particles isolated from infiltrated leaves. The N174A mutation allows genome encapsidation while N174D mutant does not appear to package RNA. (c) Negative stain EM shows N174A and N174D mutants appear to assemble as complete particles similar to WT CPMV (also shown for reference). Scale bar is 100 µm.