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. 2017 Aug 11;7:8000. doi: 10.1038/s41598-017-08010-0

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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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Gene expression levels of several markers related to plant immunity in Nicotiana benthamiana leaves transiently expressing HaSSP30 for 1, 2, and 3 days. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the pICH86988-empty (Control) or pICH86988-HaSSP30 (HaSSP30) vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity (RH). The leaves were harvested after 1, 2, and 3 days post-infiltration (dpi). Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Two reference genes were used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation (SD) of 3 independent replicates (n = 3). Two-tailed Student t-test was used to compare HaSSP30 versus Control in each day and p-values < 0.05 are indicated above the bars.