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. 2021 Jun 7;12:3335. doi: 10.1038/s41467-021-23614-x

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© The Author(s) 2021

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. 2 — a A heatmap representation of Pst avrRps4, Pst avrRpt2 or Pst (empty vector, EV) growth at 3 dpi in leaves of indicated genotypes (syringe infiltration, OD₆₀₀ = 0.0005). The significance codes are based on Tukey’s HSD test (α = 0.001, n = 12 for Pst avrRps4 and Pst avrRpt2, n = 16 for Pst). Data points were combined from three (Pst avrRps4, Pst avrRpt2) or four (Pst) independent experiments, each with four replicates. Note: significance codes are assigned based on the statistical analysis per treatment and should be read columnwise. b A heatmap of quantitative cell death assays conducted on leaves of indicated genotypes after infiltration utilising the Pseudomonas fluorescens 0–1 effector tester strain (hereafter Pf0-1) delivering avrRps4 (OD₆₀₀ = 0.2). Cell death was measured by electrolyte leakage from bacteria-infiltrated leaf discs at 6, 8, 10 and 24 hpi. Data are displayed as means from four experiments, each with four replicates (n = 16). Statistical significance codes for the difference in means are based on Tukey’s HSD test (α = 0.001). In mutants marked in red, the PAD4–ADR1s cell death branch operates in TNL^RRS1-RPS4 immunity when SAG101–NRG1s and ICS1 pathways are not functional. Note: significance codes are assigned based on the statistical analysis per timepoint and should be read columnwise. Boxplot representation of the same data is provided in Supplementary Figs. 5 and 6. c Visual cell death symptoms at 24 hpi Pf0-1 avrRps4 infiltrating into leaf halves of indicated genotypes as in b. The ratio beneath each leaf indicates number of leaves with visible tissue collapse from all infiltrated leaves in two independent experiments. White arrows mark cell death visible as tissue collapse in a manner dependent on SAG101–NRG1s. Red arrows mark cell death in lines without functional SAG101–NRG1s and ICS1. Genotypes in red have functional PAD4–ADR1s but not SAG101–NRG1s. Pst Pseudomonas syringae pv. tomato DC3000, Pf0-1 effector tester strain of Pseudomonas fluorescens 0–1, CFU colony-forming units, EV empty vector.