Cf-4/Avr4-triggered hypersensitive cell death can be suppressed by divergent pathogen effectors.
A) A schematic representation of cell death suppression assay. A. tumefaciens containing vectors carrying both NRC3 and either SPRYSEC15, AVRcap1b, or mCherry as empty vector control, were co-expressed in N. benthamiana nrc2/3/4 CRISPR lines, with the effectors suppressing the cell death response. B) Cf-4/Avr4-triggered hypersensitive cell death is suppressed by SPRYSEC15 and AVRcap1b, but not the EV control. Representative image of N. benthamiana nrc2/3/4-126.96.36.199 CRISPR line leaves which were agroinfiltrated with NRC3/suppressor constructs, as indicated above the leaf, and either autoactive NRC3D480V, Prf (Pto/AvrPto), R3a/Avr3a, or Cf-4/Avr4 as labelled on the leaf image, photographed 7–10 days after infiltration. A representative leaf of the independent nrc2/3/4-188.8.131.52 CRISPR line is shown in S11C Fig) Quantification of hypersensitive cell death. Cell death was scored based on a 0–7 scale between 7–10 days post infiltration. The results are presented as a dot plot, where the size of each dot is proportional to the count of the number of samples with the same score within each biological replicate. The experiment was independently repeated three times. The columns correspond to the different biological replicates. Significant differences between the conditions are indicated with an asterisk (*). Details of statistical analysis are presented in S11 Fig.