NRC3 E15Q can complement Cf-4/Avr4-triggered hypersensitive cell death.
A) NRC3 and the NRC3 E15Q mutant can complement Cf-4/Avr4 and Pto/AvrPto-triggered hypersensitive cell death in the N. benthamiana nrc2/3/4 CRISPR lines. Representative N. benthamiana leaves infiltrated with appropriate constructs were photographed 7–10 days after infiltration. The receptor/effector pair tested, Cf-4/Avr4 and Prf (Pto/AvrPto), as well as the NRC and NRC mutants used are labelled on the leaf image. To ensure the NRC3 E15Q mutant was not autoactive we expressed it with either Cf-4 or Pto and an EV control. B) 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. Significant differences between the conditions are indicated with an asterisk (*). C) Statistical analysis was conducted using besthr R package . The dots represent the ranked data and their corresponding means (dashed lines), with the size of each dot proportional to the number of observations for each specific value (count key below each panel). The panels on the right show the distribution of 100 bootstrap sample rank means, where the blue areas under the curve illustrate the 0.025 and 0.975 percentiles of the distribution. A difference is considered significant if the ranked mean for a given condition falls within or beyond the blue percentile of the mean distribution of the wild-type control.