Statistical analysis of Cf-4/Avr4-triggered cell death in different <i>N</i>.

benthamiana nrc mutant lines. A) Cf-4/Avr4-triggered cell death was scored on a scale of 0–7, with 0 being no response, and 7 being fully confluent cell-death in the entire infiltrated sector. Visible cell death starts appearing at a score of 4. B) Statistical analysis was conducted using the 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. C) Cf-4/Avr4-triggered hypersensitive cell death is lost in the nrc2/3 and nrc2/3/4 CRISPR lines. Representative N. benthamiana leaves infiltrated with appropriate constructs were photographed 7–10 days after infiltration. The NRC CRISPR lines, nrc2/3-209.3.3.1, nrc2/3/4-210.5.5.1, are labelled above the leaf and the receptor/effector pair tested, Cf-4/Avr4, Prf (Pto/AvrPto), Rpi-blb2/AVRblb2 or R3a/Avr3a, are labelled on the leaf image. Cf-4/EV and EV/Avr4 were also included. (TIF)