Statistical analysis of NRC MADA mutant complementation of Cf-4/Avr4-triggered cell death N.
benthamiana nrc2/3/4 CRISPR lines. A) 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. B) NRC MADA mutants cannot complement Cf-4/Avr4 or 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 NRCs tested, NRC2 and NRC3, are labelled above the leaf of NRC CRISPR line nrc2/3/4-126.96.36.199. The receptor/effector pair tested, Cf-4/Avr4 and Prf (Pto/AvrPto), are labelled on the leaf image. To ensure the NRC MADA mutants were not autoactive they were expressed with either Cf-4 or Pto and an EV control was taken along.