Fig. 5: AvrPm4⁹⁶²²⁴-induced cell death in Pm4-containing wheat protoplasts is suppressed by co-expression of SvrPm4^JIW2.

a, Protein sequence alignment of SvrPm4 candidates BgtAcSP-31023 and Bgt-51526 haplovariants found in the Pm4-virulent isolate GBR_JIW2 and in the Pm4-avirulent isolate CHE_96224. GBR_JIW2 is shown as a reference, with polymorphic residues in CHE_96224 highlighted. b, Wheat protoplast assay showing the suppression of AvrPm4-induced cell death by SvrPm4^JIW2. Protoplasts isolated from the transgenic wheat line #52, overexpressing Pm4b (used in Fig. 2), were transfected with each Svr candidate individually or cotransfected with AvrPm4. Presumed suppressor-active candidates (BgtAcSP-31023^JIW2 and Bgt-51526^JIW2) and suppressor-inactive candidates (BgtAcSP-31023⁹⁶²²⁴ and Bgt-51526⁹⁶²²⁴) were tested. The experiment was performed twice with three biological replicates each (n = 6 total). The height of the bars shows the mean value for all the replicates, and the whiskers show the standard deviation. c, HR response in N. benthamiana upon Agrobacterium-mediated co-expression of Bgt-51526^JIW2 or Bgt-51526⁹⁶²²⁴ and Pm1a-HA or a negative GUS control. Close-up images of the HR response are shown at the top. The full-size image of the corresponding N. benthamiana leaf is provided in Extended Data Fig. 10. The assay was performed three times with n = 6 leaves per experiment (n = 18 total). In each box plot, the median is shown as a central line within a box that spans from the 25th to the 75th percentile. The inner whiskers show the 95% confidence interval of the median, and the outer whiskers show the most extreme data points within 1.5 times the interquartile range. For all experiments, significance and P values (significance threshold of P < 0.05) were calculated with a two-sided ANOVA followed by Tukey’s HSD test (Supplementary Table 3). Bars or boxes with the same letter are not significantly different.