Fig. 4. Rph7 induces a cell death response when transiently expressed in Nicotiana benthamiana and Golden Promise barley protoplasts.

a Transient expression of Rph7 (YFP-tagged driven by 35S CaMV promoter) induced partial cell death. The experiment was repeated three times and included infiltration of two leaves of three to four plants with similar results. The cloned stem rust resistance NLR protein Sr27 in the presence of its corresponding avirulence protein AvrSr27 was used as a positive control for cell death, and YFP alone as a negative control. b Western blot analysis of Rph7_YFP expression in N. benthamiana. c Relative luciferase activity in barley (Hordeum vulgare L. cv. Golden Promise) protoplasts from transgenic lines were co-transformed using the same Rph7 constructs shown in a with a luciferase reporter. Values are relative to luciferase activity of cells co-transformed with GFP and the luciferase reporter, which was set to 1. The cloned stem rust resistance NLR protein Sr50 in the presence of its corresponding avirulence protein AvrSr50 were used as a positive control for cell death. The data presented captures biological replication (n = 2) where the box encompasses two middle quartiles, with central line showing mean. Whiskers extend to the furthest data point within 1.5 times the interquartile range. Samples marked by identical letters in the plots do not differ significantly (P < 0.01) using the single-tailed Tukey test. Source data are provided as a Source Data file.