Fig. 6.
BAK1 and SOBIR1 associate with RXEG1 and are required for RXEG1-mediated XEG1 recognition. a Representative leaves showing cell death induced by expression of XEG1, INF1, or NPP1 in N. benthamiana leaves treated with TRV:GFP, TRV:BAK1, or TRV:SOBIR1. XEG1, INF1, and NPP1 were transiently expressed in N. benthamiana. Infiltrated leaves (n = 6) were photographed at 3 dai. Protein expression at 2 dai was monitored by western blotting with anti-HA antibody. Protein loading is indicated by Ponceau S staining. Experiments were repeated at least three times with similar results. b Silencing efficiency of BAK1 and SOBIR1 in N. benthamiana. The transcript levels were monitored by qRT-PCR, normalized with EF-1α, and expressed as mean fold changes (±s.e.m.) relative to TRV:GFP-treated leaves, which was set as 1. * indicates significant differences (P < 0.01, Student’s t-test) in expression level when compared to that in TRV:GFP-treated plants. Experiments were repeated three times with similar results. c Ion leakage detected from N. benthamiana leaves expressing XEG1, INF1, or NPP1. Bars represent average value (±s.e.m.) of six replicates. In each treatment, different letters indicate significant differences (P < 0.01, one-way ANOVA). Experiments were repeated twice with similar results. d ROS production in N. benthamiana leaves treated with 1 μm XEG1 or EV. Mean values (±s.e.m.) of four replicates are shown. Experiments were repeated twice with similar results. e Relative expression of CYP71D20 in N. benthamiana leaves treated with 200 nm XEG1 or EV. Transcript levels were quantified by qRT-PCR and normalized with EF1-ɑ. Bars represent the mean fold changes (±s.e.m.) relative to the value in EV-treated leaves, which was set as 1. Different letters indicate significant differences (P < 0.05, one-way ANOVA). Experiments were repeated three times with similar results. f Interactions of RXEG1 with BAK1 and SOBIR1 in planta. N. benthamiana leaves were agro-infiltrated to express RXEG1-GFP, BAK1-RFP, or SOBIR1-HA and collected 2 dai after treatment with 1 μm XEG1 or EV for 10 min. Extracted proteins were subjected to Co-IP using GFP-trap® A beads and western blotting with anti-GFP, anti-RFP, and anti-HA antibodies