FIGURE 5.

NbBAK1 and NbSOBIR1 are required for Vd424Y‐induced cell death. (a) Virus‐induced gene silnecing (VIGS) technology was used to silence NbBAK1 and NbSOBIR1 by inoculation with TRV constructs (pTRV2:GFP, pTRV2:NbBAK1, and NbSOBIR1) in Nicotiana benthamiana plants. Three weeks after inoculation, GFP, BAX, Vd424Y, and Vd424Y mutations were transiently expressed in NbBAK1‐ and NbSOBIR1‐silenced N. benthamiana plant leaves. Photographs were taken 7 days after agroinfiltration. The experiment was carried out three times with five plants for each TRV construct. (b) The expression levels of NbBAK1 and NbSOBIR1 after VIGS treatment as evaluated by quantitative reverse transcription PCR. NbActin was used as the internal reference gene. Mean and SE were calculated from three independent experiments. Bars indicate ± SE. Significance level p < 0.01 is represented by *. (c) Western blot (WB) analysis of green fluorescent protein (GFP), Vd424Y, and Vd424Y mutations protein fused with HA tags after transient expression in N. benthamiana leaves. Proteins were stained with Coomassie brilliant blue (CBB) to confirm equal loading