Skip to main content
. 2021 Nov 5;23(2):237–253. doi: 10.1111/mpp.13156

FIGURE 6.

FIGURE 6

Silencing of WRKY1 impairs RepA and MMDaV‐induced cell death in Nicotiana benthamiana. (a) Effect of NbWRKY1 silencing on RepA‐induced cell death. N. benthamiana plants were first infiltrated with TRV1 and TRV2‐NbWRKY1 or TRV1 and TRV2‐GFP as a control. After 10 days, the upper leaves were infiltrated with RepA‐GFP to monitor the cell death response. Photographs were taken at 3 days postinfiltration (dpi) of RepA‐GFP. The experiment was performed three times. Six plants were used for each TRV construct per experiment. (b) Effect of NbWRKY1 silencing on MMDaV‐induced cell death in N. benthamiana. Plants were first infiltrated with TRV1 and TRV2‐NbWRKY1 or TRV1 and TRV2‐GFP as a control. After 10 days, the upper leaves were infiltrated with MMDaV to monitor the cell death response. Photographs were taken at 3 dpi of MMDaV. (c, d) Reverse transcription quantitative real‐time PCR analysis of the silencing efficiency of NbWRKY1 in plants used in (a) and (b), respectively. The expression of NbWRKY1 is represented relative to the expression of the normalizer NbGAPDH using the comparative C t method (2−ΔΔ C t). Error bars represent standard deviation of each data set. (e) Western blot analysis of RepA‐GFP fusion protein accumulation in N. benthamiana leaves used in (a) using an anti‐GFP monoclonal antibody. Ponceau staining of RuBisCO serves as a loading control. (f) PCR analysis of MMDaV accumulation in infiltrated leaf patches of plants used in (b). Specific primers were used to amplify the c.1500 bp product from the total DNA extracted from infiltrated leaf patches