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. 2019 Jan 30;20(4):575–588. doi: 10.1111/mpp.12776

Figure 1.

Figure 1

Verification of CESSNA [Controlled Expression of effectors for Synchronized and Systemic NLR (nucleotide‐binding leucine‐rich repeat) Activation]: defence gene expression, cell death and coat protein (CP) accumulation. (A) Potato virus X (PVX) CP accumulation in wild‐type (WT) plants expressing DEX::CP105 or DEX::CP106. Semi‐quantitative reverse transcription‐polymerase chain reaction (RT‐PCR) at 0, 0.5 and 1 h after dexamethasone (DEX) application (hpda); expression of EF1α serves as an internal control (bottom panel). (B) Western blot showing PVX CP accumulation in WT plants transiently expressing DEX::CP105 or DEX::CP106 at 0, 1, 2, 3 and 4 hpda. Blots were probed with horseradish peroxidase (HRP)‐conjugated goat anti‐rabbit immunoglobulin G (IgG) secondary antibody. (C) Quantitative RT‐PCR analysis of defence response and hypersensitive response marker genes. Expression pattern of PR‐1a, LOX, HIN1 ERF1 and AOX1b after DEX induction of CP105 and CP106 in Rx1:4xHA N. benthamiana. Data are means ± standard error (SE), normalized by EF1α and PP2A expression. Asterisks indicate significant differences by one‐way analysis of variance (ANOVA) (P < 0.0001). (D) Trypan blue visualizes cell death. WT and Rx1:4xHA leaves infiltrated with Agrobacterium tumefaciens carrying DEX::CP105 and DEX::CP106 constructs. Two days after infiltration, leaves were brushed with 20 µm DEX and stained the following day.