Figure 2.

The NbZAR1 MADA motif mediates XopJ4-triggered cell death in N. benthamiana. A, Cell death observed in N. benthamiana leaves after expression of ZAR1 and XopJ4. The leaves were infiltrated with Agrobacterium strains carrying ZAR1 and XopJ4 overexpression constructs at 1 d after initial agroinfiltration of RNAi constructs. Nicotiana benthamiana leaf panels expressing non-epitope-tagged wild-type and variant proteins of NbZAR1 were photographed at 5 d after the second agroinfiltration. B, Violin plots showing cell death intensity scored as an HR index based on three independent experiments. Statistical differences among the samples were analyzed with Tukey’s HSD test (P < 0.01). C, In planta protein accumulation of wild-type NbZAR1 and the variant. For anti-HA immunoblot, total proteins were prepared from N. benthamiana leaves at 2 d after the second agroinfiltration. Empty vector control is described as EV. Equal loading was checked with Reversible Protein Stain Kit (Thermo Fisher Scientific).