FIGURE 2.

Signal peptide and nuclear localization signal are required for Vd424Y‐induced cell death. (a) Schematic illustration of the Vd424Y deletion mutants: Vd424Y, the full‐length sequence of the candidate effector; Vd424N, effector variant lacking the signal peptide (SP) sequence; Vd424Y‐nls, effector variant where the predicted nuclear localization signal (NLS) was mutated to alanine residues; Vd424Y‐nls‐NES, the variant Vd424Y‐nls was fused with a NES at C‐terminus; Vd424Y‐NES, effector variant where the NES at C‐terminus was tagged to the wildtype Vd424Y allele; Vd424N‐SPNbPR1, effector variant with a fused SP of Nicotiana benthamiana pathogenesis‐related protein (NbPR1). (b) N. benthamiana leaves were infiltrated with Agrobacterium tumefaciens carrying Vd424Y, Vd424N, Vd424Y‐nls, Vd424Y‐nls‐NES, Vd424Y‐NES, Vd424N‐SP_NbPR1 , positive control BAX, and control green fluorescent protein (GFP). Representative photographs of N. benthamiana leaves were taken after 7 days. (c) Subcellular localizations of Vd424Y‐YFP, Vd424Y‐nls‐YFP, Vd424Y‐nls‐NES‐YFP, and Vd424Y‐NES‐YFP in N. benthamiana on A. tumefaciens‐mediated transient expression. Fluorescence was detected by confocal microscopy 48 hr postinfiltration. Bars, 40 µm