Figure 2.
Cytoplasmic localization of RB and IPI-O1 is required for their functions.
(A) Cytoplasmic localization of RB is required for IPI-O1-triggered HR. RB fused at the C terminus to GFP together with WT or mutated nuclear localization or export signals (NLS/nls or NES/nes) was co-expressed with either Myc-IPI-O1 or Myc-IPI-O4 in N. benthamiana.
(B) Cytoplasmic localization of IPI-O1 is required for RB-mediated HR. IPI-O1 or IPI-O4 fused at the C terminus to GFP together with WT or mutated NLS/nls or NES/nes was expressed in RB transgenic N. benthamiana plants.
(C and D) Cytoplasmic localization of both RB and IPI-O1 is required for their HR activities. RB and IPI-O1 fusion constructs from (A and B), respectively, were co-expressed in the indicated pairwise combinations in N. benthamiana plants. Myc-IPI-O1 was included as a control for the HR assays. Cell death induced at 48 hpi was visualized before (left) and after ethanol destaining (right). The infiltrated area is shown with a black circle and HR with a red circle. Scale bars correspond to 1 cm. All the experiments were repeated at least six times with similar results.