Figure 6. Activation of defense reaction in N. benthamiana leaves transiently expressingP_35S-gRCY1-HA and P_35S-cRCY1-HA under control of the CaMV 35S promoter.

HA-epitope-tagged RCY1 protein (α-HA) (A) in N. benthamiana leaves transiently expressing P_35S-gRCY1-HA, P_35S-cRCY1-HA, or pRI201-AN (Vector) as an empty-vector control was immunologically detected using anti-HA monoclonal antibody. As an internal control for protein sample quantities, the large subunit of RuBisCO was visualized by staining with CBB. The size of each band was shown at right side of the panel. In N. benthamiana leaves transiently expressing P_35S-gRCY1-HA or P_35S-cRCY1-HA, hypersensitive response (HR) cell death was visualized by trypan blue staining (B), and H₂O₂ production was detected by DAB staining (C). To evaluate HR-cell death quantitatively, electrolyte leakage (D) in N. benthamiana leaves transiently expressing P_35S-gRCY1-HA, P_35S-cRCY1-HA, or empty-vector control was measured. Expression of the defense-related genesPRB-1b, PR-2b, and PR-6 in N. benthamiana leaf tissue transiently expressing P_35S-gRCY1-HA, P_35S-cRCY1-HA, or the empty-vector control was analyzed by northern hybridization (E). As an internal control for RNA sample quantities, 18S rRNA was shown.