Figure 6.

Transient Expression of NT Mutants Induces PCD in N. benthamiana.

(A) Appearance of N. benthamiana leaves at 4 dpi (bar = 1 cm). Areas were infiltrated with Agrobacterium carrying different constructs. Areas showing cell death are circled in red.

(B) GFP confocal microscopy of leaf areas 30 hpi (bar = 10 µm) and Trypan blue staining (TB) for cell death of N. benthamiana leaves expressing wild-type or mutant CNGC12 constructs at 48 and 96 hpi, as indicated (bar = 0.1 mm). Infiltrated and uninfiltrated leaf areas are separated by a yellow line in samples showing cell death.

(C) Ion leakage of infiltrated leaf areas 4 dpi. Values shown are averages of three replicates (error bars = sd; *P < 0.05 compared with empty vector, Student’s t test). E.V., empty vector (GFP); WT, At-CNGC12; 11/12, CNGC11/12; NT^mut, CNGC12^L27E/K28E; IQ^mut, CNGC12^I572D/Q573A; CT^mut, CNGC12^V607E; Δ8, CNGC12^Δ32-39.

(D) RT-PCR of cDNA from N. benthamiana leaves 24 hpi expressing wild-type or mutant CNGC12 constructs. Nb-ACTIN was used as a loading control (26 cycles).