Figure 3.

NS_M‐dependent and NS_M‐independent hypersensitive cell death response (HR) induction by domains from Sw‐5 orthologues. (A) Co‐infiltration of agrobacteria harbouring Sw‐5b gene‐based derivatives (cloned into pK7WGF2) as depicted, with p19 (first column), BR‐01 NS_M (second column) and GRAU NS_M (third column). All constructs were always infiltrated into the same leaf per assay. Photographs were taken at 4 days post‐agroinfiltration (dpa). *As a positive control on (auto)HR triggering, the Sw‐5b full‐length gene was co‐infiltrated with p19 (first column; auto‐HR) or BR‐01‐NS_M (second column; NS_M‐dependent). (B) Co‐infiltration of Nicotiana benthamiana with agrobacteria harbouring the NB‐ARC and/or SD‐CC domains as depicted, and with p19 (from pBIN) for cell death triggering by auto‐activation. Leaves were monitored for HR at 4 dpa. Independent of the combination, each construct was infiltrated at the same agrobacterium optical density (OD). (C) Co‐infiltration of agrobacteria harbouring the NB‐ARC domains from Sw‐5a, Sw‐5b or Sw‐5a^S and p19 for auto‐HR triggering; photographs were taken at 4 dpa. (D) Western blot of the Sw‐5b‐truncated proteins expressed via pK7WGF2‐based constructs. Anti‐green fluorescent protein (anti‐GFP) was used as primary antibody. ARC, adaptor shared by Apaf‐1 (from humans), R proteins and CED‐4 (from nematodes); CC, coiled coil; LRR, leucine‐rich repeat; NB, nucleotide binding. SD, Solanaceae domain.