Investigating the functional domains of Rp1-D21 and Rp1-D to trigger a hypersensitive response (HR).

<p>(A) Schematic diagram of the Rp1-D21 and Rp1-D domain structure and the derived fragments used for agro-infiltration of <i>N</i>. <i>benthamiana</i>. The different domains are indicated using different colors: CC (purple); NB-ARC (red); LRR (dark blue). The positions of the amino acids that form the domain boundaries are indicated on the top and the abilities to induce (+) or not induce (-) HR are listed on the right of each construct. (B) The HR phenotype of the constructs fused with a C-terminal EGFP tag and transiently expressed in <i>N</i>. <i>benthamiana</i>. Representative leaves were photographed at 3 days post infiltration (dpi, left). Total protein was extracted from agro-infiltrated leaves at 30 hours post infiltration (hpi), and anti-GFP antibody was used to detect the expression of fused proteins. Equal loading of protein samples was shown by Ponceau-S staining of Rubisco subunit (right, below). The results shown were from different domains of Rp1-D21, and similar results were observed from different domains of Rp1-D. (C) The phenotype of the constructs fused with a C-terminal 3×HA tag and transiently expressed in <i>N</i>. <i>benthamiana</i>. A representative leaf was photographed at 3 dpi (left), and the same leaf was cleared by ethanol (middle). Total protein was extracted from agro-infiltrated leaves at 30 hpi, and anti-HA antibody was used to detect the expression of the fused proteins. Equal loading of protein samples was shown by Ponceau-S staining of Rubisco subunit (right). The results shown were from different domains of Rp1-D21, and similar results were observed from different domains of Rp1-D. The experiments were repeated three times with the same results.