Fig. 2: RxLR23^KM and its five allelic variants arouse various degrees of cell death responses in N. benthamiana.

A The mutations of two key amino acids (K⁹³: Lysine and M³²⁰: Methionine) in RxLR23 sequence from different virulence P. capsici strains. The RxLR23 sequence shows that these two key amino acids are K⁹³ and M³²⁰ (named as RxLR23^KM) in a highly virulent strain SD33. RxLR23 sequence contains an allelic variant of K⁹³R (named as RxLR23^RM) in the mildly virulent strain YN07. The mutations of RxLR23 sequence result in two allelic variants of K⁹³R and M³²⁰R (named as RxLR23^RR) in the weakly virulent strain Aug0202. Allelic variants of K⁹³R, M³²⁰R, and K⁹³RM³²⁰R were generated using Fast Site Mutagenesis Kit. B Typical lesions caused by ectopic expression of pBinGFP2:RxLR23^KM or five allelic variants in N. benthamiana at 4 dpi (n = 30 samples). C Trypan blue staining of agroinfiltration sites highlighted cell death by RxLR23^KM and its five isoforms at 4 dpi (n = 30 samples). D Quantification of cell death induced by RxLR23^KM and its five isoforms. Percentage of cell death in spots was analyzed from at least 30 leaves. Values are presented as mean ± SD, n = 3 independent experiments (ANOVA, ***p < 0.001). E Electrolyte leakage was measured upon expression of RxLR23^KM and its five isoforms compared with INF1. Values are presented as mean ± SD, n = 3 independent experiments (ANOVA, ***p < 0.001). F Immunoblot analysis of transiently expressed proteins in N. benthamiana leaves using anti-GFP antibodies alone. Protein loading is indicated by Ponceau staining (Ponceau S). These experiments (B, C, F) were repeated at least three times. Source data are provided as a Source Data file.