Fig. 3: Expression of RxLR23KM and its five allelic variants trigger various levels of plant defense against P. capsici infection. | Nature Communications

Fig. 3: Expression of RxLR23KM and its five allelic variants trigger various levels of plant defense against P. capsici infection.

From: A conserved oomycete effector RxLR23 triggers plant defense responses by targeting ERD15La to release NbNAC68

Fig. 3

A pBinGFP2:RxLR23KM, pBinGFP2:RxLR23RM, pBinGFP2:RxLR23RR, pBinGFP2:RxLR23K93R, pBinGFP2:RxLR23M320R, pBinGFP2:RxLR23K93RM320R, or pBinGFP2 was transiently expressed in N. benthamiana leaves and subsequently inoculated with P. capsici zoospores at 24 h (n = 30 samples). Representative lesions were taken under UV irradiation at 48 hpi. B H2O2 accumulation in N. benthamiana leaves was examined by DAB staining after inoculation with each construct at 36 h (n = 30 samples). C Mean diameter of lesions was measured in N. benthamiana leaves after inoculation with P. capsici at 48 h. Values are presented as mean ± SD, n = 30 samples. D qPCR was used to measure the relative biomass on the ratios of P. capsici to N. benthamiana leaves DNA after inoculation with P. capsici at 48 h. Pcβ-actin and NbEF1α were identified as the most suitable reference genes for normalization. The ratio expressing GFP was assigned to value 1.0. Values are presented as mean ± SD, n  = 3 independent experiments. E Expected protein sizes of RxLR23KM and its five isoforms were detected by western blots using GFP-antibodies alone. Protein loading is indicated by Ponceau staining (Ponceau S). F Relative levels of DAB staining were examined after inoculation at 36 h. The relative staining of GFP was assigned to value 1.0. Values are presented as mean ± SD, n = 30 samples. G Relative expression levels of PR1/2 were detected in N. benthamiana leaves after expression of each construct at 48 h. NbEF1α of N. benthamiana was as constitutively expressed endogenous control. Values are presented as mean ± SD, n = 3 independent experiments. The data in (C, D, F, G) were analyzed by ANOVA one-way comparison followed by least significant difference (LSD) test and different letters above the bars indicate a significant difference at p < 0.05. These experiments (A–C, E, F) were repeated at least three times. Source data are provided as a Source Data file.

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