FIGURE 1.

VdEPG1 suppresses the cell death induced by NLP1 in Verticillium dahliae. (a) Cell death assays for 11 V. dahliae genes in 6‐week‐old Nicotiana benthamiana leaves were performed by Agrobacterium‐mediated transient expression. Leaves were imaged 6 days after infiltration with Agrobacterium carrying the VdGH28 family genes. NLP1 and green fluorescent protein (GFP) were used as the positive and negative controls, respectively. Scale bar = 1 cm. (b) Semiquantitative reverse transcription‐PCR analysis of transiently expressed VdGH28 genes in N. benthamiana leaves 48 h after infiltration. NbActin was used as the control. (c) Detection of the cell death‐suppressing activity of VdEPG1. NLP1 was transiently expressed 24 h after infiltration with VdEPG1 in 6‐week‐old N. benthamiana leaves. Western blot analysis showed VdEPG1 was expressed in N. benthamiana leaves. 1, sample infiltrated with NLP1 only; 2, sample infiltrated with VdEPG1 only; 3, sample that was infiltrated with NLP1 24 h after infiltration with VdEPG1; 4, sample infiltrated with GFP only. Commassie brilliant blue (CBB) was used as the loading control. Scale bar = 1 cm. (d) Confirmation of the function of the signal peptide of VdEPG1 by yeast signal trap assay. Growth on YPRAA medium indicates the functionality of the signal peptide of VdEPG1. The signal peptide of Avr1b was used as the positive control. (e) and (f) Reverse transcription‐quantitative PCR analysis of the PAMP‐triggered immunity marker genes and pathogenesis‐related genes. N, E, and G represent NLP1, VdEPG1, and GFP, respectively. The blue, red, green, and black columns represent the samples which were infiltrated at 0, 12, 24 and 48 h, respectively. NbActin was used as a control. Values represent the mean ± standard deviation of three replicates. *p < 0.05, **p < 0.01, ***p < 0.001.