Fig. 9

a Gene expression levels of related to plant immunity in Nicotiana benthamiana leaves transiently expressing two effector genes for 8, 12, 24, 48, and 72 h. Nicotiana benthamiana leaves were infiltrated with Agrobacterium tumefaciens GV3101 carrying either the PMDC32 empty (Control) or PMDC32-smut_5844 and smut_2965 vector. The plants were incubated in a plant growth room with 12 h/12 h, night/day photoperiods at 20–24 °C with 60% relative humidity. The leaves were harvested after 8, 12, 24, 48, and 72 h post-infiltration. Total RNA was extracted and reverse transcribed into cDNA. The qPCR was performed on the cDNA using specific primers for several genes related to activation of plant immunity. Actin gene was used to normalize the data, which are presented as normalized relative quantities scaled to control. Error bars indicated the standard deviation of 3 independent replicates. b N. benthamiana leaf that infected with agrobacterium carrying GFP was dyed used DAB. c N. benthamiana leaf that infected with agrobacterium carrying smut_2965 was dyed used DAB. d N. benthamiana leaf that infected with agrobacterium carrying smut_5844 was dyed used DAB. For statistical analysis, we used analysis of variance with the R-package “ggpubr”, and significance was set at P < 0.05. *: P < 0.05, **: P < 0.01, ***: P < 0.001, ****: P < 0.0001