Making Use of Plant uORFs to Control Transgene Translation in Response to Pathogen Attack
Pathogen-inducible protein accumulation of AtLecRK-VI.2 is achieved by using the combinations of different uORFs and the NbPR1 promoter. (a) Cell death induced by indicated gene products. Leaves infiltrated with Agrobacterium harboring indicated constructs. Photos were taken at 5 dpi in white light. (b) CF-induced AtLecRK-VI.2 transcript accumulation confirmed by qRT-PCR. Transcript accumulation levels of AtLecRK-VI.2 were analyzed by qRT-PCR. P. capsici CF was infiltrated into leaves expressing the indicated plasmid. Total RNA was extracted 1 hour posttreatment. The NbELF18 gene was used as an internal reference. Bars represent standard errors from three biological replicates (;;,; Student’s -test). (c) Induced AtLecRK-VI.2 protein accumulation confirmed by Western blot assay. CF was infiltrated into leaves expressing the indicated plasmid. Proteins was extracted 2 hours posttreatment. The α-FLAG antibody was used to detect the expression of AtLecRK-VI.2 protein. Equal loading of each sample is indicated by Ponceau staining. (d) Enhanced resistance to P. capsici infection provided by ACD11 uORF-mutants coupled with the PR1 promoter and AtLecRK-VI.2 in N. benthamiana. Lesion areas were calculated from three independent experiments with at least five leaves per replicate (;;,, Student’s -test).