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Fig. 4 | Phytopathology Research

Fig. 4

From: Multiple aromatic amino acids are involved in potyvirus movement by forming π-stackings to maintain coat protein accumulation

Fig. 4

Effects of substitution retaining π-stackings on CP accumulation, replication and movement of TVBMV. a GFP fluorescence and b relative GFP fluorescence intensities in the N. benthamiana leaf patches expressing GFP-TVBMVCP, GFP-TVBMV(Y105A), GFP-TVBMV(Y105G), GFP-TVBMV(Y147A), GFP-TVBMV(Y147G), GFP-TVBMVCP (ΔY105), GFP-TVBMVCP(Y105F), GFP-TVBMVCP(Y105W), GFP-TVBMVCP (ΔY147), GFP-TVBMVCP(Y147F) and GFP-TVBMVCP(Y147W) at 5 dpai. Pictures were photographed under UV light. GFP fluorescence intensities were determined by a multi-function microplate reader. ‘Δ’ indicates the codon for the residue has been deleted. c GFP fluorescence and d viral RNA accumulation level in the systemic leaves of N. benthamiana inoculated with TVBMV-GFP, TVBMV-GFP(Y105A), TVBMV-GFP(Y105G), TVBMV-GFP(Y147A), TVBMV-GFP(Y147G), TVBMV-GFP(Y105F), TVBMV-GFP(Y105W), TVBMV-GFP(Y147F) and TVBMV-GFP(Y147W) at 7 dpai. Viral RNA accumulation was determined by RT-qPCR. Blue and green arrows indicate the infiltrated and systemic leaves, respectively. e Cell-to-cell movement of TVBMV-GFP, TVBMV-GFP(Y105A), TVBMV-GFP(Y105G), TVBMV-GFP(Y147A), TVBMV-GFP(Y147G), TVBMV-GFP(Y105F), TVBMV-GFP(Y105W), TVBMV-GFP(Y147F) and TVBMV-GFP(Y147W). GFP fluorescence was observed under a confocal microscope at 5 dpai. f The number of cells showing GFP fluorescence in each infection locus of TVBMV-GFP(Y105A), TVBMV-GFP(Y105G), TVBMV-GFP(Y147A) and TVBMV-GFP(Y147G). g The diameter of cells showing GFP fluorescence in each infection locus of TVBMV-GFP, TVBMV-GFP(Y105F), TVBMV-GFP(Y105W), TVBMV-GFP(Y147F) and TVBMV-GFP(Y147W). The values are the means ± SD from 30 infection foci per treatment. h The minus-strand (−) RNA accumulating level in the infiltrated leaf patches of N. benthamiana inoculated with TVBMV-GFP, TVBMV-GFP(Y105A), TVBMV-GFP(Y105G), TVBMV-GFP(Y147A), TVBMV-GFP(Y147G), TVBMV-GFP(Y105F), TVBMV-GFP(Y105W), TVBMV-GFP(Y147F), TVBMV-GFP(Y147W) or replication-defective mutant TVBMV-GFP (NIbΔGDD) detected by RT-qPCR at 60 hpai, with the expression of NbEF1A as an internal control. The values are the means ± SD from three biological replicates per treatment. Treatments marked with the same lowercase letters have no significant statistical difference

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