Fig. 7.

Defense activation does not increase peptide accumulation. Only the active peptide accumulates in Col cells. Defense activation does not make cells permeable to inactive peptide or dye. Leaf discs were floated on indicated mixed peptide solution overnight; 1 μM TAMRA–flg22 (T-flg22), 2 μM FAM–flg22 (F-flg22), 2 μM FAM–scrambled peptide (F-scr), 2 μM flg22 and 2 μM fluorescein were used. (A) Confocal microscopy images in red (TAMRA) and green (FAM, fluorescein) channels and a composite of fluorescence channels with or without DIC are shown. (B) TAMRA and FAM fluorescence was quantified in confocal microscopy images of the same field. Data from at least three experiments per treatment were combined and presented as percentage of fluorescence in leaf discs treated with active peptide alone. Letters show significant difference (P<0.05, ANOVA/Tukey’s test, n≥8). Dashed line is background fluorescence (mock treatment without peptides). Mixed active peptides show lower fluorescence than single peptide in each channel due to competition of TAMRA–flg22 (T-f22) and FAM–flg22 (F-f22). f22 is flg22.