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. 2016 Dec 23;7:1945. doi: 10.3389/fpls.2016.01945

FIGURE 4.

FIGURE 4

Overexpression of different CIII peroxidases phenocopies the overexpression of PER57. (A) The expression of pPER57::GUS was evaluated in 21-day-old Col-0 seedlings sprayed with 5 mM OGs or 1 μM flg22 24-hpt, alternatively the leaves were wounded by gently pressing the lamina with laboratory forceps. (B) Toluidine blue (TB) staining was used to indicate cuticle permeability in 24-day-old C24, ohy1, Col-0 and transgenic lines overexpressing the apoplastic peroxidases PER57, PER10, PER28, PER34, PER44, PER53, and PER64 (n = 4). (C) Superoxide formation was detected using NBT staining in 21-day-old C24, ohy1, Col-0 plants and plants that overexpressed apoplastic peroxidases (n = 4). (D) Disease symptoms were evaluated at 3 days post-infection (dpi) with B. cinerea in 28-day-old C24, ohy1, Col-0 plants and plants overexpressing apoplastic peroxidases (n = 3). (E) TB-stained areas were quantified in Fiji. (n = 4, ±SD, N = 12 in total from three independent experiments). ∗p < 0.05, Student’s t-test. (F) Five-microliter droplets of a suspension of B. cinerea spores (2 × 105 spores mL-1) were placed on the leaves of 28-day-old plants. Images were taken three days post inoculation. The lesion area was measured in Fiji (n = 8, ± SD). ∗p < 0.05, Student’s t-test. All experiments were repeated at least twice.