Fig. 5

Ca²⁺ signals are required for root organogenesis in the leaf explants. (A-D) Effects of EGTA on ROS accumulation. Leaf explants from the 9-day-old Col-0 seedlings were incubated on B5-agar plates containing 0.5 mM EGTA for the indicated time periods before NBT (A) or DAB (C) staining. Relative intensity of NBT (B) and DAB (D) signals in the total leaf area and near the wound site was measured using ImageJ software. Seven to ten biological replicates were averaged and statistically analyzed using Student’s t-test (*, P < 0.05; difference from Mock). Scale bars indicate 0.5 mm. Whiskers indicate SD. (E) Effects of EGTA on root organogenesis in the leaf explants. Leaf explants from the 9-day-old Col-0 seedlings were incubated on B5-agar plates containing 0, 0.5, 1, or 1.5 mM EGTA for up to 14 d. Rooting rates of three biological replicates were averaged and statistically analyzed using Student’s t-test (*, P < 0.05; difference from Mock). Each replicate contains 20–30 explants. Whiskers indicate ± SD