Figure 7.

Cuticle permeability and ROS accumulation in mutants with impaired cell wall integrity. (A,B), Rosette leaves of five-week-old Col-0 (WT), mur1, mur4, prc1, irx1-2, and fer-4 plants were stained with toluidine blue (A) to assay cuticle permeability or (B) incubated for the indicated times with 80% EtOH, and the concentration of chlorophyll leaked in the medium and of total chlorophyll was spectrophotometrically measured. Bars in (B) indicate average percentage of total chlorophyll leaked at the indicated time of incubation ± SD (n = 3). For each time point, asterisks indicate statistically significant differences with the control-treated WT samples, according to Student’s t-test (^***p < 0.01; ^*p < 0.01). (C), rosette leaves as in (A,B) were stained with DAB for ROS accumulation. These experiments were repeated three times with similar results.