Figure 6

Fluorescence microscopy analysis of SYTOX Green uptake during the membrane permeabilization assay. (A-D). Phase contrast images and (E-H) fluorescent images of untreated F. oxysporum, B. cinerea, V. dahliae and S. cerevisiae cells, respectively. (I-L) Phase contrast images and (M-P) fluorescent images of PgD5 treated F. oxysporum, B. cinerea, V. dahliae and S. cerevisiae cells respectively. Fungi were grown for 48h in the presence of PgD5 at peptide concentrations of 11 μg/mL for F. oxysporum, 4 μg/mL for B. cinerea, and 2 μg/mL for V. dahliae. S. cerevisiae cells were grown for 1h in the presence of PgD5 at 11 μg/mL. Afterwards, fungal hyphae and yeast cells were washed with 0.1 M Tris–HCl, pH 8.0, stained with 0.2 mM SYTOX for 30 min at 25°C with periodic agitation and subjected to fluorescent microscopic analysis. Bar = 20 μm.