Figure 6. Antagonistic effects of CB and phalloidin on vacuole coalescence.

Rice aleurone layers were isolated from seeds imbibed for 2 d and then incubated with distilled water, 10 μg mL⁻¹ CB and 10 μg/ml phalloidin for 5 d. Finally, the layers were examined for vacuolation after 8.5 μg mL⁻¹ AO or 2 μg L⁻¹ FDA/1 μg L⁻¹ FM4-64 staining using LSCM. (A) Live cells exhibit AO green fluorescence. (B) Statistical analyses were conducted on the vacuole number per cell. (C) Green fluorescence (FDA) stains the live cells and orange fluorescence (FM4-64) stains the dead cells. (D) Viability of the cells was quantified from at least four aleurone layers. The assay was performed at least three times and a representative aleurone layer section is shown. Data represent the mean ± SD from three independent biological replicates, and different letters indicate a significant different at p < 0.05 according to Duncan’s multiple range test. The bar represents 50 μm.