Globular vacuole appearance and cytoplasmic instability in wild-type plants treated with oryzalin.

<p><b>A.</b> Single view (top) and 3D view (bottom) of vacuoles in wild-type transition stage cells. Vacuoles were visualized following staining with 10μm BCECF staining (10 μm) of mock (0.5% DMSO) and 50μm oryzalin-treated roots. Red is propidium iodide to visualize cell walls. Bars, 10μm. <b>B.</b> Cotyledon epidermal cells from wild-type plants expressing 35S::GFP-ɣTIP, treated for 30min with 50μm oryzalin (right), or mock 0.5% DMSO (left). Bars, 10μm. <b>C.</b> Single time-point images of root epidermal division zone cells from wild-type plants expressing <i>UBQ</i>::<i>GFP-TUB6</i> treated with mock 0.5% DMSO or 50μm Oryzalin. Images were taken after 45 minutes of treatment. <b>D.</b> Time projection (standard deviation method) of cells in A showing the enhanced vacuole/cytoplasm movement in oryzalin-treated plants. White areas indicate regions that underwent positional/morphological change over the course of observation. Images correspond to 120 second time-lapses, 4 second intervals. <b>E.</b> Color merge of two time points from same time series. Green is t = 1 and magenta is t = 120. White areas indicate no movement between time points. <b>F.</b> Kymographs corresponding to yellow lines in the two-colour merged images. Enhanced dynamicity of vacuoles mirrors that observed in <i>clasp-1</i> cells. Total time is120 seconds. Bars, 10μm. <b>G.</b> Life histories of individual vacuoles from a transition stage cell over the course of 160 seconds (4 second intervals). Different coloured traces correspond to individual vacuoles. Left image shows all time-points projected onto a single image frame for reference. Right images are tilted 3D projections. The rear and side walls are kymographs for spatial and temporal reference. Bars, 10μm. <b>H.</b> Coefficients of variation (co.var.) for vacuole area in control (0.5% DMSO) and roots treated with 50 μm oryzalin. Time between compared time points 80 second. n = 30 vacuoles for each genotype. Bars indicate Standard Error.