Data supplements
Supplemental Data
Files in this Data Supplement:
- Supplemental Data -
Figure 1: Flow cytometry and epifluorescence microscopy analysis of a protoplast population isolated and purified from grape berry flesh. 3D density plot of the forward angle light scatter (FS) versus side angle light scatter (SS) of a grape cell suspension after FDA staining (A) and overlay of green fluorescence and autofluorescence histograms of the same cell suspension (B). Isolated grape cells observed under UV light (epifluorescence microscopy, C) after FDA staining.
Figure 2: Protoplasts from actively dividing Cabernet Sauvignon berry cells observed by both epifluorescence and confocal microscopy. Cells were stained with FDA (A) and with Hoechst (blue, nucleus), MitoTracker Red (red, mitochondria), and FM1-43 (green, plasma membrane) (B).
Figure 3: Mesocarp cells from fully ripened berry observed under the confocal microscope. Protoplasts were imaged by confocal laser scanning after being immersed overnight with the styryl dye FM1-43. Maximum Z projection of 20 sections covering ~30 µm (A). Intact vacuoles imaged after staining with the calcium fluorescent probe Fluo-4 AM (B). Plasma membrane integrity assessed under the fluorescence microscope with the fluorescent glucose analogue 2-NBDG (16-hr incubation) (C). Inset in C: single section of protoplast loaded with 2-NBDG observed by confocal microscopy 16 hr after incubation. (Color reproductions of figures are freely available as online supplemental data; www.ajevonline.org.)
Figure 4: Diversity in size, number, and vacuole sap composition/pH of berry-derived protoplasts as assessed after staining with the lipophilic phenazine dye Neutral Red. Vacuoles with precipitates (full cell saps; solid arrows, A, D, F, G, H) and without precipitates (empty cell saps; dashed arrows, B, C, E).
- Supplemental Data -