Fig 1.
A. Tomato Phenotypes. From left to right: ‘Black’, ‘White Beauty’, ‘Micro-Tom’ (from green stage to red stage) ‘Black’ and ‘White Beauty’ fruit fully ripens. For the 4 stages of ‘Micro-Tom’ shown, DPA (Day Post Anthesis) is 30~33 (green), 32~33 (yellow), 33~35 (orange), and 41~45 (red). B. Frozen sections of cuticle layers. From left to right: ‘Black’ mature stage, ‘White Beauty’ mature stage, ‘Micro-Tom’ (red maturity stage). The cuticle layers are found in the upper portion of each image. C. Electron microscopy of plastids in each tomato. Upper row shows Electron micrographs of plastids in each developmental stage of ‘Micro-Tom’. Lower row shows Electron micrographs of plastids in ‘Black’ and ‘White Beauty’ at mature stage. The bar in each figure represents the size scale for that figure. Red arrow shows membrane structures with high molecule density. Blue arrow shows crystal structure. Yellow arrow shows a whirled membrane structure.
Fig 2.
Isolation of plastids from tomato fruits.
A. Plastids isolated from varying stages/varieties of tomato fruits using Nycodenz density gradient centrifugation. Left side (Y-axis) indicates Nycodenz concentrations. Bands 2 and 3 are plastid fractions. B. Observation of plastids by fluorescence microscopy. In order to determine whether or not the isolated plastids were intact, we employed fluorescence microscopy (Olympus SZX9), observing plastids by differential interference, autofluorescence at 572nm, and also by detecting fluorescence using DAPI (4’, 6-diamidiano-2-phenylindole) stain. The bar in each figure represents the size scale for that figure.
Fig 3.
Carotenoid content in ‘Black’, ‘White Beauty’ and ‘Micro-Tom’ (Red).
A. Total amount of carotenoids measured in (both) immature and mature fruits. B. Amount of all-trans-violaxanthin, α-carotene, lutein and β-carotene in (both) immature and mature fruits. Sample weights of 1.5 g for mature fruit and 3.0 g for immature fruit were used to extract carotenoids. beta-Carotene, lycopene, all-trans-violaxanthin, and lutein were quantified in the pulp of tomato. Carotenoid content is reported as μg g-1 fresh weight. Carotenoid quantification was performed in three replicates.
Fig 4.
Flavonoid content in ‘Black’, ‘White Beauty’ and ‘Micro-Tom’ (Red).
The sample was 20mg (dry weight) of the pericarp portion. Flavonoid content is reported as mg g-1 dry weight. Flavonoid quantification was performed in three replicates.
Fig 5.
2D gel electrophoresis of chromoplast proteins in fruit cells of ‘Micro-Tom’ (Red), ‘Black’ and ‘White Beauty’.
Using an 11 cm IPG DryStrip (pH4-7), 100 μg of solubilized proteins were separated. Two-dimensional gel electrophoresis was performed with 12% poly-Acrylamide gel (16 cm x 16 cm). Following electrophoresis, the gel was stained with Flamingo Fluorescent Gel Stain (BIO-RAD). A list of proteins numbered in Fig 5 can be found in Table 1.
Fig 6.
Proteins of varying levels of accumulation in ‘Micro-Tom’ (Red), ‘Black’ and ‘White Beauty’.
Enlarged images of 2D-PAGE results from Fig 5 that showed significant differences in protein accumulation. Protein spots of note are indicated with broken line circles. CHRC: ‘Micro-Tom’ spot16, ‘White Beauty’ spot16, HrBP1: ‘Micro-Tom’ spots17 and 18, ‘Black’ spot13, ‘White Beauty’ spot14, TIL: ‘Micro-Tom’ spots27 and 28, ‘Black’ spot 21, ‘White Beauty’ spot25.
Table 1.
Protein list of the 2D-PAGE numbering spots in Fig 5.