Additional File 1:

Time-lapse DIC images of a maize lemma from B-I Pl plant over-accumulating anthocyanins. Images were taken every four seconds. The time-lapse series was converted into a movie at one frame per second, therefore a speedup of 4X real time. The last four images represented in Fig. 10(C, D and E) are from this time-lapse series. Note the clear round inclusions in the tubular structures and the large vacuoles at the end. Refer to the results section for a detailed description. Viewing Instructions: The movie can be visualized using either Quicktime player or Windows Media Player. The plug-ins can be downloaded from http://www.apple.com/quicktime/downlond webcite (Quicktime Player) or http://windowsmedia.com/download/download.asp webcite Player). The movie size is ~2 MB, therefore it is more efficient to view by downloading the movie to your hard drive.

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Additional File 2:

Figure 1. In situ measurement of vacuolar pH using BCECF-AM. Equal amounts (0.01 g/100 μl fresh wt) of BCECF AM loaded cells were placed in microtiter plates, and the emission measured at 535 nm, 440 nm and 490 nm excitation wavelengths (A). The 490/440 ratio was calculated (B). An in situ calibration curve was generated separately for each of the dark and light samples with various pH buffers with 0.005% digitonin (C). Vacuoles of both dark and light grown BMS cells were acidic at pH 5.8 and showed no significant pH differences (D).

Format: PPT Size: 136KB Download file

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