Fig. S1 Schematic diagram showing stages of pollen development. V, vacuole; vn,
vegetative nucleus; gn, generative nucleus
Fig. S2 GFP-SYP124 was localized in the nucleus at the microspore stage. The
microspores were stained with DAPI. a, e DAPI fluorescence was localized to the
nucleus of microspores. b, f GFP fluorescence was detected in the nucleus of
GFP-SYP124-expressing transformants but not the wild type (WT). Merged images of
GFP and DAPI fluorescence (c, g) and corresponding bright-field images (d, h). Scale
bar = 10 m
Fig. S3 Wild-type (WT) pollen grains did not emit GFP fluorescence during pollen
development. a–d Confocal microscopic images of WT pollen were captured using the
same laser power and detector levels used for GFP-SYP124 pollen in Fig.2. e Pollen
tube of WT captured with a high laser power and detector level compared with that used
for GFP-SYP124. The GFP-like fluorescence was due to autofluorescence of the pollen
wall. Scale bar = 10 m
Fig. S4 Localization patterns of GFP-SYP124, 125 and 131. a–c The pollen tube was
stained with FM4-64. Left, GFP fluorescence; middle, FM4-64 fluorescence; right,
merge. Scale bar = 10 m. Note that the intensity of GFP-SYP131 fluorescence was
uniform on the PM in the pollen tube
Fig. S5 Localization of GFP-SYP131 and 132 in the pollen grain and pollen tube. The
confocal microscopic images show of (a, d) GFP fluorescence, (b, e) autofluorescence
of the pollen wall, and (c, f) merged images of GFP fluorescence and autofluorescence.
Arrows indicate the plasma membrane of the pollen grain. Arrow heads indicate the
plasma membrane of the pollen tube. Scale bar = 10 m
Movie S1 Live cell imaging of GFP-SYP125 during pollen germination.
Movie S2 Live cell imaging of the calcium sensor protein yellow chameleon
(YC3.60).
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