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Fig. 4. ZOU expression pattern during seed development.
(A-D) Images of in situ hybridizations of digoxigenin-labelled
ZOU antisense (A-C) and sense (D) RNA probes to sections of siliques,
viewed using DIC microscopy. Signal appears purple; the dark-brown staining of
the endothelium (asterisk) does not represent signal. Embryos are indicated
with arrowheads, the surrounding endosperm with arrows. (A) Seed with early
globular stage embryo. (B) Seed with late heart stage embryo. (C) Seed with
torpedo stage embryo. (D) Control hybridization with ZOU sense probe.
(E-L) Confocal microscopy images in which autofluorescence appears red
and GFP fluorescence green. (E) Seed of transgenic plant carrying
ZOU::ZOU-GFP reporter gene fusion. The reporter is expressed in
nuclei of the embryo surrounding region (ESR) that surrounds the late heart
stage embryo. (F-L) Developing seeds of transgenic plant carrying
ZOU::H2B-YFP reporter gene. (F) Unfertilized ovule shows no transgene
expression. (G) Seed shortly after fertilization showing strong expression in
central cell. (H) Seed with endosperm at the two-nuclei stage, with uniform
expression in endosperm. (I) Seed with endosperm at the four-nuclei stage,
with uniform expression in endosperm. (J) Seed with endosperm at 12- to
16-nuclei stage; expression is becoming stronger at micropylar pole of
endosperm (arrow). (K) Seed with endosperm at 24- to 28-nuclei stage. (L) Seed
with endosperm at 44- to 48-cell stage, expression is now largely confined to
ESR. Scale bars: 100 µm.
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