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OpdCYC1C mRNA shifted to peripheries and gradually became undetectable in
the two lateral stamens, while OpdCYC1C transcripts continued to accumulate
to a high level in the dorsal and ventral staminodes (Figure 3E-F). Meanwhile,
OpdCYC1C mRNA became weak and not easily detectable in petals (Figure
3E-F). Weak mRNA signals of OpdCYC1C were also detected in the gynoecial
primordium (Figure 3E). In the middle developmental stages with flower buds
about 8 mm long, as stamens began filament elongation and anther differentia-
tion, OpdCYC1C was strongly expressed in the dorsal region (the dorsal petals
and staminode) and ventral staminode shown in RT-PCR while its weak mRNA
signal was detected in sepals, lateral and ventral petals, and lateral stamens, in
which the signal was much weaker in sepals and lateral stamens (Figure 4A, C). In
the late stages, OpdCYC1C transcripts declined in the dorsal region and ventral
staminodes, and were undetectable in other regions (Figure 4B-C). The Opd-
CYC1C mRNA was undetectable in stamens as pollen sacs began development in
the two lateral anthers while the two ventral anthers became sterile (Figure 3G).
Figure 3.
Tissue-specific expression of OpdCYC1 and OpdCYC2 during floral development in Opithandra
dinghushanensis.
A-G) RNA
in situ hybridizations with antisense probe of OpdCYC1C. A) Its mRNA is first detected in all five
petal and stamen primordia with weak signals in lateral edges and vascular tissue of sepals. B-D) Its mRNA then
weakened in two lateral stamens but strong in both dorsal and ventral staminodes with weak mRNA in the ring
meristem of corolla-tube. C-D) Successive sections from the same individual flower across base (C) and upper
parts (D) of stamens. E-F) Its expression shifts to peripheries and becomes undetectable in two enlarged lateral
stamens while remains strong in dorsal and ventral staminodes. G) Its mRNA is undetectable in stamens as pollen
sacs begin development. H-L) RNA in situ hybridizations with antisense probe of OpdCYC2A. H) Its dense
transcript accumulation first restricted to the dorsal side of the floral apex (arrow). I-K) Its strong expression
then restricted to two dorsal petals and the dorsal staminode as they are initiated. Note its mRNA later becomes
restricted to the dorsal-most parts in two dorsal petals (K). L) Its mRNA is undetectable in stamens as pollen
sacs begin development. OpdCYC1D (M) and OpdCYC2B (N) mRNA is not detected in floral tissues. As a
negative control, RNA in situ hybridizations with sense probes of OpdCYC1C (O) and OpdCYC2A (P) detect
no signal in floral tissues. dp, dorsal petal; ds, dorsal sepal; dt, dorsal staminode; g, gynoecium; lp, lateral petal;
ls, lateral sepal; lt, lateral stamen; vp, ventral petal; vs, ventral sepal; vt, ventral staminode. Scale bars, 150
ยต
m.
