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Figure 4.
Gene-specific semiquantitative RT-PCR on RNA prepared from dissected Opithandra dinghushanensis
flowers.
A) sepal
(S), dorsal petal+staminode (Dp+s), lateral/ventral petals (Lp/Vp), lateral stamens (Ls) and ventral
staminode (Vs) were dissected from flower buds of middle-stage (less than 1 cm long). B) sepal (S), dorsal
petal+staminode (Dp+s), lateral/ventral petals (Lp/Vp), lateral stamens (Ls) and ventral staminode (Vs) were
dissected from flowers of late-stage (3-4 cm long). C) Relative level of OpdCYC1C mRNA expression in middle-
stage (black) vs. late-stage (grey) compared with ACTIN. D) Relative level of OpdCYC2A mRNA expression
in middle-stage (black) vs. late-stage (grey) compared with ACTIN. E) Relative level of Opdcyclin D3a mRNA
expression in middle-stage (black) vs. late-stage (grey) compared with ACTIN. F) Relative level of Opdcyclin
D3b mRNA expression in middle-stage (black) vs. late-stage (grey) compared with ACTIN. ACTIN was used
for RT template control. The values (means ± SD) shown are determined from five independent experiments.
In contrast to OpdCYC1C, OpdCYC2A mRNA densely accumulated in the
dorsal region of the floral apex as petals and stamens became visible (Figure 3H).
Then, the OpdCYC2A expression signal was specifically concentrated in the dor-
sal petals and the dorsal staminode (Figure 3I-J). As floral organs developed, Op-
dCYC2A transcripts continued to be highly concentrated at the dorsal staminode
and the dorsal-most parts of the two dorsal petals (Figure 3J-K). In the middle
stages, the strong mRNA signal of OpdCYC2A was detected in the dorsal region
(the dorsal petals and staminode) shown in RT-PCR that declined in late stages
with no signal in other regions (Figure 4A-B, D). Its mRNA was undetectable in
stamens as pollen sacs began development in the two lateral anthers while the two
ventral anthers became sterile (Figure 3L). Even though OpdCYC1D expression
was undetectable in floral tissues using in situ hybridization (Figure 3M), its very
weak mRNA signals were detected in the dorsal region (the dorsal petals and sta-
minode) from middle to late stages using RT-PCR (Figure 4A-B). OpdCYC2B
