Brassica campestris L. ssp. chinensis M

interference had a significant role in the pollen
development of the transgenic plant and resulted
in lower pollen germination ability as well as pollen
abnormality.
Expression profile analysis of the BcMF5
promoter in Arabidopsis
We have shown that BcMF5 was specifically
expressed in the late stage of pollen development.
To understand the function of the BcMF5 promoter
in the process of its transcription, a 620-bp
sequence upstream ATG was obtained from Brassica
campestris by TAIL-PCR for the first time. A further
database search did not find its homology sequence,
showing that it is an unpublished promoter.
To identify its temporal and spatial expression
pattern, the 609+3-bp promoter-GUS fusion vector
was constructed. Subsequently, its expression
profile in Arabidopsis was studied through GUS
histochemistry staining (Figure 8). The results
indicated that the BcMF5 promoter began expres-
ARTICLE IN PRESS
Functional analysis of a novel PCP gene BcMF5 451
400
300
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21
1 2 3 4 1 2 3
Figure 5. Molecular analysis of transgenic flowering Chinese cabbage. (A) shows the PCR results of transgenic flowering
Chinese cabbage. Lane 1: GeneRuler TM 100-bp DNA ladder; lanes 2– 20: PCR products amplified from genomic DNA of
transgenic flowering Chinese cabbage; lane 21: non-transformed Chinese cabbage plant. (B) DNA gel blot analysis of
transgenic line TB-6 and TB-13 of flowering Chinese cabbage. Lane 1: the external plasmid control; lane 2: the
nontransgenic plant; lanes 3–4: the transgenic line of TB-6 and TB-13. The genomic DNA of the transgenic plants and
their controls was digested by EcoRI and hybridized with 32 P-labeled NPTII-specific probe. (C) RNA gel blot analysis of
the transgenic line TB-6 and TB-13 of pBIA9-RMF5. Electrophoresis of the floral bud total RNA through gels containing
formaldehyde (down) and its result of RNA hybridization (upper). Lane 1: floral bud total RNA of the non-transgenic
flowering Chinese cabbage; lanes 2–3: floral bud total RNA of the transgenic line TB-6 and TB-13 of pBIA9-RMF5.
sion in the early stage of anther development
(Figure 8B) and drove high levels of GUS expression
in anthers, pollen, and pollen tube in the late stage
of pollen development (Figure 8C–G). It did not,
however, drive any expression in petals, sepals, or
pistils, which was relatively consistent with the
expression pattern of BcMF5 in flowering Chinese
cabbage (Figure 3).
Discussion
Pollination in flowering plants is a highly specialized
process that culminates in the fertilization of
the ovule by a male gamete (Doughty et al., 2000).
This reproductive process depends on highly specific
interactions between pollen and the pistil with
highly discriminatory interspecific and intraspecific
recognition, both of which allow the pistil to
distinguish among genetically diverse ranges of
pollen grains arriving at the stigma (Takayama
et al., 2000a). The adhesion of pollen to the stigma
is the first step in the pollination of flowering