全 文 ：Received 21 Apr. 2004 Accepted 24 Jun. 2004
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Acta Botanica Sinica
植 物 学 报 2004, 46 (9): 1071-1074
A Mechanism Facilitates Pollination due to Stigma Behavior
in Campsis radicans (Bignoniaceae)
YANG Shu-Xiang, YANG Chun-Feng, ZHANG Tao, WANG Qing-Feng*
(College of Life Sciences, Wuhan University, Wuhan 430072, China)
Abstract: The precise factors affecting stigma behavior in Campsis radicans (L.) Seem. ex Bureau.
remain unclear up to now. In this study mechanical touch, self- and cross-pollination, and pollination with
variable amounts of pollen grains separately contacting with stigmas have been conducted to determine
the exact factor affecting the stigma behavior. Results show that mechanical touch alone cannot make
the stigmas close permanently. It is the adequate pollen (＞350) deposition that causes the stigma
permanent closure, which is in accordance with previous reports that sufficient pollen grains are necessary
for fruit development. In addition, the stigma behavior does not display differences when pollinated with
cross- or self-pollen separately; both self and cross pollen grains can germinate and grow successfully. Our
results cannot demonstrate that the stigma behavior in C. radicans is an outcrossing mechanism, but strongly
indicate it acts as a mechanism to facilitate pollination, and then enhance the reproductive success.
Key words: stigma behavior; Campsis radicans; bilobed stigma; pollination
As an important reproductive structure, stigma plays
an important role in reproductive success of flowering plants
(Guo and Huang, 1999; Yang et al., 2002). Moreover, sev-
eral recent studies show that the movement of stigma can
facilitate outcrossing and/or pollination (Fægri and van der
Pijl, 1979; Fetscher and Kohn, 1999; Li et al., 2001; Li et al.,
2004). Li et al. (2001) found that the stigma movement due
to the flexible style in Alpinia obviously encouraged
outcrossing. Fægri and van der Pijl (1979) demonstrated
that the active stigma in Iris could enhance pollination effi-
ciency by combing pollen off the pollinators. Li et al. (2004)
also found that the stigma with the “comb like” fringe hairs
in Caesalpinia crista could scrap off the pollen grains ad-
hered on the abdomen of the insects into the stigmatic
chamber. In another study, the behavior of bilobed stigma
in Mimulus aurantiacus was considered as a mechanism
to prevent the intrafloral selfing because the stigma would
quickly close in response to touch by a pollinator (Fetscher
and Kohn, 1999). The stigma in Campsis radicans
(Bignoniaceae) also owns two active lobelets. Several hy-
potheses have been addressed to the significance of stigma
closure: aiding pollen capture and receipt (Thieret, 1976);
preventing pollen grains loss from stigmas (Bertin, 1982a);
preventing intrafloral selfing ( Newcombe, 1922; 1924; Lloyd
and Yates, 1982; Ritland and Ritland, 1989); reducing inter-
ference between pollen receipt and export functions within
the flower (Webb and Lloyd, 1986); and increasing pollen
export (Fetscher et al., 2002). A series of studies on
reproductive biology of C. radicans have been reported
(Bertin, 1982a; 1982b; 1985; 1986;1990a; 1990b; Bertin and
Sullivan, 1988; Bertin et al., 1989; Bertin and Peters, 1992;
Prakash, 1994; Galetto, 1995), and the stigma behavior of C.
radicans has been noticed and evaluated (Bertin, 1982a).
However, the exact factors affecting stigma movement and
the precise role of the stigma behavior in reproductive pro-
cess have not been clearly evaluated. We hypothesize that
two potential roles should be responsible for the signifi-
cance of the stigma behavior: enhancing outcrossing and/
or facilitating pollination. In this study, we try to ascertain
the following objectives: (1) what causes the stigma open
and closure? (2) do different pollen sources (self or
outcross) show different effects on stigma behavior? (3)
whether different pollen number show different effects on
stigma behavior?
1 Materials and Methods
Campsis radicans (L.) Seem. ex Bureau. is a protandrous
hermaphrodite, sprawling or climbing woody plant, with
long, red, tubular corollas, nectary and no scent; usually
cross-pollinated by hummingbirds and bees (Bertin, 1982a;
1988; 1989). The stigma has two lobelets, which is exserted
beyond the anthers in most plants (Fig.1). Five introduced
C. radicans populations were selected in this study. All
fieldwork was conducted in Wuhan, Hubei Province be-
tween May and October in 2002 and also in 2003. At least
30 inflorescences were selected randomly from different
Acta Botanica Sinica 植物学报 Vol.46 No.9 20041072
individual plants in each population for artificial treatments.
All chosen flowers were previously bagged. Four kinds
of artificial treatments were designed to the open virgin
stigmas: (1) mechanical touch (n=150); (2) self-pollination
(n=150); (3) cross-pollination (n=150); (4) pollination with
variable amounts of pollen grains (n=450). In each treatment,
we recorded the data as follows: whether the stigma clo-
sure or reopen and the time of the stigma closure or reopen.
All pollinated stigmas were collected and fixed in FAA.
The amounts of pollen grains deposited on the stigmas in
the fourth treatment were counted under the fluorescence
microscope, a drop of 0.1% aniline blue in 0.15 mol/L
K2HPO4 (pH 9.0) was added to each stigma on a micro-
scope slide before counted. The flowers self- or cross-pol-
linated were collected separately after pollinated 0.5 h, 1 h,
2 h, 3 h, 24 h. Their pollen tube growths were observed
under the fluorescence microscope.
2 Results
During the above study, we found that the anthers have
dehisced before the flowers opened, while the bilobed stig-
mas mostly began to spread apart about two hours later. In
general, the buds opened during 7:30-9:30 (a.m.), and the
stigmas completely opened during 10:30-11:30 (a.m.). The
open stigmas always kept open if not touched, even in the
evening, till faded 3 to 4 d later.
The stigmas given the mechanical touch mostly would
close between 40 s and 150 s, all of them would reopen
within 0.5-1.0 h. The stigmas pollinated mostly would close
between 15 s and 60 s, and some of them would reopen
within 1-3 h. The reopen stigmas closed again when given
more pollen grains. Stigma behavior does not show differ-
ence between treatments of self- and cross-pollination. Our
results also indicate that there was no difference in pollen
tube growing rate between self- and cross-pollen. Results
of the stigma behavior reflection to different stigmatic pol-
len loads are summarized in Table 1. When no or few pollen
grains stimulating, the stigmas closed and most would re-
open in a short time. However, the stigma closed and only
a few would reopen in a longer time when many pollen
grains were deposited. All above stigmas were in the early
flowers, the stigmas in the late flowers that the corollas
have fallen off would not close after mechanical touch or
pollen deposition.
3 Discussion
Bertin (1989) demonstrated that C. radicans was almost
completely self-sterile following pollinations by pure self-
pollen. Our study shows that the pollen tube from self-
pollen grew as rapidly as the pollen tube from cross-pollen.
Bertin and Sullivan (1988) also have pointed out that the
ability of self pollen tubes in C. radicans to grow to full
length as rapidly as cross pollen tubes. The interference
caused by self-pollen appears to take place in the post-
pollination process. This indicates that the stigma behav-
ior could not help selecting the cross-pollen from self-pol-
len to avoid the negative influence that self-pollen caused
on reproductive success.
Stigma closure is not limited to C. radicans, it is com-
mon in several families in Scrophulariales: Scrophulariaceae,
Bignoniaceae, Martyniaceae (Fetscher and Kohn, 1999) and
Lentibulariaceae (Newcombe, 1922). Two hypotheses have
been given to explain the stigma behavior in Mimulus
aurantiacus, a species with bilobed stigma (Fetscher and
Kohn, 1999). One is that the stigma rapidly response to
touch suggests that there is something to be gained by
immediate response to pollinator visitation. The other is
that it may prevent intrafloral selfing or reduce pollen-pistil
interference. Recent study indicates that stigma closure in
monkeyflower can increase pollen export: flowers with
closed stigmas exported 2.8 times more pollen to recipient
stigmas than did flowers with open stigmas (Fetscher
et al., 2002). The precise factors affecting stigma behavior
in C. radicans remains unclear up to now.
As Bertin (1982a) mentioned, mechanical touch or pol-
len deposition can elicit the stigma closure in C. radicans.
However, mechanical touch alone cannot make the stigma
Fig.1. Flower of Campsis radicans.
YANG Shu-Xiang et al.: A Mechanism Facilitates Pollination due to Stigma Behavior in Campsis radicans (Bignoniaceae) 1073
close permanently. All closed stigmas to mechanical touch
would reopen, while hand pollinated pollen grains reduced
significantly the possibility of stigma reopening. Moreover,
detailed study shows that more than 350 pollen grains de-
posited on the stigmas can cause most stigmas permanent
closure. We therefore suggest that the pollinated stigma
secondary open may be due to the limited pollen grains
deposited on the stigmas. This argument is supported by
the result of Bertin (1982a), which also indicates that the
stigma closure only occurs if effective pollination occurs.
It is then concluded that the reopen stigmas will close per-
manently only when sufficient pollen grains accumulating
on the stigmas. Our finding supports the statement that the
closure of bilobed stigma acts as a mechanism to aid pollen
receipt and capture (Thieret, 1976). Bertin (1982a) demon-
strated that at least roughly 400 pollen grains should de-
posit on a receptive stigma to cause fruit development in C.
radicans. Moreover, a further study by Bertin (1990a) indi-
cated high pollen loads resulted in seeds that were more
likely to germinate than seeds from low pollen loads.
Therefore, sufficient stigmatic pollen loads are important
to the fruit development and offspring quality in C.
radicans. It is interesting for our results that the closure of
stigma in C. radicans is a reflection of sufficient pollen
grains deposition, which meets with the requirement of re-
productive success in post-pollination process that suffi-
cient stigmatic pollen loads are necessary for fruit develop-
ment and seed quality (Bertin, 1982a; 1990a). We then sug-
gest that as one component of plant reproductive organism,
stigma character must coevolve with the whole breeding
system (see also Guo and Huang, 1999; Yang et al., 2002).
In summary, the bilobed stigma behavior in C. radicans
shows no difference between self- and cross-pollen contact.
In other words, our results cannot demonstrate the stigma
behavior in C. radicans acts as an outcrossing mechanism.
Combined with the studies of Bertin, our finding that the
significant correlation of stigma behavior and stigmatic
pollen loads suggest that the stigma behavior in C.
radicans should be a mechanism to facilitate pollination,
and then enhance the reproductive success.
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Table 1 State of the stigma (open or closed, the time of closure or reopen) after pollinated by different pollen loads
Stigmatic pollen load First closure time (s) (n＞ 100) Reopen stigmas (%) Reopen time (min)
>350 39± 11.1 10.4 (13/124) 145.8± 22.8 (n=13)
250-350 48.3± 10.9 19.8 (20/101) 122± 23.9 (n=20)
150-250 74.9± 27.6 80.4 (82/102) 86.7± 36.4 (n=82)
0-150 83.5± 26.7 94.6 (116/123) 71.9± 28.8 (n=116)
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(Managing editor: WANG Wei)