Floral phenology,
secondary pollen presentation and pollination mechanism in Inularacemosa (Angiosperms: Asteraceae)
Peerzada Arshid Shabir 1, IrshadAhmad Nawchoo 2 & AijazAhmad Wani 3
1,2,3 Economic Botany and
Reproductive Biology Laboratory, Department of Botany, University of Kashmir,
Srinagar, Jammu and Kashmir 190006, India
1 peerzadarshid@gmail.com
(corresponding author), 2 irshadnawchoo@yahoo.co.in, 3 aijazbotku@gmail.com
doi: http://dx.doi.org/10.11609/JoTT.o3320.4498-503
Editor: Cleofas Cervancia, University of Philippines Los Baños College Laguna, Philippines. Date of publication: 26
June 2013 (online & print)
Manuscript details: Ms # o3320 | Received 26 August 2012 | Final received
08 May 2013 | Finally accepted 09 May 2013
Citation: Shabir,
P.A., I.A. Nawchoo & A.A. Wani(2013). Floral phenology, secondary pollen presentation and
pollination mechanism in Inula racemosa (Angiosperms: Asteraceae). Journal of Threatened Taxa 5(10): 4498–4503; http://dx.doi.org/10.11609/JoTT.o3320.4498-503
Copyright: © Shabiret al. 2013. Creative Commons Attribution 3.0 Unported License. JoTTallows unrestricted use of this article in any medium, reproduction and
distribution by providing adequate credit to the authors and the source of
publication.
Funding: Council of Scientific and Industrial
Research, Pusa, New Delhi.
Competing Interest: None.
Acknowledgements: The
first author is highly thankful to CSIR for providing financial assistance in
terms of JRF fellowship during this study.
Abstract: Inula racemosa Hook. f. is protandrous, discharges pollen
grains inside the anther tube and presents pollen secondarily onto the sweeping
hairs of the style. The style and stigmatic branches present the yellow clumped
pollen grains for pollination. This
study describes floral functional morphology and phenology, anther dehiscence
and pollen presentation, growth and behaviour of
style during anthesis and pollination mechanism of I.racemosa. The species is entomophilous and is
characterized by a highly asynchronous sexual phase. A large degree of asynchrony from floret
to floret in a capitulum, and capitulumto capitulum in a plant, keeps the pollen dispersed
for a longer duration. Two insect
families were represented in the pollinator survey: Hymenoptera and Diptera. A
significant correlation was observed between the number of capitulavisited per bout and foraging time. We discuss morphological features of the flowers which may enhance the pollen removal rate per bee
visit and consequently cause a high visitation and pollination rate.
Keywords: Breeding system, Inula racemosa, phenology, pollination, secondary pollen
presentation
For figures, images -- click here
Flowering plants possess
exceptionally versatile mating strategies which are
governed by distinctive features of the biology and ecology of the group. Since mating strategies employed
by a group influence the movement of genes in space and their transmission
through time, they play a fundamental role in determining spatial and temporal
patterns of genetic diversity within and between populations, and hence their
evolutionary dynamics. Because of
their immobility, plants require vectors to transfer pollen, giving rise to
diverse floral adaptations which are widely recognized to influence floral
organization, patterns of pollen dispersal, pollinating agents involved and
these reproductive traits comprise the key elements of the mating strategies of
flowering plants.
In angiosperms, individuals
potentially compete to donate pollen to receptive stigmas. Because of this, sexual selection can
shape functional floral characters (Bell 1985; Queller1997). The structural variation in
flowers and inflorescences and different mechanisms for deployment of male and
female gametes is remarkable when one considers that their sole function is to
promote reproductive success. Analysis of any plant community reveals a variety of pollination and
mating systems that coexist under apparently similar ecological conditions. Male and female gametes are deployed in
various structural and temporal combinations at the flower, inflorescence, plant or population level. Morphological and phenologicalaspects of floral design and display primarily influence the quantity and
quality of pollen dispersal during the pollination process. Several functional or adaptive floral
traits, such as protoandry, self-incompatibility, herkogamy and secondary pollen presentation, have evolved
to avoid or minimize the effects of self-interference and thus improve
outcrossing rates.
Inula racemosa Hook. f.,
commonly known as “Pushkarmoola” of family Asteraceae is distributed from temperate to sub-alpine
belts, the species has been categorized as rare according to the Red Data Book
of Indian plants (Nayar & Sastry1988.). The species is entomophilous with 10–20 capitulaper plant aggregated in racemes. Disc florets of I. racemosa are protandrous and have a peculiar mechanism of secondary
pollen presentation. In protandrous flowers male and female function is temporally
separated and pollen is presented before the stigma becomes receptive. Protandry is
common in outcrossing angiosperms (Lloyd & Webb 1986) and is often assumed to be selected for avoidance of
self-fertilization.
Secondary pollen presentation
is the developmental relocation of pollen from the anthers onto another floral
organ which then functions as the pollen presenting organ for pollination
(Howell et al. 1993) and the phenomenon has traditionally been described as a mechanism
that enhances the efficiency and accuracy of pollen exportation and/or pollen
reception, thus increasing male and/or female fitness of the plant (Ladd
1994). Diverse methods have evolved
for secondary pollen presentations with pollen being presented onto the style
(Nyman 1993), in specialized regions of the style or stigma (Westerkamp & Weber 1997), or even over the stigmatic
area (Imbert & Richards 1993), usually before
flower opening. In I. racemosa, however,
pollen is shed onto the sweeping hairs of the style prior to floret anthesis and the style subsequently elongates to display
the pollen. Thus, in I. racemosa pollen is presented by the
style and stigmatic branches before the stigma completely exposes the
two layers of receptive papillae cells which marks the entry of a floret into
the functionally female phase. To
provide information on the basic reproductive biology of I. racemosa, we initiated this study using laboratory and
field approaches to investigate: (i) floral
functional morphology; (ii) anther dehiscence and pollen presentation (iii)
growth and behaviour of style during anthesis; and (iv) pollinationmechanism (Image 1).
Material and Methods
Study area: The
study was performed during April 2009 to November 2010 on two main study sites,
i.e., Gulmarg (2650m) and Farozpora(2150m), lying at a distance of 50–60 km to the west of Srinagar
City. In addition, one transplanted
population was established and monitored at Kashmir University Botanical Garden
(KUBG-1595 m).
Floral morphology and phenology: Morphometric analyses were
performed on several capitula by collecting the
samples from flower buds to capitula at anthesis and observed under a dissecting microscope. The capitulawere examined for the spatial and temporal arrangement of male and female
sexual parts within the ray and disc florets. Various floral phenologicalstages were identified according to the development and relative location of
ray and disc florets within a capitulum. Macro images
of floral features were captured using a stereo microscope.
Stigma receptivity: Stigmas
at different developmental stages of selected plants were assessed for
receptivity by fixing stigmas of different ages in Carnoy’sfixative, followed by staining in 1% lactophenol analine blue. The stigmas with germinated pollen grains were recorded as receptive and
the number of germinated pollen grains on the stigma was taken as an index of
the degree of receptivity.
Pollination mechanism: To
unravel the pollination mechanism operative in the species, the foraging behaviour of various insects visiting the flowers for
pollen and nectar was monitored. The frequency of insect visitation was observed during different periods
of a day. The insect visiting
efficiency was calculated as:
No. of flowers visited by an
insect in one bout
IVE = -------------------------------------------------------------
Total no. of flowers available
During the foraging bout of
honeybees, three variables were recorded: the number of capitulavisited on the plant, the time spent foraging on each of these inflorescences
and the total time spent per plant and during the whole bout. Wherever possible, the plant at which
the honeybee next foraged was also recorded.
Results
Flowering phenology: The
vegetative phase of the species is followed by a highly asynchronous sexual
phase. Among the different
populations studied, the transplants growing at relatively low altitudes were
the first to enter the sexual phase during the 3rd week of May and
continued the floral bud formation up to 4th week of June. However, the high altitude natural
populations enter the sexual phase late during 3rd or 4thweek of June and the completion of bud initiation is witnessed up to 1stor 2nd week of August.
The species shows a large
asynchrony in anthesis from floret to floret in a capitulum, plant to plant in a population and across
populations growing in varying habitats. The transplants were the first to depict anthesisduring the 2nd week of June which continued
up to 2nd week of July. Among the natural populations, anthesis started in the 2nd or 3rdweek of July and continues till 3rd or 4th week of
August. Each capitulumenters anthesis with maturation of outer single whorl
of pistillate ray florets whichsurround multiple whorls of disc florets. After expansion of the ray florets that coincides with lateral expansion
of the inflorescences, the protandrous disc florets
develop distally and sequentially in whorls. Disc florets mature from the periphery
of the capitulum to the centre.
All the florets of a capitulum open within 5–8
days and all the capitula of a plant open within
18–20 days.
Floral organisation: In I. racemosaeach capitulum possesses a single outer whorl of ray
florets and multiple inner whorls of disc florets. Ray florets are yellow, gynoecious and zygomorphic. Disc florets are actinomorphic; about
2.42±0.54 cm in length, perfect, having both androecium and gynoecium. The corollais in the form five fused petals, the tips of which can be
distinguished at the upper margin of the corolla tube. The androecium is composed of five
stamens. Anthers are sagitate and fused into a tube while the filaments remain
free. The bifid stigma has long
“sweeping” trichomes on the non-receptive surface
that collect and trap pollen grains from the interior of the anther tube and
present them for pollination. Expansion of the stigmatic lobes signifies the beginning of the female
phase. The receptive surface of the
stigma is covered with stigmatic papillae. The stigma lobes were equal in length but shorter than the style.
Pollen emission and stigma receptivity: The disc florets of I. racemosa are hermaphroditic, with five anthers fused
together to form a tube and the stigma and the style grow through the centre of the tube. Before anthesis of florets, the anthers
dehisce inwardly and discharge pollen grains inside the anther tube onto the
sweeping hairs of the style but the stigma is not yet receptive. As the anther dehiscence proceeds, the
carpels develop further and begin to elongate through the tube and the pollen
grains get adhered to the sweeping hairs of the style branches. The style and stigmatic branches thus
present the yellow clumped pollen grains for pollination but the stigma
branches are still joined. From day
3–5 of anthesis, the style grows to its full
length and the two stigmatic branches completely open, while the anthers become
dry and brown and begin to wither. The stigma completely exposes the two layers of receptive papillae cells which marks the entry of a floret into the
functionally female phase. The
stigma remains receptive for three to four days after which stigmatic surface
begins to shrivel that marks the end of receptivity.
Pollination mechanism
During the flowering period,
several insect species were observed visiting the flowering heads of I. racemosa. A
large degree of asynchrony from floret to floret in a capitulumand capitulum to capitulumin a plant keeps the pollen dispersed for a longer duration and thus attracting
the insects again and again to pollinate a number of florets. Two insect families were represented in
the pollinator survey, including Hymenoptera and Diptera. However, honeybees (Apis indica) accounted for the highest number and
duration of visits for all the populations studied and thus was considered a
major pollinator for the species. The bees were observed to become active early in the morning and remain
in the field till late evening. The
mean number of inflorescence visited per plant/bee ranged from 2.53±0.71 at Gulmarg to 4.18±0.43 at Farozpora. However, the terminal capitula were significantly more visited than axillary ones
and the period of greatest insect activity was from 12:00–16:00 hr, coinciding with maximum temperatures. The mean honeybee
foraging time per capitulum was 39±3.0s, 37±7.0s and
48±6.0s and the mean foraging times per plant were 152±7.54s, 128±9.07s and
142±13.69s at Farozpora, Gulmargand KUBG respectively. Also, a
significant correlation was observed between the number of capitulavisited per bout and foraging time.
Other insect visitors that
were also observed foraging on I. racemosathough less frequently included several species of butterflies, Xylocopa spp., flies and wasps. These insects
generally have foraging behaviours similar to
honeybees but were observed to visit the plant capitulaat very low frequency (Fig. 1).
Discussion
Flowering phenology is a
developmental process crucial to determine the plant reproductive success. The
divergence of flowering times among species and populations can have important
evolutionary consequences such as reduction of interspecific pollination (McNeilly & Antonovics 1968; Waser 1978). In
the present study, the different populations of Inula racemosa studied along the altitudinal gradient
differed significantly for dates of onset and commencement of various phenophases. High altitude Gulmarg population (2650m)
emerged and started sprouting 12 days later than comparatively low altitude
populations at Farozpora (2150m) and Farozpora (2150m) populations emerged 28 days later than
low altitude populations at KUBG (1595m). Bud phenology of the plants started in 2nd 3rd or
4th week of June in natural populations and lasted for 39–48
days. In transplanted populations
bud sprouting initiated in 3rd week of May and lasts up to 35
days. The inflorescence (capitulum) enters anthesis with
maturation of the outer, single whorl of ray florets and the protandrous disc florets developed distally and
sequentially in whorls. Disc
florets matured from the periphery of the capitulumto the centre. One to two whorls of mature florets reached anthesisper day. To complete the anthesis within all florets of a capitulum,
it takes 5–8 days and within all the capitullaof a plant about 20–30 days. This asynchronous anthesis ensures the long term availability of pollen to certify effective
pollination as also supported by Wyatt (1982). Variations in degree of flowering
asynchrony within populations and consequent fitness effects have also been
shown in a number of circumstances. The timing of flowering with respect to conspecific neighboring plants
can potentially affect the seed set of individual flowers (Allen 1986), total
plant seed production (Schmitt 1983), seed quality (Marquis 1988), and timing
of seed dispersal (Lacey & Pace 1983). Flowering asynchrony within populations may also reduce intraspecific
competition for pollinators and promote outcrossing (Rogstad1994).
Flower is the structure
directly involved with pollinator attraction and its morphological and
functional traits affect the reproductive success of the plant. Thus, floral
features may have great importance in narrow endemic species (Navarro & Guitian 2002). Our observation has shown that the floral morphology of I. racemosa is especially adapted for insect attraction
and the outer colourful “petal-like” ray florets of
the capitula attract insects usually but typically
lacking an androecium and are the first to reach anthesisfollowed soon by the outermost whorl of disc florets. Pollinators are rewarded by inner disc
florets in the staminate phase (pollen and nectar) preceding the pistillate phase. This developmental sequence is a mechanism employed to prevent autogamy
(Kevan 1997). Receptive surface of the stigma get exposed by the
reflexing of the stigmatic lobes in the receptive pistillatephase. Bilobedstigmas are a typical feature of the asteracean gynoeciums (Mani & Saravanan1999) and gynoecia of I. racemosa do not
differ from this asteracean convention. I. racemosa stigmas serve a dual
purpose; in the staminate phase the stigma is utilized as a secondary pollen
presenter and in the pistillate phase the stigma
serves its more conventional purpose of a receptive surface for pollen. This type of active pollen presentation
is typical of asteracean disc florets (Ladd
1994). The stigma
lobes reflex in the pistillate phase to expose their
receptive papillae.
Secondary pollen presentation
is the result of functional and/or adaptive features of the flower
which enhance the placement of pollen onto the pollination vector
(Inouye et al. 1994; Ladd 1994). Secondary pollen presentation system is widespread in angiosperms
(Howell et al. 1993). It has been described as a strategy to improve accuracy
in pollen removal and deposition, which will result in the enhancement of the
male and female fitness (Ladd 1994) and this mechanism, seems to occur within Campanulaceae and Asteraceae by
means of stigmatic lobes curvation (Cerana 2004). Our observations show that I. racemosais protandrous and the pollen is shed from the
anthers onto the sweeping hairs of the style in the bud stage when the style
branches are joined and stigmatic surfaces are not receptive. This is consistent with the observations
on some other species of Asteraceae (Roitman 1999; Carana 2004). As the style grows out of the anther
tube, the outside of the style branches presents pollen for pollination. The receptive papillatestigmatic surface is hidden between the two appressedstyle branches, preventing self pollination. After the pollen presentation, during
the functionally female phase of the floret, the style branches separate completely
and the two layers of receptive papillae on the adaxialsurfaces become receptive by day 4–6. From day seven onwards flower enters the
senescence stage when the two style branches bend towards the centre of the floret.
In I. racemosathe florets are protandrous and consequently favour vector mediated crossing. The two families of insects documented
as pollinators in the present investigation include Hymenoptera and Diptera with a major pollinator being species of Apis in all the study sites. The Apisand Bombus pollinated members of Asteraceae are well documented by several workers like
Wyatt (1982), Lawrence (1985). Since a number of floral visitors were observed to visit the capitula of the species and this spatial and temporal
heterogeneity found in I. racemosa visitors
may suggest that, in this species, the process of generalization would be favoured instead of specialization. The different studied populations of I.racemosa differ significantly in all total
measures of foraging behaviour for honeybees, the
major pollinator. Plants varied
specifically with respect to the numbers of honeybee visits and honeybee
foraging behaviour (the number of capitulavisited, the foraging time per inflorescence and the foraging time per
plant). Furthermore, a consistent
pattern with respect to plant preferences (the plants visited most often) was
observed, e.g.,
plants inhabiting the shady habitats of Farozporareceived the lowest levels of honeybee foraging visits compared to other
populations. This is inconsistent
with the some previous studies which have reported the variation in the
availability of sunlight influence the pollinator visits and reproductive
success (Herrera 1995). Rathcke & Real (1993) also reported that fruit set of Kalmialatifolia was not limited by inadequate
pollination in sunny field sites, but was pollination limited in the shaded
forest site. This will clearly
affect the production of offspring and thus, the introduction of new individuals
into the populations each year.
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