The effects of nectar addition on pollen removal and geitonogamy in the non-rewarding orchid Anacamptis morio

It has been suggested that the absence of floral rewards in many orchid species causes pollinators to probe fewer flowers on a plant, and thus reduces geitonogamy, i.e. self-pollination between flowers, which may result in inbreeding depression and reduced pollen export. We examined the effects of nectar addition on pollinator visitation and pollen transfer by tracking the fate of colour-labelled pollen in Anacamptis morio, a non-rewarding orchid species pollinated primarily by queen bumble-bees. Addition of nectar to spurs of A. morio significantly increased the number of flowers probed by bumble-bees, the time spent on an inflorescence, pollinarium removal and the proportion of removed pollen involved in self-pollination through geitonogamy, but did not affect pollen carryover (the fraction of a pollinarium carried over from one flower to the next). Only visits that exceeded 18 s resulted in geitonogamy, as this is the time taken for removed pollinaria to bend into a position to strike the stigma. A mutation for nectar production in A. morio would result in an initial 3.8-fold increase in pollinarium removal per visit, but also increase geitonogamous self-pollination from less than 10% of pollen depositions to ca. 40%. Greater efficiency of pollen export will favour deceptive plants when pollinators are relatively common and most pollinaria are removed from flowers or when inbreeding depression is severe. These findings provide empirical support both for Darwin's contention that pollinarium bending is an anti-selfing mechanism in orchids and for the idea that floral deception serves to maximize the efficiency of pollen export.     

Doing the twist: A test of Darwin's cross-pollination hypothesis for pollinarium reconfiguration

Mating success in plants depends largely on the efficiency of pollen dispersal. For hermaphrodite plants, self-pollination, either within or among flowers, can reduce mating opportunities because of pollen and ovule discounting and inbreeding depression. Self-pollination may be particularly detrimental in plants such as orchids and asclepiads that package each flower's pollen into one or more pollinia which, together with accessory structures, comprise a pollinarium. Darwin proposed that physical reconfiguration of pollinaria serves as a mechanism for reducing the likelihood of self-pollination. To be effective, the time taken for pollinarium reconfiguration would need to exceed that spent by a pollinator on a plant. We investigated pollinarium reconfiguration (including pollinarium bending, pollinium shrinking and anther cap retention) in 19 species and found a strong positive relationship between reconfiguration time and the duration of pollinator visits. Reconfiguration times were also consistently longer than pollinator visit times. These results provide strong support for Darwin's idea that this mechanism promotes cross-pollination.     

Mechanisms and evolution of deceptive pollination in orchids

The orchid family is renowned for its enormous diversity of pollination mechanisms and unusually high occurrence of non-rewarding flowers compared to other plant families. The mechanisms of deception in orchids include generalized food deception, food-deceptive floral mimicry, brood-site imitation, shelter imitation, pseudoantagonism, rendezvous attraction and sexual deception. Generalized food deception is the most common mechanism (reported in 38 genera) followed by sexual deception (18 genera). Floral deception in orchids has been intensively studied since Darwin, but the evolution of non-rewarding flowers still presents a major puzzle for evolutionary biology. The two principal hypotheses as to how deception could increase fitness in plants are (i) reallocation of resources associated with reward production to flowering and seed production, and (ii) higher levels of cross-pollination due to pollinators visiting fewer flowers on non-rewarding plants, resulting in more outcrossed progeny and more efficient pollen export. Biologists have also tried to explain why deception is overrepresented in the orchid family. These explanations include: (i) efficient removal and deposition of pollinaria from orchid flowers in a single pollinator visit, thus obviating the need for rewards to entice multiple visits from pollinators; (ii) efficient transport of orchid pollen, thus requiring less reward-induced pollinator constancy; (iii) low-density populations in many orchids, thus limiting the learning of associations of floral phenotypes and rewards by pollinators; (iv) packaging of pollen in pollinaria with limited carry-over from flower to flower, thus increasing the risks of geitonogamous self-pollination when pollinators visit many flowers on rewarding plants. All of these general and orchid-specific hypotheses are difficult to reconcile with the well-established pattern for rewardlessness to result in low pollinator visitation rates and consequently low levels of fruit production. Arguments that deception evolves because rewards are costly are particularly problematic in that small amounts of nectar are unlikely to have a significant effect on the energy budget of orchids, and because reproduction in orchids is often severely pollen-, rather than resource-limited. Several recent experimental studies have shown that deception promotes cross-pollination, but it remains unknown whether actual outcrossing rates are generally higher in deceptive orchids. Our review of the literature shows that there is currently no evidence that deceptive orchids carry higher levels of genetic load (an indirect measure of outcrossing rate) than their rewarding counterparts. Cross-pollination does, however, result in dramatic increases in seed quality in almost all orchids and has the potential to increase pollen export (by reducing pollen discounting). We suggest that floral deception is particularly beneficial, because of its promotion of outcrossing, when pollinators are abundant, but that when pollinators are consistently rare, selection may favour a nectar reward or a shift to autopollination. Given that nectar-rewardlessness is likely to have been the ancestral condition in orchids and yet is evolutionarily labile, more attention will need to be given to explanations as to why deception constitutes an 'evolutionarily stable strategy'.     

Lack of floral nectar reduces self-pollination in a fly-pollinated orchid

One explanation for the widespread absence of floral nectar in many orchids is that it causes pollinators to visit fewer flowers on a plant, and thus reduces self-pollination. This, in turn, could increase fitness by reducing inbreeding depression in progeny and promoting pollen export. The few previous investigations of this hypothesis have all involved bee-pollinated orchids and some have given contradictory results. We studied the effects of adding artificial nectar (sucrose solution) to the spurs of a non-rewarding long-proboscid fly-pollinated orchid, Disa pulchra. Addition of nectar significantly increased the number of flowers probed by flies (2.6-fold), the time spent on a flower (5.4-fold), the number of pollinia removed per inflorescence (4.8-fold) and the proportion of removed pollen involved in self-pollination (3.5-fold). The level of self-pollination increased dramatically with the number of flowers probed by flies. Experimental self-pollination resulted in fruits with only half as many viable seeds as those arising from cross-pollination. Pollinators were more likely to fly long distances (>40 cm) when departing from non-rewarding inflorescences than when departing from rewarding ones. These findings provide support for the idea that floral deception serves to reduce pollinator-mediated self-pollination.     

Reproductive strategies of Strobilanthes kunthianus, an endemic, semelparous species in southern Western Ghats, India

Strobilanthes kunthianus (Acanthaceae) is a semelparous species with synchronized flowering and mast seeding once in 12 years. As semelparous plants have only one chance to reproduce, they are expected to develop effective strategies to prevent reproductive failure. The reproductive strategies of S. kunthianus were investigated by studying the floral traits, pollination biology, and breeding system that are critical for reproductive success. The species exhibits a series of floral traits: (1) gregarious flowering attracts a large number of Apis cerana indica , the major pollinator; (2) the stigma is sensitive to touch by the pollinator; in fresh flowers, the receptive surface faces the entry path of the incoming bee, facilitating pollen deposition; as an immediate response, the stigma curves backwards moving the receptive surface away from the path of the exiting bee, thus preventing autogamy and interference in pollen transfer; (3) flowers remain fresh for 2 days with receptive stigma and nectar and pollen reward. These traits render the species 100% pollination efficient to ensure a high seed set. As the species is self-compatible, the prevailing high degree of geitonogamous pollinations does not interfere with fruit set. The evolution of the adaptive floral traits has facilitated mast seeding in the species.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 155–163.     

Diversity, flower visitation frequency and generalism of pollinators in temperate rain forests of Chiloé Island, Chile

Species richness and taxonomic composition of pollinator assemblages are documented for 26 plant species from temperate rain forests of northern Chiloé Island, southern Chile (42°30'S). We investigated the patterns of generalism and specialization among plants and animal pollinators by comparing the flower visit frequency by different pollen vectors during the spring and summer months of three consecutive years (2000–2002). Species studied exhibited a range of floral morphologies (radial vs. zygomorphic, open vs. tubular) and rewards (nectar and/or pollen). Overall, we recorded 172 pollinator species, with an average of 6.6 species of pollen vectors/plant species. Pollinators visited an average of 15.2 plant species/pollen vector. Pollinator assemblages were dominated by Coleoptera (75 species), Diptera (56 species) and Hymenoptera (30 species), but passerine birds and hummingbirds were also important. The most specialized plants were vines, including the bee-pollinated genus Luzuriaga (Philesiaceae) and two endemic species of hummingbird-pollinated Gesneriaceae. Hymenoptera contributed 41.2% of all visits, with the bumblebee Bombus dalhbomii accounting for 22.5% of these. Plants with unspecialized flower morphology supported a higher species richness of pollinators, but visiting rates did not differ from specialized flowers.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 399–416.     

Resource limitation and pollen source (self and outcross) affecting seed production in two louseworts, Pedicularis siphonantha and P. longiflora (Orobanchaceae)

Two animal-pollinated hermaphrodite plants, Pedicularis siphonantha and P. longiflora , have been used to investigate factors limiting seed production in natural populations. To evaluate the potential seed abortion due to resources limitation, seed development has been observed and seed count conducted twice. Seed production per capsule has been compared when flowers have been removed and in a control group. Open pollination has been investigated and pollen supplementation undertaken to estimate the possibility of pollen limitation. Results show that seed abortion is frequent. Stigmatic pollen load is significantly higher than ovule number per ovary under open pollination for both species. Additional self and outcross pollen did not affect seed production. Flower removal significantly increases seed production per capsule, which indicates that seed production of the studied species is limited by available resources. To detect differences in seed production between flowers pollinated by self and outcross pollen, hand pollination of bagged flowers has also been conducted in natural populations of the two Pedicularis species. Compared with open pollination, hand-pollinating self-pollen decreases, while outcross pollen increases seed production per capsule. Such results suggest that inbreeding depression in the two self-compatible species may also result in partial seed abortion under open pollination if mixed pollen is deposited on the stigma. Our results also suggest that pollen interference plays an important role in low female fertility in the two Pedicularis species.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 83–89.     

Reproductive characters in a gynodioecious species, Silene italica (Caryophyllaceae), with attention to the gynomonoecious phenotype

Gynodioecious populations (i.e. populations with female and hermaphrodite individuals) often contain a third phenotype with an intermediate sex expression. In Silene italica , this phenotype is characterized by a mixture of pistillate and perfect flowers and is thus gynomonoecious. To characterize sexual functions of these gynomonoecious individuals and their potential influence in the maintenance of gynodioecy, reproductive characters of the three sexual phenotypes were compared over 2 years in several families of S. italica produced by crossing. We found that gynomonoecious individuals were intermediate for flower production and female fertility characters, although they did not always significantly differ from female individuals. Perfect flowers of gynomonoecious and hermaphrodite plants were similar in size and pollen production. Gynomonoecious individuals were thus intermediate in female and in male functions. Family effects were found for most of the characters. The female advantage (i.e. the fertility of females compared to the female fertility of pollen producing plants) was not dramatically different when gynomonoecious plants were taken into account.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 87 , 583–591.     

Lying to Pinocchio: floral deception in an orchid pollinated by long-proboscid flies

Plants that lack floral rewards may nevertheless attract pollinators if their flowers sufficiently resemble those of rewarding plants. Flowers of the South African terrestrial orchid Disa nervosa are similar in floral dimensions and spectral reflectance to those of a sympatric nectar-producing irid ( Watsonia densiflora s.l. ). Observations showed that the orchid and Watsonia share the same pollinator, a long-proboscid tabanid fly Philoliche aethiopica . These flies visited inflorescences of both species during their foraging bouts and most (64%) observed or captured on Watsonia inflorescences carried pollinaria of the orchid on their proboscides. They probe an average of 6.3 flowers on Watsonia inflorescences, but just 1.9 flowers on the Disa inflorescences, a behaviour which would strongly promote cross-pollination in the self-compatible orchid. The orchid generally achieves high levels of pollination success, with approximately 50% of flowers receiving or exporting pollen at some sites. Pollination success was also high at one site that lacked Watsonia plants, suggesting that the orchid does not have an obligate dependence on Watsonia . Its pollination system may therefore be characterized as intermediate between generalized food deception and specific floral mimicry.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 152 , 271–278.     

How flower biology and breeding system affect the reproductive success of the narrow endemic Polygala vayredae Costa (Polygalaceae)

Polygala vayredae is a narrow endemic species from the oriental pre-Pyrenees. Despite its conservation status and rarity, no information is available on its reproductive biology. As the flower is the structure directly involved in pollinator attraction, its morphological and functional traits have major effects on the reproductive success of the plant. In this work, the flower biology and breeding system of P. vayredae were studied to evaluate how they affect the reproductive outcome in natural populations. Flower morphology, flower rewards, and male and female functioning throughout the lifespan of the flower were assessed. Pollination experiments, involving pollinator exclusion and pollen from different sources, were conducted, and the pollen ovule index was determined. Female fitness and the occurrence of pollen limitation were assessed in three natural populations over 2 years by observing the presence of pollen on the stigma, pollen tube development, and fruit production. Polygala vayredae flowers are elaborate and long-lived with nectar rewards. The floral traits are well adapted to xenogamy and entomophily, which are in accordance with the observed breeding system and auto-incompatibility system. No mechanism of reproductive assurance was observed and P. vayredae strictly depends on pollinators to set fruit. Low fruit production was observed in the studied populations, which was largely the result of scarce, unreliable, and/or inefficient pollinators and poor pollen quality. In addition, available resources may be a limiting factor. The reproductive strategy of P. vayredae prevents inbreeding depression by a self-incompatibility system, which in years of scarce pollinators is overcome by the plant habit.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 67–81.     

Reproductive biology of the invasive exotic shrub, Rhododendron ponticum L. (Ericaceae)

The reproductive biology of an exotic species will affect its ability to become naturalized and invasive in non-native habitats. Rhododendron ponticum is an ecologically damaging exotic weed in the British Isles, which spreads predominantly by seed. I investigated how inbreeding and outcrossing affect seed production and germination in a wild population of this species in Ireland. Experimental manipulations revealed low fruit and seed set when insects were excluded from flowers, suggesting that this species has limited capability for spontaneous autogamy. Hand-pollination treatments showed that, although flowers are self-compatible (with self and same plant pollen), higher levels of seed set occur following outcrossing (xenogamy). There was no significant difference in rate of germination of seeds from inbred or outcrossed treatments. The addition of xenogamous pollen to open flowers did not increase fruit or seed set, suggesting that flowers in this population are not pollen limited: native generalist pollinators, mainly bumblebees ( Bombus spp.), are providing an adequate pollinator service. This work demonstrates that outcrossing increases seed set and pollinators are required to facilitate this. Hence, generalist native pollinators can promote invasion by exotic plants. Native pollinators can clearly play an important part in alien species invasion.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 373–381.     

Pollination of a slippery lady slipper orchid in south-west China: Cypripedium guttatum (Orchidaceae)

Cypripedium guttatum was studied in north-west Yunnan at 3490 m a.s.l. The flowers are rewardless 'kettle traps'. The structure of the lip, where pollinators are temporarily kept prisoner, and the method of their capture, are unusual in being Paphiopedilum - rather than Cypripedium -like. The deceptive orchid does not mimic any of the diverse flowers concurrently blooming in the habitat, all being visited by the polylectic pollinators of C. guttatum , viz . Lasioglossum virideglaucum, L. clypeinitens and L. sauterum , besides two additional probable pollinators and four non-pollinating visitors (all Halictidae; three new species). The bees got caught when they tried to climb onto the staminode and their forelegs slid down its slippery downward ridges, causing them to tumble to the pouch bottom. To leave, they had to climb a tunnel leading past the stigma to the anthers where a pollen smear was acquired while extruding themselves from the narrow exit. The similarities with myiophylous Paphiopedilum are discussed in view of the possibility that they may foreshadow evolutionary transitions between melittophily and myiophily found in slipper orchids.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 251–264.     

Pollination biology of the Galápagos endemic, Tournefortia rufo-sericea (Boraginaceae)

Tournefortia rufo-sericea is an endemic member of the Galápagos angiosperm flora. Although not uncommon within the archipelago, its status is presently listed as vulnerable and, as such, a complete knowledge of its pollination biology may prove useful in preventing its decline. Pollination experiments, flower-visitor observations, nectar and fluorescence studies, as well as pollen : ovule ratio and pollen size studies were included in this investigation. The small, white flowers of this species set fruit via open pollination (81%), autonomous self-pollination (80%), diurnal pollination (80%), and nocturnal pollination (85%). Ants are the most common visitor to the flowers of this species, primarily during the day, whereas beetles and moths make visits at night. A small amount of nectar is presumably produced, as moths are often seen probing the corollas. Experiments with fluorescent dust did not support interflower pollen movement. The pollen : ovule ratio was 4972 : 1, which suggests xenogamy, and the mean pollen size was 21 µm. Despite the pollen : ovule ratio, it appears that this species exhibits a breeding strategy of facultative autogamy, in which the majority of flowers set fruit via autonomous self-pollination, but limited outcrossing may occur when pollinators are available. Conservation efforts for T. rufo-sericea should focus on the control or eradication of exotic species.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 21–31.     

Reproductive effort and sex allocation strategy in Commelina benghalensis L., a common monsoon weed

Commelina benghalensis L. exhibits variability in both foliar and floral features; every plant bears three types of branches and four types of flowers. The branches are negatively geotropic, positively geotropic and diageotropic. The flowers are uni- or bisexual, chasmogamous and cleistogamous. This variability influences the breeding system as well as resource allocation to male and female functions. The plants allocate c. 15% of their total resources to reproduction, the major part of which (68.9%) is devoted to production of aerial branches. The proportion of reproductive effort (RE) allocated to various branch systems is correlated with the availability of resources at the time of their differentiation. The pollen/ovule (P/O) ratio, female : male biomass ratio and reproductive output vary between different flower and branch types; variation is more pronounced in the latter. These variations notwithstanding, the results are in line with Charnov's sex allocation theory. The cleistogamous flowers of aerial branches are, however, an exception, being male- rather than female-biased. The reason behind the deviation is, in all probability, their recent evolution from chasmogamous flowers.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 140 , 403−413.     

Invasive Africanized honey bee impact on native solitary bees: a pollen resource and trap nest analysis

Little is known of the potential coevolution of flowers and bees in changing, biodiverse environments. Female solitary bees, megachilids and Centris , and their nest pollen provisions were monitored with trap nests over a 17-year period in a tropical Mexican biosphere reserve. Invasion by feral Apis (i.e. Africanized honey bees) occurred after the study began, and major droughts and hurricanes occurred throughout. Honey bee competition, and ostensibly pollination of native plants, caused changes in local pollination ecology. Shifts in floral hosts by native bees were common and driven by plant phylogenetics, whereby plants of the same families or higher taxa were substituted for those dominated by honey bees or lost as a result of natural processes. Two important plant families, Anacardiaceae and Euphorbiaceae, were lost to competing honey bees, but compensated for by greater use of Fabaceae, Rubiaceae, and Sapotaceae among native bees. Natural disasters made a large negative impact on native bee populations, but the sustained presence of Africanized honey bees did not. Over 171 plant species comprised the pollen diets of the honey bees, including those most important to Centris and megachilids (72 and 28 species, respectively). Honey bee pollination of Pouteria (Sapotaceae) plausibly augmented the native bees' primary pollen resource and prevented their decline. Invasive generalist pollinators may, however, cause specialized competitors to fail, especially in less biodiverse environments.  No claim to original US government works. Journal compilation © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 152–160.     

Pollination ecology of Arum italicum (Araceae)

The pollination ecology of Arum italicum was studied in south-western France. This plant attracts olfactory dung-breeding flies through deceit. These insects are principally represented by Diptera, all belonging to saprophyte families. The volatilization of the odouriferous compounds, responsible for their attraction, is achieved through the production of heat by the appendix. The insects are trapped for 24 h in order to participate in both sexual phases of the protogynous inflorescence. The male flowers produce three heat events during flowering. These peaks of heat seem to be involved in the spathe movements, since they occur during the opening of the inflorescence and the liberation of the insects. The last male heat event may be linked with the liberation of pollen and its dispersion by stimulating trapped flies. According to their frequency and pollen-load, two Psychoda species appear to be the most efficient pollinators ( P. crassipenis and P. pusilla ). Nevertheless, each of the other attracted species could play a significant role under different spatio-temporal conditions. Experiments on self-pollination have shown that obligate cross-pollination is necessary for A. italicum to set seeds. Moreover, hand- and natural-pollinated plants showed similarly high abortion frequencies suggesting that seed set may be more constrained by resources rather than by pollination limitation.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 141 , 205–214.     

Transfer of pollinaria on birds’ feet: a new pollination system in orchids

We report the discovery of a new mechanism of pollination in orchids: transfer of pollinaria on the feet of birds. Observations carried out in South Africa and Malawi showed that the orchids Disa chrysostachya Sw. and Disa satyriopsis Kraenzl. are pollinated by sunbirds. Pollinaria of these orchids become attached firmly to the birds toes when they perch on the tall narrow inflorescences which are packed tightly with numerous small orange flowers. Birds typically perch on the lower part of an inflorescence while reaching up to feed on nectar in flowers on the upper part, but occasionally reverse this position to probe the lower flowers. The nectar is contained within a short bulbous spur with a narrow entrance that permits entrance of a sunbirds slender tongue. Contrary to expectation, the pollination mechanism in D. chrysostachya is remarkably efficient with about 6.1% of pollen reaching stigmas on other plants and fruit set occurring in 95% of flowers at one site. Birds seldom move their feet once perched, thus minimizing the incidence of self-pollination, either within or between flowers on an inflorescence.     

Effect of nectar supplementation on male and female components of pollination success in the deceptive orchid Dactylorhiza sambucina

Many orchids lack floral nectar rewards and therefore rely on deception to attract pollinators. To determine the effect that a mutation for nectar production would have on overall pollination success of the deceptive orchid Dactylorhiza sambucina, we recorded pollen deposition and removal in flowers of plants that had either been supplemented with an artificial nectar solution or left unmanipulated as controls. Nectar supplementation resulted in significant increases in the proportion of flowers pollinated, regardless of morph colour and the density of plants supplemented in the population. However, nectar supplementation had a significant positive effect on pollinaria removal only for the yellow morph in one experiment in which a low proportion of plants were supplemented. Thus a mutation for nectar production would have a positive effect on overall pollination success in D. sambucina, particularly the female component. The observed patterns are discussed in relation to other factors, such as cross-pollination and the reallocation of nectar resources for other plant functions, which are traditionally considered to shape the rewardless strategies of orchids.     

Reproductive biology in Anagyris foetida L. (Leguminosae), an autumn–winter flowering and ornithophilous Mediterranean shrub

As most plants of the Mediterranean region bloom in spring, there have been few studies of the reproductive biology of species with autumn–winter flowering. In this study, we investigate the breeding system of Anagyris foetida , one of the few shrubs that blooms at this time. The floral, phenological, and reproductive aspects of two populations of this Mediterranean legume from south-west Spain were studied via field and laboratory experiments. The variability of fruit and seeds was studied in another 12 Iberian populations with respect to certain meteorological parameters (temperature and rainfall). Anagyris foetida shows cauliflory, marked floral longevity, and adichogamy. The peak of flowering is in January–February. It is self-compatible, with no clear advantage of cross- over self-pollination, and with virtually no autonomous self-pollination. This is because the stigma, like some other legumes, prevents the germination of pollen if its surface is not ruptured by pollinators. The number of seeds per fruit under natural pollination was positively correlated with the total rainfall during the fruiting period (from January to May), and significantly influenced the percentage of fruit weight represented by the pericarp, in the sense that the smaller the number of viable seeds in the fruit, the greater the percentage of pericarp weight.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 519–532.