Pollinator visitation in populations of tristylous Eichhornia paniculata in northeastern Brazil

Summary The frequencies of floral morphs in populations of tristylous Eichhornia paniculata often deviate from the theoretical expectation of equality. This variation is associated with the breakdown of tristyly and the evolution of self-fertilization. Differences in morph frequencies could result from selection pressures due to variable levels of insect visitation to populations and contrasting foraging behavior among the floral morphs. We estimated pollinator densities in 16 populations and quantified visitation sequences to morphs in five populations of E. paniculata in northeastern Brazil. Foraging behavior among floral morphs was measured as the frequency of visits to morphs relative to their frequency in the population (preference) and number of flights between inflorescences of the same versus different morphs (constancy). Pollinator density (number/m²/minute) was not correlated with population size, plant density or morph diversity. Pollinator densities varied most among populations of less than 200 plants. Whether pollinators discriminated among the morphs, depended on whether they primarily collected nectar or pollen. In four populations, nectar-feeding bees (Ancyloscelis and Florilegus spp.) and butterflies showed no consistent preference or constancy among the morphs. In contrast, pollen-collecting bees (Trigona sp.) visited a lower proportion of longstyled inflorescences than expected and tended to visit more mid-and short-styled inflorescences in succession, once they were encountered. Pollinator constancy for morphs did not result from differences in inflorescence production or spatial patchiness among the morphs. Although non-random pollinator visitation to morphs in heterostylous populations could potentially affect mating and hence morph frequencies, the observed visitation patterns in this study do not provide evidence that pollinators play a major role in influencing floral morph frequencies.     

Should food‐deceptive species flower before or after rewarding species? An experimental test of pollinator visitation behaviour under contrasting phenologies

Many plant species reward their pollinators, whereas some species, particularly among orchids, do not. Similarity of floral cues between co‐flowering species influences how rapidly pollinators learn to avoid deceptive plants. This learning process, which affects the reproductive success of deceptive plants, may additionally depend on relative timing of flowering of sympatric rewarding and deceptive species. We tested the combined effects of corolla colour similarity and flowering order of rewarding and deceptive artificial inflorescences on visitation by naïve bumblebees. When deceptive inflorescences were offered after rewarding inflorescences, bumblebees visited them four times more often if both species were similar compared with when they were dissimilar. Pollinator visitation rate to deceptive inflorescences offered before rewarding inflorescences was intermediate and independent of similarity. Thus, early‐flowering deceptive species avoid the costs of dissimilarity with rewarding species. This mechanism may favour adaptive evolution of flowering phenology in deceptive species and explain why temperate deceptive orchids usually flower earlier than rewarding ones.     

Risk‐averse inflorescence departure in hummingbirds and bumble bees: could plants benefit from variable nectar volumes?

Most hermaphroditic, many-flowered plants should suffer reduced fitness from within-plant selfing (geitonogamy). Large inflorescences are most attractive to pollinators, but also promote many flower visits during a single plant visit, which may increase selfing and decrease pollen export. A plant might avoid the negative consequences of attractiveness through modification of the floral display to promote fewer flower visits, while retaining attractiveness. This report shows that increasing only the variance in nectar volume per flower results in fewer flower visits per inflorescence by wild hummingbirds ( Selasphorus rufus ) and captive bumble bees ( Bombus flavifrons ) foraging on artificial inflorescences. Inflorescences were either constant (all flowers contained the same nectar volume) or variable (half the flowers were empty, the other half contained twice as much nectar as in the constant flowers). Both types of inflorescence were simultaneously available to foragers. Risk-averse foraging behaviour was expressed as a patch departure preference: birds and bees visited fewer flowers on variable inflorescences, and this preference was expressed when resource variability could be determined only by concurrent sampling. When variance treatments were clearly labelled with colour and offered to hummingbirds, the departure effect was maintained; however, when preference was measured by inflorescence choice, birds did not consistently prefer to visit constant inflorescences. The reduced visitation lengths on variable inflorescences by both birds and bees documented in this study imply that variance in nectar production rates within inflorescences may represent an adaptive trait to avoid the costs of geitonogamy.     

Stigma�Canther reciprocity, pollinators, and pollen transfer efficiency in populations of heterostylous species of Lithodora and Glandora (Boraginaceae)

According to Darwin, the reciprocal position of sexual whorls in heterostylous plants enhances disassortative pollen transfer between different floral morphs. It is believed that greater reciprocity between morphs will promote more efficient transfer of pollen. Additionally, efficient pollination will act as a selective force in achieving greater reciprocity between floral morphs. In this study we test whether variation in reciprocity of sexual organs between morphs is related to the efficiency of pollinators in transferring pollen between them. To do this, we first describe the pollinator??s array in several populations of species of the genus formerly known as Lithodora, which have different types of stylar polymorphism and degrees of reciprocity, and determine their abundance, plant visitation rate, number of flowers visited per plant and handling time in the population. We estimate the efficiency of the pollinator arrays by use of an approximation based on qualitative (location of pollen loads on different areas of insect bodies) and quantitative (plant visitation rate) measurements. Our results show a correlation between the degree of reciprocity and the efficiency of pollinators associated with the populations. These observations suggest that pollinators are a possible selective force driving the evolution of heterostyly.     

The pollination of the gynomonoecious Bdallophytum oxylepis (Cytinaceae,Malvales)

Bdallophytum oxylepis is a rare and endemic species belonging to the Cytinaceae family, a root holoparasitic plant in which most resources are allocated to attracting pollinators. This species is gynomonoecious with intraindividual variation in flower size and sex. Moreover, the flowers exhibit sapromyophilous traits, as do other species of Bdallophytum. Firstly, this study aimed to determine whether all floral morphs can form seeds and be pollen donors (in the case of bisexual flowers). Secondly, as this species has floral traits hypothesized to adapt to particular types of pollen vectors (carrion flies), we also studied the pollination of B. oxylepis to confirm whether the syndromes correspond to what occurs in nature. Through pollination treatments, we determined that all floral morphs are functional. By monitoring the inflorescences, we found that pollination is specialized in the studied population. Stingless bees performed pollination, as they have a high visitation rate, frequency, and constancy, and they are unique visitors that deposit pollen on the stigmas. Thus, they appear to be effective pollinators rather than carrion flies, as predicted by the syndrome. As shown here, animal–plant interaction studies can help establish a basis for conserving rare species such as holoparasites. Moreover, knowledge about the reproductive aspects of B. oxylepis reveals essential clues about its life cycle and role in maintaining native pollinators with economic and cultural value, such as stingless bees.     

Are there fitness advantages in being a rewardless orchid? Reward supplementation experiments with Barlia robertiana.

The Orchidaceae characteristically contain a very large number of species that attract pollinators but do not offer them any form of reward in return for visitation. Such a strategy is highly unusual in the plant kingdom. We conducted experiments in order to manipulate the reward strategy of the rewardless bumble-bee-pollinated orchid Barlia robertiana by adding sucrose solution to inflorescences. We found that supplementation decreased the probability of a pollinator removing pollinia by approximately ten times. Despite pollinators visiting many more flowers per inflorescence on supplemented plants, eight times fewer pollinia were removed from supplemented inflorescences during each visit. Pollinia deposition patterns were not significantly affected by supplementation and no geitonogamous deposition was recorded. In populations where inflorescences were supplemented for 20 days, pollinia removal was reduced by over half for supplemented inflorescences, whereas fruit set was unmodified by supplementation. We conclude that rewardlessness would increase total seed paternity, but not change either total seed maternity or the probability that offspring were outcrossed in this species. To the authors' knowledge this is the first time that there has been an unequivocal experimental demonstration of an evolutionary advantage for rewardlessness in the Orchidaceae.     

Pollen limitation, seed predation and scape length in Primula farinosa

Floral display and reward production may affect the attractiveness of a plant to a range of interacting animals including pollinators, herbivores, and vectors of pathogenic fungi. The optimal floral phenotype should therefore depend on the relative importance of selection exerted by both mutualistic and antagonistic animals. The perennial, rosette herb Primula farinosa is polymorphic for scape length. Natural populations may include both plants with flowers displayed well above the ground (the long-scaped morph) and those with flowers positioned very close to the ground (the short-scaped morph). In this study, we conducted a field experiment to examine how the relative fitness of the two scape morphs is affected by interactions with pollinators and fruit predators in two different microhabitats (high and low vegetation). As predicted based on the difference in floral display, supplemental hand-pollination showed that fruit initiation was more strongly pollen-limited in the short-scaped than in the long-scaped morph, and that this difference was significantly larger in high than in low vegetation. Moreover, plants with a short scape experienced lower levels of fruit predation than plants with a long scape. Among open-pollinated controls, there was no significant difference in seed output between the two scape morphs. However, among plants receiving supplemental hand-pollination, short-scaped plants produced significantly more seeds than long-scaped plants. The results suggest that the positive and negative effects of a prominent floral display (increased pollination and seed predation, respectively) balance in the study population, but also that the short-scaped morph would have an advantage at higher pollination intensities. Spatial and temporal variation in pollinator activity and seed predation should result in associated variation in the relative fecundity of the two scape morphs.     

Flower-visiting bees and the breakdown of the tristylous breeding system of Eichhornia azurea (Swartz) Kunth (Pontederiaceae)

Populations of Eichhornia azurea (Swartz) Kunth (Pontederiaceae) are often composed of three floral morphs that differ in style length and anther position. Hence, E. azurea is described as tristylous. For sexual reproduction the plant requires specialized pollinators able to transfer pollen from flowers with long-, mid-, or short-level anthers to flowers with long-, mid-, or short-styles, respectively. This is because within a floral morph, self-incompatibility is the rule. Populations of E. azurea were studied from six localities in south-eastern Brazil (São Paulo and Minas Gerais). The frequencies of floral morphs, floral morphology and flower-visiting bees were recorded in each population. Ten different bee species were observed visiting plants. Four of them were considered generalist bees and not appropriate for legitimate pollination of the three floral morphs. Another three species were considered opportunists and only three species were considered to be potential pollinators. At the coastal region (Juréia), the plant population lacked the short-styled morph and the mid-styled morph and exhibited a semi-homostylous condition, in which the low-level anthers were adjacent to the stigma. This morph showed high level of self-compatibility. The disappearance of the short-styled morph from this region is likely correlated with the absence of Ancyloscelis gigas Friese, a specialized long-tongued bee that is oligolectic on this plant in other localities.  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 77 , 499–507.     

Impact of flower-dwelling crab spiders on plant-pollinator mutualisms

Indirect effects in interactions occur when a species influences a third species by modifying the behaviour of a second one. It has been suggested that indirect effects of crab spiders (Thomisidae) on pollinator behaviour can cascade down the food web and negatively affect plant fitness. However, it is poorly understood how different pollinator groups react to crab spiders and, thus, when a reduction in plant fitness is likely to occur. Using continuous video surveillance, we recorded the behaviour of pollinators on two flower species and the pollinators’ responses to three crab spider treatments: inflorescences (1) with a pinned dried spider, (2) with a spider model made of paper, and (3) without spiders (control). We found that pollinators avoided inflorescences with dried spiders only on one plant species (Anthemis tinctoria). Pollinators showed no significant avoidance of paper spiders. Honeybees and bumblebees did not react to dried spiders, but solitary bees and syrphid flies showed a strong avoidance. Finally, we found no evidence that inflorescences with dried spiders suffered from a decrease in fitness in terms of a reduced seed set. We hypothesise that top-down effects of predators on plants via pollinators depend on the degree of specialisation of pollinators and their tendency to avoid spiders.     

Flower constancy in the hoverflies Episyrphus balteatus (Degeer) and Syrphus ribesii (L.) (Syrphidae)

The causes and consequences of flower constancy have been thefocus of many studies, but almost all have examined the foragingbehavior of bumblebees, honeybees, or butterflies. We test whetherconstancy occurs in an overlooked group of pollinators, thesyrphid flies. Foraging sequences of wild flies of two species,Episyrphus balteatus and Syrphus ribesii were examined whenvisiting flowers in seminatural plant communities and in artificialarrays of two color morphs of Lobularia maritima planted ata range of frequencies. Both species exhibited marked floralconstancy when foraging in the mixed-plant community. Becauseall groups of pollinating insect so far examined exhibit constancyat least under some circumstances, we suggest that this is thepredominant strategy used by pollinators and that there is probablya common explanation. Neither syrphid species exhibited constancyto different color morphs within a plant species, in contrastto previously published studies of Hymenoptera foraging amongpolymorphic flowers, which all describe positive frequency-dependentselection. Possible explanations for this discrepancy are discussed.We argue that constancy in these syrphids is unlikely to resultfrom learning constraints on handling ability, currently themost widely accepted explanation for flower constancy, becausethey forage primarily for pollen which is easily located inmostflowers they visit.     

The effect of different chiral morphs on visitation rates and fruit set in the orchid Spiranthes spiralis

Background: The arrangement of flowers on inflorescences is important for determining the movement of pollinators within the inflorescence and, consequently, the overall mating success and fruit set of a plant.

Aims: Spiranthes spiralis is an orchid that has a spiralled inflorescence. The species has two chiral forms that show opposite coiling directions (clockwise and anti-clockwise). We tested if this arrangement of inflorescence influences pollinator attraction and behaviour.

Methods: We surveyed two natural populations, analysed the reproductive compatibility of the two morphs and estimated pollination success in natural and experimental populations.

Results: We found that the two morphs were not isolated by pre- or post-mating barriers, occurred with a similar proportion in natural populations and showed similar levels of pollination success both in natural and artificial populations. However, we found a different pattern of pollination success along the inflorescences. In the two morphs, lower flowers experienced a higher pollination rate and this rate decreased along the inflorescence faster in anti-clockwise than in clockwise individuals.

Conclusions: This finding suggests that pollinators visit the flowers sequentially from the lower part of the inflorescences and leave the anti-clockwise individuals more rapidly than the clockwise ones. However, this pollinator behaviour is not detrimental for the pollination success of either of the two morphs.     

Variable responses of hawkmoths to nectar-depleted plants in two native Petunia axillaris (Solanaceae) populations

Pollination success of deceptive orchids is affected by the density and distribution of nectar providing plant species and overall plant density. Here we extended the framework of how plant density can affect pollination to examine how it may promote the success of plant intraspecific cheaters. We compared hawkmoth behaviour in two native populations of Petunia axillaris, where we simultaneously offered rewarding and manually depleted P. axillaris. We asked whether pollinator foraging strategies change as a function of plant density and whether such changes may differentially affect nectarless plants. We observed the first choice and number of flowers visited by pollinators and found that in the dense population, pollinators visited more flowers on rewarding plants than on nectar-depleted plants. In the sparse population, such discrimination was absent. As we found no differences in nectar volume between plants of the two populations, the observed differences in plant density may be temporal. We reason that if differences were more permanent, an equivalent of the remote habitat hypothesis prevails: in a sparse population, cheating plants benefit from the absence of inter- and intraspecific competitors because pollinators tend to visit all potential resources. In a denser population, a pollinator’s optimal foraging strategy involves more selectivity. This would cause between-plant competition for pollinators in a pollinator-limited context, which applies to most hawkmoth-pollinated systems. We propose that nectar-provisioning of plants can be density-dependant, with cheaters able to persist in low density areas.     

The consequences of rewardlessness in orchids: reward-supplementation experiments with Anacamptis morio (Orchidaceae)

Pollinators are expected to respond to low reward availability in an inflorescence by visiting fewer flowers before departure, thus potentially causing reduced visitation, but also reduced geitonogamous selfing. I tested this hypothesis using Anacamptis morio, an orchid that does not reward its pollinators. Supplementation of inflorescences with artificial nectar did not result in an increase in fruit set on supplemented inflorescences compared to control inflorescences and tended to reduce pollinia removal. Supplementation resulted in reduced fruit quality, but there was no evidence that this was as a result of inbreeding depression. Behavioral experiments showed that pollinating bumble bees, as predicted, visited more flowers on supplemented inflorescences. Bumble bees also deposited more self-pollen on supplemented inflorescences, but this was marginally significant. Bumble bee queens removed significantly more pollinia from control inflorescences, while Bombus terrestris and B. lucorum workers did not. I conclude that while pollinators behaved as predicted, there was weak evidence that pollinia removal, pollen deposition, and fruit set followed the predictions of the hypothesis. I argue that this was probably because some pollinators were more efficient at removing and depositing pollen on control inflorescences, while others were not.     

The effect of nectar guides on pollinator preference: experimental studies with a montane herb

Summary Rare albino morphs of the montane larkspur Delphinium nelsonii differ from common blue-flowered morphs in overall flower color, and in the strength of a contrasting color pattern at the center of the flower that presumably guides pollinators to concealed nectar. Previous studies showed that bumblebees and hummingbirds discriminate against albinos when presented with mixtures of the 2 morphs, and that it takes these pollinators longer to fly between successive flowers on albino than on blue-flowered inflorescences. To explore the link between these observations, we measured pollinator preferences and flower-to-flower flight times (handling times) before and after painting flowers in 2 alternative ways that enhanced albino nectar guides. In all of 16 experimental replicates discrimination against albinos was reduced or eliminated after painting, and albino handling times declined toward values for blue-flowered inflorescences. This consistent result indicates that an inferior nectar guide increases the energetic cost of foraging at albinos. Increased cost in turn explains discrimination, under the reasonable assumption that hummingbirds and bumblebees are sensitive to foraging economics.     

Reduction of geitonogamy: Flower abscission for departure of pollinators

Overproduction of flowers increases the attractiveness of a plant to pollinators, but results in increased geitonogamy. In general, flower abscission has been considered to be an event subsequent to the overproduction of flowers. We observed pollinator behavior in Tilia, a self-incompatible, insect-pollinated tree. We found that pollinators sequentially visited inflorescences within a tree; however, when they met with flower abortion by abscission, they were apt to move long distances and leave the tree. We propose that plants may utilize flower abscission as a method for regulating the movements of pollinators by disturbing them in order to prevent geitonogamy.     

NEGATIVE FREQUENCY-DEPENDENT SELECTION BY POLLINATORS ON ARTIFICIAL FLOWERS WITHOUT REWARDS

Many species of nonmodel deceptively pollinated orchids are polymorphic for corolla color. These species are pollinated by naive insects searching for nectar, and are not mimics. It has been suggested that the foraging behavior of insect pollinators during the avoidance learning process results in these stable corolla color polymorphisms; for this to occur pollinators must induce negative frequency-dependent selection on corolla color. Therefore the hypothesis that pollinator behavior results in a preference for rare color morphs of deceptive species was tested experimentally. Bumblebees (Bombus terrestris) foraged in the laboratory on arrays of artificial flowers with different corolla color morphs. Morphs were varied in frequency, and bumblebee preferences were recorded on arrays where morphs did and did not contain sucrose solution rewards. Bumblebees preferred the most common color morph when flowers contained sucrose solution rewards, but overvisited rare morphs when sampling flowers that contained no rewards. Bumblebees also tended to move between unlike color morphs when these were unrewarding, suggesting that a probabilistic sampling strategy was adopted. Thus experiments demonstrated that pollinator behavior could result in a selective advantage for rare color morphs of plant species that are pollinated by deception without mimicry, which would induce negative frequency-dependent selection on corolla color. The observed pollinator behavior could allow stable corolla color polymorphisms to be maintained by selection in nonmodel deceptively pollinated species.     

Geitonogamy in rewarding and unrewarding inflorescences: modelling pollen transfer on actual foraging sequences

Many orchid species are unusual in that they provide no nectar or pollen rewards for their pollinators. Absence of reward is expected to have a fundamental effect on pollinator visitation patterns. In particular the number of flowers visited per inflorescence is expected to be affected in both unrewarding and co-flowering rewarding species. We used arrays of artificial inflorescences, which could be either rewarding or unrewarding and were differentiated by their colour, to test how many flowers bumblebees visit in each type of inflorescence. The frequency of the two colours was varied, thus modelling the case where different frequencies of both an unrewarding and rewarding species were present in a patch. We found that bumblebees visited more flowers per rewarding inflorescence after they have experienced unrewarding or partially emptied rewarding inflorescences. We used these results to simulate pollen transfer and thus predict selfing rates on rewarding inflorescences. We found these increased when nectar depleted or when there was a greater proportion of unrewarding inflorescences in the patch. Conversely, we found that the number of flowers bumblebees visited on each unrewarding inflorescence did not significantly change through experiments. Selfing rates for unrewarding inflorescences were predicted to depend principally on the number of these inflorescences bumblebees visited rather than on the number of flowers they visit per inflorescence. This was because most visitors to orchids are supposed to be naive, and pollinators that commence foraging carrying no pollen will necessarily self any flower they pollinate on the first inflorescence they visit. Thus the average selfing rate is expected to increase as the sequence of inflorescences visited decreases in length.     

Pollinator community responses to the spatial population structure of wild plants: A pan-European approach

Land-use changes can alter the spatial population structure of plant species, which may in turn affect the attractiveness of flower aggregations to different groups of pollinators at different spatial scales. To assess how pollinators respond to spatial heterogeneity of plant distributions and whether honeybees affect visitation by other pollinators we used an extensive data set comprising ten plant species and their flower visitors from five European countries. In particular we tested the hypothesis that the composition of the flower visitor community in terms of visitation frequencies by different pollinator groups were affected by the spatial plant population structure, viz. area and density measures, at a within-population (‘patch’) and among-population (‘population’) scale. We found that patch area and population density were the spatial variables that best explained the variation in visitation frequencies within the pollinator community. Honeybees had higher visitation frequencies in larger patches, while bumblebees and hoverflies had higher visitation frequencies in sparser populations. Solitary bees had higher visitation frequencies in sparser populations and smaller patches. We also tested the hypothesis that honeybees affect the composition of the pollinator community by altering the visitation frequencies of other groups of pollinators. There was a positive relationship between visitation frequencies of honeybees and bumblebees, while the relationship with hoverflies and solitary bees varied (positive, negative and no relationship) depending on the plant species under study. The overall conclusion is that the spatial structure of plant populations affects different groups of pollinators in contrasting ways at both the local (‘patch’) and the larger (‘population’) scales and, that honeybees affect the flower visitation by other pollinator groups in various ways, depending on the plant species under study. These contrasting responses emphasize the need to investigate the entire pollinator community when the effects of landscape change on plant–pollinator interactions are studied.     

The function and adaptive significance of tristyly in Pontederia cordata L. (Pontederiaceae)

Darwin proposed that the adaptive significance of tristyly is to promote insect-mediated pollination among floral morphs with anthers and stigmas at equivalent levels (legitimate pollination). Strong pollen trimorphism in Pontederia cordata enables an evaluation of this hypothesis. In an investigation of pollen flow patterns in different parts of the North American range of the species, legitimate pollination of some morphs was observed in all but one population investigated. However at only one site (Ft. McCoy, Florida) was Darwin's hypothesis confirmed for the three floral morphs. The long-styled morph most frequently exhibited legitimate pollination, the mid-styled morph was intermediate, and the short-styled morph rarely experienced legitimate pollination.
Temporal variation in pollen flow was observed during the flowering season of P. cordata . Total stigmatic pollen loads decreased as the density of inflorescences and activity of pollinators declined. In spite of variable total loads the legitimate component was relatively constant in the long- and mid-styled forms, while in the short-styled form this measure increased as the season progressed. Little variation was detected between stigmatic samples collected at different times during a single day.
Legitimate pollination may be inhibited by local foraging of pollinators and the spatial segregation of floral morphs. At Paugh Lake (Ontario) over 75% of all bumble bee flights are among the five nearest neighbours of a particular inflorescence. In the same population, there is a probability of over 70% that the nearest three neighbours of an inflorescence are of the same floral morph. Despite these influences, legitimate pollination occurs in P. cordata , suggesting substantial pollen carryover.     

Combined effects of inflorescence architecture, display size, plant density and empty flowers on bumble bee behaviour: experimental study with artificial inflorescences

Pollen dispersal by pollinators is governed by the extent to which diverse effects on pollinator behaviour act independently or augment or moderate each other. Using artificial inflorescences, we assessed the behavioural responses of bumble bees to inflorescence architecture (raceme, panicle, and umbel), inflorescence size (7 or 13 flowers), inter-inflorescence distance and the proportion of empty flowers per inflorescence. The advantage of large inflorescences in terms of attractiveness was larger for racemes and umbels than for panicles, whereas the effect of inter-inflorescence distance on the number of successive probes was smaller for racemes than for panicles and umbels. The number of flowers probed per visit increased almost proportionally with display size when fewer flowers were empty, whereas the number increased less when many flowers were empty. Our results suggest that display size and the spatial arrangement of flowers and nectar within inflorescences can contribute to efficient pollination by affecting pollinator behaviour interactively.