Pollinator-mediated plant coexistence requires high levels of pollinator specialization.

In pollen-limited plant communities, the foraging behavior of pollinators might mediate coexistence and competitive exclusion of plant species by determining which plants receive conspecific pollen. A key question is whether realistic pollinator foraging behavior promotes coexistence or exclusion of plant species. We use a simulation model to understand how pollinator foraging behavior impacts the coexistence dynamics of pollen-limited plants. To determine whether pollinators are likely to provide a biologically important coexistence mechanism, we compare our results to bee foraging data from the literature and from a novel experimental analysis. Model results indicate that strong specialization at the level of individual foraging paths is required to promote coexistence. However, few empirical studies have robustly quantified within-bout specialization. Species-level data suggest that foraging behavior is sufficient to permit pollinator-mediated coexistence in species-poor plant communities and possibly in diverse communities where congeneric plants co-occur. Our experiments using bumblebees show that individual-level specialization does exist, but not at levels sufficient to substantially impact coexistence dynamics. The literature on specialization within natural foraging paths suffers from key limitations, but overall suggests that pollinator-mediated coexistence should be rare in diverse plant communities.

Mimicking orchids lure bees from afar with exaggerated ultraviolet signals.

Flowers have many traits to appeal to pollinators, including ultraviolet (UV) absorbing markings, which are well-known for attracting bees at close proximity (e.g., <1 m). While striking UV signals have been thought to attract pollinators also from far away, if these signals impact the plant pollinia removal over distance remains unknown. Here, we report the case of the Australian orchid Diuris brumalis, a nonrewarding species, pollinated by bees via mimicry of the rewarding pea plant Daviesia decurrens. When distant from the pea plant, Diuris was hypothesized to enhance pollinator attraction by exaggeratedly mimicking the floral ultraviolet (UV) reflecting patterns of its model. By experimentally modulating floral UV reflectance with a UV screening solution, we quantified the orchid pollinia removal at a variable distance from the model pea plants. We demonstrate that the deceptive orchid Diuris attracts bee pollinators by emphasizing the visual stimuli, which mimic the floral UV signaling of the rewarding model Daviesia. Moreover, the exaggerated UV reflectance of Diuris flowers impacted pollinators' visitation at an optimal distance from Da. decurrens, and the effect decreased when orchids were too close or too far away from the model. Our findings support the hypothesis that salient UV flower signaling plays a functional role in visual floral mimicry, likely exploiting perceptual gaps in bee neural coding, and mediates the plant pollinia removal at much greater spatial scales than previously expected. The ruse works most effectively at an optimal distance of several meters revealing the importance of salient visual stimuli when mimicry is imperfect.

Comparative psychophysics of Western honey bee (Apis mellifera) and stingless bee (Tetragonula carbonaria) colour purity and intensity perception.

Bees play a vital role as pollinators worldwide and have influenced how flower colour signals have evolved. The Western honey bee, Apis mellifera (Apini), and the Buff-tailed bumble bee, Bombus terrestris (Bombini) are well-studied model species with regard to their sensory physiology and pollination capacity, although currently far less is known about stingless bees (Meliponini) that are common in pantropical regions. We conducted comparative experiments with two highly eusocial bee species, the Western honey bee, A. mellifera, and the Australian stingless bee, Tetragonula carbonaria, to understand their colour preferences considering fine-scaled stimuli specifically designed for testing bee colour vision. We employed stimuli made of pigment powders to allow manipulation of single colour parameters including spectral purity (saturation) or colour intensity (brightness) of a blue colour (hue) for which both species have previously shown innate preferences. Both A. mellifera and T. carbonaria demonstrated a significant preference for spectrally purer colour stimuli, although this preference is more pronounced in honey bees than in stingless bees. When all other colour cues were tightly controlled, honey bees receiving absolute conditioning demonstrated a capacity to learn a high-intensity stimulus significant from chance expectation demonstrating some capacity of plasticity for this dimension of colour perception. However, honey bees failed to learn low-intensity stimuli, and T. carbonaria was insensitive to stimulus intensity as a cue. These comparative findings suggest that there may be some common roots underpinning colour perception in bee pollinators and how they interact with flowers, although species-specific differences do exist.

Dynamic monitoring of vital functions and tissue re-organization in Saturnia pavonia (Lepidoptera, Saturniidae) during final metamorphosis by non-invasive MRI.

Magnetic resonance imaging (MRI) is the key whole-body imaging technology for observing processes within a living object providing excellent resolution and contrast between soft tissues. In the present work, we exploited the non-destructive properties of MRI to track longitudinally the dynamic changes that take place in developing pupae of the Emperor Moth (Saturnia pavonia) during the last days before eclosion. While in diapause pupae, body fluid was almost homogeneously distributed over the internal compartments, as soon as wings, legs, flight muscles and the head region were fully developed, a significant redistribution of water levels occurred between thoracic and abdominal regions. During the last two days before eclosion, the developing moths transferred substantial amounts of liquid into the gut and the labial gland, and in case of females, into developing eggs. Concomitantly, the volume of the air sacs increased drastically and their expansion/compression became clearly visible in time-resolved MR images. Furthermore, besides ventilation of the tracheal system, air sacs are likely to serve as volume reservoir for liquid transfer during development of the moths inside their pupal case. In parallel, we were able to monitor noninvasively lipid consumption, cardiac activity and haemolymph circulation during final metamorphosis.

Flower Color Evolution and the Evidence of Pollinator-Mediated Selection.

The evolution of floral traits in animal-pollinated plants involves the interaction between flowers as signal senders and pollinators as signal receivers. Flower colors are very diverse, effect pollinator attraction and flower foraging behavior, and are hypothesized to be shaped through pollinator-mediated selection. However, most of our current understanding of flower color evolution arises from variation between discrete color morphs and completed color shifts accompanying pollinator shifts, while evidence for pollinator-mediated selection on continuous variation in flower colors within populations is still scarce. In this review, we summarize experiments quantifying selection on continuous flower color variation in natural plant populations in the context of pollinator interactions. We found that evidence for significant pollinator-mediated selection is surprisingly limited among existing studies. We propose several possible explanations related to the complexity in the interaction between the colors of flowers and the sensory and cognitive abilities of pollinators as well as pollinator behavioral responses, on the one hand, and the distribution of variation in color phenotypes and fitness, on the other hand. We emphasize currently persisting weaknesses in experimental procedures, and provide some suggestions for how to improve methodology. In conclusion, we encourage future research to bring together plant and animal scientists to jointly forward our understanding of the mechanisms and circumstances of pollinator-mediated selection on flower color.

A bee's eye view of remarkable floral colour patterns in the south-west Australian biodiversity hotspot revealed by false colour photography.

BACKGROUND AND AIMS: Colour pattern is a key cue of bee attraction selectively driving the appeal of pollinators. It comprises the main colour of the flower with extra fine patterns, indicating a reward focal point such as nectar, nectaries, pollen, stamens and floral guides. Such advertising of floral traits guides visitation by the insects, ensuring precision in pollen gathering and deposition. The study, focused in the Southwest Australian Floristic Region, aimed to spot bee colour patterns that are usual and unusual, missing, accomplished by mimicry of pollen and anthers, and overlapping between mimic-model species in floral mimicry cases. METHODS: Floral colour patterns were examined by false colour photography in 55 flower species of multiple highly diverse natural plant communities in south-west Australia. False colour photography is a method to transform a UV photograph and a colour photograph into a false colour photograph based on the trichromatic vision of bees. This method is particularly effective for rapid screening of large numbers of flowers for the presence of fine-scale bee-sensitive structures and surface roughness that are not detectable using standard spectrophotometry. KEY RESULTS: Bee- and bird-pollinated flowers showed the expected but also some remarkable and unusual previously undetected floral colour pattern syndromes. Typical colour patterns include cases of pollen and flower mimicry and UV-absorbing targets. Among the atypical floral colour patterns are unusual white and UV-reflecting flowers of bee-pollinated plants, bicoloured floral guides, consistently occurring in Fabaceae spp., and flowers displaying a selective attractiveness to birds only. In the orchid genera (Diuris and Thelymitra) that employ floral mimicry of model species, we revealed a surprising mimicry phenomenon of anthers mimicked in turn by model species. CONCLUSION: The study demonstrates the applicability of 'bee view' colour imaging for deciphering pollinator cues in a biodiverse flora with potential to be applied to other eco regions. The technique provides an exciting opportunity for indexing floral traits on a biome scale to establish pollination drivers of ecological and evolutionary relevance.

Visibility and attractiveness of Fritillaria (Liliaceae) flowers to potential pollinators.

Visual floral characters play an important role in shaping plant-pollinator interactions. The genus Fritillaria L. (Liliaceae), comprising approximately 140 species, is described as displaying a remarkable variety of flower colours and sizes. Despite this variation in visual floral traits of fritillaries, little is known about the potential role of these features in shaping plant-pollinator interactions. Here, we seek to clarify the role of visual attraction in species offering a robust food reward for pollinators early in the spring, which is the case for Fritillaria. We also searched for potential tendencies in the evolution of floral traits crucial for plant-pollinator communication. The generality of species with green and purple flowers may indicate an influence of environmental factors other than pollinators. The flowers of the studied species seem to be visible but not very visually attractive to potential pollinators. The food rewards are hidden within the nodding perianth, and both traits are conserved among fritillaries. Additionally, visual floral traits are not good predictors of nectar properties. When in the flowers, pollinators are navigated by nectar guides in the form of contrasting nectary area colouration. Flower colour does not serve as a phenotypic filter against illegitimate pollinators-red and orange bird-pollinated fritillaries are visible to bees.

Salvage of floral resources through re-absorption before flower abscission.

Plants invest floral resources, including nectar and pigment, with likely consequent reproductive costs. We hypothesized that plants, whose flowers abscise with age, reabsorb nectar and pigment before abscission. This was tested with flowers of Rhododendron decorum, which has large, conspicuous white flowers that increasingly abscise corollas as flowers age. As this species is pollinated by bees, we also hypothesized that nectar concentration would be relatively high (i.e., > 30% wt/vol) and petals would contain UV-absorbing pigment. Floral nectar volume and concentration were sampled on successive days until abscission (up to ten days old, peak at five days) and for sub-sample of four-day-old flowers. Flowers just abscised were similarly sampled. Flower colours were measured using a modified camera, with recordings of spectral reflectance for abscised and open non-abscised flowers. Pigment content was summed values of red, green, blue channels of false color photos. As expected, flowers reabsorbed almost all nectar before abscission, separately reabsorbing nectar-sugar and nectar-water, and petals contained UV-absorbing pigment. However, flowers did not reabsorb pigment and nectar-concentration was < 30% wt/vol. That flowers reabsorb nectar, not pigment, remains unexplained, though possibly pigment reabsorption is uneconomical. Understanding floral resource reabsorption therefore requires determination of biochemical mechanisms, plus costs/benefits for individual plants.

Changes in floral nectar are unlikely adaptive responses to pollinator flight sound.

Under noiseless experimental conditions, sugar concentration of secreted floral nectar may increase after flower exposure to nearby sounds of pollinator flight (Veits et al. 2019). However, we reject the argument that this represents adaptive plant behaviour, and consider that the appealing analogy between a flower and human ear is unjustified.

A Comparative Study of Food Source Selection in Stingless Bees and Honeybees: Scent Marks, Location, or Color.

In social bees, the choice of food sources is based on several factors, including scent marks, color, and location of flowers. Here, we used similar setups, in which two stingless bee species, Melipona subnitida and Plebeia flavocincta, and the Western honeybee, Apis mellifera, were tested regarding the importance of chemical cues, color cues, and location-dependent cues for foraging behavior. It was determined whether workers chose food sources according to (1) scent marks deposited by conspecifics, (2) the color hue of a food source, (3) the trained location or the proximity of a food source to the hive. All three species preferred the scent-marked over an unmarked feeder that was presented simultaneously, but M. subnitida showed a weaker preference compared to the other species. When trained to blue feeders all three bee species preferred blue, but A. mellifera showed the strongest fidelity. The training to yellow feeders led to less distinct color choices. Only workers of M. subnitida mostly orientated at the training position and the close proximity to the nest. Whether the distance of a feeding site influenced the choice was dependent on the tested parameter (color or scent marks) and the species. Workers of M. subnitida preferably visited the feeder closer to the nest during the scent mark trials, but choose randomly when tested for color learning. Worker honeybees preferred the closer feeding site if trained to yellow, but not if trained to blue, and preferred the more distant feeder during the scent mark trials. Workers of P. flavocincta preferred the closer feeder if trained to blue or yellow, and preferred the more distant feeder during the scent mark trials. The disparity among the species corresponds to differences in body size. Smaller bees are known for reduced visual capabilities and might rely less on visual parameters of the target such as color hue, saturation, or brightness but use scent cues instead. Moreover, the dim-light conditions in forest habitats might reduce the reliability of visual orientation as compared to olfactory orientation. Honeybees showed the most pronounced orientation at floral color cues.

Nectar mimicry: a new phenomenon.

Nectar is the most common floral reward for flower-visiting flies, bees, bats and birds. Many flowers hide nectar in the floral tube and preclude sensing of nectar by flower-visitors from a distance. Even in those flowers that offer easily accessible nectar, the nectaries are mostly inconspicuous to the human eye and the amount of nectar is sparse. It is widely accepted that many flowers display nectar guides in order to direct flower-visitors towards the nectar. Using false colour photography, covering ultraviolet, blue and green ranges of wavelength, revealed a yet unknown conspicuousness of nectar, nectaries and false nectaries for bees due to concordant reflection in the ultraviolet range of wavelength. Nectars, many nectaries and false nectaries have glossy surfaces and reflect all incident light including UV-light. In most cases, this is not particularly conspicuous to the human eye, but highly visible for UV-sensitive insects, due to the fact that the glossy areas are often positioned in UV-absorbing central flower parts and thus produce a strong UV-signal. The optical contrast produced by the glossiness of small smooth areas in close proximity to nectar holders represents a widespread yet overlooked floral cue that nectarivorous flower-visitors might use to locate the floral nectar.

High innate preference of black substrate in the chive gnat, Bradysia odoriphaga (Diptera: Sciaridae).

The chive gnat, Bradysia odoriphaga, is a notorious pest of Allium species in China. Colour trapping is an established method for monitoring and control of Bradysia species. In order to clarify the effect of colour preference of B. odoriphaga for the perched substrate, multiple-choice tests were used to assess the response of the chive gnat to different colour hues and brightness levels under different intensities of white illumination and two spectrally different illuminations. Given the choice among four colours differing in hue under different intensities of white illumination and two spectrally different illuminations, chive gnat adults significant preferred the black substrate, a lesser preference to brown and green substrates, and the least preference to orange substrate irrespective of illumination. Given the choice among four levels of brightness under the same illumination conditions as those in the previous experiment (different intensities of white illumination and two spectrally different illuminations), chive gnats preferred black substrate over dark grey, light grey and white substrates. Meanwhile, both virgin and mated adults significantly preferred black over other colour hues and brightness. Based on our results, we conclude that the chive gnat adults significantly prefer black substrates irrespective of colour hues and brightness. This behaviour does not alter with ambient light condition changes. No difference observed between choices of female and male adults. Our results provide new insight for understanding the colour choice behaviour in chive gnat and pave a way to improve monitoring and control of chive gnats and management.

Bumblebees are able to perceive amino acids via chemotactile antennal stimulation.

Like all animals, bees need to consume essential amino acids to maintain their body's protein synthesis. Perception and discrimination of amino acids are, however, still poorly understood in bees (and insects in general). We used chemotactile conditioning of the proboscis extension response (PER) to examine (1) whether Bombus terrestris workers are able to perceive amino acids by means of their antennae and (if so) which ones, (2) whether they are able to differentiate between different amino acids, and (3) whether they are able to differentiate between different concentrations of the same amino acid. We found that workers perceived asparagine, cysteine, hydroxyproline, glutamic acid, lysine, phenylalanine, and serine, but not alanine, leucine, proline, or valine by means of their antennae. Surprisingly, they were unable to differentiate between different (perceivable) amino acids, but they distinguished between different concentrations of lysine. Consequently, bumblebees seem to possess amino acid receptors at the tip of their antennae, which enable a general perception of those solute amino acids that have an additional functional group (besides the common amino and carboxylic groups). They may thus have the ability to assess the overall amino acid content of pollen and nectar prior to ingestion.

Pollen grain morphology is not exclusively responsible for pollen collectability in bumble bees.

Bee-pollinated plants face a dilemma in that bees both passively transport pollen grains among conspecific flowers and actively collect pollen to feed their larvae. Therefore, mechanisms that reduce pollen collection by bees have evolved in melittophilous plants. Malvaceae pollen is uncollectable for corbiculate bees which has previously been ascribed to pollen size, spines, and pollenkitt. We analysed the influence of pollen grain properties (diameter, spine length, spine density) on the collectability of echinate (spiny) pollen by bumble bees (Bombus terrestris). Workers individually foraging on one of eight plant species from six families performed significantly less pollen foraging on plants which have large, echinate pollen grains. Nevertheless, neither pollen grain size, spine length, nor spine density prove to be an absolute disqualifier for collectability. While pollen foragers did not shift to nectar collection but seized visiting flowers with uncollectable pollen, nectar foragers performed regular foraging bouts on these plants. Pollen that is uncollectable for corbiculate bees limits pollen depletion by generalist bumble bees and probably also honey bees while maintaining them as pollinators, which is an effective solution to the pollen dilemma. As previous assumptions about the impact of pollen morphology on its collectability are disproved, potentially determining factors are discussed.

Functional significance of the optical properties of flowers for visual signalling.

Background: Flower coloration is a key enabler for pollinator attraction. Floral visual signals comprise several components that are generated by specific anatomical structures and pigmentation, and often have different functions in pollinator attraction. Anatomical studies have advanced our understanding of the optical properties of flowers, and evidence from behavioural experiments has elucidated the biological relevance of different components of floral visual signals, but these two lines of research are often considered independently. Scope: Here, we review current knowledge about different aspects of the floral visual signals, their anatomical and optical properties, and their functional significance in plant-pollinator visual signalling. We discuss common aspects, such as chromatic and achromatic contrast, hue, saturation and brightness, as well as less common types of visual signals, including gloss, fluorescence, polarization and iridescence in the context of salience of floral colour signals and their evolution, and highlight promising avenues for future research.

How flower colour signals allure bees and hummingbirds: a community-level test of the bee avoidance hypothesis.

Colour signals are the main floral trait for plant-pollinator communication. Owing to visual specificities, flower visitors exert different selective pressures on flower colour signals of plant communities. Although they evolved to attract pollinators, matching their visual sensitivity and colour preferences, floral signals may also evolve to avoid less efficient pollinators and antagonistic flower visitors. We evaluated evidence for the bee avoidance hypothesis in a Neotropical community pollinated mainly by bees and hummingbirds, the campo rupestre. We analysed flower reflectance spectra, compared colour variables of bee-pollinated flowers (bee-flowers; 244 species) and hummingbird-pollinated flowers (hummingbird-flowers; 39 species), and looked for evidence of bee sensorial exclusion in hummingbird-flowers. Flowers were equally contrasting for hummingbirds. Hummingbird-flowers were less conspicuous to bees, reflecting mainly long wavelengths and avoiding red-blind visitors. Bee-flowers reflected more short wavelengths, were more conspicuous to bees (higher contrasts and spectral purity) than hummingbird-flowers and displayed floral guides more frequently, favouring flower attractiveness, discrimination and handling by bees. Along with no phylogenetic signal, the differences in colour signal strategies between bee- and hummingbird-flowers are the first evidence of the bee avoidance hypothesis at a community level and reinforce the role of pollinators as a selective pressure driving flower colour diversity.

The yellow specialist: dronefly Eristalis tenax prefers different yellow colours for landing and proboscis extension.

Droneflies, imagoes of the hoverfly Eristalis tenax, are known to possess a preference for yellow flowers, i.e. they prefer to visit yellow flowers and prefer to extend the proboscis to yellow colours. In this study we disentangle these colour preferences by investigating the landing reaction and proboscis reflex with particular reference to intensity, spectral purity and dominant wavelength of colour stimuli and their UV reflection properties. In multiple-choice tests, naïve and non-trained flies prefer to land on yellow colours independent of their UV reflection characteristics, but also accept blue, white and pink colours if they absorb UV and are of sufficient brightness. Flies trained to land on colours other than yellow still prefer yellow colours to some extent. Moreover, the flies prefer bright over dark yellow colours even if trained to dark yellow ones. The flies refuse to land on dark colours of all hues. Naïve flies exhibit the proboscis reflex only to pure yellow pollen. These experiments show for the first time that landing in droneflies is triggered by a yellow colour independent of its UV reflection properties, but proboscis extension is triggered by yellow colours strongly absorbing blue and UV. The ability to discriminate colours is better than predicted by the categorical colour vision model. The colour preferences in E. tenax represent a fine-tuned ability to visit yellow flowers displaying a UV bull's-eye pattern.

Spectral purity, intensity and dominant wavelength: Disparate colour preferences of two Brazilian stingless bee species.

Bees use floral colour as a major long distance orientation cue. While it is known for bumblebees and honeybees that dominant wavelength (≙ colour hue), colour contrast and spectral purity (≙ saturation) are crucial for flower detection and discrimination, only little is known about colour preferences in stingless bees (Meliponini). In this experiment freely flying workers of two Brazilian species of stingless bees-Partamona helleri and Melipona bicolor-were tested for colour preferences concerning the colour parameters dominant wavelength, spectral purity and intensity (≙ brightness). Each individual bee had to perform 57 tests, in which a definite series of dual choices between colour stimuli varying in intensity, spectral purity or dominant wavelength were presented. The results show that P. helleri chose colours of higher spectral purity and preferred bluish colours, while M. bicolor made generalized colour choices. Intensity did not influence the colour choice of any bee species. The results of P. helleri are consistent with findings for honeybees and bumblebees, while colour preferences in M. bicolor seem to be absent.