Ancient insect vision tuned for flight among rocks and plants underpins natural flower colour diversity.

Understanding the origins of flower colour signalling to pollinators is fundamental to evolutionary biology and ecology. Flower colour evolves under pressure from visual systems of pollinators, like birds and insects, to establish global signatures among flowers with similar pollinators. However, an understanding of the ancient origins of this relationship remains elusive. Here, we employ computer simulations to generate artificial flower backgrounds assembled from real material sample spectra of rocks, leaves and dead plant materials, against which to test flowers' visibility to birds and bees. Our results indicate how flower colours differ from their backgrounds in strength, and the distributions of salient reflectance features when perceived by these key pollinators, to reveal the possible origins of their colours. Since Hymenopteran visual perception evolved before flowers, the terrestrial chromatic context for its evolution to facilitate flight and orientation consisted of rocks, leaves, sticks and bark. Flowers exploited these pre-evolved visual capacities of their visitors, in response evolving chromatic features to signal to bees, and differently to birds, against a backdrop of other natural materials. Consequently, it appears that today's flower colours may be an evolutionary response to the vision of diurnal pollinators navigating their world millennia prior to the first flowers.

Intrafloral patterns of color and scent in Capparis spinosa L. and the ghosts of its selection past.

PREMISE: Capparis spinosa is a widespread charismatic plant, in which the nocturnal floral habit contrasts with the high visitation by diurnal bees and the pronounced scarcity of hawkmoths. To resolve this discrepancy and elucidate floral evolution of C. spinosa, we analyzed the intrafloral patterns of visual and olfactory cues in relation to the known sensory biases of the different visitor guilds (bees, butterflies, and hawkmoths). METHODS: We measured the intrafloral variation of scent, reflectance spectra, and colorimetric properties according to three guilds of known visitors of C. spinosa. Additionally, we sampled visitation rates using a motion-activated camera. RESULTS: Carpenter bees visited the flowers eight times more frequently than nocturnal hawkmoths, at dusk and in the following morning. Yet, the floral headspace of C. spinosa contained a typical sphingophilous scent with high emission rates of certain monoterpenes and amino-acid derived compounds. Visual cues included a special case of multisensory nectar guide and color patterns conspicuous to the visual systems of both hawkmoths and bees. CONCLUSIONS: The intrafloral patterns of sensory stimuli suggest that hawkmoths have exerted strong historical selection on C. spinosa. Our study revealed two interesting paradoxes: (a) the flowers phenotypically biased towards the more inconsistent pollinator; and (b) floral display demands an abundance of resources that seems maladaptive in the habitats of C. spinosa. The transition to a binary pollination system accommodating large bees has not required phenotypic changes, owing to specific eco-physiological adaptations, unrelated to pollination, which make this plant an unusual case in pollination ecology.

Different mechanisms underlie implicit visual statistical learning in honey bees and humans.

The ability of developing complex internal representations of the environment is considered a crucial antecedent to the emergence of humans' higher cognitive functions. Yet it is an open question whether there is any fundamental difference in how humans and other good visual learner species naturally encode aspects of novel visual scenes. Using the same modified visual statistical learning paradigm and multielement stimuli, we investigated how human adults and honey bees (Apis mellifera) encode spontaneously, without dedicated training, various statistical properties of novel visual scenes. We found that, similarly to humans, honey bees automatically develop a complex internal representation of their visual environment that evolves with accumulation of new evidence even without a targeted reinforcement. In particular, with more experience, they shift from being sensitive to statistics of only elemental features of the scenes to relying on co-occurrence frequencies of elements while losing their sensitivity to elemental frequencies, but they never encode automatically the predictivity of elements. In contrast, humans involuntarily develop an internal representation that includes single-element and co-occurrence statistics, as well as information about the predictivity between elements. Importantly, capturing human visual learning results requires a probabilistic chunk-learning model, whereas a simple fragment-based memory-trace model that counts occurrence summary statistics is sufficient to replicate honey bees' learning behavior. Thus, humans' sophisticated encoding of sensory stimuli that provides intrinsic sensitivity to predictive information might be one of the fundamental prerequisites of developing higher cognitive abilities.

Fly pollination drives convergence of flower coloration.

Plant-pollinator interactions provide a natural experiment in signal evolution. Flowers are known to have evolved colour signals that maximise their ease of detection by the visual systems of important pollinators such as bees. Whilst most angiosperms are bee pollinated, our understanding on how the second largest group of pollinating insects, flies, may influence flower colour evolution is limited to the use of categorical models of colour discrimination that do not reflect the small colour differences commonly observed between and within flower species. Here we show by comparing flower signals that occur in different environments including total absence of bees, a mixture of bee and fly pollination within one plant family (Orchidaceae) from a single community, and typical flowers from a broad taxonomic sampling of the same geographic region, that perceptually different colours, empirically measured, do evolve in response to different types of insect pollinators. We show evidence of both convergence among fly-pollinated floral colours but also of divergence and displacement of colour signals in the absence of bee pollinators. Our findings give an insight into how both ecological and agricultural systems may be affected by changes in pollinator distributions around the world.

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.

Comparative psychophysics of colour preferences in two species of non-eusocial Australian native halictid bees.

Colour signalling by flowers appears to be the main plant-pollinator communication system observed across many diverse species and locations worldwide. Bees are considered one of the most important insect pollinators; however, native non-eusocial bees are often understudied compared to managed eusocial species, such as honeybees and bumblebees. Here, we tested two species of native Australian non-eusocial halictid bees on their colour preferences for seven different broadband colours with bee-colour-space dominant wavelengths ranging from 385 to 560 nm and a neutral grey control. Lasioglossum (Chilalictus) lanarium demonstrated preferences for a UV-absorbing white (455 nm) and a yellow (560 nm) stimulus. Lasioglossum (Parasphecodes) sp. showed no colour preferences. Subsequent analyses showed that green contrast and spectral purity had a significant positive relationship with the number of visits by L. lanarium to stimuli. Colour preferences were consistent with other bee species and may be phylogenetically conserved and linked to how trichromatic bees processes visual information, although the relative dearth of empirical evidence on different bee species currently makes it difficult to dissect mechanisms. Past studies and our current results suggest that both innate and environmental factors might both be at play in mediating bee colour preferences.

Einstein, von Frisch and the honeybee: a historical letter comes to light.

The work of the Nobel Laureate Karl von Frisch, the founder of this journal, was seminal in many ways. He established the honeybee as a key animal model for experimental behavioural studies on sensory perception, learning and memory, and first correctly interpreted its famous dance communication. Here, we report on a previously unknown letter by the Physicist and Nobel Laureate Albert Einstein that was written in October 1949. It briefly addresses the work of von Frisch and also queries how understanding animal perception and navigation may lead to innovations in physics. We discuss records proving that Einstein and von Frisch met in April 1949 when von Frisch visited the USA to present a lecture on bees at Princeton University. In the historical context of Einstein's theories and thought experiments, we discuss some more recent discoveries of animal sensory capabilities alien to us humans and potentially valuable for bio-inspired design improvements. We also address the orientation of animals like migratory birds mentioned by Einstein 70 years ago, which pushes the boundaries of our understanding nature, both its biology and physics.

Spontaneous choices for insect-pollinated flower shapes by wild non-eusocial halictid bees.

The majority of angiosperms require animal pollination for reproduction, and insects are the dominant group of animal pollinators. Bees are considered one of the most important and abundant insect pollinators. Research into bee behaviour and foraging decisions has typically centred on managed eusocial bee species, including Apis mellifera and Bombus terrestris. Non-eusocial bees are understudied with respect to foraging strategies and decision making, such as flower preferences. Understanding whether there are fundamental foraging strategies and preferences that are features of insect groups can provide key insights into the evolution of flower-pollinator co-evolution. In the current study, Lasioglossum (Chilalictus) lanarium and Lasioglossum (Parasphecodes) sp., two native Australian generalist halictid bees, were tested for flower shape preferences between native insect-pollinated and bird-pollinated flowers. Each bee was presented with achromatic images of either insect-pollinated or bird-pollinated flowers in a circular arena. Both native bee species demonstrated a significant preference for images of insect-pollinated flowers. These preferences are similar to those found in A. mellifera, suggesting that flower shape preference may be a deep-rooted evolutionary occurrence within bees. With growing interest in the sensory capabilities of non-eusocial bees as alternative pollinators, the current study also provides a valuable framework for further behavioural testing of such species.

Use of temporal and colour cueing in a symbolic delayed matching task by honey bees.

Honey bees (Apis mellifera) are known for their capacity to learn arbitrary relationships between colours, odours and even numbers. However, it is not known whether bees can use temporal signals as cueing stimuli in a similar way during symbolic delayed matching-to-sample tasks. Honey bees potentially process temporal signals during foraging activities, but the extent to which they can use such information is unclear. Here, we investigated whether free-flying honey bees could use either illumination colour or illumination duration as potential context-setting cues to enable their subsequent decisions for a symbolic delayed matching-to-sample task. We found that bees could use the changing colour context of the illumination to complete the subsequent spatial vision task at a level significantly different from chance expectation, but could not use the duration of either a 1 or 3 s light as a cueing stimulus. These findings suggest that bees cannot use temporal information as a cueing stimulus as efficiently as other signals such as colour, and are consistent with previous field observations suggesting a limited interval timing capacity in honey bees.

Spontaneous quantity discrimination of artificial flowers by foraging honeybees.

Many animals need to process numerical and quantity information in order to survive. Spontaneous quantity discrimination allows differentiation between two or more quantities without reinforcement or prior training on any numerical task. It is useful for assessing food resources, aggressive interactions, predator avoidance and prey choice. Honeybees have previously demonstrated landmark counting, quantity matching, use of numerical rules, quantity discrimination and arithmetic, but have not been tested for spontaneous quantity discrimination. In bees, spontaneous quantity discrimination could be useful when assessing the quantity of flowers available in a patch and thus maximizing foraging efficiency. In the current study, we assessed the spontaneous quantity discrimination behaviour of honeybees. Bees were trained to associate a single yellow artificial flower with sucrose. Bees were then tested for their ability to discriminate between 13 different quantity comparisons of artificial flowers (numeric ratio range: 0.08-0.8). Bees significantly preferred the higher quantity only in comparisons where '1' was the lower quantity and where there was a sufficient magnitudinal distance between quantities (e.g. 1 versus 12, 1 versus 4, and 1 versus 3 but not 1 versus 2). Our results suggest a possible evolutionary benefit to choosing a foraging patch with a higher quantity of flowers when resources are scarce.

Improved color constancy in honey bees enabled by parallel visual projections from dorsal ocelli.

How can a pollinator, like the honey bee, perceive the same colors on visited flowers, despite continuous and rapid changes in ambient illumination and background color? A hundred years ago, von Kries proposed an elegant solution to this problem, color constancy, which is currently incorporated in many imaging and technological applications. However, empirical evidence on how this method can operate on animal brains remains tenuous. Our mathematical modeling proposes that the observed spectral tuning of simple ocellar photoreceptors in the honey bee allows for the necessary input for an optimal color constancy solution to most natural light environments. The model is fully supported by our detailed description of a neural pathway allowing for the integration of signals originating from the ocellar photoreceptors to the information processing regions in the bee brain. These findings reveal a neural implementation to the classic color constancy problem that can be easily translated into artificial color imaging systems.

Symbolic representation of numerosity by honeybees ( Apis mellifera): matching characters to small quantities.

The assignment of a symbolic representation to a specific numerosity is a fundamental requirement for humans solving complex mathematical calculations used in diverse applications such as algebra, accounting, physics and everyday commerce. Here we show that honeybees are able to learn to match a sign to a numerosity, or a numerosity to a sign, and subsequently transfer this knowledge to novel numerosity stimuli changed in colour properties, shape and configuration. While honeybees learned the associations between two quantities (two; three) and two signs (N-shape; inverted T-shape), they failed at reversing their specific task of sign-to-numerosity matching to numerosity-to-sign matching and vice versa (i.e. a honeybee that learned to match a sign to a number of elements was not able to invert this learning to match the numerosity of elements to a sign). Thus, while bees could learn the association between a symbol and numerosity, it was linked to the specific task and bees could not spontaneously extrapolate the association to a novel, reversed task. Our study therefore reveals that the basic requirement for numerical symbolic representation can be fulfilled by an insect brain, suggesting that the absence of its spontaneous emergence in animals is not due to cognitive limitation.

Climate is a strong predictor of near-infrared reflectance but a poor predictor of colour in butterflies.

Colour variation across climatic gradients is a common ecogeographical pattern; yet there is long-standing contention over underlying causes, particularly selection for thermal benefits. We tested the evolutionary association between climate gradients and reflectance of near-infrared (NIR) wavelengths, which influence heat gain but are not visible to animals. We measured ultraviolet (UVA), visible (Vis) and NIR reflectance from calibrated images of 372 butterfly specimens from 60 populations (49 species, five families) spanning the Australian continent. Consistent with selection for thermal benefits, the association between climate and reflectance was stronger for NIR than UVA-Vis wavelengths. Furthermore, climate predicted reflectance of the thorax and basal wing, which are critical to thermoregulation; but it did not predict reflectance of the entire wing, which has a variable role in thermoregulation depending on basking behaviour. These results provide evidence that selection for thermal benefits has shaped the reflectance properties of butterflies.

Colour preferences of Tetragonula carbonaria Sm. stingless bees for colour morphs of the Australian native orchid Caladenia carnea.

Innate colour preferences promote the capacity of pollinators to find flowers, although currently there is a paucity of data on how preferences apply to real flowers. The Australian sugarbag bee (Tetragonula carbonaria Sm.) has innate preferences for colours, including UV-absorbing white. Sugarbag bees are pollinators of the terrestrial orchid Caladenia carnea R.Br., which has both white and pink morphs. In laboratory conditions, we tested flower-naïve bees with the white and pink flower morphs revealing a significant preference for the white morph, consistent with experiments using artificial stimuli. In experiments to understand how bees may select food-deceptive orchids following habituation to a particular colour morph, we observed a significant increase in choices towards novel white flowers. We also observed that the presence of a UV-reflecting dorsal sepal signal significantly increased bee choices compared to flowers that had the UV signal blocked. Our findings demonstrate that innate preference testing of insect pollinators with artificial stimuli is replicated in a biologically significant scenario with flowers. The findings also underscore how food-deceptive orchids can receive sufficient pollinator visits to ensure pollination by having different morphs that draw on the innate preferences of bees and their ability to make decisions in a complex ecological setting.

Surpassing the subitizing threshold: appetitive-aversive conditioning improves discrimination of numerosities in honeybees.

Animals including humans, fish and honeybees have demonstrated a quantity discrimination threshold at four objects, often known as subitizing elements. Discrimination between numerosities at or above the subitizing range is considered a complex capacity. In the current study, we trained and tested two groups of bees on their ability to differentiate between quantities (4 versus 5 through to 4 versus 8) when trained with different conditioning procedures. Bees trained with appetitive (reward) differential conditioning demonstrated no significant learning of this task, and limited discrimination above the subitizing range. In contrast, bees trained using appetitive-aversive (reward-aversion) differential conditioning demonstrated significant learning and subsequent discrimination of all tested comparisons from 4 versus 5 to 4 versus 8. Our results show conditioning procedure is vital to performance on numerically challenging tasks, and may inform future research on numerical abilities in other animals.

Floral colour structure in two Australian herbaceous communities: it depends on who is looking.

BACKGROUND AND AIMS: Pollinator-mediated interactions between plant species may affect the composition of angiosperm communities. Floral colour signals should play a role in these interactions, but the role will arise from the visual perceptions and behavioural responses of multiple pollinators. Recent advances in the visual sciences can be used to inform our understanding of these perceptions and responses. We outline the application of appropriate visual principles to the analysis of the annual cycle of floral colour structure in two Australian herbaceous communities. METHODS: We used spectrographic measurements of petal reflectance to determine the location of flowers in a model of hymenopteran colour vision. These representations of colour perception were then translated to a behaviourally relevant metric of colour differences using empirically calibrated colour discrimination functions for four hymenopteran species. We then analysed the pattern of colour similarity in terms of this metric in samples of co-flowering plants over the course of a year. We used the same method to analyse the annual pattern of phylogenetic relatedness of co-flowering plants in order to compare colour structure and phylogenetic structure. KEY RESULTS: Co-flowering communities at any given date seldom had colour assemblages significantly different from random. Non-random structure, both dispersion and clustering, occurred occasionally, but depended on which bee observer is considered. The degree of colour similarity was unrelated to phylogenetic similarity within a co-flowering community. CONCLUSIONS: Perceived floral colour structure varied with the sensory capabilities of the observer. The lack of colour structure at most sample dates, particularly the rarity of strong dispersion, suggests that plants do not use chromatic signals primarily to enable bees to discriminate between co-flowering species. It is more likely that colours make plants detectable in a complex landscape.

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.

Perception of contextual size illusions by honeybees in restricted and unrestricted viewing conditions.

How different visual systems process images and make perceptual errors can inform us about cognitive and visual processes. One of the strongest geometric errors in perception is a misperception of size depending on the size of surrounding objects, known as the Ebbinghaus or Titchener illusion. The ability to perceive the Ebbinghaus illusion appears to vary dramatically among vertebrate species, and even populations, but this may depend on whether the viewing distance is restricted. We tested whether honeybees perceive contextual size illusions, and whether errors in perception of size differed under restricted and unrestricted viewing conditions. When the viewing distance was unrestricted, there was an effect of context on size perception and thus, similar to humans, honeybees perceived contrast size illusions. However, when the viewing distance was restricted, bees were able to judge absolute size accurately and did not succumb to visual illusions, despite differing contextual information. Our results show that accurate size perception depends on viewing conditions, and thus may explain the wide variation in previously reported findings across species. These results provide insight into the evolution of visual mechanisms across vertebrate and invertebrate taxa, and suggest convergent evolution of a visual processing solution.

The path to colour discrimination is S-shaped: behaviour determines the interpretation of colour models.

Most of our current understanding on colour discrimination by animal observers is built on models. These typically set strict limits on the capacity of an animal to discriminate between colour stimuli imposed by physiological characteristics of the visual system and different assumptions about the underlying mechanisms of colour processing by the brain. Such physiologically driven models were not designed to accommodate sigmoidal-type discrimination functions as those observed in recent behavioural experiments. Unfortunately, many of the fundamental assumptions on which commonly used colour models are based have been tested against empirical data for very few species and many colour vision studies solely rely on physiological measurements of these species for predicting colour discrimination processes. Here, we test the assumption of a universal principle of colour discrimination only mediated by physiological parameters using behavioural data from four closely related hymenopteran species, considering two frequently used models. Results indicate that there is not a unique function describing colour discrimination by closely related bee species, and that this process is independent of specific model assumptions; in fact, different models produce comparable results for specific test species if calibrated against behavioural data.

Why background colour matters to bees and flowers.

Flowers are often viewed by bee pollinators against a variety of different backgrounds. On the Australian continent, backgrounds are very diverse and include surface examples of all major geological stages of the Earth's history, which have been present during the entire evolutionary period of Angiosperms. Flower signals in Australia are also representative of typical worldwide evolutionary spectral adaptations that enable successful pollination. We measured the spectral properties of 581 natural surfaces, including rocks, sand, green leaves, and dry plant materials, sampled from tropical Cairns through to the southern tip of mainland Australia. We modelled in a hexagon colour space, how interactions between background spectra and flower-like colour stimuli affect reliable discrimination and detection in bee pollinators. We calculated the extent to which a given locus would be conflated with the loci of a different flower-colour stimulus using empirically determined colour discrimination regions for bee vision. Our results reveal that whilst colour signals are robust in homogeneous background viewing conditions, there could be significant pressure on plant flowers to evolve saliently-different colours to overcome background spectral noise. We thus show that perceptual noise has a large influence on how colour information can be used in natural conditions.