Possible Mechanisms for the Formation of Flower Size Preferences by Foraging Bumblebees

Large flowers often contain larger nectar rewards, and receive more pollinator visits, than small flowers. We studied possible behavioral mechanisms underlying the formation of flower size preferences in bumblebees, using a two-phase laboratory experiment. Experimentally naive Bombus terrestris (L.) foraged on artificial flowers that bore either a large (3.8 cm diameter) or a small (2.7 cm diameter) display of a uniform color. Only flowers of one display size contained nectar rewards. We changed the display color and the locations of large and small flowers in the second experimental phase. We recorded the bees' choices in both phases. Almost half of the bees (41%) made their first visit to a small flower. The bees learned to associate display size with food reward, and chose rewarding flowers with >85% accuracy by the end of each experimental phase. Some learning occurred within the bees' first three flower visits. Learning of the size–reward association was equally good for large and small displays in the first experimental phase, but better for small displays in the second phase. Formation of size–reward associations followed a similar course in both phases. This suggests that the bees did not apply their experience from the first learning phase to the new situation of the second phase. Rather, they treated each phase of the experiment as an independent learning task. Our results suggest that associative learning is involved in the formation of preferences for large displays by bees. Moreover, bees that had learned to prefer large displays in one foraging situation may not transfer this preference to a novel situation that is sufficiently different. We propose that this feature of the bees' behavior can select for honest advertising in flowers.     

Honeybee (Apis mellifera ligustica) Response to Differences in Handling Time, Rewards and Flower Colours

Free flying honeybees were tested outdoors on blue–white and blue–yellow dimorphic artificial flower patches to examine the influence of reward difference, flower handling‐time difference and flower colour choice on foraging decisions. We employed different flower‐well depths to vary handling times (costs), and differences in sucrose molarity to vary reward quality. Tests were performed with 2 and 6 μl rewards to vary quantity. We show that when handling time is correlated with flower‐colour morphs on a pedicellate artificial flower patch, a honeybee's foraging behaviour is dependent on the flower colours used in the choice tests. This supports a honeybee foraging model where constraints are a significant factor in decision making. Bees visiting blue–yellow flower patches exhibited flower constancy to colour, where they restricted most visits to a single flower colour, some bees to blue and others to yellow, irrespective of handing time differences. When offered a choice of equally rewarding blue or white flowers, bees were not constrained by flower colour and chose to visit flowers with a lower handling time. When reward molarity varied with well depth between blue and white flowers, foragers chose shallow‐well flowers (short‐handling time) with a smaller net harvest rate over deep‐well flowers (long‐handling time) with a greater net harvest rate. Results using the blue–white dimorphic flower patch suggest that when foraging options simultaneously involve reward and handling‐time choices, honeybee forager behaviour is inconsistent with an absolute method of evaluating profit.     

Volume and density of microglomeruli in the honey bee mushroom bodies do not predict performance on a foraging task

The mushroom bodies (MBs) are insect brain regions important for sensory integration, learning, and memory. In adult worker honey bees (Apis mellifera ), the volume of neuropil associated with the MBs is larger in experienced foragers compared with hive bees and less experienced foragers. In addition, the characteristic synaptic structures of the calycal neuropils, the microglomeruli, are larger but present at lower density in 35‐day‐old foragers relative to 1‐day‐old workers. Age‐ and experience‐based changes in plasticity of the MBs are assumed to support performance of challenging tasks, but the behavioral consequences of brain plasticity in insects are rarely examined. In this study, foragers were recruited from a field hive to a patch comprising two colors of otherwise identical artificial flowers. Flowers of one color contained a sucrose reward mimicking nectar; flowers of the second were empty. Task difficulty was adjusted by changing flower colors according to the principle of honey bee color vision space. Microglomerular volume and density in the lip (olfactory inputs) and collar (visual inputs) compartments of the MB calyces were analyzed using anti‐synapsin I immunolabeling and laser scanning confocal microscopy. Foragers displayed significant variation in microglomerular volume and density, but no correlation was found between these synaptic attributes and foraging performance. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1057–1071, 2017     

Flower Choice and Learning in Foraging Bumblebees: Effects of Variation in Nectar Volume and Concentration

Bees collect food from flowers that differ in morphology, color, and scent. Nectar‐seeking foragers can rapidly associate a flower's cues with its profitability, measured as caloric value or ‘net energy gain,’ and generally develop preferences for more profitable species. If two flower types are equally easy to discover and feed from, differences in profitability will arise from differences in the volume or the sugar concentration of their nectar crops. Although there has been much study of how bees respond to one or the other of these two kinds of nectar variation, few studies have considered both at once. We presented free‐foraging bumblebees with two different types of equally rewarding artificial flowers. After a period of familiarization, we made one type more rewarding than the other by increasing its nectar concentration, volume, or both. Bees responded more rapidly to a change in the reward's sugar concentration than to a change in its volume, even if the profitability differences were approximately equal. Sucrose concentration differences (40% vs. 13%) caused bees to virtually abandon the more dilute flower type, whether both types offered the same volume (2 μl) or the less concentrated reward offered higher volume (7 μl vs. 0.85 μl). When the two types of flower differed only in nectar volume (7 μl vs. 0.85 μl), the less rewarding type continued to receive 22% of the visits. We propose three different hypotheses to explain the stronger response of the bees to changes in sugar concentration: (i) their response threshold to sucrose concentration might change; (ii) less time is needed to assess the concentration of a reward than its volume; and (iii) a smaller sample size may be needed for reliable estimation of profitability when flowers differ in concentration.     

Reward Tracking and Memory Decay in the Monarch Butterfly,Danaus plexippus L. (Lepidoptera: Nymphalidae)

Due to their long‐distance migration routes and high longevity, monarch butterflies (Danaus plexippus) are likely to benefit from learning how to discriminate and remember suitable feeding resources. In this study, we assessed monarchs’ abilities to track changing nectar sources over time and to retain learned information presented in two conditioning schedules. Non‐preferred (blue and red) and preferred (yellow) artificial flowers were concomitantly offered to monarchs in a three‐phase experiment. In each phase, flowers of only one color contained sucrose solution, while the others contained water. The rewarding color was changed in each phase. Instantaneous observations were made to assess butterfly visits to each color during each phase; continuous observations over the first 90 min of a new phase allowed us to look in more detail at the transition process. Overall, monarchs tracked sucrose availability, visiting the rewarding flowers more often than the unrewarding ones, regardless of innate preferences. However, butterflies reverted to innate color preferences when the newly rewarding color was different from the initial trained color. In a second experiment, memory decay was compared for butterflies trained according to two schedules: ‘single training’ (sucrose solution in red vs. water in blue artificial flowers in one 15‐min session per day) or ‘intermittent training’ (as above, but in two 7.5‐min sessions per day). Afterwards, butterflies were tested on alternate days for a week in arrays containing unrewarding models of both colors. Following either training schedule, memory persisted for at least 3 d after reinforcement ceased. Our findings reveal that monarchs are able to change their feeding responses according to the flowers’ reward status despite innate preferences, as well as to retain flower information for about half a week regardless of the conditioning dynamics.     

Bumblebees Do Not Respond to Variance in Nectar Concentration

Worker bumblebees (Bombus fervidus) were given repeated binary choices between two colors of artificial flowers with the same associated mean nectar concentration (X? = 20%), but with different variances in nectar concentration. Flowers of one color, yellow or blue, rewarded a bee with 1 μl of 20% sucrose solution (low-variance flower type) on each visit (p = 1) and flowers of the other color rewarded a bee on each visit with 1 μl of either 10% or 30% sucrose (p = 0.5; high-variance flower type). Of the 10 bees tested, nine showed no preference for either the high- or low-variance flowers (indifferent or risk-insensitive). This result is similar to honeybee responses to variation in nectar concentration, despite differences in foraging ecology between bumblebees and honeybees. Flower-choice behaviour in the presence of variance in nectar concentration is a response to the expected concentration of the alternative flower types. Possible mechanisms of risk-sensitive foraging behaviour in bees are discussed.     

Nectarivore foraging ecology: rewards differing in sugar types

Abstract.

• 1 Honey bees, visiting artificial flower patches, were used as a model system to study the effects of sugar type (sucrose, glucose, fructose, and mixed monosaccharide), caloric reward, and floral colour on nectarivore foraging behaviour. Observed behaviour was compared to the predictions of various (sometimes contradictory) foraging models.
• 2 Bees drank indiscriminately from flowers in patches with a blue-white flower dimorphism when caloric values of rewards were equal (e.g. 1M sucrose in both colours; 1 M sucrose versus 2 M monosaccharide of either type), but when nectar caloric rewards were unequal, they switched to the flower colour with the calorically greater reward.
• 3 In yellow-blue dimorphic flower patches, on the other hand, bees did not maximize caloric reward. Rather, bees were individually constant, some to blue, others to yellow, regardless of the sugar types or energy content of the rewards provided in the two flower morphs.
• 4 The results suggest that optimal foraging theory (maximization of net caloric gain per unit time) is a robust predictor of behaviour with regard to the sugar types common to nectars; such optimal foraging is, however, limited by a superstructure of individual constancy.

     

The learning abilities of the white cabbage butterfly,Pieris rapae,foraging for flowers

This study examines the role of learning and memory in the butterflyPieris rapae crucivora Boisduval during foraging for flowers. In an outdoor cage with 6 flower species,P. rapae showed various visiting patterns: some visited only one species, while others visited several species in a day. The foraging process for flowers ofErigeron annuus (L.) Pers. could be divided into two successive steps: (1) landing on the nectaring caputs, and (2) finding the source of nectar in the caput. Butterflies learned to proceed through the two steps more efficiently with successive attempts: they gradually decreased landings on nectarless caputs and probings on the nectarless petals of ligulate flowers respectively. As a result, handling time per unit caputs became shorter, and apparent rewards per unit time, i.e. the efficiency of collecting nectar, increased. In addition, once learned,P. rapae could remember a rewarding flower color for 3 days, which was not interfered with by learning another flower color. This indicates thatP. rapae keeps memory for a period longer than 3 days, and that they can remember at least two flower species as suitable flower resources. Furthermore, data indicated that they sometimes can apply the foraging skills obtained on other flower species to a novel one. These abilities could enable butterflies to easily switch flower species, or to enhance labile preference. It has been known thatP. rapae also shows flower constancy, which may be due to memory constraints. Therefore, they may appropriately use two foraging tactics: visit consistency and labile preference, to get enough nectar according to their circumstances.     

Dynamics of forager arrivals and nectar renewal in flowers of Anchusa strigosa

Summary Long-tongued Anthophora spp. bees collecting nectar from flowers of Anchusa strigosa (Boraginaceae) exhibit systematic foraging. Successive forager arrivals at individual flowers are not independent, and the time elapsed between successive arrivals at a particular flower is distributed more uniformly than be expected on the basis of a random arrival process. Distributions of inter-arrival time for individual flowers show standard deviation/mean ratios of 0.44–0.79, a range which is consistent with results obtained for two other plant-pollinator systems. The rate at which nectar is renewed between successive forager arrivals is independent of the amount of nectar in the flower, and the renewal process is strongly linear. Practical and theoretical implications of these results are discussed.     

Does the Flower Constancy of Bumble Bees Reflect Foraging Economics?

We examined the effects of floral reward level and spatial arrangement on the propensity of bumble bees to exhibit flower constancy. In three separate experiments, we compared the flower constancy of bees on dimorphic arrays of blue and yellow flowers that differed either in reward concentration, reward volume, or inter‐flower distance. Overall, flower choice patterns varied among bees, ranging from random selection to complete constancy. When flowers contained greater reward volumes and were spaced farther apart, bees showed less flower constancy and more moves to closely neighbouring flowers. Changes in reward concentration had no effect on flower constancy; however, more dilute rewards produced shorter flight times between flowers. In addition, there was a strong positive relationship between degree of flower constancy and net rate of energy gain when flowers were spaced farther apart, indicating that constant bees were more economic foragers than inconstant bees. Together, these results support the view that the flower constancy of pollinators reflects an economic foraging decision.     

Foraging Response of Turkish Honey Bee Subspecies to Flower Color Choices and Reward Consistency

Foraging behavior of Apis mellifera caucasica, A.m. carnica and A.m. syriaca in Turkey was studied for intrinsic subspecies-based differences. Models of forager flower-color fidelity, risk sensitive behavior and maximizing net gain were tested. Foragers were presented artificial flower patches containing blue, white and yellow flowers. Some bees of each subspecies showed high fidelity to yellow flowers, while others favored blue and white flowers. The degree of fidelity, however, differed among subspecies and was dependent upon which color was favored. Bees of all subspecies demonstrated risk indifferent behavior regardless of whether they favored yellow flowers or blue and white flowers. Flower handling time differed among subspecies and increased with reward quantity, and when a reward was present. Flight time between consecutive flowers also differed among honey bee subspecies. Foragers of all subspecies had a higher net gain when visiting flowers with consistent rewards.     

Reversal Learning and Risk‐Averse Foraging Behavior in the Monarch Butterfly,Danaus plexippus (Lepidoptera: Nymphalidae)

Learning ability allows insects to respond to a variable environment, and to adjust their behaviors in response to positive or negative experiences. Pollinating insects readily learn to associate floral characteristics, such as color, shape, or pattern, with appetitive stimuli, such as the presence of a nectar reward. However, in nature pollinators may also encounter flowers that contain distasteful or toxic nectar, or offer highly variable nectar volumes, providing opportunities for aversive learning or risk‐averse foraging behavior. Whereas some bees learn to avoid flowers with unpalatable or unreliable nectar rewards, little is known about how Lepidoptera respond to such stimuli. We used a reversal learning paradigm to establish that monarch butterflies learn to discriminate against colored artificial flowers that contain salt solution, decreasing both number of probes and probing time on flowers of a preferred color and altogether avoiding artificial flowers of a non‐preferred color. In addition, when we offered butterflies artificial flowers of two different colors, both of which contained the same mean nectar volume but which differed in variance, the monarchs exhibited risk‐averse foraging: they probed the constant flowers significantly more than the variable ones, regardless of flower color or butterfly sex. Our results add to our understanding of butterfly foraging behavior, as they demonstrate that monarchs can respond to aversive as well as appetitive stimuli, and can also adjust their foraging behavior to avoid floral resources with high variance rewards.     

Can honey bees change foraging patterns?

Abstract. 1. Foraging patterns were studied using honey bees on artificial flower patches to determine if given individuals could change behaviours under differing conditions.
2. Two types of flower patches were used; those simulating a population of flowers, dimorphic for colour, and grids simulating a single colour-dimorphic inflorescence.
3. In the simulated population of flowers bees were individually constant to colour over a range of reward volumes and flower patch sizes.
4. Each bee remained individually constant to a flower morph when visiting a population-type grid but changed to random visitation on the simulated inflorescence.
5. On the simulated inflorescence, with morphs providing unequal qualities of reward, most bees foraged on the higher molarity morph.
6. Most, but not all bees, failed to minimize uncertainty on the simulated inflorescence.
7. On the simulated inflorescence, bees failed to optimize when one morph provided a greater reward volume than did the other.
8. In the population of flowers bees flew from flower to flower, whereas, they walked on the simulated inflorescence.     

Flower reshaping in the transition to hummingbird pollination in Loasaceae subfam. Loasoideae despite absence of corolla tubes or spurs

Many angiosperm lineages present transitions from bee to hummingbird pollination. The flower design in most of these lineages includes either corolla tubes or nectar spurs, structures that commonly experienced an elongation with the acquisition of hummingbird pollination. It is proposed that this increases the fit between the bird head and flower structures, and isolates or partially blocks bees from the interaction. But can this transition occur if the ancestral flower design lacks tubes or spurs? Here we focus on the transition from bee to hummingbird pollination in the Loasaceae subfamily Loasoideae. Loasoideae flowers have radial corollas with separated petals; therefore, they do not display corolla tubes nor nectar spurs. These flowers also present a whorl of nectar scales and staminodes, unique to the subfamily, which is involved in flower–pollinator fit and in nectar harvesting. To explore flower shape adaptation to hummingbird pollination, we tested for correspondence between pollinators and flower shape in Loasoideae. In order to achieve this, we first compared the evolutionary history of flower phenotype and pollination mode, and then used stochastic character mapping and geometric-morphometric variables in a comparison of alternative evolutionary models. The results of our study suggest that the transition from bee to bird pollination was accompanied by changes in the shape of the staminodial complex, along with the evolution of relatively closed corollas. Moreover, while bird pollination seems to be the end point in the evolution of pollination syndromes in many angiosperm lineages, rodent pollinated flowers probably evolved from ancestral bird pollinated flowers in Loasoideae. Our findings suggest that the evolution of bird pollinated flowers from ancestral bee pollinated flowers does not require the presence of corolla tubes or spurs, and can take place as long as the flower design includes structures participating in flower–pollinator fit.     

Floral Temperature and Optimal Foraging: Is Heat a Feasible Floral Reward for Pollinators?

As well as nutritional rewards, some plants also reward ectothermic pollinators with warmth. Bumble bees have some control over their temperature, but have been shown to forage at warmer flowers when given a choice, suggesting that there is some advantage to them of foraging at warm flowers (such as reducing the energy required to raise their body to flight temperature before leaving the flower). We describe a model that considers how a heat reward affects the foraging behaviour in a thermogenic central-place forager (such as a bumble bee). We show that although the pollinator should spend a longer time on individual flowers if they are warm, the increase in total visit time is likely to be small. The pollinator's net rate of energy gain will be increased by landing on warmer flowers. Therefore, if a plant provides a heat reward, it could reduce the amount of nectar it produces, whilst still providing its pollinator with the same net rate of gain. We suggest how heat rewards may link with plant life history strategies.     

Floral reward production is timed by an insect pollinator

Interval timing--sensitivity to elapsing durations--has recently been found to occur in an invertebrate pollinator, the bumble-bee (Bombus impatiens). Here, bumble-bees were required to time the interval between the start of foraging in a patch of low-quality artificial flowers providing 25% sucrose and the availability of a high-quality flower (HQF) that provided 50% sucrose after a fixed delay. The delay changed after every 20 foraging bouts in the order 30-150-30 s. Bees visited the HQF sooner when the delay was 30 s than when it was 150 s, and visits to the HQF peaked near the end of both delays. When the delay changed to 150 s, bees appeared to time both the previous 30 s delay and the new delay. To examine whether bees also learned what kind of reward was provided at the HQF, its usual reward was replaced with 25% sucrose in a final foraging bout. Bumble-bees rejected the HQF on the reward-replacement test. These results show that bumble-bees remembered both when reward was produced by the HQF and what type of reward was produced. These findings indicate that bumble-bees can learn both the timing and content of reward production.     

Colour learning when foraging for nectar and pollen: bees learn two colours at once

Bees are model organisms for the study of learning and memory, yet nearly all such research to date has used a single reward, nectar. Many bees collect both nectar (carbohydrates) and pollen (protein) on a single foraging bout, sometimes from different plant species. We tested whether individual bumblebees could learn colour associations with nectar and pollen rewards simultaneously in a foraging scenario where one floral type offered only nectar and the other only pollen. We found that bees readily learned multiple reward–colour associations, and when presented with novel floral targets generalized to colours similar to those trained for each reward type. These results expand the ecological significance of work on bee learning and raise new questions regarding the cognitive ecology of pollination.     

Honey bee handling behaviour on the papilionate flower of Robinia pseudoacacia L.

Papilionate flowers, such as those of Robinia pseudoacacia L., show tripping mechanisms that prevent pollen release: only those bees which apply the right force on petals induce pollen to be deposited on their bodies. Apis mellifera is considered a poor visitor of such flowers, since individuals are usually too weak to trip the mechanism. Despite this, the honey bee pays frequent visits to flowers of R. pseudoacacia and produces a much appreciated unifloral honey. We investigated how bees manipulate R. pseudoacacia flowers, whether they contact the plant’s reproductive core and if there is any appreciable difference related to the manipulation of individual flowers. Honey bees showed two strategies for resource collection, namely legitimate visits and robberies. Legitimate visits were more frequent and about 63 % entailed contact with the flower’s reproductive core. We distinguished two behaviours, one to achieve successful positioning on the flower and the other for nectar intake. These behaviours were clearly perceptible and described by different curves of time frequency distribution. From the beginning to the end of anthesis, flowers were classified into four types on the basis of their morphological and phenological traits. Positioning time differed significantly depending on the flower type, with less time needed for more ageing flowers. Time spent in nectar intake was instead highly variable and independent of flower ageing. Selecting the right flower type would appear to lead to obtaining the R. pseudoacacia reward, overcoming species-specific physical inability. Moreover, the role of honey bees as pollinators of R. pseudoacacia is considered. Finally, the relations between petal characteristics and strength needed to trip the mechanism in papilionate flowers is also discussed in the light of nectar foragers.     

Flower choice copying in bumblebees

We tested a hypothesis originating with Darwin that bees outside the nest exhibit social learning in flower choices. Naive bumblebees, Bombus impatiens, were allowed to observe trained bees or artificial bees forage from orange or green flowers. Subsequently, observers of bees on green flowers landed more often on green flowers than non-observing controls or observers of models on orange flowers. These results demonstrate that bumblebees can change flower choice by observations of non-nest mates, a novel form of social learning in insects that could provide unique benefits to the colony.     

Flower color change accelerated by bee pollination in Tibouchina (Melastomataceae)

Floral color changes are common among Melastomataceae and have been interpreted as a warning mechanism for bees to avoid old flowers, albeit increasing long-distance flower display. Here the reproductive systems of Tibouchina pulchra and T. sellowiana were investigated by controlled pollinations. Their pollinators were identified, and experiments on floral color and fragrance changes were conduced to verify if those changes affect the floral visitation. Both Tibouchina species are self compatible. The flowers lasted three days or more, and the floral color changed from white in the 1st day to pink in the following days. Pollen deposition on stigma induced floral color change. The effectiveness of the pollination is dependent on bees’ size; only large bees were regarded as effective pollinators. In experimental tests, the bees in T. pulchra preferred the natural white flowers while the visitors of T. sellowiana were attracted by both natural and mimetic 1st-day flowers (2nd-day flowers with experimentally attached 1st-day flower petals). During the experiments on floral fragrance, the bees visited both natural and mimetic 1st-day flowers (2nd-day flowers with 1st-day flower scents). In both experiments, the bees avoided natural 2nd-day flowers, but seldom visited modified 2nd-day flowers. The attractiveness of T. pulchra and T. sellowiana flowers cannot be attributed exclusively to the color or the fragrance separately, both factors seemingly act together.