Bee Preference Research Articles

Foraging of Honeybees from Different Ecological Areas Determined through Melissopalynological Analysis and DNA Metabarcoding.

The environment significantly impacts the lives of bees and their feeding. This study aimed to investigate bee foraging using melissopalynological analysis and DNA metabarcoding in intensive farming, reserved, and urbanized areas. The highest alpha diversity was observed in the reserved and intensive farming areas. The urbanized area had less diversity. In the intensive farming area, Sinapis, Helianthus, and Fagopyrum predominated; in the reserved area, Melilotus, Helianthus, and Brassica predominated. In the urbanized area, garden plants, namely radish (Raphanus sativus) and cucumber (Cucumis hystrix), and agricultural plants, namely soybean (Glycine max) and melon (Cucumis melo), were often found. The most significant agreement was between the rbcL and the melissopalynological analysis. The ITS2 revealed equal matches with both rbcL and melissopalynology, but this marker missed or underestimated some genera. Trifolium pretense and Brassica nigra were identified simultaneously by the melissopalinology method and two genetic markers in DNA metabarcoding. The species Convolvulus arvensis, Melilotus officinalis, Echium vulgare, Brassica rapa, Helianthus divaricatus, and Onobrychis viciifolia were found in all ecological areas. Imperfect databases impose some limits in the identification of some taxa using metabarcoding. The further research and expansion of plant databases is needed. Studying the food preferences of bees in different environmental conditions and landscapes is necessary to develop measures to preserve their populations.

Climatic fluctuations alter the preference of stingless bees (Apidae, Meliponini) towards food contaminated with acephate and glyphosate

The global decline of pollinators has become a major concern for the scientific community, policymakers, and the general public. Among the main drivers of diminishing bee populations is the widespread use of agrochemicals. To gain a comprehensive understanding of the foraging dynamics of bees at agrochemical-contaminated areas, it is essential to consider both environmental conditions and the specific foraging ecology of bee species. For the first time, we conducted a semi-field study to investigate whether stingless bees exhibit a preference for food contaminated with agrochemicals compared to non- contaminated food, under natural weather conditions. Colonies of Plebeia lucii Moure, 2004 were placed in a greenhouse and subjected to a preference test, where bees were given the freedom to choose between contaminated or non-contaminated food sources following a preliminary training period. Within the greenhouse, we placed feeders containing realistic concentrations of an insecticide (acephate: 2 mg a.i./L), a herbicide (glyphosate: 31.3 mg a.i./L), or a mixture of both, alongside non-contaminated food. Environmental variables (temperature, humidity, and light intensity) were monitored throughout the experiment. At higher temperatures, the foragers preferred food containing the mixture of both agrochemicals or uncontaminated food over the other treatments. At lower temperatures, by contrast, the bees preferred food laced with a single agrochemical (acephate or glyphosate) over uncontaminated food or the agrochemical mixture. Our findings indicate that agrochemical residues in nectar pose a significant threat to P. lucii colonies, as foragers do not actively avoid contaminated food, despite the detrimental effects of acephate and glyphosate on bees. Furthermore, we demonstrate that even minor, natural fluctuations in environmental conditions can alter the colony exposure risk. Despite the interplay between temperature and bees' preference for contaminated food, foragers consistently collected contaminated food containing both agrochemicals, whether isolated or in combination, throughout the whole experiment.

A multifunctional role for riboflavin in the yellow nectar of Capsicum baccatum and Capsicum pubescens.

A few Capsicum (pepper) species produce yellow-colored floral nectar, but the chemical identity and biological function of the yellow pigment are unknown. A combination of analytical biochemistry techniques was used to identify the pigment that gives Capsicum baccatum and Capsicum pubescens nectars their yellow color. Microbial growth assays, visual modeling, and honey bee preference tests for artificial nectars containing riboflavin were used to assess potential biological roles for the nectar pigment. High concentrations of riboflavin (vitamin B2) give the nectars their intense yellow color. Nectars containing riboflavin generate reactive oxygen species when exposed to light and reduce microbial growth. Visual modeling also indicates that the yellow color is highly conspicuous to bees within the context of the flower. Lastly, field experiments demonstrate that honey bees prefer artificial nectars containing riboflavin. Some Capsicum nectars contain a yellow-colored vitamin that appears to play roles in (1) limiting microbial growth, (2) the visual attraction of bees, and (3) as a reward to nectar-feeding flower visitors (potential pollinators), which is especially interesting since riboflavin is an essential nutrient for brood rearing in insects. These results cumulatively suggest that the riboflavin found in some Capsicum nectars has several functions.

Floral Trait Preferences of Three Common wild Bee Species.

The interaction between bees and flowering plants is mediated by floral cues that enable bees to find foraging plants. We tested floral cue preferences among three common wild bee species: Lasioglossum villosulum, Osmia bicornis, and Bombus terrestris. Preferences are well studied in eusocial bees but almost unknown in solitary or non-eusocial generalist bee species. Using standardized artificial flowers altered in single cues, we tested preferences for color hue, achromatic contrast, scent complexity, corolla size, and flower depth. We found common attractive cues among all tested bees. Intensively colored flowers and large floral displays were highly attractive. No preferences were observed in scent complexity experiments, and the number of volatiles did not influence the behavior of bees. Differing preferences were found for color hue. The specific behaviors were probably influenced by foraging experience and depended on the flower choice preferences of the tested bee species. In experiments testing different flower depths of reward presentation, the bees chose flat flowers that afforded low energy costs. The results reveal that generalist wild bee species other than well-studied honeybees and bumblebees show strong preferences for distinct floral cues to find potential host plants. The diverse preferences of wild bees ensure the pollination of various flowering plants.

Fields of flowers with few strikes: how oligolectic bees manage their foraging behavior on Calibrachoa elegans (Solanaceae).

In specialized plant-pollinator associations, partners may exhibit adaptive traits, which favor the maintenance of the interaction. The association between Calibrachoa elegans (Solanaceae) and its oligolectic bee pollinator, Hexantheda missionica (Colletidae), is mutualistic and forms a narrowly specialized pollination system. Flowers of C. elegans are pollinated exclusively by this bee species, and the bees restrict their pollen resources to this plant species. The pollen presentation schedules of C. elegans were evaluated at the population level to test the hypothesis that H. missionica females adjust their foraging behavior to the resource offering regime of C. elegans plants. For this, the number of new flowers and anthers opened per hour (as a proxy for pollen offering) was determined, and pollen advertisement was correlated with the frequency of flower visits during the day. Preferences of female bees for flowers of different stages were also investigated, and their efficiency as pollinators was evaluated. Pollen offering by C. elegans was found to be partitioned throughout the day through scattered flower openings. Females of H. missionica indeed adjusted their foraging activity to the most profitable periods of pollen availability. The females preferred new, pollen-rich flowers over old ones and gathered pollen and nectar selectively according to flower age. Such behaviors must optimize female bee foraging efficiency on flowers. Female bees set 93% of fruit after a single visit. These findings guarantee their importance as pollinators and the persistence of the specialized plant-pollinator association.

Drought stress influences foraging preference of a solitary bee on two wildflowers.

Pollinators provide critical ecosystem services, maintaining biodiversity and benefiting global food production. However, plants, pollinators, and their mutualistic interactions may be affected by drought, which has increased in severity and frequency under climate change. Using two annual, insect-pollinated wildflowers (Phacelia campanularia and Nemophila menziesii), we asked how drought impacts floral traits and foraging preferences of a solitary bee (Osmia lignaria) and explore potential implications for plant reproduction. In greenhouses, we experimentally subjected plants to drought to induce water stress, as verified by leaf water potential. To assess the impact of drought on floral traits, we measured flower size, floral display size, nectar volume, and nectar sugar concentration. To explore how drought-induced effects on floral traits affected bee foraging preferences, we performed choice trials. Individual female bees were placed into foraging arenas with two conspecific plants, one droughted and one non-droughted, and were allowed to forage freely. We determined that P. campanularia is more drought-tolerant than N. menziesii based on measures of turgor loss point, and confirmed that droughted plants were more drought-stressed than non-droughted plants. For droughted plants of both species, floral display size was reduced, and flowers were smaller and produced less, more-concentrated nectar. We found that bees preferred non-droughted flowers of N. menziesii. However, bee preference for non-droughted P. campanularia flowers depended on the time of day and was detected only in the afternoon. Our findings indicate that bees prefer visiting non-droughted flowers, likely reducing pollination success for drought-stressed plants. Lack of preference for non-droughted P. campanularia flowers in the morning may reflect the higher drought tolerance of this species. This work highlights the potentially intersecting, short-term physiological and pollinator behavioral responses to drought and suggests that such responses may reshape plant-pollinator interactions, ultimately reducing reproductive output for less drought-tolerant wildflowers.

Protein/Lipid ratio of pollen biases the visitation of bumblebees (Bombus ignitus Smith) to male-fertile cultivars of the Japanese pear (Pyrus pyrifolia Nakai).

Bees have been known to visit the male-fertile cultivars of self-incompatible flowering plants more frequently than the male-sterile cultivars, but the origin of this preference is poorly understood. Here, we demonstrate that this preference is driven by the higher protein/lipid ratio of male-fertile pollen compared with male-sterile pollen by way of two caged-behavioral assays with six cultivars. In the first assay, flower-naïve bumblebees (Bombus ignitus Smith) showed a significantly higher flower-visitation rate to male-fertile cultivars (pollen germination rate > 55%; > 14 visits/10 min) of the Japanese pear (Pyrus pyrifolia Nakai) than male-sterile cultivars (pollen germination rate ≤ 20%; > 6 visits/10 min). In the second, bees still preferred the anthers of male-fertile cultivars (5-9 visits/10 min) more than those of male-sterile ones (less than 1 visit in 10 min) even in the absence of all other organs (i.e., petals, pistil, nectar), indicating that pollen is responsible for the preference. We then analyzed the macronutrient content of the pollen and its visual cues, and found that the bee preference was highly correlated with the protein/lipid ratio (0.3-1.6) but not color variables such as (a)chromatic contrast, intensity, and spectral purity. We conclude that the protein/lipid ratio influences the foraging behavior of the bumblebees likely by serving as (1) a chemotactile cue while antennating, (2) a gustatory cue after intake, and (3) an olfactory cue. In addition, the low bee visitation rate to poorly viable pollen could be due to its low protein/lipid ratio.

Strawberry varieties differ in pollinator-relevant floral traits.

A rising global population will need more food, increasing demand for insect pollination services. However, general insect declines conflict with this demand. One way to mitigate this conflict is to grow crop flowers that are easier for insects to find and more rewarding to those that visit them. This study quantifies variation in the pollinator-relevant traits of nectar and pollen production, flower size and flower shape in commercial strawberry, finding significant variation between varieties in all traits. Bumblebees could learn to distinguish between the extremes of variation in flower shape, but this learning is very slow, indicating that this variation is at the limit of that which can be detected by bumblebees. Bee preferences for nectar of differing sugar concentrations at field-realistic volumes were consistent with previous observations at larger volumes, suggesting that it is valid to translate lab findings to the field. This study builds on our knowledge of the range of pollinator reward present in a single cultivated species and of the impact of field-realistic levels of variation in floral traits on bumblebee preferences.

Unraveling the floral preference: Bee pollen identification and characterization of Tetragonula laeviceps

The stingless bee (Tetragonula laeviceps) is capable of yielding products such as honey, propolis, and bee pollen, which hold promise as potential sources of nutrition or alternatives in the field of medicine. The physical and chemical attributes of these products are influenced by intrinsic factors, including dietary sources and bee species, as well as extraneous variables such as management techniques, seasonal variations, and environmental parameters. The Campus of the Biology Faculty UGM encompasses a rich repository of flowering plant species that present a viable resource for the sustenance of stingless bees. Consequently, this research endeavor seeks to delineate and characterize the plant species that serve as the sustenance of stingless bees by elucidating the pollen types discovered within their nest. Pollen specimens were directly procured from the nests and subjected to be analyzed with light microscopy and scanning electron microscopy (SEM). At least four distinct varieties of pollen have been identified, originating from divergent plant species, namely those belonging to the Pinaceae, Fabaceae, Myrtaceae, and Brassicaceae families. The dietary preferences of stingless bees, as discerned through the identification of pollen types, exhibit congruence with the plant species in the vicinity of their nesting sites.

Marmara Adasında Bulunan Bal Arılarının (Apis mellifera L.) Polen Tercihleri

Islands have a special importance in terms of biodiversity. At the same time, it is important to follow the feeding behavior of pollinators that ensure the continuation of biodiversity throughout the island. In this study, it was aimed to follow the pollen collection activities of honey bees (Apis mellifera L.) on Marmara Island and to determine the pollen sources. Pollen sampling was carried out weekly during the 14 weeks from the first week of April to the first week of July when honey bees intensively carried out pollen collection activities. The sampled pollen grains were classified according to their color and examined by light microscopy. A total of 34 taxa were identified from the pollen grains. The taxa represented in significant proportions were; Rosaceae, Fabaceae, and Centaurea, Ericaceae, Brassicaceae, Cistus, Helianthemum, Vitis and Salix. They constitute a total of 79.37%. Rosaceae family pollen has been found as the most preferred taxa by honey bees for 8 weeks. The first two weeks of May was the most productive period in terms of pollen species in Marmara Island and the amount of pollen brought to the hives decreased in early July. In this study, it was tried to shed light on the pollen collection preferences of honey bees and pollination periods of honey bee used plants under conditions of Marmara Island.

Blueberry floral probiotics: nectar microbes inhibit the growth of Colletotrichum pathogens.

To identify whether microorganisms isolated from blueberry flowers can inhibit the growth of Colletotrichum, an opportunistic plant pathogen that infects flowers and threatens yields, and to assess the impacts of floral microbes and Colletotrichum pathogens on artificial nectar sugars and honey bee consumption. The growth inhibition of Colletotrichum (Colletotrichum acutatum, Colletotrichum fioriniae, and Colletotrichum gloeosporioides) was screened using both artificial nectar co-culture and dual culture plate assays. All candidate nectar microbes were screened for antagonism against a single C. acutatum isolate. Then, the top four candidate nectar microbes showing the strongest inhibition of C. acutatum (Neokomagataea thailandica, Neokomagataea tanensis, Metschnikowia rancensis, and Symmetrospora symmetrica) were evaluated for antagonism against three additional C. acutatum isolates, and single isolates of both C. fioriniae and C. gloeosporioides. In artificial nectar assays, single and three-species cultures inhibited the growth of two of four C. acutatum isolates by ca. 60%, but growth of other Colletotrichum species was not affected. In dual culture plate assays, inhibition was observed for all Colletotrichum species for at least three of four selected microbial antagonists (13%‒53%). Neither honey bee consumption of nectar nor nectar sugar concentrations were affected by any microbe or pathogen tested. Selected floral microbes inhibited growth of all Colletotrichum species in vitro, although the degree of inhibition was specific to the assay and pathogen examined. In all microbial treatments, nectar sugars were preserved, and honey bee preference was not affected.

Leafcutter bee preference of plant saplings in plant nurseries: context for future research and conservation

Leafcutter bee preference of plant saplings in plant nurseries: context for future research and conservation

Floral preferences of carpenter bees (Apidae: Xylocopini: Xylocopa) from Maranhão, Northeast Brazil

ABSTRACT The current study aimed to recognize the floral resources used by bees of the genus Xylocopa in transition physiognomies. We compiled the information available from biological collections and analyzed metrics of dystrophic interaction. We listed 2547 records of Xylocopa in flowers, across twelve identified bee species. The bees were collected from 109 plant species, belonging to 77 genera and 36 families. An interaction network was constructed based on 193 interactions. In which there was a significant correlation between the abundance of bees collected and the richness of plants visited. The most generalist bees were X. cearensis and X. frontalis, and the plants with the most records were Chamaecrista hispidula and C. ramosa. Significant seasonality was observed in the total number of individuals. On the other hand, there was a weak similarity among Xylocopa species according to plant species, despite the clear relationship of the bees according to physiognomies. The interaction network proved to be highly nested, indicating a functional partitioning that allows several species to occur in the same environment. The contribution made by the current work for its regional scope stands out, dealing with areas that have distinct characteristics.

Chemical Composition of Four Industrial Hemp (Cannabis sativa L.) Pollen and Bee Preference.

Apart from its economic value, industrial hemp (Cannabis sativa L.) is a prolific pollen producer, serving as a food source for bees. However, little is known regarding the extent to which varietal differences in hemp pollen chemistry influences bee preference. Here, we report the chemical profile of pollen from four hemp varieties (Canda, CFX-2, Henola, and Joey) and bee abundance and diversity, using direct visual counts and pan traps. The number and type of bees on each variety was recorded and the chemical composition (proximate analysis and mineral, amino, and fatty acid profiles) of pollen from each hemp variety was determined. During the entire sampling period, three bee types (bumble bees, honey bees, and sweat bees) were recorded, with a combined total of 1826. Among these, sweat bees and bumble bees were the most prevalent and were highest on the Joey variety. The four varieties expressed protein content ranging from 6.05% to 6.89% and the highest in Henola. Seventeen amino acids were expressed in all varieties, with leucine recording the highest content ranging from 4.00 mg/g in Canda to 4.54 mg/g in Henola. In general, Henola expressed high protein, amino acid, and saturated and monosaturated fatty acid contents and recorded significantly fewer bees compared with Joey, which had a low content of these components and a high content of polyunsaturated fatty acids. Our findings suggest that, while industrial hemp offers abundant and accessible pollen that would promote bee health and sustainability of their ecosystem services, the nutritional quality may not be adequate for bee growth and development as an exclusive pollen source.

Honey bees exhibit greater patch fidelity than bumble bees when foraging in a common environment

AbstractAnimals commonly exhibit a tendency to return to previously visited locations. Such tendency is manifested at different scales, for example, fidelity to a site or fidelity to a specific patch within a site. Although patch fidelity has important implications for the pollinators and the plants they visit, our understanding of patch fidelity, and the extent to which it varies among bee species, remains limited. Here, we used a mark–reobservation approach to compare patch fidelity and patch size preference between one bumble bee and one honey bee species foraging on patches of Medicago sativa L. Honey bees exhibited greater patch fidelity (76%) than bumble bees (47%); they were more likely to return to the patch where they were marked. Patch size affected the level of patch fidelity for bumble bees but not for honey bees. Bumble bees were more likely to return to larger rather than smaller patches. In addition to patch fidelity, we detected preference of bees for the larger patches. Bees visited the larger patches more often than the smaller patches. These results add patch fidelity to the already known repertoire of differences in foraging strategies between a bumble bee and a honey bee species. It also indicates how the simultaneous study of distinct species in a similar environment can reveal important previously undetected information about their behavioral ecology. Observed differences in patch fidelity and patch size preferences may have important implications for crop pollination and conservation habitat design.

Complex preference relationships between native and non-native angiosperms and foraging insect visitors in a suburban greenspace under field and laboratory conditions

The introduction and spread of non-native flora threatens native pollinators and plants. Non-native angiosperms can compete with native plants for pollinators, space, and other resources which can leave native bees without adequate nutritional or nesting resources, particularly specialist species. In the current study, we conducted flower preference experiments through field observations and controlled binary choice tests in an artificial arena to determine the impact of field vs. laboratory methods on flower preferences of native bees for native or non-native flowers within their foraging range. We conducted counts of insect pollinators foraging on the flowers of three plant species in a suburban green belt including one native (Arthropodium strictum) and two non-native (Arctotheca calendula and Taraxacum officinale) plant species. We then collected native halictid bees foraging on each of the three plant species and conducted controlled binary tests to determine their preferences for the flowers of native or non-native plant species. In the field counts, halictid bees visited the native plant significantly more than the non-native species. However, in the behavioural assays when comparing A. strictum vs. A. calendula, Lasioglossum (Chilalictus) lanarium (Family: Halictidae), bees significantly preferred the non-native species, regardless of their foraging history. When comparing A. strictum vs. T. officinale, bees only showed a preference for the non-native flower when it had been collected foraging on the flowers of that plant species immediately prior to the experiment; otherwise, they showed no flower preference. Our results highlight the influence that non-native angiosperms have on native pollinators and we discuss the complexities of the results and the possible reasons for different flower preferences under laboratory and field conditions.

Melıssopalynologıcal study of Apıs Mellıfera L. honey sourced from dıfferent localıtıes in the mıddle belt of Nıgerıa

Melissopalynological studies are useful to determine the floral contents and geographical origin of honey samples. The great diversity of plants allows bees to produce honey that is highly valuable to the consumers. This study was aimed to determine pollen types in honey samples collected from the Middle Belt region to provide information on botanical origin of honeys. Six honey samples were collected from Benue and Kogi States in North Central, Nigeria. According to their pollen spectra, they were multiflora honeys. The honey samples collected from Benue State had a total pollen count of 8, 932, thirty nine pollen types in twenty five (25) families of plant and samples from Kogi State recorded 3,859 pollen, 29 pollen types in nineteen 19 plant families. A total of thirteen pollen types were identified to generic level, fifteen to species and nine to family level in honey samples collected from Benue State whereas in Kogi State samples, eleven pollen types were identified to generic level, eight to species and nine to family level. The major pollen types in Benue honey samples were Sarcocephalus latifolius, Phyllanthus sp., Lannea sp., Hymenocardia acida, Syzygium guineense, Solanum sp., Parkia biglobosa, members of Anacardiaceae, Combretaceae/Melastomataceae, Euphorbiaceae and Sapindaceae whereas Hymenocardia acida, Elaeis guineensis, Solanum sp., Cassia sp., Sarcocephalus latifolius, members of Combretaeaeceae/Melastomataceae and Fabaceae were dominant in samples from Kogi State. The pollen analyses provided important information on the floral source preferences of bees and on honey contents, which can be used by traditional beekeepers and the public in general in promoting the production and consumption of high quality honey in the Middle Belt region of Nigeria.

Honey Bees Prefer Pollen Substitutes Rich in Protein Content Located at Short Distance from the Apiary.

The availability of floral resources is crucial for honey bee colonies because it allows them to obtain protein from pollen and carbohydrates from nectar; typically, they consume these nutrients in the form of bee bread, which has undergone fermentation. However, the intensification of agriculture, urbanization, changes to the topography, and harsh environmental conditions are currently impacting foraging sites due to habitat loss and scarcity of food resources. Thus, this study aimed to assess honey bee preference for various pollen substitute diet compositions. Bee colonies perform poorly because of specific environmental problems, which ultimately result in pollen scarcity. Pollen substitutes located at various distance from the bee hive were also investigated in addition to determining the preferences of honey bees for various pollen substitute diets. The local honey bee (Apis mellifera jemenitica) colonies and different diets (four main treatments, namely, chickpea flour, maize flour, sorghum flour, wheat flour; each flour was further mixed with cinnamon powder, turmeric powder, flour only, flour mixed with both cinnamon and turmeric powder) were used. Bee pollen was used as a control. The best performing pollen substitutes were further placed at 10, 25, and 50 m distances from the apiary. Maximum bee visits were observed on bee pollen (210 ± 25.96) followed by chickpea flour only (205 ± 19.32). However, there was variability in the bee visits to the different diets (F (16,34) = 17.91; p < 0.01). In addition, a significant difference in diet consumption was observed in control (576 ± 58.85 g) followed by chickpea flour only (463.33 ± 42.84 g), compared to rest of the diets (F (16,34) = 29.75; p < 0.01). Similarly, foraging efforts differed significantly (p < 0.01) at the observed time of 7-8 A.M., 11-12 A.M., and 4-5 P.M. at the distance of 10, 25, and 50 m away from the apiary. Honey bees preferred to visit the food source that was closest to the hive. This study should be very helpful for beekeepers in supplementing their bee colonies when there is a shortage or unavailability of pollens, and it is much better to keep the food source near the apiary. Future research needs to highlight the effect of these diets on bee health and colony development.

Western honey bee (Apis mellifera L.) attraction to commercial pollen substitutes and wildflower pollen in vitro

AbstractMany beekeepers feed their western honey bee (Apis mellifera) colonies artificial pollen substitutes to provide colonies with adequate nutrition during times of limited pollen quantity or quality. We provided caged worker bees commercially available pollen substitutes (AP23, MegaBee, UltraBee) and wildflower pollen in a choice‐test to determine their relative attraction to/preference for the diets. We measured diet consumption by honey bees and observed honey bee behaviour to evaluate bee preferences for certain diets. The bees interacted with and consumed more wildflower pollen than they did any of the commercially available pollen substitutes. Our data suggest that bees have a strong preference for wildflower pollen over commercially available pollen substitutes.

Single gene mutation in a plant MYB transcription factor causes a major shift in pollinator preference

Understanding the molecular basis of reproductive isolation and speciation is a key goal of evolutionary genetics. In the South American genus Petunia, the R2R3-MYB transcription factor MYB-FL regulates the biosynthesis of UV-absorbing flavonol pigments, a major determinant of pollinator preference. MYB-FL is highly expressed in the hawkmoth-pollinated P.axillaris, but independent losses of its activity in sister taxa P.secreta and P.exserta led to UV-reflective flowers and associated pollinator shifts in each lineage (bees and hummingbirds, respectively). We created a myb-fl CRISPR mutant in P.axillaris and studied the effect of this single gene on innate pollinator preference. The mutation strongly reduced the expression of the two key flavonol-related biosynthetic genes but only affected the expression of few other genes. The mutant flowers were UV reflective as expected but additionally contained low levels of visible anthocyanin pigments. Hawkmoths strongly preferred the wild-type P.axillaris over the myb-fl mutant, whereas both social and solitary bee preference depended on the level of visible color of the mutants. MYB-FL, with its specific expression pattern, small number of target genes, and key position at the nexus of flavonol and anthocyanin biosynthetic pathways, provides a striking example of evolution by single mutations of large phenotypic effect.