Residue dynamics of a contact and a systemic fungicide in pollen, nectar, and other plant matrices of courgette (Cucurbita pepo L.).

Pollen and nectar can be contaminated with a range of pesticides, including insecticides, fungicides, and herbicides. Since these matrices are important food sources for pollinators and other beneficial insects, their contamination can represent a key route of exposure. However, limited knowledge exists with respect to pesticide residue levels and their dynamics in these matrices for many crops and active ingredients (AIs). We used controlled glasshouse studies to investigate the residue dynamics of a systemic (cyprodinil) and a contact (fludioxonil) fungicide in the floral matrices and other plant parts of courgette/zucchini (Cucurbita pepo L.). We aimed to better understand the processes behind residue accumulation and decline in pollen and nectar. Each AI was applied to plants, either by spraying whole plants or by targeted spraying onto leaves only. Samples of pollen, nectar, anthers, flowers, and leaves were taken on the day of application and each subsequent morning for up to 13 days and analysed for residues using LC-MS/MS. Significant differences in residue levels and dynamics were found between AIs and floral matrices. The present study allowed for the identification of potential routes by which residues translocate between tissues and to link those to the physicochemical properties of each AI, which may facilitate the prediction of residue levels in pollen and nectar. Residues of the contact AI declined more quickly than those of the systemic AI in pollen and nectar. Our results further suggest that the risk of oral exposure for pollinators may be considerably reduced by using contact AIs during the green bud stage of plants, but application of systemic compounds could still result in a low, but continuous long-term exposure for pollinators with limited decline.

Global assessment of honeybee exposure to pesticides through guttation consumption: An indicator approach.

Guttation consumption is a potential pathway of pesticide residue exposure in honeybees. However, modeling tools for assessing honeybee exposure to pesticide residues in guttation drops are lacking. In this study, we propose an indicator-based approach for qualitatively or quantitatively analyzing the guttation-based exposure pathway, allowing us to conduct region-specific pesticide residue exposure assessments for honeybees. Exposure scores (the product of guttation production and residue level scores) were established to compare or rank honeybee exposure to pesticide residues via guttation intake across locations using three specified indicators (i.e., air temperature, relative humidity, and precipitation intensity). Warm, dry regions had high residue level scores (indicating high residue levels in guttation), whereas cold, wet regions had high guttation production scores (indicating high possibilities of guttation formation on leaf surfaces); their exposure scores were a combination of these two values. We evaluated and ranked honeybee exposure to imidacloprid residue across regions in Brazil, China, the United States, and selected European Union member states, revealing that pesticide application in many Brazilian federative units may raise honeybee risks due to high exposure scores. We also compared the guttation pathway to other common exposure pathways (nectar and pollen), suggesting that for some moderately lipophilic compounds, the guttation exposure pathway may not be ignored and should be further evaluated.

A critical review on the accumulation of neonicotinoid insecticides in pollen and nectar: Influencing factors and implications for pollinator exposure.

Neonicotinoids are a class of neuro-active insecticides widely used to protect major crops, primarily because of their broad-spectrum insecticidal activity and low vertebrate toxicity. Owing to their systemic nature, plants readily take up neonicotinoids and translocate them through roots, leaves, and other tissues to flowers (pollen and nectar) that serve as a critical point of exposure to pollinators foraging on treated plants. The growing evidence for potential adverse effects on non-target species, especially pollinators, and persistence has raised serious concerns, as these pesticides are increasingly prevalent in terrestrial and aquatic systems. Despite increasing research efforts, our understanding of the potential toxicity of neonicotinoids and the risks they pose to non-target species remains limited. Therefore, this critical review provides a succinct evaluation of the uptake, translocation, and accumulation processes of neonicotinoids in plants and the factors that may affect the eventual build-up of neonicotinoids in pollen and nectar. The role of plant species, as well as the physicochemical properties and application methods of neonicotinoids is discussed. Potential knowledge gaps are identified, and questions meriting future research are suggested for improving our understanding of the relationship between neonicotinoid residues in plants and exposure to pollinators.

Pesticide mixtures detected in crop and non-target wild plant pollen and nectar.

Cultivation of mass flowering entomophilous crops benefits from the presence of managed and wild pollinators, who visit flowers to forage on pollen and nectar. However, management of these crops typically includes application of pesticides, the presence of which may pose a hazard for pollinators foraging in an agricultural environment. To determine the levels of potential exposure to pesticides, their presence and concentration in pollen and nectar need assessing, both within and beyond the target crop plants. We selected ten pesticide compounds and one metabolite and analysed their occurrence in a crop (Brassica napus) and a wild plant (Rubus fruticosus agg.), which was flowering in field edges. Nectar and pollen from both plants were collected from five spring and five winter sown B. napus fields in Ireland, and were tested for pesticide residues, using QuEChERS and Liquid Chromatography tandem mass spectrometry (LC-MS/MS). Pesticide residues were detected in plant pollen and nectar of both plants. Most detections were from fields with no recorded application of the respective compounds in that year, but higher concentrations were observed in recently treated fields. Overall, more residues were detected in B. napus pollen and nectar than in the wild plant, and B. napus pollen had the highest mean concentration of residues. All matrices were contaminated with at least three compounds, and the most frequently detected compounds were fungicides. The most common compound mixture was comprised of the fungicides azoxystrobin, boscalid, and the neonicotinoid insecticide clothianidin, which was not recently applied on the fields. Our results indicate that persistent compounds like the neonicotinoids, should be continuously monitored for their presence and fate in the field environment. The toxicological evaluation of the compound mixtures identified in the present study should be performed, to determine their impacts on foraging insects that may be exposed to them.

Some bee-pollinated plants provide nutritionally incomplete pollen amino acid resources to their pollinators.

For pollinators such as bees, nectar mainly provides carbohydrates and pollen provides proteins, amino acids, and lipids to cover their nutritional needs. Here, to examine differences in pollinator resources, we compared the amino acid profiles and total amino acid contents of pollen from 32 common entomophilous plants in seven families. Our results showed that the amino acid profiles and contents in pollen samples differed according to the plant family and the chromatography method used, i.e., high-performance liquid chromatography (HPLC) versus ion exchange chromatography (IEX). Pollen from Boraginaceae species had the highest total amino acid contents (361.2-504 µg/mg) whereas pollen from the Malvaceae family had the lowest total amino acid contents (136-243.1 µg/mg). Calculating an amino acid score (AAS) that reflects pollen nutritional quality showed that slightly less than half of the species (19 out of 32) had the maximum nutritional score (AAS = 1) and offered high nutritional quality pollen amino acids for bee pollinators. Though they had high total amino acid contents, the amino acid composition of the studied Boraginaceae species and several members of the Fabaceae was not optimal, as their pollen was deficient in some essential amino acids, resulting in suboptimal amino acid scores (AAS < 0.7). Except for cysteine, the measured amino acid contents were higher using IEX chromatography than using HPLC. IEX chromatography is more robust and is to be preferred over HPLC in future amino acid analyses. Moreover, our observations show that some bee-pollinated species fail to provide complete amino acid resources for their pollinators. Although the implications for pollinator behavior remain to be studied, these deficiencies may force pollinators to forage from different species to obtain all nutritionial requirements.

Modeling pesticide residues in nectar and pollen in support of pesticide exposure assessment for honeybees: A generic modeling approach.

Pesticide residues in nectar and pollen of plants can damage honeybees; however, few modeling approaches have simulated residue levels in nectar and pollen in support of exposure assessment for honeybees. This study introduced a generic modeling approach based on plant uptake models and simple partitioning rules that specifies soil incorporation and foliar spray application scenarios of pesticides and is flexible for conducting variability analysis for various environmental conditions, pesticide application patterns, chemical individuals, and plant varieties. The results indicated that, in general, systemic or moderate lipophilicity (log KOW of ~2.5) pesticides have relatively high simulated residue levels in nectar and pollen because of the enhanced residue uptake process from soil. For non-systemic or highly lipophilic pesticides, the residue uptake via leaf surface deposition pathway can be enhanced, and more residues will be bioaccumulated in pollen than nectar due to a relatively high lipid content of pollen (as compared to nectar), but the overall residue levels in nectar and pollen are lower than systemic or moderately lipophilic pesticides. The variability analysis showed that environmental conditions, pesticide application patterns, chemical properties, and plant varieties cause considerable variations in simulated residue levels in nectar and pollen, indicating that spatiotemporal, chemical, and plant-related factors must be considered in pesticide exposure assessment for honeybees. Moreover, the comparison between the simulated and measured data showed a high degree of consistency, indicating that the proposed model could be used to conduct a screening-level pesticide exposure assessment for honeybees.

Is there a risk to honeybees from use of thiamethoxam as a sugar beet seed treatment?

The ban imposed by the European Union on the use of neonicotinoids as sugar beet seed treatments was based on the exposure of bees to residues of neonicotinoids in pollen and nectar of succeeding crops. To address this concern, residues of thiamethoxam (TMX) and clothianidin (CTD) were analyzed in soil collected from fields planted in at least the previous year with thiamethoxam-treated sugar beet seed. This soil monitoring program was conducted at 94 sites across Germany in two separate years. In addition, a succeeding crop study assessed residues in soil, guttation fluid, pollen, and nectar sampled from untreated succeeding crops planted in the season after thiamethoxam seed-treated sugar beet at eight field sites across five countries. The overall mean residues observed in soil monitoring were 8.0 ± 0.5 µg TMX + CTD/kg in the season after the use of treated sugar beet seed. Residue values decreased with increasing time interval between the latest thiamethoxam or clothianidin application before sugar beet drilling and with lower application frequency. Residues were detected in guttation fluid (2.0-37.7 µg TMX/L); however, the risk to pollinators from this route of exposure is likely to be low, based on the reported levels of consumption. Residues of thiamethoxam and clothianidin in pollen and nectar sampled from the succeeding crops were detected at or below the limit of quantification (0.5-1 µg a.i./kg) in 86.7% of pollen and 98.6% of nectar samples and, unlike guttation fluid residues, were not correlated with measured soil residues. Residues in pollen and nectar are lower than reported sublethal adverse effect concentrations in studies with honeybee and bumble bee individuals and colonies fed only thiamethoxam-treated sucrose, and are lower than those reported to result in no effects in honeybees, bumble bees, and solitary bees foraging on seed-treated crops. Integr Environ Assess Manag 2022;18:709-721. © 2021 SYNGENTA. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Pesticide residues in honeybee-collected pollen: does the EU regulation protect honeybees from pesticides?

Researchers globally identify pesticides as one of the main reasons for pollinator decline. In the European Union (EU), extensive legislation is implemented to protect pollinators from harmful pesticide exposure. The aim of our study was to discover whether the pesticide residue levels in honeybee matrices, such as nectar and pollen, exceeded the chronic or acute toxicity levels when beehives were located next to fields treated with specific insecticides. The insecticides were used according to the EU legislation and its national implementation. The experiments were conducted in turnip rape, oilseed rape, and caraway fields in southern Finland during the years 2019 and 2020. The pesticides used in the experiments contained the active substances lambda-cyhalothrin (2019), esfenvalerate (2020), and tau-fluvalinate (2020). However, the honeybee-collected pollen and nectar were analyzed for residues of more than 100 active substances. The results showed that the pesticide residue levels clearly remained under the oral acute toxicity for honeybees, although we found high levels of thiacloprid residues in the pollen collected in 2019. The pesticide residues in nectar were below LOQ values, which was most likely due to the rainy weather conditions together with the chosen sampling method. No statistically significant differences were observed between the insecticide-treated and untreated fields. In light of our research, the EU legislation protected honeybees from oral acute toxicity during the years 2019 and 2020. However, potential sublethal effects of thiacloprid and other pesticide compounds found in the collected pollen cannot be ruled out. In the future, constant monitoring of pesticide exposure of honeybees and wild pollinators should be established to ensure that pesticide legislation, and its implementation across the EU successfully protects pollinators and their services in agricultural environments.

A chemometric approach for the differentiation of 15 monofloral honeys based on physicochemical parameters.

BACKGROUND: Although the main method for authentication of monofloral honey is pollen analysis, other classification approaches have been also applied. However, the majority of the existing classification models so far have utilized a few honey types or a few honey samples of each honey type, which can lead to inaccurate results. Aiming at addressing this, the goal of the present study was to create a classification model by analysing in total 250 honey samples from 15 different monofloral honey types in ten physicochemical parameters and then, multivariate analysis [multivariate analysis of variance (MANOVA), principal component analysis (PCA) and multi-discriminant analysis (MDA)] was applied in an effort to distinguish and classify them. RESULTS: Electrical conductivity and colour were found to have the highest discriminative power, allowing the classification of monofloral honey types, such as oak, knotgrass and chestnut honey, as well as the differentiation between honeydew and nectar honeys. The classification model had a high predictive power, as the 84.4% of the group cases was correctly classified, while for the cases of chestnut, strawberry tree and sunflower honeys the respective prediction was correct by 91.3%, 95% and 100%, allowing further determination of unknown honey samples. CONCLUSION: It seems that the characterization of monofloral honeys based on their physicochemical parameters through the proposed model can be achieved and further applied on other honey types. The results could contribute to the development of methodologies for the determination of honey's botanical origin, based on simple techniques, so that these can be applied for routine analysis. © 2021 Society of Chemical Industry.

Essential role of papillae flexibility in nectar capture by bees.

Many bees possess a tongue resembling a brush composed of a central rod (glossa) covered by elongated papillae, which is dipped periodically into nectar to collect this primary source of energy. In vivo measurements show that the amount of nectar collected per lap remains essentially constant for sugar concentrations lower than 50% but drops significantly for a concentration around 70%. To understand this variation of the ingestion rate with the sugar content of nectar, we investigate the dynamics of fluid capture by Bombus terrestris as a model system. During the dipping process, the papillae, which initially adhere to the glossa, unfold when immersed in the nectar. Combining in vivo investigations, macroscopic experiments with flexible rods, and an elastoviscous theoretical model, we show that the capture mechanism is governed by the relaxation dynamics of the bent papillae, driven by their elastic recoil slowed down through viscous dissipation. At low sugar concentrations, the papillae completely open before the tongue retracts out of nectar and thus, fully contribute to the fluid capture. In contrast, at larger concentrations corresponding to the drop of the ingestion rate, the viscous dissipation strongly hinders the papillae opening, reducing considerably the amount of nectar captured. This study shows the crucial role of flexible papillae, whose aspect ratio determines the optimal nectar concentration, to understand quantitatively the capture of nectar by bees and how physics can shed some light on the degree of adaptation of a specific morphological trait.

Overview of Analytical Methods for the Determination of Neonicotinoid Pesticides in Honeybee Products and Honeybee.

Neonicotinoid pesticides are widely applied for controlling pests in a variety of agriculture crops. Due to the systemic distribution in plants, neonicotinoid pesticides have been found in nectar and pollen, which are the main source of food for the important pollinator honeybee. The risk of neonicotinoid residues in honeybee products and honeybee has caused great attention since their impacts on the environment, ecology, and food safety issues. These concerns require the accurate and sensitive determination of neonicotinoids and their metabolites in the honeybee products and honeybee. Since the trace residue level of neonicotinoid and the complexity of the samples, analysis of neonicotinoid targets in these important matrices is still a great challenge. The present review provides general overview of analytical methods for the determination of neonicotinoid pesticides and their metabolites in honeybee products and honeybee.

Reduced Water Negatively Impacts Social Bee Survival and Productivity Via Shifts in Floral Nutrition.

Pollinators provide a key ecosystem service vital for the survival and stability of the biosphere. Identifying factors influencing the plant-pollinator mutualism and pollinator management is necessary for maintaining a healthy ecosystem. Since healthy beehives require substantial amounts of carbohydrates (nectar) and protein (pollen) from forage plants such as clover, we must assess how resources offered by plants change under limited water conditions in order to fully understand how drought modifies the pollination mutualism. Here we document how reduced water availability leads to decreased nectar quality and quantity and decreased protein quality of pollen. Furthermore, we provide conclusive evidence that these lower quality resources lead to decreased survival and productivity in both developing honey bees (Hymenoptera: Apidae) and bumble bees (Hymenoptera: Apidae). The results emphasize the importance of the nutritional effects of reduced water on bees when predicting shifts of pollination mutualisms under climate change.

Low anthocyanin plum nectar does not impact cognition, blood pressure and gut microbiota in healthy older adults: A randomized crossover trial.

Queen Garnet plum (QGP), known for its high levels of anthocyanins, is a hybrid of the Japanese plum developed in Queensland, Australia. Anthocyanins provide the red, blue, and purple pigments in plants with demonstrated beneficial health effects. This study hypothesized that low-dose anthocyanin QGP intake will have a significant positive effect on cognition, blood pressure, and gut microbiota in healthy older adults. A randomized crossover trial was conducted to determine the effect and within subject variance on cognition and 24 hr. ambulatory blood pressure in older adults without cognitive impairment following daily consumption of 200 mL low-dose anthocyanin (5 mg/100 g) QGP nectar (intervention) or raspberry cordial (control). Secondary outcomes included inflammatory markers (C-reactive protein), nerve growth factor (BDNF), and gut microbiota (16S rRNA gene sequencing). Twenty-eight participants (55+ years) were recruited. Each randomized treatment arm lasted for 8 weeks with a 4-week washout period. Cognition, blood pressure, and urine samples were measured at each visit (5 total) while blood and fecal samples were collected at baseline, 8 weeks, and 20 weeks. Repeated-measures ANOVA was used to analyze the data. Across the treatments, no significant difference was observed for the different domains of cognition, blood pressure, or anti-inflammatory biomarkers. No intervention effect was found for genera or class of gut microbes. Low anthocyanin nectar derived from the QGP did not have any significant effects on cognition, blood pressure, or gut microbiota in healthy older adults.

Herbivory and Time Since Flowering Shape Floral Rewards and Pollinator-Pathogen Interactions.

Herbivory can induce chemical changes throughout plant tissues including flowers, which could affect pollinator-pathogen interactions. Pollen is highly defended compared to nectar, but no study has examined whether herbivory affects pollen chemistry. We assessed the effects of leaf herbivory on nectar and pollen alkaloids in Nicotiana tabacum, and how herbivory-induced changes in nectar and pollen affect pollinator-pathogen interactions. We damaged leaves of Nicotiana tabacum using the specialist herbivore Manduca sexta and compared nicotine and anabasine concentrations in nectar and pollen. We then pooled nectar and pollen by collection periods (within and after one month of flowering), fed them in separate experiments to bumble bees (Bombus impatiens) infected with the gut pathogen Crithidia bombi, and assessed infections after seven days. We did not detect alkaloids in nectar, and leaf damage did not alter the effect of nectar on Crithidia counts. In pollen, herbivory induced higher concentrations of anabasine but not nicotine, and alkaloid concentrations rose and then fell as a function of days since flowering. Bees fed pollen from damaged plants had Crithidia counts 15 times higher than bees fed pollen from undamaged plants, but only when pollen was collected after one month of flowering, indicating that both damage and time since flowering affected interaction outcomes. Within undamaged treatments, bees fed late-collected pollen had Crithidia counts 10 times lower than bees fed early-collected pollen, also indicating the importance of time since flowering. Our results emphasize the role of herbivores in shaping pollen chemistry, with consequences for interactions between pollinators and their pathogens.

Male flowers of Aconitum compensate for toxic pollen with increased floral signals and rewards for pollinators.

Many plants require animal pollinators for successful reproduction; these plants provide pollinator resources in pollen and nectar (rewards) and attract pollinators by specific cues (signals). In a seeming contradiction, some plants produce toxins such as alkaloids in their pollen and nectar, protecting their resources from ineffective pollinators. We investigated signals and rewards in the toxic, protandrous bee-pollinated plant Aconitum napellus, hypothesizing that male-phase flower reproductive success is pollinator-limited, which should favour higher levels of signals (odours) and rewards (nectar and pollen) compared with female-phase flowers. Furthermore, we expected insect visitors to forage only for nectar, due to the toxicity of pollen. We demonstrated that male-phase flowers emitted more volatile molecules and produced higher volumes of nectar than female-phase flowers. Alkaloids in pollen functioned as chemical defences, and were more diverse and more concentrated compared to the alkaloids in nectar. Visitors actively collected little pollen for larval food but consumed more of the less-toxic nectar. Toxic pollen remaining on the bee bodies promoted pollen transfer efficiency, facilitating pollination.

Analysis on safety assessment of Tephrosia vogelii Hook to Apis cerana cerana.

Tephrosia vogelii Hook was excellent insecticidal plant, it was introduced into China and planted over a large area in Guangdong province. The main active components of T. vogelii was rotenone and it widely found in leaves and pods of T. vogelii. This paper study of the safety assessment of T. vogelii flowers to worker bees. In this paper, the content of rotenone in T. vogelii petal, nectar, pollen, pistil, and stamen samples were investigated by HPLC, and tested the toxicity of T. vogelii flowers for Apis cerana cerana during 24 h. The dissipation and dynamic of rotenone in A. c. cerana different biological compartments were investigated under indoor conditions during 24 h. The results showed, The LT50 of T. vogelii flowers to worker bees were collected from the eastern, western, southern, northern and top were 13.95, 24.17, 12.55, 26.48, and 18.84 h, the haemolymph of worker bees have the highest content of rotenone, the least accumulation of rotenone in workers bee's thorax, and the rate of dissipation was slowly during the whole study. In conclusion, the results showed the T. vogelii create security risks to worker bees under some ecosystems.

Nectar quality changes the ecological costs of chemically defended pollen.

Plants often compete in a marketplace that involves the exchange of floral rewards for pollination service [1]. This marketplace is frequently viewed as revolving around a single currency, typically nectar. While this focus has established pollinators such as bees as classic models in foraging ecology, in reality many plants provide both pollen and nectar, which vary in composition within and across species [2]. How this complexity impacts interactions between plants, pollinators, and co-flowering competitors is unknown. We explored how variation in two axes of reward chemistry - nectar sugar and pollen alkaloid content - impacted competition for bumblebee visits. The effect of variation in one reward depended on the presence and quality of the other - bees discriminated against flowers with more defended pollen when all flowers offered the same quality nectar. However, bees preferred flowers with highly defended pollen when they offered higher quality nectar, suggesting that attractive nectar can overcome the ecological costs of defended pollen. Recognizing the interdependence of these floral currencies may help identify traits that drive indirect interactions between plants and clarify broader evolutionary patterns of floral reward phenotypes.

Flowers and climate change: a metabolic perspective.

Adverse climatic conditions at the time of flowering severely hinder crop yields and threaten the interactions between plants and their pollinators. These features depend on a common trait: the metabolism of flowers. In this Viewpoint article, we aim to provide insight into the metabolic changes that occur in flowers in response to changes in climate and emphasize that these changes severely impact the fitness of autogamous and allogamous species, plant-pollinator interactions, and overall ecosystem health. We review the biochemical processes that lead to failure of gamete development and to alterations of color, scent and nectar secretion. Then, making use of open access expression data, we examine the expression of genes that may drive these changes in response to heat and drought. Finally, we present measurements of metabolites from flowers exposed to a heat wave and discuss how the results of this short-term experiment may give rise to misleading conclusions regarding the positive effect of heat on flower fitness. We hope this article draws attention to this often-neglected dynamic and its important consequences.

A matter of taste: the adverse effect of pollen compounds on the pre-ingestive gustatory experience of sugar solutions for honeybees.

In addition to sugars, nectar contains multiple nutrient compounds in varying concentrations, yet little is known of their effect on the reward properties of nectar and the resulting implications for insect behaviour. We examined the pre-ingestive responses of honeybees to sucrose solutions containing a mix of pollen compounds, the amino acids proline or phenylalanine, or known distasteful substances, quinine and salt. We predicted that in taste and learning assays, bees would respond positively to the presence of nutrient compounds in a sucrose solution. However, bees' proboscis extension responses decreased when their antennae were stimulated with pollen- or amino acid-supplemented sucrose solutions. Compared to pure sucrose, bees exhibited worse acquisition when conditioned to an odour with pollen-supplemented sucrose as the unconditioned stimulus. Such learning impairment was also observed with quinine-containing sucrose solutions. Our results suggest that bees can use their antennae to detect pollen compounds in floral nectars. Depending on the type and concentrations of compounds present, this may result in nectar being perceived as distasteful by bees, making it less effective in reinforcing the learning of floral cues. Such reward devaluation might be adaptive in cases where plants benefit from regulating the frequency of bee visitation.

Bumblebee Rejection of Toxic Pollen Facilitates Pollen Transfer.

Many bees are effective pollen collectors; however, pollen grains collected by bees for larval food are lost for plant sexual reproduction. Recognition of these conflicting interests between bees and flowers is essential for understanding of reproduction for both bees and flowers [1-3]. Plant defense compounds in pollen may function to reduce pollen waste by deterring ineffective pollinators [4-6], but this hypothesis remains unexamined. Here, we provide evidence that secondary metabolites in pollen function as chemical defense by deterring some bees from gathering pollen. In two Dipsacus species, a defense compound, dipsacus saponin [7], occurs in pollen but not in nectar. We observed that bumblebees disliked grooming bitter-tasting pollen with a high saponin content. Manipulation of saponin concentrations in nectar and measurements of corbicular pollen showed that the bumblebee species differed in their tolerance to saponin. Those species susceptible to saponin groomed little Dipsacus pollen into their pollen loads, and their ungroomed pollen was observed to be effectively delivered to stigmas. By rewarding bees with edible nectar, but not pollen, plants solve the conflict of pollen partitioning between sexual and reward functions. Ungroomed toxic pollen on the bee body promotes pollen transfer efficiency, facilitating pollination.