Pistil Mating Type and Morphology Are Mediated by the Brassinosteroid Inactivating Activity of the S-Locus Gene BAHD in Heterostylous Turnera Species.

Heterostyly is a breeding system that promotes outbreeding through a combination of morphological and physiological floral traits. In Turnera these traits are governed by a single, hemizygous S-locus containing just three genes. We report that the S-locus gene, BAHD, is mutated and encodes a severely truncated protein in a self-compatible long homostyle species. Further, a self-compatible long homostyle mutant possesses a T. krapovickasii BAHD allele with a point mutation in a highly conserved domain of BAHD acyl transferases. Wild type and mutant TkBAHD alleles were expressed in Arabidopsis to assay for brassinosteroid (BR) inactivating activity. The wild type but not mutant allele caused dwarfism, consistent with the wild type possessing, but the mutant allele having lost, BR inactivating activity. To investigate whether BRs act directly in self-incompatibility, BRs were added to in vitro pollen cultures of the two mating types. A small morph specific stimulatory effect on pollen tube growth was found with 5 µM brassinolide, but no genotype specific inhibition was observed. These results suggest that BAHD acts pleiotropically to mediate pistil length and physiological mating type through BR inactivation, and that in regard to self-incompatibility, BR acts by differentially regulating gene expression in pistils, rather than directly on pollen.

Productivity performance of peach trees, insecticidal and antibacterial bioactivities of leaf extracts as affected by nanofertilizers foliar application.

The current study was performed on eight years old peach (Prunus persica L. Batsch) trees cv. Florida prince to study the influence of spraying of commercial nano fertilizer on vegetative growth, pollen grain viability, yield, and fruit quality of the "Florida prince" peach cultivar. Furthermore, extracts from the nanofertilizer treated leaves were studied for their bioactivity as insecticidal or bactericidal activities against some stored grain insects and plant bacterial pathogens. Seventy uniform peach trees were sprayed three time as follow: before flowering; during full bloom, and one month later in addition using the water as a control. Commercial silver particales (Ag NPs) at 10, 12.5, and 15 mL/L and zinc particales (Zn NPs) at 2.5, 5 and 7.5 mL/L as recommended level in a randomized complete block design in ten replicates/trees. Spraying Ag NP at 15 mL/L increased shoot diameter, leaf area, total chlorophyll, flower percentage, fruit yield and fruit physical and chemical characteristics, followed by Ag NPs at 12.5 mL/L and Zn NPs at 7.5 mL/L. Moreover, Zn and Ag NPs caused a highly significant effect on pollen viability. Different type of pollen aberrations were detected by Zn NPs treatment. The commercial Ag NPs showed a high increase in pollen viability without any aberrations. The Ag NPs significantly increased the pollen size, and the spores also increased and separated in different localities, searching about the egg for pollination and fertilization. Peach leaves extract was examined for their insecticidal activity against rice weevil (Sitophilus oryzea L.) and the lesser grain borer (Rhyzopertha dominica, Fabricius) by fumigation method. The antibacterial activity of all treatments was also performed against molecularly identified bacteria. Ag NPs treated leaves extract at concentration 3000 µg/mL were moderate sufficient to inhibit all the bacterial isolates with inhibition zone (IZ) ranged 6-8.67 mm with high efficiency of acetone extracts from leaves treated with Ag NPs compared with Zn NPs. Also, S. oryzae was more susceptible to acetone extracts from leaves treated with both nanomaterials than R. dominica.

Potential Risk to Pollinators from Nanotechnology-Based Pesticides.

The decline in populations of insect pollinators is a global concern. While multiple factors are implicated, there is uncertainty surrounding the contribution of certain groups of pesticides to losses in wild and managed bees. Nanotechnology-based pesticides (NBPs) are formulations based on multiple particle sizes and types. By packaging active ingredients in engineered particles, NBPs offer many benefits and novel functions, but may also exhibit different properties in the environment when compared with older pesticide formulations. These new properties raise questions about the environmental disposition and fate of NBPs and their exposure to pollinators. Pollinators such as honey bees have evolved structural adaptations to collect pollen, but also inadvertently gather other types of environmental particles which may accumulate in hive materials. Knowledge of the interaction between pollinators, NBPs, and other types of particles is needed to better understand their exposure to pesticides, and essential for characterizing risk from diverse environmental contaminants. The present review discusses the properties, benefits and types of nanotechnology-based pesticides, the propensity of bees to collect such particles and potential impacts on bee pollinators.

Floral Odors and the Interaction between Pollinating Ceratopogonid Midges and Cacao.

Most plant species depend upon insect pollination services, including many cash and subsistence crops. Plants compete to attract those insects using visual cues and floral odor which pollinators associate with a reward. The cacao tree, Theobroma cacao, has a highly specialized floral morphology permitting pollination primarily by Ceratopogonid midges. However, these insects do not depend upon cacao flowers for their life cycle, and can use other sugar sources. To understand how floral cues mediate pollination in cacao we developed a method for rearing Ceratopogonidae through several complete lifecycles to provide material for bioassays. We carried out collection and analysis of cacao floral volatiles, and identified a bouquet made up exclusively of saturated and unsaturated, straight-chain hydrocarbons, which is unusual among floral odors. The most abundant components were tridecane, pentadecane, (Z)-7-pentadecene and (Z)-8-heptadecene with a heptadecadiene and heptadecatriene as minor components. We presented adult midges, Forcipomyia sp. (subgen. Forcipomyia), Culicoides paraensis and Dasyhelea borgmeieri, with natural and synthetic cacao flower odors in choice assays. Midges showed weak attraction to the complete natural floral odor in the assay, with no significant evidence of interspecific differences. This suggests that cacao floral volatiles play a role in pollinator behavior. Midges were not attracted to a synthetic blend of the above four major components of cacao flower odor, indicating that a more complete blend is required for attraction. Our findings indicate that cacao pollination is likely facilitated by the volatile blend released by flowers, and that the system involves a generalized odor response common to different species of Ceratopogonidae.

Chronic oral exposure to field-realistic pesticide combinations via pollen and nectar: effects on feeding and thermal performance in a solitary bee.

Pesticide use is one of the main causes of pollinator declines in agricultural ecosystems. Traditionally, most laboratory studies on bee ecotoxicology test acute exposure to single compounds. However, under field conditions, bees are often chronically exposed to a variety of chemicals, with potential synergistic effects. We studied the effects of field-realistic concentrations of three pesticides measured in pollen and nectar of commercial melon fields on the solitary bee Osmia bicornis L. We orally exposed females of this species throughout their life span to 8 treatments combining two neonicotinoid insecticides (acetamiprid, imidacloprid) and a triazole fungicide (myclobutanil) via pollen and sugar syrup. We measured pollen and syrup consumption, longevity, ovary maturation and thermogenesis. Pesticide intake was three orders of magnitude higher via syrup than pollen. At the tested concentrations, no synergistic effects emerged, and we found no effects on longevity and ovary maturation. However, all treatments containing imidacloprid resulted in suppressed syrup consumption and drastic decreases in thoracic temperature and bee activity. Our results have important implications for pesticide regulation. If we had measured only lethal effects we would have wrongly concluded that the pesticide combinations containing imidacloprid were safe to O. bicornis. The incorporation of tests specifically intended to detect sublethal effects in bee risk assessment schemes should be an urgent priority. In this way, the effects of pesticide exposure on the dynamics of bee populations in agroecosystems will be better assessed.

A review of the factors that influence pesticide residues in pollen and nectar: Future research requirements for optimising the estimation of pollinator exposure.

In recent years, the impact of Plant Protection Products (PPPs) on insect pollinator decline has stimulated significant amounts of research, as well as political and public interest. PPP residues have been found in various bee-related matrices, resulting in governmental bodies worldwide releasing guidance documents on methods for the assessment of the overall risk of PPPs to different bee species. An essential part of these risk assessments are PPP residues found in pollen and nectar, as they represent a key route of exposure. However, PPP residue values in these matrices exhibit large variations and are not available for many PPPs and crop species combinations, which results in inaccurate estimations and uncertainties in risk evaluation. Additionally, residue studies on pollen and nectar are expensive and practically challenging. An extrapolation between different cropping scenarios and PPPs is not yet justified, as the behaviour of PPPs in pollen and nectar is poorly understood. Therefore, this review aims to contribute to a better knowledge and understanding of the fate of PPP residues in pollen and nectar and to outline knowledge gaps and future research needs. The literature suggests that four primary factors, the crop type, the application method, the physicochemical properties of a compound and the environmental conditions have the greatest influence on PPP residues in pollen and nectar. However, these factors consist of many sub-factors and initial effects may be disguised by different sampling methodologies, impeding their exact characterisation. Moreover, knowledge about these factors is ambiguous and restricted to a few compounds and plant species. We propose that future research should concentrate on identifying relationships and common features amongst various PPP applications and crops, as well as an overall quantification of the described parameters; in order to enable a reliable estimation of PPP residues in pollen, nectar and other bee matrices.

Toxicity and side effects of some insecticides applied in cotton fields on Apis mellifera.

Honeybee (Apis mellifera L.) provides not only bee products of immense value but also render invaluable free service as cross-pollination and propagation of several cultivated and wild species, thereby, maintaining biological diversity. Bee larvae and adults might be killed or suffer various sublethal effects when placed in contact with pollen and nectar contaminated with insecticides. The present work was conducted to investigate the toxicity of seven insecticides on laboratory using oral toxicity test and their side effects on A. mellifera in cotton fields. Results indicated that lambda-cyhalothrin was the most toxic-tested pesticide, recording the lowest LC50 and LC90 values at all tested periods and the lowest LT50 and LT90 at all tested concentrations, followed by abamectin, spinosad, chlorpyrifos, and emamectin benzoate. On the other side, dipel and pyridalyl recording the highest LC50 and LC90 at all tested periods and the highest LT50 and LT90 at all tested concentrations. As for the application of pesticides in cotton fields, the tested pesticides significantly increased the number of dead workers in comparison with control. The tested pesticides significantly decreased bee foraging activities, i.e., number of foraging workers, number of worker collecting nectar, number of worker gathering pollen grains, area of broad workers, and honey bee yields. Dipel and pyridalyl were the most safety pesticides on honey bee workers in laboratory and field, so it could be introduced as a component in IPM programs of cotton pests.

Endocrine disruption and chronic effects of plant protection products in bees: Can we better protect our pollinators?

Exposure to plant protection products (PPPs) is one of the causes for the population decline of pollinators. In addition to direct exposure, pollinators are exposed to PPPs by pollen, nectar and honey that often contain residues of multiple PPPs. While in legislation PPPs are regarded mainly for their acute toxicity in bees, other effects such as neurotoxicity, immunotoxicity, behavioural changes, stress responses and chronic effects that may harm different physiologically and ecologically relevant traits are much less or not regarded. Despite the fact that endocrine disruption by PPPs is among key effects weakening survival and thriving of populations, pollinators have been poorly investigated in this regard. Here we summarize known endocrine disruptive effects of PPPs in bees and compare them to other chronic effects. Endocrine disruption in honey bees comprise negative effects on reproductive success of queens and drones and behavioural transition of nurse bees to foragers. Among identified PPPs are insecticides, including neonicotinoids, fipronil, chlorantraniliprole and azadirachtin. So far, there exists no OECD guideline to investigate possible endocrine effects of PPPs. Admittedly, investigation of effects on reproduction success of queens and drones is rarely possible under laboratory conditions. But the behavioural transition of nurse bees to foragers could be a possible endpoint to analyse endocrine effects of PPPs under laboratory conditions. We identified some genes, including vitellogenin, which regulate this transition and which may be used as biomarkers for endocrine disruptive PPPs. We plea for a better implementation of the adverse outcome pathway concept into bee's research and propose a procedure for extending and complementing current assessments, including OECD guidelines, with additional physiological and molecular endpoints. Consequently, assessing potential endocrine disruption in pollinators should receive much more relevance.

Neonicotinoid pesticide limits improvement in buzz pollination by bumblebees.

Neonicotinoid pesticides have been linked to global declines of beneficial insects such as bumblebees. Exposure to trace levels of these chemicals causes sub-lethal effects, such as reduced learning and foraging efficiency. Complex behaviours may be particularly vulnerable to the neurotoxic effects of neonicotinoids. Such behaviours may include buzz pollination (sonication), in which pollinators, usually bees, use innate and learned behaviours to generate high-frequency vibrations to release pollen from flowers with specialised anther morphologies. This study assesses the effect of field-realistic, chronic exposure to the widely-used neonicotinoid thiamethoxam on the development of sonication buzz characteristics over time, as well as the collection of pollen from buzz-pollinated flowers. We found that the pollen collection of exposed bees improved less with increasing experience than that of unexposed bees, with exposed bees collecting between 47% and 56% less pollen by the end of 10 trials. We also found evidence of two distinct strategies for maximising pollen collection: (1) extensions to the duration of individual buzzes and (2) extensions of the overall time spent buzzing. We find new complexities in buzz pollination, and conclude that the impacts of field-realistic exposure to a neonicotinoid pesticide may seriously compromise this important ecosystem service.

Translocation of heavy metals from soils into floral organs and rewards of Cucurbita pepo: Implications for plant reproductive fitness.

Metals and metalloids in soil could be transferred into reproductive organs and floral rewards of hyperaccumulator plants and influence their reproductive success, yet little is known whether non-hyperaccumulator plants can translocate heavy metals from soil into their floral organs and rewards (i.e., nectar and pollen) and, if so, whether plant reproduction will be affected. In our studies, summer squash (Cucurbita pepo L. cv. Golden Apple) was exposed to heavy-metal treatments during bud stage to investigate the translocation of soil-supplemented zinc, copper, nickel and lead into its floral organs (pistil, anther and nectary) and rewards (nectar and pollen) as well as floral metal accumulation effects on its reproduction. The results showed that metals taken up by squash did translocate into its floral organs and rewards, although metal accumulation varied depending on different metal types and concentrations as well as floral organ/reward types. Mean foraging time of honey bees to each male and female flower of squash grown in metal-supplemented soils was shorter relative to that of plants grown in control soils, although the visitation rate of honeybees to both male and female flowers was not affected by metal treatments. Pollen viability, pollen removal and deposition as well as mean mass per seed produced by metal-treated squash that received pollen from plants grown in control soils decreased with elevated soil-supplemented metal concentrations. The fact that squash could translocate soil-supplemented heavy metals into floral organs and rewards indicated possible reproductive consequences caused either directly (i.e., decreasing pollen viability or seed mass) or indirectly (i.e., affecting pollinators' visitation behavior to flowers) to plant fitness.

Ornamental plants on sale to the public are a significant source of pesticide residues with implications for the health of pollinating insects.

Garden centres frequently market nectar- and pollen-rich ornamental plants as "pollinator-friendly", however these plants are often treated with pesticides during their production. There is little information on the nature of pesticide residues present at the point of purchase and whether these plants may actually pose a threat to, rather than benefit, the health of pollinating insects. Using mass spectrometry analyses, this study screened leaves from 29 different 'bee-friendly' plants for 8 insecticides and 16 fungicides commonly used in ornamental production. Only two plants (a Narcissus and a Salvia variety) did not contain any pesticide and 23 plants contained more than one pesticide, with some species containing mixtures of 7 (Ageratum houstonianum) and 10 (Erica carnea) different agrochemicals. Neonicotinoid insecticides were detected in more than 70% of the analysed plants, and chlorpyrifos and pyrethroid insecticides were found in 10% and 7% of plants respectively. Boscalid, spiroxamine and DMI-fungicides were detected in 40% of plants. Pollen samples collected from 18 different plants contained a total of 13 different pesticides. Systemic compounds were detected in pollen samples at similar concentrations to those in leaves. However, some contact (chlorpyrifos) and localised penetrant pesticides (iprodione, pyroclastrobin and prochloraz) were also detected in pollen, likely arising from direct contamination during spraying. The neonicotinoids thiamethoxam, clothianidin and imidacloprid and the organophosphate chlorpyrifos were present in pollen at concentrations between 6.9 and 81 ng/g and at levels that overlap with those known to cause harm to bees. The net effect on pollinators of buying plants that are a rich source of forage for them but simultaneously risk exposing them to a cocktail of pesticides is not clear. Gardeners who wish to gain the benefits without the risks should seek uncontaminated plants by growing their own from seed, plant-swapping or by buying plants from an organic nursery.

Does particulate matter along roadsides interfere with plant reproduction? A comparison of effects of different road types on Cichorium intybus pollen deposition and germination.

The roadside habitat can be challenging for plants, which must maintain normal biological processes despite an influx of airborne pollutants. While the effects of many gases on plants have been quantified, the impacts of particulate pollutants have been relatively less studied. This is especially true of field experiments where particle dispersion may be influenced by meteorology and roadway use. We examined chicory (Cichorium intybus L.) along roadsides in the Cincinnati, Ohio metropolitan area to assess particulate influence on plant pollination through stigmatic clogging. We compared flowers collected from plants situated along interstates, U.S. highways, state highways, and county roads as these different road-types vary in motor vehicle usage and thus should have varying levels of particulate deposition on flowers. We examined floral stigmas for total particulates, total pollen, and percentage of pollen tube germination to determine whether particulates may interfere with early reproductive processes. Our results suggest that there was minimal variation of particulate matter found on chicory stigmas among road-types. Furthermore, the deposition of particulates on stigmas based on road-type did not show a strong link to variation in pollen deposition and pollen germination. There was also no significant relationship between total particulate levels and pollen germination rates across all road types. Future studies should investigate other plant species that may be more sensitive to roadside pollution, such as economically important crops. Locations in which vehicle use is increasing and where pollutants are not regulated strictly should also be examined as the effects of airborne particulates in early plant reproduction would be expected to be more substantial in these areas.

Telipogon peruvianus (Orchidaceae) Flowers Elicit Pre-Mating Behaviour in Eudejeania (Tachinidae) Males for Pollination.

Several neotropical orchid genera have been proposed as being sexually deceptive; however, this has been carefully tested in only a few cases. The genus Telipogon has long been assumed to be pollinated by male tachinid flies during pseudocopulatory events but no detailed confirmatory reports are available. Here, we have used an array of methods to elucidate the pollination mechanism in Telipogon peruvianus. The species presents flowers that have a mean floral longevity of 33 days and that are self-compatible, although spontaneous self-pollination does not occur. The flowers attract males of four tachinid species but only the males of an undescribed Eudejeania (Eudejeania aff. browni; Tachinidae) species are specific pollinators. Males visit the flowers during the first few hours of the day and the pollination success is very high (42% in one patch) compared with other sexually deceptive species. Female-seeking males are attracted to the flowers but do not attempt copulation with the flowers, as is usually described in sexually deceptive species. Nevertheless, morphological analysis and behavioural tests have shown an imperfect mimicry between flowers and females suggesting that the attractant stimulus is not based only on visual cues, as long thought. Challenging previous conclusions, our chemical analysis has confirmed that flowers of Telipogon release volatile compounds; however, the role of these volatiles in pollinator behaviour remains to be established. Pollinator behaviour and histological analyses indicate that Telipogon flowers possess scent-producing structures throughout the corolla. Our study provides the first confirmed case of (i) a sexually deceptive species in the Onciidinae, (ii) pollination by pre-copulatory behaviour and (iii) pollination by sexual deception involving tachinid flies.

Large-scale monitoring of effects of clothianidin-dressed oilseed rape seeds on pollinating insects in northern Germany: residues of clothianidin in pollen, nectar and honey.

This study was part of a large-scale monitoring project to assess the possible effects of Elado® (10 g clothianidin & 2 g ß-cyfluthrin/kg seed)-dressed oilseed rape seeds on different pollinators in Northern Germany. Firstly, residues of clothianidin and its active metabolites thiazolylnitroguanidine and thiazolylmethylurea were measured in nectar and pollen from Elado®-dressed (test site, T) and undressed (reference site, R) oilseed rape collected by honey bees confined within tunnel tents. Clothianidin and its metabolites could not be detected or quantified in samples from R fields. Clothianidin concentrations in samples from T fields were 1.3 ± 0.9 µg/kg and 1.7 ± 0.9 µg/kg in nectar and pollen, respectively. Secondly, pollen and nectar for residue analyses were sampled from free flying honey bees, bumble bees and mason bees, placed at six study locations each in the R and T sites at the start of oilseed rape flowering. Honey samples were analysed from all honey bee colonies at the end of oilseed rape flowering. Neither clothianidin nor its metabolites were detectable or quantifiable in R site samples. Clothianidin concentrations in samples from the T site were below the limit of quantification (LOQ, 1.0 µg/kg) in most pollen and nectar samples collected by bees and 1.4 ± 0.5 µg/kg in honey taken from honey bee colonies. In summary, the study provides reliable semi-field and field data of clothianidin residues in nectar and pollen collected by different bee species in oilseed rape fields under common agricultural conditions.

Honeybees Produce Millimolar Concentrations of Non-Neuronal Acetylcholine for Breeding: Possible Adverse Effects of Neonicotinoids.

The worldwide use of neonicotinoid pesticides has caused concern on account of their involvement in the decline of bee populations, which are key pollinators in most ecosystems. Here we describe a role of non-neuronal acetylcholine (ACh) for breeding of Apis mellifera carnica and a so far unknown effect of neonicotinoids on non-target insects. Royal jelly or larval food are produced by the hypopharyngeal gland of nursing bees and contain unusually high ACh concentrations (4-8 mM). ACh is extremely well conserved in royal jelly or brood food because of the acidic pH of 4.0. This condition protects ACh from degradation thus ensuring delivery of intact ACh to larvae. Raising the pH to ≥5.5 and applying cholinesterase reduced the content of ACh substantially (by 75-90%) in larval food. When this manipulated brood was tested in artificial larval breeding experiments, the survival rate was higher with food supplemented by 100% with ACh (6 mM) than with food not supplemented with ACh. ACh release from the hypopharyngeal gland and its content in brood food declined by 80%, when honeybee colonies were exposed for 4 weeks to high concentrations of the neonicotinoids clothianidin (100 parts per billion [ppb]) or thiacloprid (8,800 ppb). Under these conditions the secretory cells of the gland were markedly damaged and brood development was severely compromised. Even field-relevant low concentrations of thiacloprid (200 ppb) or clothianidin (1 and 10 ppb) reduced ACh level in the brood food and showed initial adverse effects on brood development. Our findings indicate a hitherto unknown target of neonicotinoids to induce adverse effects on non-neuronal ACh which should be considered when re-assessing the environmental risks of these compounds. To our knowledge this is a new biological mechanism, and we suggest that, in addition to their well documented neurotoxic effects, neonicotinoids may contribute to honeybee colony losses consecutive to a reduction of the ACh content in the brood food.

Effects of the herbicide dicamba on nontarget plants and pollinator visitation.

Nearly 80% of all pesticides applied to row crops are herbicides, and these applications pose potentially significant ecotoxicological risks to nontarget plants and associated pollinators. In response to the widespread occurrence of weed species resistant to glyphosate, biotechnology companies have developed crops resistant to the synthetic-auxin herbicides dicamba and 2,4-dichlorophenoxyacetic acid (2,4-D); and once commercialized, adoption of these crops is likely to change herbicide-use patterns. Despite current limited use, dicamba and 2,4-D are often responsible for injury to nontarget plants; but effects of these herbicides on insect communities are poorly understood. To understand the influence of dicamba on pollinators, the authors applied several sublethal, drift-level rates of dicamba to alfalfa (Medicago sativa L.) and Eupatorium perfoliatum L. and evaluated plant flowering and floral visitation by pollinators. The authors found that dicamba doses simulating particle drift (≈1% of the field application rate) delayed onset of flowering and reduced the number of flowers of each plant species; however, plants that did flower produced similar-quality pollen in terms of protein concentrations. Further, plants affected by particle drift rates were visited less often by pollinators. Because plants exposed to sublethal levels of dicamba may produce fewer floral resources and be less frequently visited by pollinators, use of dicamba or other synthetic-auxin herbicides with widespread planting of herbicide-resistant crops will need to be carefully stewarded to prevent potential disturbances of plant and beneficial insect communities in agricultural landscapes.

Neonicotinoid pesticide exposure impairs crop pollination services provided by bumblebees.

Recent concern over global pollinator declines has led to considerable research on the effects of pesticides on bees. Although pesticides are typically not encountered at lethal levels in the field, there is growing evidence indicating that exposure to field-realistic levels can have sublethal effects on bees, affecting their foraging behaviour, homing ability and reproductive success. Bees are essential for the pollination of a wide variety of crops and the majority of wild flowering plants, but until now research on pesticide effects has been limited to direct effects on bees themselves and not on the pollination services they provide. Here we show the first evidence to our knowledge that pesticide exposure can reduce the pollination services bumblebees deliver to apples, a crop of global economic importance. Bumblebee colonies exposed to a neonicotinoid pesticide provided lower visitation rates to apple trees and collected pollen less often. Most importantly, these pesticide-exposed colonies produced apples containing fewer seeds, demonstrating a reduced delivery of pollination services. Our results also indicate that reduced pollination service delivery is not due to pesticide-induced changes in individual bee behaviour, but most likely due to effects at the colony level. These findings show that pesticide exposure can impair the ability of bees to provide pollination services, with important implications for both the sustained delivery of stable crop yields and the functioning of natural ecosystems.

A new pollinating seed-consuming mutualism between Rheum nobile and a fly fungus gnat, Bradysia sp., involving pollinator attraction by a specific floral compound.

Pollinating seed-consuming mutualisms are regarded as exemplary models for studying coevolution, but they are extremely rare. In these systems, olfactory cues have been thought to play an important role in facilitating encounters between partners. We present a new pollinating seed-consuming mutualism from the high Himalayas between the endemic herb, Rheum nobile, and a fly fungus gnat, Bradysia sp. Seed production resulting from pollination by Bradysia flies and seed consumption by their larvae were measured to determine the outcome of this interaction. Floral scent analyses and behavioural tests were conducted to investigate the role of olfactory cues in pollinator attraction. Rheum nobile is self-compatible, but it depends mainly on Bradysia sp. females for pollination. Seed production resulting from pollination by adult flies is substantially higher than subsequent seed consumption by their larvae. Behavioural tests showed that an unusual floral compound, 2-methyl butyric acid methyl ester, emitted by plants only during anthesis, was attractive to female flies. Our results indicate that the R. nobile-Bradysia sp. interaction represents a new pollinating seed-consuming mutualism, and that a single unusual compound is the specific signal in the floral scent of R. nobile that plays a key role in attracting its pollinator.

Insecticide use in hybrid onion seed production affects pre- and postpollination processes.

Research on threats to pollination service in agro-ecosystems has focused primarily on the negative impacts of land use change and agricultural practices such as insecticide use on pollinator populations. Insecticide use could also affect the pollination process, through nonlethal impacts on pollinator attraction and postpollination processes such as pollen viability or pollen tube growth. Hybrid onion seed (Allium cepa L., Alliaceae) is an important pollinator-dependent crop that has suffered yield declines in California, concurrent with increased insecticide use. Field studies suggest that insecticide use reduces pollination service in this system. We conducted a field experiment manipulating insecticide use to examine the impacts of insecticides on 1) pollinator attraction, 2) pollen/stigma interactions, and 3) seed set and seed quality. Select insecticides had negative impacts on pollinator attraction and pollen/stigma interactions, with certain products dramatically reducing pollen germination and pollen tube growth. Decreased pollen germination was not associated with reduced seed set; however, reduced pollinator attraction was associated with lower seed set and seed quality, for one of the two female lines examined. Our results highlight the importance of pesticide effects on the pollination process. Overuse may lead to yield reductions through impacts on pollinator behavior and postpollination processes. Overall, in hybrid onion seed production, moderation in insecticide use is advised when controlling onion thrips, Thrips tabaci, on commercial fields.

[Role of ethylene in the control of gametophyte-sporophyte interactions in the course of the progamic phase of fertilization].

We investigated dynamics of the content of 1-aminocyclopropane-l-carboxylic acid (ACC) and ethylene production in male gametophyte development and germination in fertile (self-compatible and self-incompatible) and sterile clones of petunia. Fertile male gametophyte development was accompanied by two peaks of ethylene production by anther tissues. The first peak occurred during the microspore development simultaneously with the degeneration of both the tapetal tissues and the middle layers of the anther wall. The second peak coincided with dehydration and maturation of pollen grains. In the anther tissues of the sterile line of petunia, tenfold higher ethylene production was observed at the meiosis stage compared with that in fertile male gametophytes. This fact correlated with the degeneration of both microsporocytes and tapetal tissues. Exogenously applied ethylene (1-100 ppm) induced a degradation of the gametophytic generation at the meiosis stage. According to the obtained data, ethylene synthesis in germinating male gametophyte is provided by a 100-fold ACC accumulation in mature pollen grains. The male gametophyte germination, both in vitro, on the culture medium, and in vivo, on the stigma surface, was accompanied by an increase in ethylene production. Depending on the type of pollination, germination of pollen on the stigma surface and the pollen tube growth in the tissues of style were accompanied by various levels ofACC and ethylene release. The male gametophyte germination after self-compatible pollination was accompanied by higher content of ACC as compared with the self-incompatible clone, whereas, after the self-incompatible pollination, we observed a higher level of ethylene production compared with compatible pollination. For both types of pollination, ACC and ethylene were predominantly produced in the stigma tissues. Inhibitor of ethylene action, 2,5-norbornadiene (NBN), blocked both the development and germination of the male gametophyte. These results suggest that ethylene is an important factor in male gametophyte development, germination, and growth at the progamic phase of fertilization.