Macroevolution of floral scent chemistry across radiations of male euglossine bee-pollinated plantsMacroevolución de olores florales a través de radiaciones de plantas polinizadas por abejas euglosinas machosMacroevolução dos voláteis florais em radiações de plantas polinizadas por machos de abelhas Euglossini.

Floral volatiles play key roles as signaling agents that mediate interactions between plants and animals. Despite their importance, few studies have investigated broad patterns of volatile variation across groups of plants that share pollinators, particularly in a phylogenetic context. The "perfume flowers," Neotropical plant species exhibiting exclusive pollination by male euglossine bees in search of chemical rewards, present an intriguing system to investigate these patterns due to the unique function of their chemical phenotypes as both signaling agents and rewards. We leverage recently developed phylogenies and knowledge of biosynthesis, along with decades of chemical ecology research, to characterize axes of variation in the chemistry of perfume flowers, as well as understand their evolution at finer taxonomic scales. We detect pervasive chemical convergence, with many species across families exhibiting similar volatile phenotypes. Scent profiles of most species are dominated by compounds of either the phenylpropanoid or terpenoid biosynthesis pathways, while terpenoid compounds drive more subtle axes of variation. We find recapitulation of these patterns within two independent radiations of perfume flower orchids, in which we further detect evidence for the rapid evolution of divergent floral chemistries, consistent with the putative importance of scent in the process of adaptation and speciation.

Male Emphorini (Hymenoptera: Apidae) bees use flowers of Ipomoea carnea (Convolvulaceae) as overnight resting sites / Machos de abelhas Emphorini (Hymenoptera: Apidae) utilizam flores de Ipomoea carnea (Convolvulaceae) como locais de pernoite

Abstract The present study describes the behavior of male Emphorini bees in exploiting Convolvulaceae flowers as sleeping-sheltering place. The observations occurred inadvertently during a field study in an Agroforestry System in the Caatinga region, near the banks of the São Francisco River in Petrolina, Pernambuco - NE-Brazil. Males were observed and collected in wilted flowers of Ipomoea carnea (Convolvulaceae) during the evening. The sampling revealed nine Emphorini bees, belonging to three oligolectic species specialized on Convolvulaceae. We propose that the infundibuliform morphology of Ipomoea carnea flowers and the varied timing of anthesis could influence the choice of these flowers as overnight resting places. Furthermore, we endorse the idea present in the literature that males of oligolectic bees generally engage in patrolling flowers utilized by females, thereby enhancing the chances of encountering potential mates, making these flowers rendezvous places. The observation of this behavior highlights the interdependence between bees and plants and raises the possibility that the loss of these flowers could negatively impact bees not only in a nutrition aspect but also regarding shelter and reproduction.

Resumo O presente estudo documenta o comportamento de abelhas machos do grupo Emphorini em explorar flores da família Convolvulaceae como local para dormir e se abrigar. As observações foram realizadas inadvertidamente durante um estudo de campo em um Sistema Agroflorestal na região da Caatinga, próximo às margens do Rio São Francisco, em Petrolina, Pernambuco - Nordeste do Brasil. Machos foram observados e coletados em flores fechadas de Ipomoea carnea (Convolvulaceae) durante o entardecer. Em nossa amostragem, encontramos nove machos da tribo Emphorini, pertencentes a três espécies oligoléticas especializadas em Convolvulaceae. Propomos que a morfologia infundibuliforme das flores de Ipomoea carnea e os diferentes períodos de antese das flores podem influenciar a escolha dessas flores como locais de descanso durante à noite. Além disso, nossos dados endossam a ideia de que machos de abelhas oligoléticas geralmente patrulham flores utilizadas por fêmeas, aumentando assim as chances de encontrar possíveis parceiras, tornando essas flores locais de encontro para acasalamento. O registro desse comportamento destaca a interdependência entre abelhas e plantas e levanta a possibilidade de que a perda dessas flores possa impactar negativamente as abelhas não apenas em relação a sua nutrição, mas também em relação ao abrigo e reprodução.

The iconic cactus of the Caatinga dry forest, Cereus jamacaru (Cactaceae) has high sphingophily specialization and pollinator dependence.

Cereus jamacaru is a cactus distributed in Northeastern Brazil, with high symbolic value to this region. However, the interaction, behavior and the role of pollinators remains poorly understood. Here, we investigate the reproductive biology, addressing the ecological significance of floral attributes, including details about floral signaling. The study was carried at three areas of the Caatinga, in 2015, 2017 and 2021. We analyzed the floral morphometry, volume and concentration of the nectar, and characterized the colour and scent of flowers. Additionally, we described the pollinator behavior and performed controlled pollination experiments. The 'Mandacaru' is self-incompatible, has nocturnal anthesis and the nectar is accumulated as droplets in a long hypanthial tube. The flowers have a reflective pattern with a dark outer surface and a white inner surface. (E)-nerolidol is the major component (87.4%) of its floral perfume. We registered the sphingid moth Cocytius antaeus visiting the flowers. The floral attributes, attractants and rewards drives to a sphingophily, and the pollination treatments showed the dependence to fruit set by C. antaeus, the pollinator registered. In this case, if the apparent lack of pollinator diversity encompasses its entire range, the loss of the hawkmoth could severely impact the reproductive success of the cactus.

Mixed pollination system and floral signals of Paepalanthus (Eriocaulaceae): insects and geitonogamy ensure high reproductive success.

BACKGROUND AND AIMS: Eriocaulaceae exhibit a great variety of floral traits associated with insect (e.g. nectariferous structures) and wind pollination (unisexual flowers, exposed sexual organs and small pollen grains), as well as the 'selfing syndrome' (small flowers, short distance between stigma and anthers, and temporal overlap of male and female phases). Paepalanthus bifidus, P. subtilis and P. tortilis are related species that differ in form, size and colour of floral structures. We aimed to investigate the pollination and reproductive biology of these three species. METHODS: We analysed the floral biology, floral visitors, pollinator behaviour, and the contribution of insects, wind and spontaneous geitonogamy to fruit set. We also evaluated the floral colour and scent of the species. Colour reflectance of capitula of each species was measured and plotted in models of insect vision. Floral scent samples were extracted and the compounds were compared to vegetative scent samples. KEY RESULTS: In all species, the staminate and pistillate flowers are arranged in alternating cycles with a temporal overlap between these phases. Ants were the most frequent floral visitors and were effective pollinators in P. bifidus and P. tortilis, while flies were occasional pollinators in P. tortilis. Floral visitors were not observed in P. subtilis. In all species, fruits were produced by spontaneous geitonogamy, with no evidence of wind pollination. According to the models of insect vision, the colours of the capitula of P. bifidus and P. subtilis are the most inconspicuous for ants and flies. We found no difference between the emission of volatiles of inflorescences and vegetative structures. CONCLUSIONS: This study suggests that ant pollination might be more widespread in Eriocaulaceae than currently assumed. Furthermore, for small monocarpic plants, mixed mating strategies are most favourable, by ensuring reproduction either by outcrossing when pollinators are abundant or by spontaneous geitonogamy when pollinations are scarce/absent.

Floral Scent Chemistry and Pollinators of a Sexually Dimorphic Neotropical Orchid.

Catasetum is a speciose Neotropical orchid genus of which male and female flowers emit scents acting both as attractant and reward for their exclusive pollinators, male orchid bees (Euglossini: Apidae). In Catasetum, it is well known that flowers display a remarkably morphological sexual dimorphism. However, it remains poorly investigated whether this is also true for floral scents. Here, we investigated the pollination ecology and floral scent traits (chemistry and total emission) of C. maranhense, a species endemic to the Brazilian N/NE region. Males of Euglossa securigera are the only pollinators of C. maranhense. The floral scent of C. maranhense is composed of 29 volatile compounds, with eucalyptol, indole, (E)-Methyl p-methoxycinnamate, and (Z)-Methyl p-methoxycinnamate accounting for more than 80% of the scent bouquet. No sexual dimorphism was detected in any of the traits investigated. We discuss the ecological and evolutionary significance of our findings to Catasetum species and other unisexual perfume plants.

Differential Evolutionary History in Visual and Olfactory Floral Cues of the Bee-Pollinated Genus Campanula (Campanulaceae).

Visual and olfactory floral signals play key roles in plant-pollinator interactions. In recent decades, studies investigating the evolution of either of these signals have increased considerably. However, there are large gaps in our understanding of whether or not these two cue modalities evolve in a concerted manner. Here, we characterized the visual (i.e., color) and olfactory (scent) floral cues in bee-pollinated Campanula species by spectrophotometric and chemical methods, respectively, with the aim of tracing their evolutionary paths. We found a species-specific pattern in color reflectance and scent chemistry. Multivariate phylogenetic statistics revealed no influence of phylogeny on floral color and scent bouquet. However, univariate phylogenetic statistics revealed a phylogenetic signal in some of the constituents of the scent bouquet. Our results suggest unequal evolutionary pathways of visual and olfactory floral cues in the genus Campanula. While the lack of phylogenetic signal on both color and scent bouquet points to external agents (e.g., pollinators, herbivores) as evolutionary drivers, the presence of phylogenetic signal in at least some floral scent constituents point to an influence of phylogeny on trait evolution. We discuss why external agents and phylogeny differently shape the evolutionary paths in floral color and scent of closely related angiosperms.

Bioactive Male-Produced Volatiles from Anastrepha obliqua and their Role in Attraction of Conspecific Females.

The study of insect semiochemicals, especially pheromones, is of fundamental importance for the development of strategies for controlling agricultural pests. In this study, volatile compounds involved in the communication between males and females of the fruit fly, Anastrepha obliqua (Diptera: Tephritidae), for mating purposes were characterized to develop attractant formulations for females of this species. Extracts containing volatile compounds released by males of A. obliqua were obtained by the dynamic headspace technique and analyzed by gas chromatography coupled with an electroantennographic detector (GC-EAD) and gas chromatography coupled with mass spectrometry (GC-MS). Twenty-one volatile compounds were identified in the aeration extracts of males. Five of them caused EAD responses from the antennae of females: 1-heptanol, linalool, (Z)-3-nonen-1-ol, (E,Z)-3,6-nonadien-1-ol, and (Z,E)-α-farnesene. Six synthetic mixtures of these compounds, including the five-component blend and all possible four-component blends, were formulated in a biopolymer and used in behavioral bioassays conducted in the laboratory arena with conspecific virgin females. One blend of 1-heptanol, linalool, (Z)-3-nonen-1-ol, and (Z,E)-α-farnesene attracted more females than the collection of volatiles from virgin males used as control. The other mixtures were as attractive to A. obliqua females as the control treatment. This study indicates potential for use of these compounds in monitoring and control strategies for this pest.

A Semivolatile Floral Scent Marks the Shift to a Novel Pollination System in Bromeliads.

Perfume flowers (sensu Vogel1) produce intense scents that function both as attractants and as the sole rewards for pollinators. The scent is collected exclusively by male euglossine bees and used during pre-mating behavior.2-5 Perfume flowers have evolved independently in 15 angiosperm families, with over 1,000 reported species across the Neotropical region.6 Members of Cryptanthus (Bromeliaceae) represent a puzzling exception among perfume flowers, as flowers produce nectar and do not emit a noticeable scent yet still attract euglossine males.7 Here, we studied the pollination ecology of Cryptanthus burle-marxii and decode the chemical communication between its flowers and euglossine males. Field observations revealed euglossine males and hummingbirds as potential pollinators. The bees always contacted anthers/stigma of C. burle-marxii while scraping the petals to obtain chemicals, whereas nectar-seeking hummingbirds normally only contacted the anthers. Based on gas chromatography-mass spectrometry/nuclear magnetic resonance analyses of flower scent samples and bioassays, we identified the diterpene copalol as the only floral scent compound triggering scent-gathering behavior in euglossine males. Unlike euglossine-bee-mediated pollination, hummingbird pollination is ancestral in the Cryptanthus clade, suggesting a case of an ongoing pollinator shift8-10 mediated by the evolution of perfume as a reward. Copalol was previously unknown as a floral scent constituent and represents the heaviest and least-volatile compound known to attract euglossine males. Our study provides the first experimental evidence that semivolatile floral compounds can mediate euglossine bee interactions. Male euglossine pollination in other plant species lacking noticeable floral scents11-13 suggests that semivolatile-mediated pollinator attraction is more widespread than currently appreciated.

Subtle Chemical Variations with Strong Ecological Significance: Stereoselective Responses of Male Orchid Bees to Stereoisomers of Carvone Epoxide.

Different enantiomers of chiral compounds within floral perfumes usually trigger distinct responses in insects; however, this has frequently been neglected in studies investigating semiochemicals in plant-pollinator interactions. Approximately 1000 neotropical plants produce floral perfumes as the only reward for pollinators, i.e. male euglossine bees. The chiral compound carvone epoxide is a key component of the scent bouquet of many perfume-rewarding plants that are pollinated by males of Eulaema. Here, we tested the biological activity of the four carvone epoxide stereoisomers to four Eulaema species occurring in the Atlantic Rainforest of NE-Brazil. We determined the stereochemistry of carvone epoxide in the floral scent of several Catasetum species, tested whether the antennae of bees respond differentially to these stereoisomers and investigated if there is a behavioural preference for any of the stereoisomers. We found that 1) Catasetum species emit only the (-)-trans-stereoisomer of carvone epoxide, 2) for E. atleticana and E. niveofasciata antennal responses to the (-)-trans-carvone epoxide were significantly stronger than those to (-)-cis-carvone epoxide, 3) the strength and pattern of antennal responses to all 4 stereoisomers (separately tested) did not differ among Eulaema species, and 4) there were significant differences in attractiveness of the four stereoisomers to the bees species with the (-)-trans-stereoisomer being particularly attractive. We assume (-)-trans-carvone epoxide to be the dominant isomer in perfume-rewarding plants pollinated by Eulaema. The universal occurrence of carvone epoxide in Catasetum species pollinated by Eulaema, suggests that this compound has evolved in perfume-rewarding as a specific attractant for Eulaema bees as pollinators.

Flower Visitors of Campanula: Are Oligoleges More Sensitive to Host-Specific Floral Scents Than Polyleges?

The pollen diet provided by adult bees to their offspring varies immensely. While some species collect pollen on several plants irrespective of their phylogenetic relatedness (polyleges), others collect only on plants within a genus or family (oligoleges). Floral scents play a central role in bee-plant interactions. To locate flowers, polyleges are assumed to rely on compounds commonly found as floral scent constituents, whereas oligoleges rely on unusual compounds to recognize host flowers unambiguously. Campanula flowers are visited by both polylectic and oligolectic species, and their scent bouquets consist of common and unusual (e.g., spiroacetals) volatiles. In a comparative approach, we performed electroantennographic analyses to investigate the antennal responses of three polyleges and three oligoleges to three common volatiles and four spiroacetals. We hypothesized that: 1) oligoleges and polyleges should respond similarly to common flower volatiles, and 2) Campanula oligoleges should be more sensitive to spiroacetals than are polyleges. In corroboration, we found that antennal sensitivity to common volatiles was similar among bees irrespective of pollen diet, whereas oligoleges of Campanula were more sensitive to spiroacetals than polyleges. Newly emerged bees of the Campanula oligolege Chelostoma rapunculi rely on spiroacetals for recognizing host-flowers, and our results suggest that this might also be true for other Campanula oligoleges, since Chelostoma campanularum and Hoplitis mitis also were able to perceive these specific volatiles at very low concentrations. Together, our results provide interesting insights into the significance of olfactory adaptations in oligolectic and polylectic bee species.

Finding flowers in the dark: nectar-feeding bats integrate olfaction and echolocation while foraging for nectar.

Nectar-feeding bats depend mainly on floral nectar to fulfil their energetic requirements. Chiropterophilous flowers generally present strong floral scents and provide conspicuous acoustic echoes to attract bats. While floral scents are assumed to attract bats over long distances, acoustic properties of flower structures may provide detailed information, thus supporting the localization of a flower at close ranges. So far, to our knowledge, there is no study trying to understand the relative importance as well as the combination of these generally coupled cues for detection (presence) and localization (exact position) of open flowers in nature. For a better comprehension of the significance of olfaction and echolocation in the foraging behaviour of nectar-feeding bats, we conducted two-choice experiments with Leptonycteris yerbabuenae. We tested the bats' behaviour in three experimental scenarios with different cues: (i) olfaction versus echolocation, (ii) echolocation versus echolocation and olfaction, and (iii) olfaction versus echolocation and olfaction. We used the floral scent of the bat-pollinated cactus Pachycereus pringlei as olfactory cue and an acrylic paraboloid as acoustic cue. Additionally, we recorded the echolocation behaviour of the bats and analysed the floral scent of P. pringlei. When decoupled cues were offered, bats displayed no preference in choice for any of the two cues. However, bats reacted first to and chose more often the coupled cues. All bats echolocated continuously and broadcast a long terminal group before a successful visit. The floral scent bouquet of P. pringlei is composed of 20 compounds, some of which (e.g. methyl benzoate) were already reported from chiropterophilous plants. Our investigation demonstrates for the first time to our knowledge, that nectar-feeding bats integrate over different sensory modes for detection and precise localization of open flowers. The combined information from olfactory and acoustic cues allows bats to forage more efficiently.

Host choice in a bivoltine bee: how sensory constraints shape innate foraging behaviors.

BACKGROUND: Many insects have multiple generations per year and cohorts emerging in different seasons may evolve their own phenotypes if they are subjected to different selection regimes. The bivoltine bee Andrena bicolor is reported to be polylectic and oligolectic (on Campanula) in the spring and summer generations, respectively. Neurological constraints are assumed to govern pollen diet in bees. However, evidence comes predominantly from studies with oligolectic bees. We have investigated how sensory constraints influence the innate foraging behavior of A. bicolor and have tested whether bees of different generations evolved behavioral and sensory polyphenism to cope better with the host flowers available in nature when they are active. RESULTS: Behavioral and sensory polyphenisms were tested in choice assays and electroantennographic analyses, respectively. In the bioassays, we found that females of both generations (1) displayed a similar innate relative reliance on visual and olfactory floral cues irrespective of the host plants tested; (2) did not prefer floral cues of Campanula to those of Taraxacum (or vice versa) and (3) did not display an innate preference for yellow and lilac colors. In the electroantennographic analyses, we found that bees of both generations responded to the same set of compounds. CONCLUSION: Overall, we did not detect seasonal polyphenism in any trait examined. The finding that bees of both generations are not sensory constrained to visit a specific host flower, which is in strict contrast to results from studies with oligolectic bees, suggest that also bees of the second generation have a flexibility in innate foraging behavior and that this is an adaptive trait in A. bicolor. We discuss the significance of our findings in context of the natural history of A. bicolor and in the broader context of host-range evolution in bees.

Evaluation of the Activity of the Essential Oil from an Ornamental Flower against Aedes aegypti: Electrophysiology, Molecular Dynamics and Behavioral Assays.

Dengue fever has spread worldwide and affects millions of people every year in tropical and subtropical regions of Africa, Asia, Europe and America. Since there is no effective vaccine against the dengue virus, prevention of disease transmission depends entirely on regulating the vector (Aedes aegypti) or interrupting human-vector contact. The aim of this study was to assess the oviposition deterrent activity of essential oils of three cultivars of torch ginger (Etlingera elatior, Zingiberaceae) against the dengue mosquito. Analysis of the oils by gas chromatography (GC)-mass spectrometry revealed the presence of 43 constituents, of which α-pinene, dodecanal and n-dodecanol were the major components in all cultivars. Solutions containing 100 ppm of the oils exhibited oviposition deterrent activities against gravid Ae. aegypti females. GC analysis with electroantennographic detection indicated that the oil constituents n-decanol, 2-undecanone, undecanal, dodecanal, trans-caryophyllene, (E)-ß-farnesene, α-humulene, n-dodecanol, isodaucene and dodecanoic acid were able to trigger antennal depolarization in Ae. aegypti females. Bioassays confirmed that solutions containing 50 ppm of n-dodecanol or dodecanal exhibited oviposition deterrent activities, while a solution containing the alcohol and aldehyde in admixture at concentrations representative of the oil presented an activity similar to that of the 100 ppm oil solution. Docking and molecular dynamics simulations verified that the interaction energies of the long-chain oil components and Ae. aegypti odorant binding protein 1 were quite favorable, indicating that the protein is a possible oviposition deterrent receptor in the antenna of Ae. aegypti.

(E)-Caryophyllene and α-Humulene: Aedes aegypti Oviposition Deterrents Elucidated by Gas Chromatography-Electrophysiological Assay of Commiphora leptophloeos Leaf Oil.

Aedes aegypti is responsible for the transmission of dengue, a disease that infects millions of people each year. Although essential oils are well recognized as sources of compounds with repellent and larvicidal activities against the dengue mosquito, much less is known about their oviposition deterrent effects. Commiphora leptophloeos, a tree native to South America, has important pharmacological properties, but the chemical profile and applicability of its essential oil in controlling the spread of the dengue mosquito have not been investigated. The aim of this study was to determine the composition of C. leptophloeos leaf oil and to evaluate its larvicidal and oviposition deterrent effects against A. aegypti. Fifty-five components of the essential oil were detected by gas chromatography (GC)-mass spectrometry, with α-phellandrene (26.3%), (E)-caryophyllene (18.0%) and ß-phellandrene (12.9%) identified as the major constituents. Bioassays showed that the oil exhibited strong oviposition deterrent effects against A. aegypti at concentrations between 25 and 100 ppm, and possessed good larvicidal activity (LC50 = 99.4 ppm). Analysis of the oil by GC coupled with electroantennographic detection established that seven constituents could trigger antennal depolarization in A. aegypti gravid females. Two of these components, namely (E)-caryophyllene and α-humulene, were present in substantial proportions in the oil, and oviposition deterrence assays confirmed that both were significantly active at concentrations equivalent to those present in the oil. It is concluded that these sesquiterpenes are responsible, at least in part, for the deterrent effect of the oil. The oviposition deterrent activity of the leaf oil of C. leptophloeos is one of the most potent reported so far, suggesting that it could represent an interesting alternative to synthetic insecticides. The results of this study highlight the importance of integrating chemical and electrophysiological methods for screening natural compounds for their potential in combating vectors of insect-borne diseases.

Visual and Olfactory Floral Cues of Campanula (Campanulaceae) and Their Significance for Host Recognition by an Oligolectic Bee Pollinator.

Oligolectic bees collect pollen from a few plants within a genus or family to rear their offspring, and are known to rely on visual and olfactory floral cues to recognize host plants. However, studies investigating whether oligolectic bees recognize distinct host plants by using shared floral cues are scarce. In the present study, we investigated in a comparative approach the visual and olfactory floral cues of six Campanula species, of which only Campanula lactiflora has never been reported as a pollen source of the oligolectic bee Ch. rapunculi. We hypothesized that the flowers of Campanula species visited by Ch. rapunculi share visual (i.e. color) and/or olfactory cues (scents) that give them a host-specific signature. To test this hypothesis, floral color and scent were studied by spectrophotometric and chemical analyses, respectively. Additionally, we performed bioassays within a flight cage to test the innate color preference of Ch. rapunculi. Our results show that Campanula flowers reflect the light predominantly in the UV-blue/blue bee-color space and that Ch. rapunculi displays a strong innate preference for these two colors. Furthermore, we recorded spiroacetals in the floral scent of all Campanula species, but Ca. lactiflora. Spiroacetals, rarely found as floral scent constituents but quite common among Campanula species, were recently shown to play a key function for host-flower recognition by Ch. rapunculi. We conclude that Campanula species share some visual and olfactory floral cues, and that neurological adaptations (i.e. vision and olfaction) of Ch. rapunculi innately drive their foraging flights toward host flowers. The significance of our findings for the evolution of pollen diet breadth in bees is discussed.

Pollination biology in the dioecious orchid Catasetum uncatum: How does floral scent influence the behaviour of pollinators?

Catasetum is a neotropical orchid genus that comprises about 160 dioecious species with a remarkable sexual dimorphism in floral morphology. Flowers of Catasetum produce perfumes as rewards, which are collected only by male euglossine bees. Currently, floral scents are known to be involved in the selective attraction of specific euglossine species. However, sexual dimorphism in floral scent and its eventual role in the pollination of Catasetum species have never been investigated. Here, we have investigated the pollination of Catasetum uncatum and asked: (1) Is floral scent a sexual dimorphic trait? (2) Does pollinarium removal/deposition affect scent emission? (3) Does sexual dimorphism in floral scent and changed scent emission have implications with regard to the behaviour of the pollinators? The frequency and behaviour of floral visitors were observed in non-manipulated flowers (both flower sexes) and in manipulated flowers (pistillate only) in which pollinaria were deposited. Scents of staminate and pistillate flowers (both manipulated and non-manipulated) were collected by using dynamic headspace methods and analysed chemically. Electrophysiological analyses were performed to detect compounds triggering antennal depolarisation in the euglossine species. C. uncatum is pollinated mainly by males of Euglossa nanomelanotricha. Pollinators were more frequent in pistillate than in staminate inflorescences. Bees approaching staminate flowers frequently flew away without visiting them, a behavioural pattern not observed in pistillate flowers. In the chemical analyses, we recorded 99 compounds, 31 of which triggered antennal depolarisation in pollinators. Multivariate analyses with the electrophysiological-active compounds did not detect differences between the scent composition of staminate and pistillate flowers. Pollinarium removal or deposition resulted in diminished scent emission within 24h in staminate and pistillate flowers, respectively. Surprisingly, bees discriminated pollinated from non-pollinated pistillate flowers as early as 2h after pollination. The rapid loss in the attractiveness of flowers following pollinarium removal/deposition can be interpreted as a strategy to direct pollinators to non-pollinated flowers. We have found no evidence that euglossine males discriminate staminate from pistillate flowers by means of floral scent. Instead, we speculate that bees use visual cues, such as sex dimorphic traits, to discriminate flowers of different sexes. Together, our results provide interesting insights into the evolution of floral signals in gender-dimorphic species and into its significance in plant reproductive biology.

The chemical basis of host-plant recognition in a specialized bee pollinator.

Many pollinators specialize on a few plants as food sources and rely on flower scents to recognize their hosts. However, the specific compounds mediating this recognition are mostly unknown. We investigated the chemical basis of host location/recognition in the Campanula-specialist bee Chelostoma rapunculi using chemical, electrophysiological, and behavioral approaches. Our findings show that Ca. trachelium flowers emit a weak scent consisting of both widespread and rare (i.e., spiroacetals) volatiles. In electroantennographic analyses, the antennae of bees responded to aliphatics, terpenes, aromatics, and spiroacetals; however, the bioassays revealed a more complex response picture. Spiroacetals attracted host-naive bees, whereas spiroacetals together with aliphatics and terpenes were used for host finding by host-experienced bees. On the intrafloral level, different flower parts of Ca. trachelium showed differences in the absolute and relative amounts of scent, including spiroacetals. Scent from pollen-presenting flower parts elicited more feeding responses in host-naive bees as compared to a scentless control, whereas host-experienced bees responded more to the nectar-presenting parts. Our study demonstrates the occurrence of learning (i.e., change in the bee's innate chemical search-image) after bees gain foraging experience on host flowers. We conclude that highly specific floral volatiles play a key role in host-flower recognition by this pollen-specialist bee, and discuss our findings into the broader context of host-recognition in oligolectic bees.

Mutual reproductive dependence of distylic Cordia leucocephala (Cordiaceae) and oligolectic Ceblurgus longipalpis (Halictidae, Rophitinae) in the Caatinga.

BACKGROUND AND AIMS: The close relationship between distylic Cordia leucocephala and the bee Ceblurgus longipalpis, both endemic to the Caatinga, north-east Brazil, was investigated, emphasizing reproductive dependence, morphological adaptations of the partners, and pollen flow. METHODS: In the municipality of Pedra, in the Caatinga of Pernambuco, the breeding system and reproductive success of C. leucocephala, its interaction with flower visitors and inter- and intramorph pollen flow were determined. KEY RESULTS: The bee Ceblurgus longipalpis, the unique flower visitor and effective pollinator of self-incompatible Cordia leucocephala, presents morphological features adapted to exploit hidden pollen and nectar in the long and narrow corolla tubes. Pollen of low-level anthers is collected with hairs on prolonged mouthparts and pollen of high-level anthers with clypeus, mandibles, and labrum, showing pollen removal from both levels with the same effectiveness. In both morphs, this results in similar legitimate, i.e. intermorph cross-pollen flow. Illegitimate pollen flow to stigmas of pin flowers, however, was much higher than to stigmas of thrum flowers. Moreover, more illegitimate pollen was transported to stigmas of pin and less to those of thrum flowers when compared with legitimate pollen flow. CONCLUSIONS: The study reveals a one-to-one reproductive inter-dependence between both partners. Data indicate that this relationship between bee species and plant species is one of the rare cases of monolecty among bees. Monotypic Ceblurgus longipalpis, the only rophitine species of Brazil, evolved prolonged mouthparts rare among short-tongued bees that enable them to access pollen from flowers with short-level anthers hidden for bees of other species, and nectar at the base of the flower tube.

Comunidade de abelhas (Hymenoptera, Apoidea) e plantas em uma área do Agreste pernambucano, Brasil / Community of bees (Hymenoptera, Apoidea) and plants in an area of Agreste in Pernambuco, Brazil

O Agreste é uma região de transição entre floresta tropical úmida e caatinga no nordeste brasileiro. Nessa região, grande parte da vegetação nativa foi desmatada para a implantação de pastagens. Não é sabido se áreas degradadas mantém uma apifauna e flora melitófila diversificada, ou quais são associações entre abelhas e plantas que ocorrem nessas áreas. A cobertura vegetal atual é composta por pastos, vegetação ruderal e restos da vegetação nativa. Abelhas e plantas por elas visitadas foram coletadas mensalmente entre agosto de 2001 e julho de 2002, durante dois dias consecutivos entre 5h30 e 17h30. Foram coletados 1.004 indivíduos de abelhas pertencentes a 79 espécies. Apidae foi a família mais abundante e com maior riqueza de espécies (732 indivíduos e 43 espécies), seguida por Halictidae (194 indivíduos e 20 spp.), Megachilidae (47 indivíduos e 13 spp.), Colletidae (16 indivíduos e 2 spp.) e Andrenidae (15 indivíduos e 1 sp.). Foram registradas apenas três espécies de abelhas eussocais e cinco de Euglossini, dois grupos altamente diversificados nas florestas neotropicais. A ausência de abelhas sem ferrão nativas dos gêneros Plebeia, Frieseomelitta, Partamona, Scaptotrigona e Trigonisca, assim como de outras espécies de Euglossini, deve estar relacionada à falta de sítios de nidificação e à escassez de fontes de pólen e néctar nessa área degradada. Foram registradas 87 espécies de plantas melitófilas, a maioria ervas e arbustos. Árvores nativas isoladas, assim como plantas ornamentais e frutíferas cultivadas contribuem para manter parte da diversidade da comunidade de abelhas nativas.

The Agreste is a transition region of tropical rainforest and Caatinga in northeastern Brazil. In this region, the majority of the native Atlantic Rainforest was destroyed to give place to livestock farming. It is not known whether degraded areas maintain a diversified bee-plant community or not and which kinds of bee-plant interactions occur. The vegetation at the study site comprises pastures, ruderal vegetation and minute remanents of rainforest vegetation. Both bees and associated plants were collected monthly, from August 2001 to July 2002, sampling during two consecutive days from 5:30 am to 5:30 pm. We collected a total of 1004 bees from 79 species. Apidae was the most abundant and species-rich bee family (732 individuals, 43 species) followed by Halictidae (194 individuals and 20 spp.), Megachilidae (47 individuals and 13 spp.), Colletidae (16 individuals and 2 spp.) and Andrenidae (15 individuals and 1 sp.). Only three species of eusocial bees and five of euglossine bees were recorded, even though both groups are diversified in Neotropical rainforests. The absence of native stingless bees of the genera Plebeia, Frieseomelitta, Partamona, Scaptotrigona and Trigonisca, as well as of other euglossine bee species, is probably due to the lack of nesting sites and to the paucity of pollen and nectar resources in this disturbed area. The bees visited flowers of 87 plant species, mainly of herbs and small shrubs. Isolated rainforest trees in the pasture as well as cultivated fruit crops contributed to some extent to maintain a diversified native bee community.