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

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PREMISE: Flowering plants with poricidal anthers are commonly visited by buzzing bees, which vibrate flowers to extract pollen. However, not all flower visitors are in fact pollinators, and features such as body size and duration of flower visits are important factors in determining pollination effectiveness. We tested whether bee-to-flower size relationships predict the pollination effectiveness of flower visitors of a buzz-pollinated species (Chamaecrista ramosa, Fabaceae). METHODS: We sorted 13 bee taxa into three groups: smaller than, equivalent to ("fit-size"), and larger than flower herkogamy (spatial separation between anthers and stigma). We expected the latter two groups to touch the stigmas, which would be an indicator of pollination effectiveness, more frequently than the first group. To test this hypothesis, we assessed contact with stigmas, foraging behavior, and duration of visits for the three size groups of bees. RESULTS: Our data reveal that small bees scarcely touched the stigmas, while large and fit-size bees were the most efficient pollinators, achieving high stigma-touching rates, conducting much shorter flower visits, and visiting flowers and conspecific plants at high rates during foraging bouts. CONCLUSIONS: The results did not show size-matching among bees and flowers, as expected, but rather a minimum size threshold of efficient pollinators. The finding of such a threshold is a nonarbitrary approach to predicting pollination effectiveness of visitors to herkogamous flowers with poricidal anthers.

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The foraging activity of diurnal bees often relies on flower availability, light intensity and temperature. We do not know how nocturnal bees, which fly at night and twilight, cope with these factors, especially as light levels vary considerably from night to day and from night to night due to moon phase and cloud cover. Given that bee apposition compound eyes function at their limits in dim light, we expect a strong dependence of foraging activity on light intensity in nocturnal bees. Besides being limited by minimum light levels to forage, nocturnal bees should also avoid foraging at brighter intensities, which bring increased competition with other bees. We investigated how five factors (light intensity, flower availability, temperature, humidity, and wind) affect flower visitation by Neotropical nocturnal bees in cambuci (Campomanesia phaea, Myrtaceae). We counted visits per minute over 30 nights in 33 cambuci trees. Light intensity was the main variable explaining flower visitation of nocturnal bees, which peaked at intermediate light levels occurring 25 min before sunrise. The minimum light intensity threshold to visit flowers was 0.00024 cd/m2. Our results highlight the dependence of these nocturnal insects on adequate light levels to explore resources.

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Four isolates of two novel ascosporogenous species belonging to the clade Starmera were obtained from cactus tissues and rotting wood in Brazil. Results of analyses of the sequences of the ITS and D1/D2 domains of the large subunit rRNA gene indicated that the two isolates of the cactophilic species are related to Starmera caribaea and Starmera pilosocereana, yeasts that are associated with cacti and require an organic source of sulfur for growth. We propose the novel species Starmera foglemanii sp. nov. (CBS 16113T; MycoBank number: MB 834400) to accommodate these isolates. The other two isolates are phylogenetically related to Candida dendrica, Candida laemsonensis and Candida berthetii, also in the Starmera clade. The novel species name Starmera ilhagrandensis sp. nov. (CBS 16316T; MycoBank number: MB 834402) is proposed for this species.

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The specialised mutualism between Tococa guianensis and ants housed in its leaf domatia is a well-known example of myrmecophily. A pollination study on this species revealed that flowers in the bud stage exude a sugary solution that is collected by ants. Given the presence of this unexpected nectar secretion, we investigated how, where, and when floral buds of T. guianensis secret nectar and what function it serves. We studied a population of T. guianensis occurring in a swampy area in the Cerrado of Brazil by analyzing the chemical composition and secretion dynamics of the floral-bud nectar and the distribution and ultrastructure of secretory tissues. We also measured flower damage using ant-exclusion experiments. Floral bud nectar was secreted at the tip of the petals, which lack a typical glandular structure but possess distinctive mesophyll due to the presence of numerous calcium oxalate crystals. The nectar, the production of which ceased after flower opening, was composed mainly of sucrose and low amounts of glucose and fructose. Nectar was consumed by generalist ants and sporadically by stingless bees. Ant exclusion experiments resulted in significantly increased flower damage. The floral nectar of T. guianensis is produced during the bud stage. This bud-nectar has the extranuptial function of attracting generalist ants that reduce florivory. Pollen is the unique floral resource attracting pollinators during anthesis. Tococa guianensis, thus, establishes relationships with two functional groups of ant species: specialist ants acting against herbivory and generalist ants acting against florivory.

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Communication between plants and nocturnal pollinators in low light conditions is mainly guided by floral scents, which is well documented for plants pollinated by bats, moths, and beetles. Just recently, nocturnal bees have been added to the list of pollinators known to respond to floral scents of their host plants. Little is known about the floral scent chemistry of plants visited and pollinated by nocturnal bees. Among these plants are economically important fruit crops of the family Myrtaceae. We aimed to analyze the nocturnal floral scent profiles of 10 species of Myrtaceae (only diurnal P. cattleianum was sampled after sunrise) and address the following questions: i) What are the main floral scent compounds emitted by the species? ii) Are the floral scent profiles similar to those described for other species pollinated by nocturnal bees? Floral scents were collected by dynamic headspace and analyzed by GC-MS (gas chromatography - mass spectrometry). The total amount of scent trapped ranged from 74 ng/flower/hour for Syzygium malaccense to 7556 ng/flower/hour for Eugenia dysenterica. A total of 46 floral scent compounds were detected in the samples with the most abundant compounds being the aromatics benzaldehyde, benzyl alcohol, 2-phenylethanol, methyl salicylate, 2-phenylethyl acetate, and benzyl acetate; the aliphatic compound 1-octanol; and the monoterpene linalool. The different species exhibited different relative scent patterns. Overall, the nocturnal scents of the studied species of Myrtaceae are dominated by aromatic compounds, which is in contrast to the scent profiles described for other plants pollinated by nocturnal bees.

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Caryocar brasiliense is a flagship species of the Brazilian Cerrado. It produces flowers with a strong peculiar scent, which are pollinated by bats and occasionally moths with short mouthparts. However, the cues responsible for attracting these nocturnal pollinators remain unknown. We aimed to identify osmophores of C. brasiliense, describe the ultrastructure of the cells involved in the synthesis and release of floral odour, and identify the constituents of the floral bouquet. We performed field observations and histochemical and ultrastructural analyses of flowers focusing on the androecium. Gas chromatography-mass spectrometry was used to analyse the scents emitted. Filament epidermal cells were found to possess an unusual shape and be responsible for the main production and release of odour. These cells, called foraminous cells, are elongate and possess pores where their cell walls are abruptly thin. The cuticle is practically absent over the pores, which facilitates odour emission. The foraminous cells have conspicuous nuclei and organelle-rich cytoplasm where oil droplets can be seen prior to anthesis. The features of these cells remain similar during anthesis, but many vesicles fuse with the plasma membrane and the number of oil droplets in the cytosol decreases. Twenty-nine components were found in the scent, especially fatty acid derivatives and N- and S-bearing compounds. Our analyses revealed that the androecium of C. brasiliense has a particular structure that acts as an osmophore. The scent from the androecium resembles that of the entire flower, which is an unprecedented finding for a plant with single flowers as the pollination unit.

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Floral scent is an important component of the trait repertoire of flowering plants, which is used to attract and manipulate pollinators. Despite advances during the last decades about the chemicals released by flowers, there is still a large gap in our understanding of chemical communication between flowering plants and their pollinators. We analyzed floral scents of guarana (Paullinia cupana, Sapindaceae), an economically important plant of the Amazon, using chemical analytical approaches, and determined the attractiveness of the scent to its nocturnal bee pollinators using behavioral assays in the field. Pollen loads of attracted bees were also analyzed. Inflorescences of guarana emit strong scents, both during day and at night, with some semi-quantitative differences between day- and night-time scents. Synthetic scent mixtures containing some of the identified floral scent components, including the most abundant ones, i.e., linalool and (E)-ß-ocimene, successfully attracted the nocturnal Megalopta bee pollinators. Pollen analyses revealed that many of the attracted bees had pollen grains from previous visits to guarana flowers on their bodies. Overall, our data show that guarana flowers attract nocturnal bee visitors by their strong scents and suggest that the chemical communication between this plant and its pollinators is a key component in crop production of this economically important plant species.

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Pollinators search for multiple flora resources throughout their life cycle. Most studies, however, only assess how bees discriminate floral cues in the context of nectar foraging. In the present study, we sought to elucidate whether oil-collecting bees discriminate flowers of Byrsonima variabilis (Malpighiaceae) with petals of different colours when foraging for pollen or oil. As the colour of the standard petal changes during anthesis, we characterised the spectral reflectance patterns of flowers throughout anthesis and modelled chromatic perceptual space to determine how these colour patterns are perceived by bees. Through the quantification of flower pollen in the different phases, we found that the colour of the standard petal is an honest cue of the presence of pollen. Centridine bees preferentially visited flowers with a yellow (bee's green) colour when searching for pollen, but indiscriminately visited flowers with different petal colours when searching for floral oil. We suggest that standard petals, in the species studied and others of the genus, like nectar guides, act as pollen guides, which oil-collecting females use to detect pollen-rich flowers. Moreover, they use different floral clues during foraging for different resources in the same host plant.

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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.

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