Antibodies to Pseudogymnoascus destructans are not sufficient for protection against white‐nose syndrome

White‐nose syndrome (WNS) is a fungal disease caused by Pseudogymnoascus destructans (Pd) that affects bats during hibernation. Although millions of bats have died from WNS in North America, mass mortality has not been observed among European bats infected by the fungus, leading to the suggestion that bats in Europe are immune. We tested the hypothesis that an antibody‐mediated immune response can provide protection against WNS by quantifying antibodies reactive to Pd in blood samples from seven species of free‐ranging bats in North America and two free‐ranging species in Europe. We also quantified antibodies in blood samples from little brown myotis (Myotis lucifugus) that were part of a captive colony that we injected with live Pd spores mixed with adjuvant, as well as individuals surviving a captive Pd infection trial. Seroprevalence of antibodies against Pd, as well as antibody titers, was greater among little brown myotis than among four other species of cave‐hibernating bats in North America, including species with markedly lower WNS mortality rates. Among little brown myotis, the greatest titers occurred in populations occupying regions with longer histories of WNS, where bats lacked secondary symptoms of WNS. We detected antibodies cross‐reactive with Pd among little brown myotis naïve to the fungus. We observed high titers among captive little brown myotis injected with Pd. We did not detect antibodies against Pd in Pd‐infected European bats during winter, and titers during the active season were lower than among little brown myotis. These results show that antibody‐mediated immunity cannot explain survival of European bats infected with Pd and that little brown myotis respond differently to Pd than species with higher WNS survival rates. Although it appears that some species of bats in North America may be developing resistance to WNS, an antibody‐mediated immune response does not provide an explanation for these remnant populations.     

Pollination syndromes in a specialised plant‐pollinator interaction: does floral morphology predict pollinators in Calceolaria?

Pollination syndromes are defined as suites of floral traits evolved in response to selection imposed by a particular group of pollinators (e.g., butterflies, hummingbirds, bats). Although numerous studies demonstrated their occurrence in plants pollinated by radically different pollinators, it is less known whether it is possible to identify them within species pollinated by one functional pollinator group. In such a framework, we expect floral traits to evolve also in response to pollinator subgroups (e.g., species, genera) within that unique functional group. On this, specialised pollination systems represent appropriate case studies to test such expectations. Calceolaria is a highly diversified plant genus pollinated by oil‐collecting bees in genera Centris and Chalepogenus. Variation in floral traits in Calceolaria has recently been suggested to reflect adaptations to pollinator types. However, to date no study has explicitly tested that observation. In this paper, we quantitatively test that hypothesis by evaluating the presence of pollination syndromes within the specialised pollination system formed by several Calceolaria and their insect pollinators. To do so, we use multivariate approaches and explore the structural matching between the morphology of 10 Calceolaria taxa and that of their principal pollinators. Our results identify morphological matching between floral traits related to access to the reward and insect traits involved in oil collection, confirming the presence of pollinator syndromes in Calceolaria. From a general perspective, our findings indicate that the pollination syndrome concept can be also extended to the intra‐pollinator group level.     

Does intraspecific behavioural variation of pollinator species influence pollination? A quantitative study with hummingbirds and a Neotropical shrub

• Floral visitors differ in their efficacy as pollinators, and the impact of different pollinator species on pollen flow and plant reproduction has been frequently evaluated. In contrast, the impact of intraspecific behavioural changes on their efficacy as pollinators has seldom been quantified.
• We studied a self‐incompatible shrub Palicourea rigida (Rubiaceae) and its hummingbird pollinators, which adjust their behaviour according to floral resource availability. Fluorescence microscopy was used to access pollen tube growth and incompatibility reaction in pistils after a single visit of territorial or intruder hummingbirds in two populations. To characterise the plant populations and possible differences in resource availability between areas we used a three‐term quadrat variance method to detect clusters of floral resources.
• Within‐species variation in foraging behaviour, but not species identity, affected pollinator efficacy. Effectively, hummingbirds intruding into territories deposited more compatible pollen grains on P. rigida stigmas than territory holders in both study areas. Additionally, territory holders deposited more incompatible than compatible pollen grains.
• Our results imply that intraspecific foraging behaviour variation has consequences for pollination success. Quantifying such variation and addressing the implications of intraspecific variability contribute to a better understanding of the dynamics and consequences of plant–pollinator interactions.

     

Do artificial nectar feeders affect bat–plant interactions in an Ecuadorian cloud forest?

Plant–pollinator interactions are critical to ecosystems. However, when artificial nectar feeders are available in an area, they could draw pollinators away from plants. We tested the effects of artificial nectar feeders in an Ecuadorian cloud forest on four aspects of bat–plant interactions: (1) bat relative abundance; (2) bat pollen loads; (3) flower visitation rates, and (4) breeding success of a bat‐pollinated species (Burmeistera glabrata). We divided the study site into areas close to (~30 m) and far from (~500 m) three different feeder sites. At each distance, we captured nectar bats (Anoura caudifer, Anoura cultrata, and Lonchophylla robusta) to estimate their relative abundance and to collect pollen from fur and fecal samples. We also videotaped flowers to estimate bat visitation rates and recorded different breeding success variables of B. glabrata. We found that areas close to feeders have higher relative bat abundance by a factor of 40. In spite of this, the presence of feeders did not affect bat pollen loads, nor the flower visitation rates and breeding success of B. glabrata. Interestingly, there were differences in pollen loads between the three bat species, in that L. robusta individuals rarely carried pollen and were only captured near feeders.     

Pollination efficiency of cockroaches and other floral visitors of Clusia blattophila

• Cockroaches have rarely been documented as pollinators. In this paper we examine whether this is because they might be inefficient at pollination compared to other pollinators. Clusia blattophila, a dioecious shrub growing on isolated rocky outcrops in French Guiana, is pollinated by Amazonina platystylata cockroaches and provides a valuable system for the study of cockroach pollination efficiency.
• We examined the species composition of the visitor guild and visitation rates by means of camcorder recordings and visitor sampling. Then, we investigated the capacity for pollen transfer of principal visitors and found correlations between visitation rates and pollen loads on stigmas. In an exclusion experiment we determined the contributions of individual species to pollination success.
• Amazonina platystylata, crickets and two species of Diptera transferred pollen, but the number of transferred pollen grains was only related to visitation rates in the case of cockroaches. Crickets visited and rarely carried pollen. Dipterans were as frequent as cockroaches, carried similar pollen loads, but transferred much less pollen. An estimated 41% and 17% of ovules were pollinated by cockroaches and dipterans, respectively. The remaining ovules were not pollinated. There was no spatial variation in pollinator guild composition, but cockroaches visited flowers less frequently at the smaller study site.
• We demonstrate that cockroaches pollinate a large proportion of ovules. Their pollination service is not confined to one study site and, unlike that provided by dipterans, is not limited to certain years. We suggest that cockroach pollination has been overlooked and that cockroach‐pollinated plants, which share certain floral features, possess adaptations to pollination by cockroaches.

     

Comparative phylogeography and asymmetric hybridization between cryptic bat species

Cryptic speciation and hybridization are two key processes that affect the origin and maintenance of biodiversity and our ability to understand and estimate it. To determine how these two processes interact, we studied allopatric and sympatric colonies of two cryptic bat species (Eptesicus serotinus and Eptesicus isabellinus) with parapatric distribution in the Iberian Peninsula. These species are the main reservoir for the most commonly rabies virus found in bats in Europe: the European bat Lyssavirus type 1 (EBLV‐1). We used mtDNA and nuclear microsatellite markers to confirm the taxonomic status of both species and to show a more pronounced and geographically based genetic structure in E. isabellinus than in its sibling E. serotinus. Using approximate Bayesian computation (ABC), we inferred rapid range expansion in both species after the Last Glacial Maximum until reaching their present distributions. ABC analysis also supported interspecific differences in genetic diversity and structure, pointing to an earlier expansion of E. isabellinus northward. We found no evidence of mitochondrial introgression between species, but nuclear markers identified a male‐mediated ongoing asymmetric hybridization from E. isabellinus to E. serotinus (28% hybrids in E. serotinus and 5% in E. isabellinus) in the contact zone. Although none of the bats studied tested positive for Lyssavirus RNA, the asymmetric hybridization supports the potential for the recently suggested interspecific transmission of EBLV‐1 from E. isabellinus into E. serotinus.     

Differences in seasonal survival suggest species‐specific reactions to climate change in two sympatric bat species

Long‐lived animals with a low annual reproductive output need a long time to recover from population crashes and are, thus, likely to face high extinction risk, if the current global environmental change will increase mortality rates. To aid conservation of those species, knowledge on the variability of mortality rates is essential. Unfortunately, however, individual‐based multiyear data sets that are required for that have only rarely been collected for free‐ranging long‐lived mammals. Here, we used a five‐year data set comprising activity data of 1,445 RFID‐tagged individuals of two long‐lived temperate zone bat species, Natterer's bats (Myotis nattereri) and Daubenton's bats (Myotis daubentonii), at their joint hibernaculum. Both species are listed as being of high conservation interest by the European Habitats Directive. Applying mixed‐effects logistic regression, we explored seasonal survival differences in these two species which differ in foraging strategy and phenology. In both species, survival over the first winter of an individual's life was much lower than survival over subsequent winters. Focussing on adults only, seasonal survival patterns were largely consistent with higher winter and lower summer survival but varied in its level across years in both species. Our analyses, furthermore, highlight the importance of species‐specific time periods for survival. Daubenton's bats showed a much stronger difference in survival between the two seasons than Natterer's bats. In one exceptional winter, the population of Natterer's bats crashed, while the survival of Daubenton's bats declined only moderately. While our results confirm the general seasonal survival pattern typical for hibernating mammals with higher winter than summer survival, they also show that this pattern can be reversed under particular conditions. Overall, our study points toward a high importance of specific time periods for population dynamics and suggests species‐, population‐, and age class‐specific responses to global climate change.     

Pollen transfer in fragmented plant populations: insight from the pollen loads of pollinators and stigmas in a mass‐flowering species

Pollinator and/or mate scarcity affects pollen transfer, with important ecological and evolutionary consequences for plant reproduction. However, the way in which the pollen loads transported by pollinators and deposited on stigmas are affected by pollination context has been little studied. We investigated the impacts of plant mate and visiting insect availabilities on pollen transport and receipt in a mass‐flowering and facultative autogamous shrub (Rhododendron ferrugineum). First, we recorded insect visits to R. ferrugineum in plant patches of diverse densities and sizes. Second, we analyzed the pollen loads transported by R. ferrugineum pollinators and deposited on stigmas of emasculated and intact flowers, in the same patches. Overall, pollinators (bumblebees) transported much larger pollen loads than the ones found on stigmas, and the pollen deposited on stigmas included a high proportion of conspecific pollen. However, comparing pollen loads of emasculated and intact flowers indicated that pollinators contributed only half the conspecific pollen present on the stigma. At low plant density, we found the highest visitation rate and the lowest proportion of conspecific pollen transported and deposited by pollinators. By contrast, at higher plant density and lower visitation rate, pollinators deposited larger proportion of conspecific pollen, although still far from sufficient to ensure that all the ovules were fertilized. Finally, self‐pollen completely buffered the detrimental effects on pollination of patch fragmentation and pollinator failure. Our results indicate that pollen loads from pollinators and emasculated flowers should be quantified for an accurate understanding of the relative impacts of pollinator and mate limitation on pollen transfer in facultative autogamous species.     

Asymmetric and frequency‐dependent pollinator‐mediated interactions may influence competitive displacement in two vernal pool plants

A plant species immigrating into a community may experience a rarity disadvantage due to competition for the services of pollinators. These negative reproductive interactions have the potential to lead to competitive displacement or exclusion of a species from a site. In this study, we used one‐ and two‐species arrays of potted plants to test for density and frequency dependence in pollinator‐mediated and above‐ground intraspecific and interspecific competition between two species of Limnanthes that have overlapping ranges, but rarely occur in close sympatry. There were asymmetric competitive effects; the species responded differently to their frequency within 16‐plant replacement series arrays. Limnanthes douglasii rosea experienced stronger reductions in lifetime and per‐flower fertility, likely due to pollinator‐mediated competition with Limnanthes alba. This effect may be linked to asymmetrical competition through heterospecific pollen transfer. This study demonstrates that pollinator‐mediated competition may discourage establishment of L. d. rosea in sites already occupied by its congener.     

Negative Effects of Conspecific Floral Density on Fruit Set of Two Neotropical Understory Plants

Plant reproductive success is usually positively related to conspecific floral density, but neutral or negative effects of floral density on reproduction have also been reported. Differences in the relationship between reproduction and floral density largely originate from a trade‐off between increasing attractiveness versus increasing competition for pollinators at high floral densities. Although floral densities strongly vary in the understory of tropical forests, for instance, due to variation in light availability, little is known about the density dependence of reproduction in tropical understory plants. We used path analyses to disentangle direct and indirect effects of canopy openness and floral density on fruit set and analyzed the relationship between pollen load and floral density for two Neotropical understory plants, Heliconia metallica and Besleria melancholica. In both species, fruit set was not directly related to canopy openness, but decreased with increasing floral density. In H. metallica, canopy openness had an indirect negative effect on reproduction mediated by its effects on floral density. Effects of floral density on pollen loads were species‐specific. In B. melancholica, pollen loads linearly decreased with increasing floral density, indicating competition for pollinators at high densities. In H. metallica, pollen loads were reduced at both low and high densities, indicating an interplay of facilitative and competitive effects of floral density on pollen deposition. In contrast to other studies, we found negative density dependence of reproduction in both understory species. Negative effects of floral density on reproduction appear to be related to pollinator‐mediated effects on reproduction rather than to variation in abiotic conditions.     

Ornithophilous and Chiropterophilous Pollination in Musa itinerans (Musaceae), a Pioneer Species in Tropical Rain Forests of Yunnan, Southwestern China1

The role of bats and sunbirds in the pollination ecology of Musa itinerans Cheesman (Musaceae) was studied in the tropical seasonal rain forests of Xishuangbanna, southern Yunnan, China. It was found that both long–tongued fruit bats (Macroglossus sobrinus) and sunbirds (Arachnothera longirostris) were effective pollinators of M. itinerans. Nectar production had two peaks, one during the day and one during night (0800–1200 h and 2000–2400 h), which allowed the two different foragers to visit at specific times. The visitation patterns of the two foragers coincided with both flowering time and nectar production. By measuring the differences in fruit weight and seed production among different bagging experiments, we found that birds and bats were equally effective as pollinators of this species.     

Flowering biology of Aspidistra (Asparagaceae): new data on pollination by dipteran insects

The genus Aspidistra comprises about 160 species of herbaceous plants. In previous investigations, different authors suggested that Aspidistra is pollinated by flies, fungus gnats, slugs, amphipods and even collembolans. The vast majority of data was based on only one species, A. elatior, and was not confirmed by direct observations. Recent field observations of anthetic plants confirm the idea of pollination by flies for two other species of the genus. The present study summarizes direct observations of flowering phenology and pollination of five previously unstudied species of Aspidistra. Flowering biology was studied using visual observations as well as video and photo monitoring. Flowers and captured pollinators were investigated using scanning electron microscopy. As a result, knowledge of the flowering biology of Aspidistra has been increased. The absence of heat production and an ultraviolet pattern were verified. The present data show that flowers of Aspidistra can serve as a food resource for animals. The investigated species of Aspidistra are pollinated by dipteran insects: A. formosa, A. marasmioides and A. subrotata are pollinated by fungus gnats; A. multiflora and A. oviflora are pollinated by flies. The occurrence of fungus‐gnat pollination was directly demonstrated for the first time in the genus Aspidistra. The present study confirmed the idea of myiophily in the genus Aspidistra. Apparently, myiophily is common and diverse in Aspidistra. More likely, pollinators are attracted by scent, which is undetectable by humans, but the attraction mechanism is still not completely clear.     

Floral traits of mammal‐pollinated Mucuna macrocarpa (Fabaceae): Implications for generalist‐like pollination systems

Floral traits are adapted by plants to attract pollinators. Some of those plants that have different pollinators in different regions adapt to each pollinator in each region to maximize their pollination success. Mucuna macrocarpa (Fabaceae) limits the pollinators using its floral structure and is pollinated by different mammals in different regions. Here, we examine the relationships between floral traits of M. macrocarpa and the external morphology of mammalian pollinators in different regions of its distribution. Field surveys were conducted on Kyushu and Okinawajima Island in Japan, and in Taiwan, where the main pollinators are the Japanese macaque Macaca fuscata, Ryukyu flying fox Pteropus dasymallus, and red‐bellied squirrel Callosciurus erythraeus, respectively. We measured the floral shapes, nectar secretion patterns, sugar components, and external morphology of the pollinators. Results showed that floral shape was slightly different among regions and that flower sizes were not correlated with the external morphology of the pollinators. Volume and sugar rate of nectar were not significantly different among the three regions and did not change throughout the day in any of the regions. However, nectar concentration was higher in Kyushu than in the other two regions. These results suggest that the floral traits of M. macrocarpa are not adapted to each pollinator in each region. Although this plant limits the number of pollinators using its flower structure, it has not adapted to specific mammals and may attract several species of mammals. Such generalist‐like pollination system might have evolved in the Old World.     

Habitat usage of Daubenton's bat (Myotis daubentonii), common pipistrelle (Pipistrellus pipistrellus), and soprano pipistrelle (Pipistrellus pygmaeus) in a North Wales upland river catchment

Distributions of Daubenton's bat (Myotis daubentonii), common pipistrelle, (Pipistrellus pipistrellus), and soprano pipistrelle (Pipistrellus pygmaeus) were investigated along and altitudinal gradient of the Lledr River, Conwy, North Wales, and presence assessed in relation to the water surface condition, presence/absence of bank‐side trees, and elevation. Ultrasound recordings of bats made on timed transects in summer 1999 were used to quantify habitat usage. All species significantly preferred smooth water sections of the river with trees on either one or both banks; P. pygmaeus also preferred smooth water with no trees. Bats avoided rough and cluttered water areas, as rapids may generate high‐frequency echolocation‐interfering noise and cluttered areas present obstacles to flight. In lower river regions, detections of bats reflected the proportion of suitable habitat available. At higher elevations, sufficient habitat was available; however, bats were likely restricted due to other factors such as a less predictable food source. This study emphasizes the importance of riparian habitat, bank‐side trees, and smooth water as foraging habitat for bats in marginal upland areas until a certain elevation, beyond which bats in these areas likely cease to forage. These small‐scale altitudinal differences in habitat selection should be factored in when designing future bat distribution studies and taken into consideration by conservation planners when reviewing habitat requirements of these species in Welsh river valleys, and elsewhere within the United Kingdom.     

The evolution of bat pollination: a phylogenetic perspective

Background

Most tropical and subtropical plants are biotically pollinated, and insects are the major pollinators. A small but ecologically and economically important group of plants classified in 28 orders, 67 families and about 528 species of angiosperms are pollinated by nectar-feeding bats. From a phylogenetic perspective this is a derived pollination mode involving a relatively large and energetically expensive pollinator. Here its ecological and evolutionary consequences are explored.

Scope and Conclusions

This review summarizes adaptations in bats and plants that facilitate this interaction and discusses the evolution of bat pollination from a plant phylogenetic perspective. Two families of bats contain specialized flower visitors, one in the Old World and one in the New World. Adaptation to pollination by bats has evolved independently many times from a variety of ancestral conditions, including insect-, bird- and non-volant mammal-pollination. Bat pollination predominates in very few families but is relatively common in certain angiosperm subfamilies and tribes. We propose that flower-visiting bats provide two important benefits to plants: they deposit large amounts of pollen and a variety of pollen genotypes on plant stigmas and, compared with many other pollinators, they are long-distance pollen dispersers. Bat pollination tends to occur in plants that occur in low densities and in lineages producing large flowers. In highly fragmented tropical habitats, nectar bats play an important role in maintaining the genetic continuity of plant populations and thus have considerable conservation value.     

Colour vision variation in leaf‐nosed bats (Phyllostomidae): Links to cave roosting and dietary specialization

Bats are a diverse radiation of mammals of enduring interest for understanding the evolution of sensory specialization. Colour vision variation among species has previously been linked to roosting preferences and echolocation form in the suborder Yinpterochiroptera, yet questions remain about the roles of diet and habitat in shaping bat visual ecology. We sequenced OPN1SW and OPN1LW opsin genes for 20 species of leaf‐nosed bats (family Phyllostomidae; suborder Yangochiroptera) with diverse roosting and dietary ecologies, along with one vespertilionid species (Myotis lavali). OPN1LW genes appear intact for all species, and predicted spectral tuning of long‐wavelength opsins varied among lineages. OPN1SW genes appear intact and under purifying selection for Myotis lavali and most phyllostomid bats, with two exceptions: (a) We found evidence of ancient OPN1SW pseudogenization in the vampire bat lineage, and loss‐of‐function mutations in all three species of extant vampire bats; (b) we additionally found a recent, independently derived OPN1SW pseudogene in Lonchophylla mordax, a cave‐roosting species. These mutations in leaf‐nosed bats are independent of the OPN1SW pseudogenization events previously reported in Yinpterochiropterans. Therefore, the evolution of monochromacy (complete colour blindness) has occurred in both suborders of bats and under various evolutionary drivers; we find independent support for the hypothesis that obligate cave roosting drives colour vision loss. We additionally suggest that haematophagous dietary specialization and corresponding selection on nonvisual senses led to loss of colour vision through evolutionary sensory trade‐off. Our results underscore the evolutionary plasticity of opsins among nocturnal mammals.     

Pollinator‐mediated assemblage processes in California wildflowers

Community assembly is the result of multiple ecological and evolutionary forces that influence species coexistence. For flowering plants, pollinators are often essential for plant reproduction and establishment, and pollinator‐mediated interactions may influence plant community composition. Here, we use null models and community phylogenetic analyses of co‐occurrence patterns to determine the role of pollinator‐mediated processes in structuring plant communities dominated by congeners. We surveyed three species‐rich genera (Limnanthes, Mimulus and Clarkia) with centres of diversity in the Sierra Nevada of California. Each genus contains species that co‐flower and share pollinators, and each has a robust phylogeny. Within each genus, we surveyed 44–48 communities at three spatial scales, measured floral and vegetative traits and tested for segregation or aggregation of: (i) species, (ii) floral traits (which are likely to be influenced by pollinators), and (iii) vegetative traits (which are likely affected by other environmental factors). We detected both aggregation and segregation of floral traits that were uncorrelated with vegetative trait patterns; we infer that pollinators have shaped the community assembly although the mechanisms may be varied (competition, facilitation, or filtering). We also found that mating system differences may play an important role in allowing species co‐occurrence. Together, it appears that pollinators influence community assemblage in these three clades.     

Phyllostomid Bat Assemblage Structure in Amazonian Flooded and Unflooded Forests

In Amazonia, the assemblages of several taxa differ significantly between upland terra firme and white‐water flooded várzea forests, but little is known about the diversity and distribution of bats in these two forest types. We compare the spatio‐temporal patterns of bat assemblage composition and structure in adjacent terra firme and várzea forests in the lower Purus River region of central Brazilian Amazonia. Bats were sampled using mist nets at five sites in each forest type during 40 nights (2400 net‐hours). We captured 1069 bats representing 42 species and Phyllostomidae bats comprised 99.3 percent of all captures. The bat assemblages in várzea and terra firme forests were significantly different, mainly due to a marked dissimilarity in species composition and in the number of captures during high‐water season. In addition, bat assemblages within forest types differed significantly between seasons for both terra firme and várzea. Frugivores dominated the bat assemblages in both forest types. Overall guild structure did not change between várzea and terra firme or between seasons, but frugivore and animalivore abundance increased significantly in várzea forest during the inundation. The difference in assemblage structure observed in the high‐water season is probably caused by the annual várzea flooding, which provides an effective barrier to the persistence of many understory bats. We also hypothesize that some bat species may undertake seasonal movements between forest types in response to fruit abundance, and our results further underline the importance of floodplain habitats for the conservation of species in the Amazon.     

Secondary pollen presentation and the temporal dynamics of stylar hair retraction and style elongation in Campanula trachelium (Campanulaceae)

To increase the accuracy of pollen capture and transfer by pollinators some plant species have developed secondary pollen presentation structures. Because the presence of secondary pollen presentation structures at the pistil may reduce the spatial separation between the sexual functions and increase the risk of self‐interference and selfing, temporal segregation of the sexual organs, triggered by visiting insects, can be expected to occur. We investigated secondary pollen presentation and the temporal dynamics of the sexual phases in combination with the physiological self‐incompatibility system in Campanula trachelium, a protandrous insect‐pollinated herb. Stylar hair retraction (male function) and curling of the stigmatic lobes (female function) were modelled using Gompertz growth functions. Finally, we performed pollination experiments in the lab and field to assess seed set and pollen limitation under natural conditions. About 68% of the total pollen load was captured by stylar hairs. Manual manipulation of the stylar hairs, mimicking pollinator visitation, significantly shortened the male phase and accelerated the female phase, resulting in a significant decline in temporal overlap between the two sexual functions. Conversely, when pollinators and/or manual manipulations were lacking, the male phase was substantially prolonged and sexual overlap was maximal. This suggests that spreading of the sexual phases and thus the risk of sexual interference are largely determined by the interaction between stylar hairs and visiting pollinators. Natural seed set was high and not pollinator limited. Overall, these results indicate that secondary pollen presentation and partial protandry resulted in efficient pollen capture, transfer and deposition.     

Floral volatile sesquiterpenes of Elsholtzia rugulosa (Lamiaceae) selectively attract Asian honey bees

There are few floral volatiles of compounds that have been properly assessed for attracting pollinators. The intense plant odour of Elsholtzia rugulosa (Lamiaceae) to humans may be attributed to the presence of high concentrations of β‐caryophyllene. In a previous study, Zhang, Yang, and Zhang (Scientific Reports, 6, 2016a, 276161) speculated that the presence of β‐caryophyllene attracts pollinators (e.g., honey bees) to its flowers, an assumption that was assessed through the exploration of the functional significance of specific floral volatile compounds and the evaluation of their effects on the behaviour of Apis cerana (Asian honey bees; a known pollinator) and two non‐pollinators, a hornet (Vespa velutina) and a bumblebee (Bombus sp.). The results from these behavioural experiments indicated that both β‐caryophyllene and β‐elemene (both naturally found in flowers of E. rugulosa) were individually attractive to the Asian honey bees. Moreover, the combination of the two was more effective than either of them separately. In contrast, neither compound, nor a blend of the two at different concentrations, was attractive to the hornet and bumblebee species. These results demonstrate that β‐caryophyllene and β‐elemene play a key signalling role in attracting Asian honey bees to E. rugulosa.