Floral scent in bird- and beetle-pollinated Protea species (Proteaceae): Chemistry,emission rates and function

Evolutionary shifts between pollination systems are often accompanied by modifications of floral traits, including olfactory cues. We investigated the implications of a shift from passerine bird to beetle pollination in Protea for floral scent chemistry, and also explored the functional significance of Protea scent for pollinator attraction. Using headspace sampling and gas chromatography–mass spectrometry, we found distinct differences in the emission rates and chemical composition of floral scents between eight bird- and four beetle-pollinated species. The amount of scent emitted from inflorescences of beetle-pollinated species was, on average, about 10-fold greater than that of bird-pollinated species. Floral scent of bird-pollinated species consists mainly of small amounts of “green-leaf volatiles” and benzenoid compounds, including benzaldehyde, anisole and benzyl alcohol. The floral scent of beetle-pollinated species is dominated by emissions of linalool, a wide variety of other monoterpenes and the benzenoid methyl benzoate, which imparts a fruity odour to the human nose. The number of compounds recorded in the scent of beetle-pollinated species was, on average, greater than in bird-pollinated species (45 versus 29 compounds, respectively). Choice experiments using a Y-maze showed that a primary pollinator of Protea species, the cetoniine beetle Atrichelaphinis tigrina, strongly preferred the scent of inflorescences of the beetle-pollinated Protea simplex over those of the bird-pollinated sympatric congener, Protea roupelliae. This study shows that a shift from passerine bird- to insect-pollination can be associated with marked up-regulation and compositional changes in floral scent emissions.     

Variation in scent emission among floral parts and inflorescence developmental stages in beetle-pollinated Protea species (Proteaceae)

Floral fragrances are an important component for pollinator attraction in beetle-pollinated flowers. Several genera in the Proteaceae contain beetle-pollinated species. However, there is no information on the floral scent chemistry of beetle-pollinated members of the family. In this paper we report on the spatial variation and differences between developmental stages in emission of inflorescence (flowerhead) volatiles of four South African Protea species (P. caffra, P. dracomontana, P. simplex, and P. welwitschii) that are pollinated by cetoniine beetles. The scents from different inflorescence parts (bracts, perianth, styles, and nectar) and from successive anthesis stages of whole inflorescences were sampled using dynamic headspace collection and identified using GC–MS. Although the four species shared many scent compounds, possibly reflecting their close phylogenetic relationships and common pollinators, they showed significant differences in overall scent composition due to various species-specific compounds, such as the unique tiglate esters found in the scent of P. welwitschii. The strongest emissions and largest number of volatiles, especially monoterpenes, were from inflorescences at full pollen dehiscence. Senescing inflorescences of two species and nectars of all species emitted proportionally high amounts of acetoin (3-hydroxy-2-butanone) and aromatic alcohols, typical fermentation products. As a consequence, the scent composition of nectar was much more similar among species than was the scent composition of other parts of the inflorescence. These results illustrate how the blends of compounds that make up the overall floral scent are a dynamic consequence of emissions from various plant parts.     

Chemical diversity of floral volatiles in Asclepiadoideae-Asclepiadeae (Apocynaceae)

Floral volatiles of 15 plant species from 8 genera (Cynanchum, Funastrum, Gonolobus, Metastelma, Oxypetalum, Orthosia, Sarcostemma, and Vincetoxicum) of the Apocynaceae-Asclepiadoideae tribe Asclepiadeae were investigated to get a better understanding of the odor chemistry, and of the role of odors in pollinator attraction. Floral volatiles were collected via headspace adsorption and analysed by GC–MS (gas chromatography–mass spectrometry). Of the 170 volatile compounds detected, 128 were identified, most of which are widespread floral scent compounds known from many different plant species. The species can be divided into two main groups: (1) species dominated by monoterpenoids (linalool, Z/E-ocimene), and (2) species dominated by benzenoids (benzaldehyde, phenylacetaldehyde, 2-phenylethyl acetate). Species of both chemotypes are pollinated mainly by Hymenoptera and Lepidoptera, and possibly these insects are effectively attracted by both monoterpenoids and benzenoids; alternatively, benzenoid-dominated plant species might attract a different subset of pollinators than the monoterpenoid-dominated plants.     

Evolution and diversity of floral scent chemistry in the euglossine bee-pollinated orchid genus Gongora

•Background and Aims Animal-pollinated angiosperms have evolved a variety of signalling mechanisms to attract pollinators. Floral scent is a key component of pollinator attraction, and its chemistry modulates both pollinator behaviour and the formation of plant–pollinator networks. The neotropical orchid genus Gongora exhibits specialized pollinator associations with male orchid bees (Euglossini). Male bees visit orchid flowers to collect volatile chemical compounds that they store in hind-leg pouches to use subsequently during courtship display. Hence, Gongora floral scent compounds simultaneously serve as signalling molecules and pollinator rewards. Furthermore, because floral scent acts as the predominant reproductive isolating barrier among lineages, it has been hypothesized that chemical traits are highly species specific. A comparative analysis of intra- and inter-specific variation of floral scent chemistry was conducted to investigate the evolutionary patterns across the genus.•Methods Gas chromatography–mass spectrometry (GC-MS) was used to analyse the floral scent of 78 individuals belonging to 28 different species of Gongora from two of the three major lineages sampled across the neotropical region. Multidimensional scaling and indicator value analyses were implemented to investigate the patterns of chemical diversity within and among taxonomic groups at various geographic scales. Additionally, pollinator observations were conducted on a sympatric community of Gongora orchids exhibiting distinct floral scent phenotypes.•Key Results A total of 83 floral volatiles, mainly terpenes and aromatic compounds, were detected. Many of the identified compounds are common across diverse angiosperm families (e.g. cineole, eugenol, β-ocimene, β-pinene and terpinen-4-ol), while others are relatively rare outside euglossine bee-pollinated orchid lineages. Additionally, 29 volatiles were identified that are known to attract and elicit collection behaviour in male bees. Floral scent traits were less variable within species than between species, and the analysis revealed exceptional levels of cryptic diversity. Gongora species were divided into 15 fragrance groups based on shared compounds. Fragrance groups indicate that floral scent variation is not predicted by taxonomic rank or biogeographic region.•Conclusions Gongora orchids emit a diverse array of scent molecules that are largely species specific, and closely related taxa exhibit qualitatively and quantitatively divergent chemical profiles. It is shown that within a community, Gongora scent chemotypes are correlated with near non-overlapping bee pollinator assemblies. The results lend support to the hypothesis that floral scent traits regulate the architecture of bee pollinator associations. Thus, Gongora provides unique opportunities to examine the interplay between floral traits and pollinator specialization in plant–pollinator mutualisms.     

Electrophysiological and behavioural responses of the housefly to “sweet” volatiles of the flowers of Caralluma europaea (Guss.) N.E. Br.

In sapromyiophilous plants, up to date, long range attraction of fly pollinators has been thoroughly investigated and attributed to “fetid” floral compounds, while the “sweet” floral scent fraction has not been specifically investigated and its role has received little attention. The aim of the present study was to verify if terpenoids, which are the main compounds of the floral bouquet of Caralluma europaea, play a role in the attraction of its pollinator Musca domestica. Terpinolene, α-terpinene and linalool, described as the three main volatiles of the flowers of C. europaea, were evaluated in electrophysiological investigations and blends of these compounds as well as the whole fresh flowers were used in behavioural assays. Antennae of housefly adults showed positive dose-dependent responses to all the chemicals tested. Houseflies were attracted by the odour of the fresh flowers and by the reconstructed terpenoid blend at the dose of 100 μg. At the dose of 10 μg, the blend did not produce any attraction. The results of the present study support the hypothesis that terpinolene, α-terpinene and linalool emitted by C. europaea flowers are involved in pollinator attraction and demonstrate the importance of the “sweet” scent in this sapromyiophilous species.     

Speciation in sexually deceptive orchids: pollinator-driven selection maintains discrete odour phenotypes in hybridizing species

Ophrys orchids mimic the female sex pheromones of their pollinator species to attract males for pollination. Reproductive isolation in Ophrys is based on the selective attraction of only a single pollinator species. A change of floral odour can result in the attraction of a new pollinator species that acts as an isolation barrier towards other sympatrically occurring Ophrys species. Ophrys lupercalis, Ophrys bilunulata, and Ophrys fabrella grow sympatrically and bloom consecutively on Majorca and are pollinated by three species of Andrena. We investigated variation of phenotypic and genotypic flower traits, aiming to study the role of the floral odour for reproductive isolation and speciation. Using chemical and electrophysiology (gas chromatography coupled with an electroantennographic detector) methods, we show that the three Ophrys species use the same odour compounds for pollinator attraction, but in different proportions. A comparison of the floral odour bouquets in a multivariate analysis revealed a clear grouping of plants from the same species, although with an overlap between species. A comparison of the same plants using molecular markers gave a contrasting result. Although O. lupercalis and O. fabrella were genetically well separated, plants of O. bilunulata did not form a distinct group but were similar to either O. lupercalis or O. fabrella. Our data indicate gene flow and hybridization to occur between O. bilunulata and O. lupercalis as well as between O. bilunulata and O. fabrella. All plants of O. bilunulata, despite having different genotypes, showed a very similar floral odour. This reflects a strong selective pressure by the pollinating males. The overlap of genotypes of O. bilunulata and O. fabrella supports our hypothesis that O. fabrella diverged from O. bilunulata by scent variation and the attraction of a new pollinator species, Andrena fabrella. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98 , 439–451.     

Floral scents of chafer-pollinated asclepiads and a potential hybrid

Floral scent is a key functional trait for pollinator attraction to flowers, but is poorly documented in many plant lineages and pollination systems. In South African grasslands, chafer beetles (Scarabaeidae: Cetoniinae), particularly Atrichelaphinis tigrina, Cyrtothyrea marginalis and Leucoscelis spp., are common floral visitors and specialized pollination by these beetles has recently been established in several asclepiad, orchid and protea species. Chafer beetles are known to be attracted by a variety of floral volatile compounds and scent has been suggested to be an important signal in these chafer-operated pollination systems. In this study, we used dynamic headspace extraction methods and coupled gas chromatography–mass spectrometry (GC–MS) to examine the chemical composition of the floral scents of seven putatively chafer-pollinated asclepiad species in the genera Asclepias, Pachycarpus and Xysmalobium. We identified 15–57 compounds in the scents of these species, of which seven were common to all species examined. The scent profiles of each species separate into discrete clusters in two dimensional space based on non-metric multidimensional scaling (NMDS), indicating clear distinctions between species and suggesting that plants may use different combinations of volatiles to attract beetles. Two plants suspected to be intergeneric hybrids were also examined. Data on pollination systems, morphology and scent chemistry are consistent with the hypothesis that these plants are hybrids between the chafer-pollinated species Asclepias woodii and Pachycarpus concolor. The results of this study are discussed in relation to the role of chafer beetles as generalist pollinators of specialized asclepiads.     

Cockroaches as pollinators of Clusia aff. sellowiana (Clusiaceae) on inselbergs in French Guiana

Background and Aims

A report is made on a new species of Clusia related to C. sellowiana that dominates the vegetation of the Nouragues inselberg in French Guiana. The focus is on the pollination biology and on the remarkable relationship of this plant species to Amazonina platystylata, its cockroach pollinator. This appears to be only the second record of pollination by cockroaches.

Methods

Pollination ecology was investigated by combining morphological studies, field observations and additional experiments. Floral scent was analysed by gas chromatography–mass spectrometry. The role of acetoin, the major component of the scent of this species of Clusia, in attracting pollinators was examined in field attraction experiments. The ability of cockroaches to perceive acetoin was investigated by electroantennography (EAG).

Key Results

The Clusia species studied produces seeds only sexually. Its nocturnal flowers are visited by crickets, ants, moths and cockroaches. A species of cockroach, Amazonina platystylata, is the principal pollinator. The reward for the visit is a liquid secretion produced by tissues at the floral apex and at the base of the ovary. Although the cockroaches have no structures specialized for pollen collection, their body surface is rough enough to retain pollen grains. The cockroaches show significant EAG reactions to floral volatiles and acetoin, suggesting that the floral scent is a factor involved in attracting the cockroaches to the flowers.

Conclusions

The results suggest that the plant–cockroach interaction may be quite specialized and the plant has probably evolved a specific strategy to attract and reward its cockroach pollinators. Acetoin is a substance involved in the chemical communication of several other cockroach species and it seems plausible that the plant exploits the sensitivity of cockroaches to this compound to attract them to the flowers as part of the pollination syndrome of this species.Key words: Clusia, cockroaches, acetoin, pollination, floral scent, floral reward, plant–animal interaction, inselberg, French Guiana, Amazonina platystylata     

Testing the effect of individual scent compounds on pollinator attraction in nature using quasi-isogenic Capsella lines

Premise

Floral scent, usually consisting of multiple compounds, is a complex trait, and its role in pollinator attraction has received increasing attention. However, disentangling the effect of individual floral scent compounds is difficult due to the complexity of isolating the effect of single compounds by traditional methods.

Methods

Using available quasi-isogenic lines (qILs) that were generated as part of the original mapping of the floral scent volatile-related loci CNL1 (benzaldehyde) and TPS2 (β-ocimene) in Capsella, we generated four genotypes that should only differ in these two compounds. Plants of the four genotypes were introduced into a common garden outside the natural range of C. rubella or C. grandiflora, with individuals of a self-compatible C. grandiflora line as pollen donors, whose different genetic background facilitates the detection of outcrossing events. Visitors to flowers of all five genotypes were compared, and the seeds set during the common-garden period were collected for high-throughput amplicon-based sequencing to estimate their outcrossing rates.

Results

Benzaldehyde and β-ocimene emissions were detected in the floral scent of corresponding genotypes. While some pollinator groups showed specific visitation preferences depending on scent compounds, the outcrossing rates in seeds did not vary among the four scent-manipulated genotypes.

Conclusions

The scent-manipulated Capsella materials constructed using qILs provide a powerful system to study the ecological effects of individual floral scent compounds under largely natural environments. In Capsella, individual benzaldehyde and β-ocimene emission may act as attractants for different types of pollinators.

     

Pollinator convergence and the nature of species' boundaries in sympatric Sardinian Ophrys (Orchidaceae)

Background and Aims

In the sexually deceptive Ophrys genus, species isolation is generally considered ethological and occurs via different, specific pollinators, but there are cases in which Ophrys species can share a common pollinator and differ in pollen placement on the body of the insect. In that condition, species are expected to be reproductively isolated through a pre-mating mechanical barrier. Here, the relative contribution of pre- vs. post-mating barriers to gene flow among two Ophrys species that share a common pollinator and can occur in sympatry is studied.

Methods

A natural hybrid zone on Sardinia between O. iricolor and O. incubacea, sharing Andrena morio as pollinator, was investigated by analysing floral traits involved in pollinator attraction as odour extracts both for non-active and active compounds and for labellum morphology. The genetic architecture of the hybrid zone was also estimated with amplified fragment length polymorphism (AFLP) markers, and pollination fitness and seed set of both parental species and their hybrids in the sympatric zone were estimated by controlled crosses.

Key Results

Although hybrids were intermediate between parental species in labellum morphology and non-active odour compounds, both parental species and hybrids produced a similar odour bouquet for active compounds. However, hybrids produced significantly lower fruit and seed set than parental species, and the genetic architecture of the hybrid zone suggests that they were mostly first-generation hybrids.

Conclusions

The two parental species hybridize in sympatry as a consequence of pollinator overlap and weak mechanical isolation, but post-zygotic barriers reduce hybrid frequency and fitness, and prevent extensive introgression. These results highlight a significant contribution of late post-mating barriers, such as chromosomal divergence, for maintaining reproductive isolation, in an orchid group for which pre-mating barriers are often considered predominant.Key words: AFLP markers, floral scent variation, hybrid zone, hybrid fitness, Ophrys iricolor, Ophrys incubacea, reproductive isolation, sexual deception     

Floral scent is different between sexual phases within individuals in a synchronously dichogamous shrub (Canella winterana) but there is no distinct female or male scent profile across individuals

Floral scent is a highly complex and variable floral characteristic that is involved in pollinator attraction. One possible cause for variation in floral scent can be sexual identity of the flower. Here, we examine the floral scent bouquet of a synchronously dichogamous shrub, Canella winterana (L.) Gaertn.We used dynamic headspace extraction and gas chromatography – mass spectroscopy, followed by statistical analysis using non-metric multidimensional scaling, SIMPER, MANOVA, and PERMANOVA to identify and compare the scent profiles of Canella winterana in its female and male phase for multiple individuals as well as multiple inflorescences of a single tree over one cycle of its entire sexual phase transition. The scent profile of C. winterana is composed of 49 volatile organic compounds and dominated by five compounds. We found no evidence for distinct male or female scent profiles; however, there were significant differences in scent emission between different sexes within some individuals. Two compounds explained over half of the variation between sexual stages within individuals. Our exploration of a single tree's sexual phase transition, including neuter phase, found that five compounds dominated the female phase scent bouquet and that female phase was distinct from male and neuter phase. This study offers new insight into the role that variability in floral scent between sexual phases might play in variable pollinator behavioral responses. These results suggest partial support for two distinct hypotheses regarding the differences between the sex phases (1) honest signaling and (2) sexual mimicry.     

Floral scent emission and pollinator attraction in two species of <Emphasis Type="Italic">Gymnadenia</Emphasis> (Orchidaceae)

We investigated scent composition and pollinator attraction in two closely related orchids, Gymnadenia conopsea (L.) R.Br. s.l. and Gymnadenia odoratissima (L.) Rich. in four populations during the day and night. We collected pollinators of both species using hand nets and sampled floral odour by headspace sorption. We analysed the samples by gas chromatography with mass spectrometry to identify compounds and with electroantennographic detection to identify compounds with physiological activity in pollinators. In order to evaluate the attractiveness of the physiologically active compounds, we carried out trapping experiments in the field with single active odour substances and mixtures thereof. By collecting insects from flowers, we caught eight pollinators of G. conopsea, which were members of four Lepidoptera families, and 37 pollinators of G. odoratissima, from five Lepidopteran families. There was no overlap in pollinator species caught from the two orchids using nets. In the scent analyses, we identified 45 volatiles in G. conopsea of which three (benzyl acetate, eugenol, benzyl benzoate) were physiologically active. In G. odoratissima, 44 volatiles were identified, of which seven were physiologically active (benzaldehyde, phenylacetaldehyde, benzyl acetate, 1-phenyl-2,3-butandione, phenylethyl acetate, eugenol, and one unknown compound). In field bioassays using a mixture of the active G. odoratissima compounds and phenylacetaldehyde alone we caught a total of 25 moths, some of which carried Gymnadenia pollinia. A blend of the active G. conopsea volatiles placed in the G. odoratissima population did not attract any pollinators. The two orchids emitted different odour bouquets during the day and night, but G. odoratissima showed greater temporal differences in odour composition, with phenylacetaldehyde showing a significant increase during the night. The species differed considerably in floral odour emission and this differentiation was stronger in the active than non-active compounds. This differentiation of the two species, especially in the emission of active compounds, appears to have evolved under selection for attraction of different suites of Lepidopteran pollinators.     

Floral volatiles play a key role in specialized ant pollination

Chemical signals emitted by plants are crucial to understand the ecology and evolution of plant–animal interactions. Scent is an important component of floral phenotype and represents a decisive communication channel between plants and floral visitors. Floral volatiles promote attraction of mutualistic pollinators and, in some cases, serve to prevent flower visitation by antagonists such as ants. Despite ant visits to flowers have been suggested to be detrimental to plant fitness, in recent years there has been a growing recognition of the positive role of ants in pollination. Nevertheless, the question of whether floral volatiles mediate mutualisms between ants and ant-pollinated plants still remains largely unexplored. Here we review the documented cases of ant pollination and investigate the chemical composition of the floral scent in the ant-pollinated plant Cytinus hypocistis. By using chemical-electrophysiological analyses and field behavioural assays, we examine the importance of olfactory cues for ants, identify compounds that stimulate antennal responses, and evaluate whether these compounds elicit behavioural responses. Our findings reveal that floral scent plays a crucial role in this mutualistic ant–flower interaction, and that only ant species that provide pollination services and not others occurring in the habitat are efficiently attracted by floral volatiles. 4-oxoisophorone, (E)-cinnamaldehyde, and (E)-cinnamyl alcohol were the most abundant compounds in Cytinus flowers, and ant antennae responded to all of them. Four ant pollinator species were significantly attracted to volatiles emitted by Cytinus inflorescences as well as to synthetic mixtures and single antennal-active compounds. The small amount of available data so far suggest that there is broad interspecific variation in floral scent composition among ant-pollinated plants, which could reflect differential responses and olfactory preferences among different ant species. Many exciting discoveries will be made as we enter into further research on chemical communication between ants and plants.     

Evolution of floral scent in relation to self-incompatibility and capacity for autonomous self-pollination in the perennial herb Arabis alpina

Background and AimsThe transition from outcrossing to selfing is a frequent evolutionary shift in flowering plants and is predicted to result in reduced allocation to pollinator attraction if plants can self-pollinate autonomously. The evolution of selfing is associated with reduced visual floral signalling in many systems, but effects on floral scent have received less attention. We compared multiple populations of the arctic–alpine herb Arabis alpina (Brassicaceae), and asked whether the transition from self-incompatibility to self-compatibility has been associated with reduced visual and chemical floral signalling. We further examined whether floral signalling differ between self-compatible populations with low and high capacity for autonomous self-pollination, as would be expected if benefits of signalling decrease with reduced dependence on pollinators for pollen transfer.MethodsIn a common garden we documented flower size and floral scent emission rate and composition in eight self-compatible and nine self-incompatible A. alpina populations. These included self-compatible Scandinavian populations with high capacity for autonomous self-pollination, self-compatible populations with low capacity for autonomous self-pollination from France and Spain, and self-incompatible populations from Italy and Greece.Key ResultsThe self-compatible populations produced smaller and less scented flowers than the self-incompatible populations. However, flower size and scent emission rate did not differ between self-compatible populations with high and low capacity for autonomous self-pollination. Floral scent composition differed between self-compatible and self-incompatible populations, but also varied substantially among populations within the two categories.ConclusionsOur study demonstrates extensive variation in floral scent among populations of a geographically widespread species. Contrary to expectation, floral signalling did not differ between self-compatible populations with high and low capacity for autonomous self-pollination, indicating that dependence on pollinator attraction can only partly explain variation in floral signalling. Additional variation may reflect adaptation to other aspects of local environments, genetic drift, or a combination of these processes.     

Spatial and temporal patterns of floral scent emission in Dianthus inoxianus and electroantennographic responses of its hawkmoth pollinator

Scent emission is important in nocturnal pollination systems, and plant species pollinated by nocturnal insects often present characteristic odor compositions and temporal patterns of emission. We investigated the temporal (day/night; flower lifetime) and spatial (different flower parts, nectar) pattern of flower scent emission in nocturnally pollinated Dianthusinoxianus, and determined which compounds elicit physiological responses on the antennae of the sphingid pollinator Hyles livornica.The scent of D.inoxianus comprises 68 volatile compounds, but is dominated by aliphatic 2-ketones and sesquiterpenoids, which altogether make up 82% of collected volatiles. Several major and minor compounds elicit electrophysiological responses in the antennae of H. livornica. Total odor emission does not vary along day and night hours, and neither does along the life of the flower. However, the proportion of compounds eliciting physiological responses varies between day and night. All flower parts as well as nectar release volatiles. The scent of isolated flower parts is dominated by fatty acid derivatives, whereas nectar is dominated by benzenoids. Dissection (= damage) of flowers induced a ca. 20-fold increase in the rate of emission of EAD-active volatiles, especially aliphatic 2-ketones.We suggest that aliphatic 2-ketones might contribute to pollinator attraction in D. inoxianus, even though they have been attributed an insect repellent function in other plant species. We also hypothesize that the benzenoids in nectar may act as an honest signal (‘nectar guide’) for pollinators.     

Pollination biology of Eulophia alta (Orchidaceae) in Amazonia: effects of pollinator composition on reproductive success in different populations

Background and Aims

Spatial variation in pollinator composition and abundance is a well-recognized phenomenon. However, a weakness of many studies claiming specificity of plant–pollinator interactions is that they are often restricted to a single locality. The aim of the present study was to investigate pollinator effectiveness of the different flower visitors to the terrestrial orchid Eulophia alta at three different localities and to analyse whether differences in pollinator abundance and composition effect this plant''s reproductive success.

Methods

Natural pollination was observed in vivo, and manipulative experiments were used to study the pollination biology and breeding system of E. alta at three sites near Manaus, Brazil. To gain a better understanding of the underlying mechanisms of pollinator attraction, nectar composition and secretion patterns were also studied, floral scent composition was analysed and a bioassay was conducted.

Key Results

Flower visitors, pollinator composition, pollinia transfer efficiency of particular pollinator species and natural fruit set differed among the investigated populations of E. alta. Flowers were self-compatible, partially autogamous and effectively pollinated by five bee species (four Centris species and Xylocopa muscaria). Visiting insects appeared to imbibe small amounts of hexose-rich nectar. Nectar sugar content was highest on the third day after flower opening. Floral fragrance analyses revealed 42 compounds, of which monoterpenes and benzenoids predominated. A bioassay using floral parts revealed that only floral tissue from the labellum chamber and labellum tip was attractive to flower visitors.

Conclusions

The data suggest that observed differences in reproductive success in the three populations cannot be explained by absolute abundance of pollinators alone. Due to behavioural patterns such as disturbance of effective pollinators on flowers by male Centris varia bees defending territory, pollinia transfer efficiencies of particular pollinator species also vary between study sites and result in differing reproductive success.     

Flower Scent and Pollination in Selected Neotropical Palms

Abstract: The flower scents of 14 palm species were collected in the field in Ecuador and Puerto Rico by head-space adsorption and analysed by gas chromatography-mass spectrometry. Insect visitors were recorded in seven of the species in Ecuador. The floral scent of the different species was dominated by a variety of compounds, e.g., the fatty-acid derived 3-pentanone and the hydrocarbon series dodecane to pentadecane, the benzenoid compound 1,4-dimethoxybenzene, the isoprenoids ( E )-ocimene, myrcene, linalool, and ( E )-α-farnesene and the nitrogen-containing compound 2-methoxy- sec -butylpyrazine. Rather than mirroring the systematics of the studied palm species, the chemical composition of the floral scent reflected the pollination mode. The scent of beetle-pollinated species was characterized by large amounts of one or a few dominant compounds, whereas fly- and bee-pollinated species contained a mixture of several compounds in smaller total amounts. We suggest that specific scent compounds, as found in the beetle-pollinated species, have evolved as a response to pollinator preferences. The importance of olfactory cues in relation to visual cues is higher in beetle-pollinated species than in species pollinated by flies and bees.     

Integrating floral scent, pollination ecology and population genetics

1 . Floral scent is a key factor in the attraction of pollinators. Despite this, the role of floral scent in angiosperm speciation and evolution remains poorly understood. Modern population genetic approaches when combined with pollination ecology can open new opportunities for studying the evolutionary role of floral scent.
2 . A framework of six hypotheses for the application of population genetic tools to questions about the evolutionary role of floral scent is presented. When floral volatile chemistry is linked to pollinator attraction we can analyse questions such as: Does floral volatile composition reflect plant species boundaries? Can floral scent facilitate or suppress hybridization between taxa? Can the attraction of different pollinators influence plant mating systems and pollen-mediated gene flow? How is population genetic structure indirectly influenced by floral scent variation?
3 . The application of molecular tools in sexually deceptive orchids has confirmed that volatile composition reflects species boundaries, revealed the role of shared floral odour in enabling hybridization, confirmed that the sexual attraction mediated by floral odour has implications for pollen flow and population genetic structure and provided examples of pollinator-mediated selection on floral scent variation. Interdisciplinary studies to explore links between floral volatile variation, ecology and population genetics are rare in other plant groups.
4 . Ideal study systems for future floral scent research that incorporate population genetics will include closely related taxa that are morphologically similar, sympatric and co-flowering as well as groups that display wide variation in pollination mechanisms and floral volatiles.     

Volatiles associated with different flower stages and leaves of Acacia cyclops and their potential role as host attractants for Dasineura dielsi (Diptera: Cecidomyiidae)

Acacia cyclops (Fabaceae) is an Australian species which was introduced into South Africa in the nineteenth century. Because of its invasive status in South Africa, a gall midge, Dasineura dielsi (Diptera: Cecidomyiidae), was released in 2001 in order to impact its reproduction by inducing galls on the flowers and thereby preventing seed set. Nothing is known about the cues used by D. dielsi for locating its host flowers. As part of an initial investigation into whether or not chemical cues might play a role in host finding, we analysed headspace samples of Acacia cyclops volatiles from leaves and reproductive parts at different stages (early bud, late bud, early flowering, and senescing flowering stages) using gas chromatography–mass spectrometry (GC–MS). In total, 72 different compounds were detected of which 62 were identified. The analyses showed that open flowers, the stage used by D. dielsi for oviposition, and yellow buds had similar odour compositions with (Z)-3-hexen-1-ol acetate, 4-oxoisophorone, (Z)-β-ocimene, an unknown aliphatic compound, heptadecane, and nonadecane dominating in open flowers. Leaf volatiles were distinct from those in the reproductive plant parts by their high relative amount of (Z)-β-ocimene. (Z)-3-Hexen-1-ol acetate had its maximum relative amount in the green bud samples and was much lower in the later floral stages. In contrast, 4-oxoisophorone peaked in yellow buds and open flowers with little or none of it found in younger or older stages. The volatile compounds of the different flower stages and leaves are discussed in relation to their potential role as attractants used by the biocontrol agent D. dielsi to locate its host plant.     

Pollination of the “carrion flowers” of an African stapeliad (<Emphasis Type="Italic">Ceropegia mixta</Emphasis>: Apocynaceae): the importance of visual and scent traits for the attraction of flies

Flowers that mimic carrion or faeces exhibit unusual traits, the evolution and functional significance of which remain poorly understood. Odour is an important pollinator attractant, but visual traits and interactions between visual and scent traits have seldom been considered. We studied pollination of the “carrion flowers” of Ceropegia mixta [= Orbea variegata], analysed floral traits and used manipulative experiments to explore the contributions of visual and scent traits to pollinator attraction. Flowers were pollinated primarily by Musca domestica (Muscidae), with lesser contributions by Calliphoridae and Sarcophagidae flies. The floral odour (analysed using gas chromatography–mass spectrometry) was dominated by oligosulphides and phenol. Comparison of floral and abiotic background colours (analysed using reflectance spectrometry) using a fly colour vision model suggested that flowers would be chromatically indistinguishable from the background. Comparison of fly arrival rates at concealed (but still scented) versus exposed flowers showed that flies can locate flowers without visual cues, but visitation was higher when the flowers were visible. Experiments using model flowers with odour supplied by real flowers (to explore the significance of dark flowers and dark spots on a pale background, which both occur frequently in flowers that mimic carrion or faeces) showed that scented black flowers attracted significantly more flies than similarly scented human-yellow flowers, while the presence or size of black spots on the corolla had no effect on the attraction of flies. Our results suggest that there is a visual component to fly attraction, but some traits, such as the mottled patterning, may not have evolved to enhance pollinator attraction.