Role In Pollinator Attraction Research Articles

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

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. 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. 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. 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.

Antennae of psychodid and sphaerocerid flies respond to a high variety of floral scent compounds of deceptive Arum maculatum L.

Insect-pollinated plants often release complex mixtures of floral scents to attract their pollinators. Yet scent compounds eliciting physiological or behavioural responses in pollinators have only been identified in few plant species. The sapromyiophilous aroid Arum maculatum releases a highly diverse dung-like scent with overall more than 300 different compounds recorded so far to attract its psychodid and other fly pollinators. The volatiles’ role in pollinator attraction is mostly unknown. To identify potential behaviourally active compounds, we recorded electroantennographic responses of four Psychodidae and one Sphaeroceridae species to (1) inflorescence scents of A. maculatum and (2) the scents released by cow dung, likely imitated by the plant species. Here we show that these flies are sensitive to 78 floral volatiles of various chemical classes, 18 of which were also found in cow dung. Our study, which for the first time determined physiologically active compounds in the antennae of Psychoda spp. and Sphaeroceridae, identified various volatiles not known to be biologically active in any floral visitors so far. The obtained results help deciphering the chemical basis that enables A. maculatum and other plants, pollinated by psychodids and sphaerocerids, to attract and deceive their pollinators.

Micro- and Macroscale Patterns of Petal Morphogenesis in Nigella damascena (Ranunculaceae) Revealed by Geometric Morphometrics and Cellular Analyses.

Petals, the inner organs in a differentiated perianth, generally play an important role in pollinator attraction. As such they exhibit an extraordinary diversity of shapes, sizes, and colors. Being involved in pollinator attraction and reward, they are privileged targets of evolution. The corolla of the Ranunculaceae species Nigella damascena consists of elaborate nectariferous petals, made of a stalk, upper, and lower lips forming a nectar pouch, shiny pseudonectaries, and pilose ears. While the main events of petal development are properly described, a few is known about the pattern of organ size and shape covariation and the cellular dynamics during development. In this study, we investigated the relationships between morphogenesis and growth of N. damascena petals using geometric morphometrics coupled with the study of cell characteristics. First, we found that petal shape and size dynamics are allometric during development and that their covariation suggests that petal shape change dynamics are exponentially slower than growth. We then found that cell proliferation is the major driver of shape patterning during development, while petal size dynamics are mostly driven by cell expansion. Our analyses provide a quantitative basis to characterize the relationships between shape, size, and cell characteristics during the development of an elaborate floral structure. Such studies lay the ground for future evo-devo investigations of the large morphological diversity observed in nectariferous structures, in Ranunculaceae and beyond.

Pollination and Floral Biology of a Rare Morning Glory Species Endemic to Thailand, Argyreia siamensis.

Argyreia siamensis is extremely rare, and very little is known about its reproduction. The species has colorful flowers that seem likely to attract pollinators, but population sizes are typically small (<30 individuals). To determine whether poor reproduction contributes to its rarity, we investigated its mating system and potential pollinators in two populations. We also examined the staminal trichomes and floral nectary to investigate their role in pollinator attraction. The mating system was assessed with a bagging experiment and pollinator visits were recorded with action cameras. Additionally, we tested the staminal trichomes and floral nectary for terpenes and flavonoids and examined floral nectary micromorphology via scanning electron microscope and compound light microscope. Our results reveal that A. siamensis is self-incompatible and dependent on pollinators; the western population was pollinated by bees (Meliponini and Amegilla), while the eastern population was mainly pollinated by skipper butterflies (Hesperiidae). Both staminal trichomes and the floral nectary appear to contribute to pollinator attraction through the presence of terpenes and flavonoids (in both secretory structures) and nectariferous tissue and nectarostomata (in the nectary). Our results indicate that A. siamensis has reliable and effective pollinators and that insufficient pollination is likely not a primary cause of its rarity.

Pollination ecology and floral scent chemistry of Philodendron fragrantissimum (Araceae)

ABSTRACT In French Guiana, Philodendron fragrantissimum (Hook.) G.Don was specifically pollinated by a single species of night-active cyclocephaline scarab, Cyclocephala simulatrix Höhne. Its inflorescences exhibited short-lasting anthesis (~30 h) and characteristic floral traits such as floral thermogenesis, edible/nutritious floral tissues, and profuse floral scent emission. Our insect exclusion experiments indicate that entomophilous cross-pollination is obligatory for P. fragrantissimum, with olfactory signalling playing a pivotal role in pollinator attraction. Three volatile organic compounds – methyl benzoate, (Z)-jasmone, and dehydrojasmone – dominate the floral scent of pistillate phase inflorescences (84–99%), but their relative proportions were different according to the headspace sampling method used, dynamic or static. Local pollination services provided by both female and male C. simulatrix are effective, as evidenced by a high visitation rate (>70%) that resulted in 55% of the inflorescences maturing into infructescences. Moreover, pollination was highly efficient, since 2–3 beetles sufficed to pollinate an average 94% of the pistillate flowers.

TCP and MADS-Box Transcription Factor Networks Regulate Heteromorphic Flower Type Identity in Gerbera hybrida.

The large sunflower family, Asteraceae, is characterized by compressed, flower-like inflorescences that may bear phenotypically distinct flower types. The CYCLOIDEA (CYC)/TEOSINTE BRANCHED1-like transcription factors (TFs) belonging to the TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) protein family are known to regulate bilateral symmetry in single flowers. In Asteraceae, they function at the inflorescence level, and were recruited to define differential flower type identities. Here, we identified upstream regulators of GhCYC3, a gene that specifies ray flower identity at the flower head margin in the model plant Gerbera hybrida We discovered a previously unidentified expression domain and functional role for the paralogous CINCINNATA-like TCP proteins. They function upstream of GhCYC3 and affect the developmental delay of marginal ray primordia during their early ontogeny. At the level of single flowers, the Asteraceae CYC genes show a unique function in regulating the elongation of showy ventral ligules that play a major role in pollinator attraction. We discovered that during ligule development, the E class MADS-box TF GRCD5 activates GhCYC3 expression. We propose that the C class MADS-box TF GAGA1 contributes to stamen development upstream of GhCYC3 Our data demonstrate how interactions among and between the conserved floral regulators, TCP and MADS-box TFs, contribute to the evolution of the elaborate inflorescence architecture of Asteraceae.

Lip morphology and ultrastructure of osmophores in Cyclopogon (Orchidaceae) reveal a degree of morphological differentiation among species.

Floral fragrances play an important role in pollinator attraction; they serve as signals for primary rewards such as nectar. The presence of osmophores (a specialized glandular tissue that produces fragrance) in the terrestrial orchid Cyclopogon has only been described in a single species. There are fragrant and odorless species within this genus, but no data on the similarities or differences between them has been published. Here, we present new data on the lip morphology and ultrastructure from eight Cyclopogon species to better understand the mechanisms of pollinator attraction of this group. To achieve this goal, we used light, scanning, and transmission electron microscopy. Five out of eight species presented globular trichomes associated with fragrance production on the abaxial side of the labellum. Together with subepidermal cells, they compose the secretory tissue. Despite the absence of globular trichomes, C. apricus also presented cells associated with fragrance production. Lipid droplets, plastids containing plastoglobuli, and starch were indicators of secretory activity present in these species. This same pattern of fragrance production and release, associated with flower morphology, may result in the restriction of pollination by halictid bees.

Aroma components of Cephalanthera orchids

The SPME-GC-MS analysis of the aroma components of Cephalanthera damasonium, Cephalanthera rubra, and Cephalanthera longifolia were reported. The main components found in C. damasonium were pentadecane, hexadecane, heptadecane and α-farnesene. The same compounds were found in the scent of C. rubra. Although C. damasonium and C. rubra have different pollination mechanisms, their spectrum of volatile compounds surprisingly does not differ significantly. In C. longifolia pentadecane and heptadecane were found together with cis-β-farnesene. As C. longifolia is allogamous, the differences in scent composition may play a role in pollinator attraction.

Nectar dynamics and reproductive biology of Passiflora actinia Hook. (Passifloraceae) in Araucaria Forest

ABSTRACT Nectar production has an important role in pollinator attraction and successful fruit production in many self-incompatible angiosperm groups. The reproductive biology of Passiflora actinia was studied here and related to nectar dynamics. Passiflora actinia presented a temporal segregation of male and female functions at the beginning of anthesis. Due to the movements of floral verticils, the anthers were positioned in a way that favors pollination two hours before the stigmas reached the same position. The nectary consisted of an epidermis with stomata and a parenchyma rich in starch, which was hydrolyzed during anthesis. The nectary organization is probably associated with the continuous production of nectar during anthesis as well as with the high mean nectar concentration. Hand pollination tests indicated that Passiflora actinia is obligately xenogamous, depending on large bees for pollination, specifically the carpenter bee Xylocopa augusti. The continuous production of nectar may increase the number of bee visits, thus favoring pollen flow.

Pollinator specialization in the enigmatic Rafflesia cantleyi: A true carrion flower with species-specific and sex-biased blow fly pollinators

The plants of the enigmatic genus Rafflesia are well known for their gigantic flowers and their floral features such as pungent floral scent and vivid dark color, which mimics the food/brood sites of carrion. However, information on the pollination biology of this plant group remains limited and mostly anecdotal. In the present paper, we studied the floral volatiles of R. cantleyi Solms-Laubach and their role in pollinator attraction. To achieve these aims, the floral scent was collected in situ in the field using a dynamic headspace method followed by chemical analysis via GC-MS. The olfactory preferences of pollinators to the identified chemical compounds, were tested singly and in blends, in flight tunnel bioassays and compared with responses to headspace floral extracts. In addition, flower-visiting calliphorid flies and the local carrion fly community were sampled and identified. Five species of calliphorid flies (subfamilies of Chrysomyinae and Calliphorinae), all females, were found on the flowers, whereas nine species were found in the traps that were baited with tainted meat in the surrounding habitat. However, only flower visitors of one blow fly species, Chrysomya chani Kurahashi, were observed to carry R. cantleyi pollen after visiting male flowers. The floral volatiles emitted by male flowers in full bloom were dominated by two sulphur-containing compounds, dimethyl disulfide (DMDS) and dimethyl trisulfide (DMTS). These were accompanied by other minor compounds such as benzenoids (4), monoterpenoids (4), trace amounts of aliphatic compounds (1), and sesquiterpenes (1). In flight-tunnel bioassays, a female-specific positive response of C. chani flies to individual DMDS, DMTS, and a blend of DMDS and DMTS was evident. Our findings suggest that R. cantleyi biochemically mimics carrion and that relative ratio of oligosulfides in the floral scent play a key role in sex-biased pollinator specialization, attracting only female C. chani carrion flies to the flowers.

Orchid floral volatiles: Biosynthesis genes and transcriptional regulations

Abstract Floral scent and colour are key trait for many floricultural crops. Floral volatiles are biologically and economically important plant derived compounds and they play vital role in pollinator attraction, plant defense and interaction with surrounding the environment. Orchidaceae is one of the largest and most widespread families of flowering plants, with more than 25,000 species. Orchids are one of the most significant plants in ecologically adopted every habitation on earth. Floral volatiles is not limited in its role to pollinator attraction, it’s widely used for perfumes, cosmetics, flavourings and therapeutic applications. Even though biochemistry of floral volatiles is still new field for investigation.Due to importance of the plant biology orchid floral scent research were deeply needed. Analysis of the biosynthesis mechanisms involved in floral scent is necessary to understand the fine-scale molecular functions and to breed to new cultivars through floral volatile genetic engineering approaches. Now a days many researchers developed floral scent models in plants. However, little is known about the pathways responsible for floral scent in orchids, due to the long life cycle and large genome size. Only some terpenoid pathways were reported in orchids. In this review, we mainly concentrated orchid floral volatile regulation and compounds synthesis responsible pathways. Moreover,we mentioned emission of orchid floral volatiles and their function in pollination ecology. This information will provide a basic information on orchid floral scent research.

Looks matter: changes in flower form affect pollination effectiveness in a sexually deceptive orchid.

Many species of the sexually deceptive genus Ophrys are characterized by insect-like flowers. Their form has been traditionally considered to play an important role in pollinator attraction and manipulation. Yet, the evolution of the floral form remains insufficiently understood. We hypothesize that pollinator-mediated selection is essential for driving floral form evolution in Ophrys, but that form components are being subjected to varying selection pressures depending on their role in mediating interactions with pollinators. By using the Eucera-pollinated Ophrys leochroma as a model, our aim has been to assess whether and in what manner pollination effectiveness is altered by experimental manipulation of the flower form. Our results show that floral form plays an essential and, so far, underestimated role in ensuring effective pollination by mechanically guiding pollinators towards the reproductive structures of the flower. Pollinators are significantly less effective in interacting with flowers having forms altered to resemble those of species pollinated by different hymenopteran genera. Further, those components used by pollinators as gripping points were found to be more effective in ensuring pollinia transfer than those with which pollinators do not directly interact. Thus, mechanically active and inactive components appear to be under different selection pressures. As a consequence, mechanically active components of the flower form could reflect adaptations to the interaction with particular pollinator groups, whereas mechanically inactive components can vary more freely. Disentangling selection patterns between the functionally different components of flower form may provide valuable insights into the mechanisms driving the morphological diversification of sexually deceptive pollination systems.

Floral scent chemistry and pollination in the Neotropical aroid genus Xanthosoma (Araceae)

The Neotropical aroid genus Xanthosoma (Caladieae, Araceae) embraces about 140 species of tuberous herbs whose night-blooming inflorescences emit strong, pleasant floral scents, presumably involved in the atttaction of pollinators. The number of studies on the floral scent chemistry of aroids, as well as on their role in pollinator attraction, has undergone a considerable increase in the last decade. In Xanthosoma, however, neither have never been properly assessed. In the present study, we provide an overview on the reproductive and pollination biology of Xanthosoma species and add new reports on the pollinators of some species. Furthermore, we investigate the floral scent chemistry of 13 species through dynamic headspace sample collection and analysis by gas chromatography coupled to mass spectrometry (GC–MS). A thorough literature survey and novel data from our own personal findings revealed that floral visitors are known for only 12 species of Xanthosoma, which represent less than 10% of its currently documented species diversity. Cyclocephaline scarabs (Melolonthidae, Cyclocephalini) were reported in inflorescences of 11 of the 12 species and are assumed to be the main pollinators associated with them. A total of 76 VOCs, mainly terpenes and benzenoids, were detected. Floral scent bouquets diverge considerably among analyzed species of Xanthosoma mainly due to the presence of unique prominent compounds that ensure originality of the blends rather than because of their quantitative complexity (number of constituents), as analyzed samples were typically dominated by few (2–4) VOCs that together accounted for >80% of the total relative scent discharge. A number of these dominant VOCs had already been isolated in other species of aroids and reported as effective attractants of cyclocephaline scarabs in field biossays, namely (Z)-jasmone, p-vinylanisole and dihydro-β-ionone and we speculate that along with other dominant compounds they are directly involved in pollinator selection in Xanthosoma.

Flowering Phenology and Floral Visitors in Distylous Populations ofPsychotria carthagenensis(Rubiaceae) in Brazilian Cerrado

Flowering synchrony has a relevant role in pollinator attraction. In distylous species, synchronous blooming between individuals of both morphs means that the chances of intermorph crosses are increased, which is important for the maintenance of the heterostylous system. The aim of the present study was to describe the flowering phenology and the behavior of the floral visitors in three populations of Psychotria carthagenensis Jacq. (Rubiaceae) between January 2008 and January 2010. Flower and pollinator censuses were conducted at three urban forest fragments in Brazil. Psychotria carthagenensis flowering occurred between October and December, with a high degree of synchrony within populations as well as between floral morphs. The exotic Apis mellifera L. (Apidae) was the main pollinator in the three populations. A high degree of similarity was found in the behavior of floral visitors as well as in nectar production. Visitation rates did not differ between floral morphs. Similar floral recompenses (flower availability and nectar) as well as no preference in pollinators' visiting are thought to be contributing to a greater intermorph visitation to pin and thrum flowers in P. carthagenensis.

Flower power and the mustard bomb: Comparative analysis of gene and genome duplications in glucosinolate biosynthetic pathway evolution in Cleomaceae and Brassicaceae.

Glucosinolates (GS) are a class of plant secondary metabolites that provide defense against herbivores and may play an important role in pollinator attraction. Through coevolution with plant-interacting organisms, glucosinolates have diversified into a variety of chemotypes through gene sub- and neofunctionalization. Polyploidy has been of major importance in the evolutionary history of these gene families and the development of chemically separate GS types. Here we study the effects of polyploidy in Tarenaya hassleriana (Cleomaceae) on the genes underlying GS biosynthesis. We established putative orthologs of all gene families involved in GS biosynthesis through sequence comparison and their duplication method through calculation of synonymous substitution ratios, phylogenetic gene trees, and synteny comparison. We drew expression data from previously published work of the identified genes and compared expression in several tissues. We show that the majority of gene family expansion in T. hassleriana has taken place through the retention of polyploid duplicates, together with tandem and transpositional duplicates. We also show that the large majority (>75%) is actively expressed either globally or in specific tissues. We show that MAM and CYP83 gene families, which are crucial to GS diversification in Brassicaceae, are also recruited into specific tissue expression pathways in Cleomaceae. Many GS genes have expanded through polyploidy, gene transposition duplication, and tandem duplication in Cleomaceae. Duplicate retention through these mechanisms is similar to A. thaliana, but based on the expression of GS genes, Cleomaceae-specific diversification of GS genes has taken place.

Hawk-moth pollination and elaborate petals in Cucurbitaceae: The case of the Caribbean endemic Linnaeosicyos amara

Linnaeosicyos amara is a little known cucurbit endemic to the Caribbean island of Hispaniola. The species possesses conspicuously fringed petals which are known in just four other cucurbit genera and hypothesised to play a role in pollinator attraction. A population of flowering L. amara plants was observed for a total of 78h, which revealed that Sphingid moths are the primary pollinators. Floral scent was dominated by the terpenoid (E)-β-ocimene. The elaborate petal fringes almost doubled the apparent diameter of L. amara flowers and the maximal extension of petal fringes was found to coincide with the peak visitation period of hawk-moths, with withering starting soon after. We conclude that petal fringes likely play a role in the visual attraction of hawk-moths by exploiting their preference for large flowers with deeply-divided petal margins, whilst limiting energy costs to the plant.

The role of preadaptations or evolutionary novelties for the evolution of sexually deceptive orchids.

The role of preadaptations or evolutionary novelties for the evolution of sexually deceptive orchids.

Reproductive ecology of Ameroglossum pernambucense (Scrophulariaceae): is this ornithophilous and threatened shrub highly adapted to a naturally fragmented habitat?

Many rare plants exist in a naturally fragmented distribution and are expected to exhibit reproductive adaptations to isolation. Thus, understanding the reproduction of these plants might be important in predicting the future of artificially fragmented species. Ameroglossum pernambucense is a threatened ornithophilous shrub with naturally fragmented populations on granite outcrops in north-eastern Brazil. The current research studied two populations of A. pernambucense and determined if three reproductive features (mating system, flowering phenology and nectar dynamics) facilitate the colonisation of new areas and the gene flow among the outcrops. To verify the species’ reproductive efficiency, pollinator efficiency and pollen limitation were tested. Ameroglossum pernambucense has a mixed mating system, and pollinators increased seed production. The pollinator efficiency test revealed that hummingbirds can supply the entire pollen demand. No pollen limitation was detected. Variation in sunset time was the main factor to explain the flowering pattern of A. pernambucense. Nectar was abundantly produced after successive removals. The reproductive traits observed in A. pernambucense were interpreted as highly adapted to its fragmented condition. Phenological photoperiodic regulation may increase the flowering synchrony and chances of gene flow among outcrops in environmentally distinct microsites. The mixed mating system favours both the colonisation of isolated outcrops and allogamy. High flower production and its high energetic value may have an important role in pollinator attraction and may increase long-distance outcrossing. Our data suggest that artificially fragmented plants with reproductive features similar to those observed here might be reproductively less susceptible to fragmentation.

The influence of pigmentation patterning on bumblebee foraging from flowers of Antirrhinum majus

Patterns of pigmentation overlying the petal vasculature are common in flowering plants and have been postulated to play a role in pollinator attraction. Previous studies report that such venation patterning is significantly more attractive to bee foragers in the field than ivory or white flowers without veins. To dissect the ways in which venation patterning of pigment can influence bumblebee behaviour, we investigated the response of flower-naïve individuals of Bombus terrestris to veined, ivory and red near-isogenic lines of Antirrhinum majus. We find that red venation shifts flower colour slightly, although the ivory background is the dominant colour. Bees were readily able to discriminate between ivory and veined flowers under differential conditioning but showed no innate preference when presented with a free choice of rewarding ivory and veined flowers. In contrast, both ivory and veined flowers were selected significantly more often than were red flowers. We conclude that advantages conferred by venation patterning might stem from bees learning of their use as nectar guides, rather than from any innate preference for striped flowers.

Possible chemical mimicry of the European lady’s slipper orchid (Cypripedium calceolus)

Pollination based on insect deception has been debated in the scientific community since it was first reported over two hundred years ago. A vast majority of deceptive syndromes occur within the orchid family. While many cheating flowers have been described and are well known, there are still many curious cases that need further investigation. One prime example of such a case is Cypripedium calceolus, known as European lady’s slipper orchid. While the flower has been of interest to many prominent scientists for over a century, its pollination is still not fully understood. Both visual and olfactory cues seem to play an important role in pollinator attraction. In this study we focussed on the olfactory cues in order to explore their relationship (in future experiments) with floral visual cues, including the unique asymmetry of these flowers. Some of the plants’ floral fragrances were used in Electroantennography experiments. Eleven chemical compounds were applied to the antennae of Bombus terrestris and Apis mellifera. Even though these species are not regular visitors of C. calceolus, we were interested to see whether there were common principles in their responses to the flowers’ scent that might justify extrapolating to other pollinator species such as sand bees that get trapped in these orchids and fly out of the flowers afterwards with pollen smeared on their body. The results show that while both species react similarly to most of the odours, some of the tested acetates induced a significantly greater reaction in B. terrestris antennae. These acetates play an important role in bumblebee pheromones, but their relevance for the natural pollinators of C. calceolus remains to be confirmed to see whether chemical mimicry by these flowers is deliberately employed to attract pollinators.