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Pollination and parasitism in functionally dioecious figs.

Fig wasps (Agaonidae: Hymenoptera) are seed predators and their interactions with Ficus species (Moraceae) range from mutualism to parasitism. Recently considerable attention has been paid to conflicts of interest between the mutualists and how they are resolved in monoecious fig species. However, despite the fact that different conflicts can arise, little is known about the factors that influence the persistence of the mutualism in functionally dioecious Ficus. We studied the fig pollinator mutualism in 14 functionally dioecious fig species and one monoecious species from tropical lowland rainforests near Madang, Papua New Guinea. Observations and experiments suggest that (i) pollinating wasps are monophagous and attracted to a particular host species; (ii) pollinating and non-pollinating wasps are equally attracted to gall (male) figs and seed (female) figs in functionally dioecious species; (iii) differing style lengths between gall figs and seed figs may explain why pollinators do not develop in the latter; (iv) negative density dependence may stabilize the interaction between pollinating wasps and their parasitoids; and (v) seed figs may reduce the search efficiency of non-pollinators. This increased pollinator production without a corresponding decrease in seed production could provide an advantage for dioecy in conditions where pollinators are limiting.

Animals↗

Pollination-Induced Ethylene in Carnation (Role of Stylar Ethylene in Corolla Senescence).

In carnation (Dianthus caryophyllus L. cv White Sim) cell to cell communication between the pollen and pistil induces ovary development and corolla senescence. The production of elevated ethylene by the style is the first measurable postpollination response. This is followed by a wave of ethylene production from the other floral organs. To investigate the regulation of ethylene biosynthesis in pollinated flowers we measured ethylene production and the expression of 1-aminocyclopropane-1-carboxylate synthase and 1-aminocyclopropane-1-carboxylate oxidase transcripts in individual floral organs after pollination. Ethylene production by pollinated styles can be defined temporally by three distinct peaks. By pollinating a single style from a multistyle gynoecium, it was determined that the unpollinated style produces ethylene that corresponds to the first and third peaks observed from a pollinated style. Inhibition of ethylene action in the pollinated style by diazocyclopentadiene treatment prevented both pollination-induced corolla senescence and ethylene production from the ovaries and petals. Treatment with diazocyclopentadiene decreased stylar ethylene production during the second peak and completely inhibited the third peak of ethylene in both pollinated and unpollinated styles. This later auto-catalytic ethylene in styles is likely responsible for pollination-induced corolla senescence and ovary development.

Journal Article↗

Three 1-aminocyclopropane-1-carboxylate synthase genes regulated by primary and secondary pollination signals in orchid flowers.

The temporal and spatial expression patterns of three 1-aminocyclopropane-1-carboxylate (ACC) synthase genes were investigated in pollinated orchid (Phalaenopsis spp.) flowers. Pollination signals initiate a cascade of development events in multiple floral organs, including the induction of ethylene biosynthesis, which coordinates several postpollination developmental responses. The initiation and propagation of ethylene biosynthesis is regulated by the coordinated expression of three distinct ACC synthase genes in orchid flowers. One ACC synthase gene (Phal-ACS1) is regulated by ethylene and participates in amplification and interorgan transmission of the pollination signal, as we have previously described in a related orchid genus. Two additional ACC synthase genes (Phal-ACS2 and Phal-ACS3) are expressed primarily in the stigma and ovary of pollinated orchid flowers. Phal-ACS2 mRNA accumulated in the stigma within 1 h after pollination, whereas Phal-ACS1 mRNA was not detected until 6 h after pollination. Similar to the expression of Phal-ACS2, the Phal-ACS3 gene was expressed within 2 h after pollination in the ovary. Exogenous application of auxin, but not ACC, mimicked pollination by stimulating a rapid increase in ACC synthase activity in the stigma and ovary and inducing Phal-ACS2 and Phal-ACS3 mRNA accumulation in the stigma and ovary, respectively. These results provide the basis for an expanded model of interorgan regulation of three ACC synthase genes that respond to both primary (Phal-ACS2 and Phal-ACS3) and secondary (Phal-ACS1) pollination signals.

Base Sequence↗

A phylogenetic study of pollinator conservatism among sexually deceptive orchids.

Orchids of the genus Chiloglottis are pollinated through the sexual deception of male wasps mainly from the genus Neozeleboria (Tiphiidae: Thynninae). The orchids mimic both the appearance and sex pheromones of wingless female thynnines but provide no reward to the deceived males. Despite the asymmetry of this interaction, strong pollinator specificity is typical. Such plant-pollinator interactions would seem to be relatively flexible in the plant's adaptive response to variation in the local pollinator resource. However, we present DNA sequence data on both orchids and wasps that demonstrate a pattern of pollinator conservatism operating at a range of taxonomic levels. Sequence data from the wasps indicate 15 of 16 Chiloglottis pollinators are closely related members of one clade of Thynninae. A pattern of congruence between orchid and wasp phylogenies is also demonstrated below the generic level, such that related orchids tend to use related thynnine wasps as specific pollinators. Comparative physiological data on the wasp responses to the floral scents of two Chiloglottis species and one outgroup, Arthrochilus, indicate similar attractive volatile chemicals are used by related orchid taxa. By extension, we infer a similarity of sex pheromone signals among related thynnines. Thus, the conservative pattern of pollinator change in sexually deceptive orchids may reflect phylogenetic patterns in the sex pheromones of their pollinators.

Base Sequence↗

Pollination systems and floral traits in cerrado woody species of the Upper Taquari region (central Brazil).

Plant species present flowers with varied morphological and functional features, which may be associated to pollination systems, including species pollinated by wind, beetles, moths, bees, small insects, birds, or bats. We calculated the frequencies of the pollination systems among woody species in five cerrado fragments in central-western Brazil and tested whether the pollination systems were indeed related to floral traits. We sampled 2,280 individuals, belonging to 121 species, ninety-nine of which were described in relation to all floral traits. Most species had diurnal anthesis, pale colors, and open flowers. The most frequent groups were those composed by the species pollinated by bees, small insects, and moths. A Principal Component Analysis of the species and floral traits showed that there was a grouping among species with some pollination systems, such as those pollinated mainly by beetles, moths, birds, and bats, for which inferences based on the floral traits are recommended in cerrado sites. For the species pollinated mainly by bees or small insects, inferences based on the floral traits are not recommended, due to the large dispersion of the species scores and overlapping between these two groups, which probably occurred due to the specificity absence in plant-pollinator relationships.

Animals↗

Pollination of greenhouse tomatoes by the Australian bluebanded bee Amegilla (Zonamegilla) holmesi (Hymenoptera: Apidae).

The pollination effectiveness of bluebanded bees of the species Amegilla (Zonamegilla) holmesi Rayment (Hymenoptera: Apidae) was evaluated in tomato plants, Lycopersicon esculentum Miller (Solanaceae), cultivated in two greenhouse chambers. Bluebanded bee pollination was compared with mechanical pollination and no supplementary pollination. Pollination effectiveness was compared between treatments by using the percentage of fruit set, fruit weight, fruit diameter, fruit roundness, and the number of seeds per fruit. Both the bluebanded bee pollination and the mechanical pollination treatments significantly increased fruit set, individual fruit weight, and diameter compared with the control treatment. Fruit were also significantly rounder and contained significantly more seeds. Positive correlations were found for fruit weight versus seed number, maximum diameter versus seed number and minimum diameter versus seed number. We conclude that the use of A. holmesi for pollinating greenhouse tomatoes in Australia may be an effective alternative to the use of mechanical pollination.

Animals↗

Effects of local density on pollination and reproduction in Delphinium nuttallianum and Aconitum columbianum (Ranunculaceae).

Plant populations vary in density both naturally and as a consequence of anthropogenic impacts. Density in turn can influence pollination by animals. For example, plants in dense populations might enjoy more frequent visitation if pollinators forage most efficiently in such populations. We explored effects of plant density on pollination and seed set in the larkspur Delphinium nuttallianum and monkshood Aconitum columbianum. At our site in the Colorado Rocky Mountains, flowers of D. nuttallianum are pollinated primarily by queen bumble bees, solitary bees, and hummingbirds, whereas those of A. columbianum are pollinated primarily by queen and worker bumble bees. We found that the quantity of pollination service to both species (pollinator visitation rate and pollen deposition) was at best weakly related to density. In contrast, seed set declined by approximately one-third in sparse populations relative to nearby dense populations. This decline may stem from the receipt of low-quality pollen, for example, inbred pollen. Alternatively, sparsity may indicate poor environmental conditions that lower seed set for reasons unrelated to pollination. Our results demonstrate the value of simultaneously exploring pollinator behavior, pollen receipt, and seed set in attempting to understand how the population context influences plant reproductive success.

Journal Article↗

Pollination by flies, bees, and beetles of Nuphar ozarkana and N. advena (Nymphaeaceae).

Nuphar comprises 13 species of aquatic perennials distributed in the temperate Northern Hemisphere. The European species N. lutea and N. pumila in Norway, the Netherlands, and Germany are pollinated by bees and flies, including apparent Nuphar specialists. This contrasts with reports of predominant beetle pollination in American N. advena and N. polysepala. We studied pollination in N. ozarkana in Missouri and N. advena in Texas to assess whether (1) there is evidence of pollinator shifts associated with floral-morphological differences between Old World and New World species as hypothesized by Padgett, Les, and Crow (American Journal of Botany 86: 1316-1324. 1999) and (2) whether beetle pollination characterizes American species of Nuphar. Ninety-seven and 67% of flower visits in the two species were by sweat bees, especially Lasioglossum (Evylaeus) nelumbonis. Syrphid fly species visiting both species were Paragus sp., Chalcosyrphus metallicus, and Toxomerus geminatus. The long-horned leaf beetle Donacia piscatrix was common on leaves and stems of N. ozarkana but rarely visited flowers. Fifteen percent of visits to N. advena flowers were by D. piscatrix and D. texana. The beetles' role as pollinators was investigated experimentally by placing floating mesh cages that excluded flies and bees over N. advena buds about to open and adding beetles. Beetles visited 40% of the flowers in cages, and flowers that received visits had 69% seed set, likely due to beetle-mediated geitonogamy of 1st-d flowers. Experimentally outcrossed 1st-d flowers had 62% seed set, and open-pollinated flowers 76%; 2nd-d selfed or outcrossed flowers had low seed sets (9 and 12%, respectively). Flowers are strongly protogynous and do not self spontaneously. Flowers shielded from pollinators set no seeds. A comparison of pollinator spectra in the two Old World and three New World Nuphar species studied so far suggests that the relative contribution of flies, bees, and beetles to pollen transfer in any one population depends more on these insects' relative abundances (and in the case of Donacia, presence) and alternative food sources than on stamen length differences between Old World and New World pond-lilies.

Journal Article↗

Effect of zearalenone treatment on the production of wheat haploids via the maize pollination system.

The ability of zearalenone (ZEN) to stimulate the growth of haploid wheat embryos formed following the pollination of wheat spikes with maize pollen was tested. The maize pollination system was used as a model to compare the activity of ZEN with that of auxin analogues. Three solutions, each with a different concentration of ZEN (6.0, 3.0 or 1.5 microM), and a solution of 2,4-dichlorophenoxyacetic acid (control) were tested for their effect on ovary swelling, frequency of embryo formation and the ability to regenerate plants. In total, 3,105 florets of 282 spikes from five different cultivars of hexaploid winter wheat ( Triticum aestivum L.) were pollinated with maize ( Zea mays L. cv. Gama) pollen and treated with the ZEN solutions. The highest concentration of ZEN (6.0 microM) was the most effective in inducing ovary swelling (84 swollen ovaries/100 pollinated florets) and increasing the frequency of embryo induction (18.9 embryos/100 pollinated florets), but these embryos were severely deformed. They had low capability to germinate in vitro, while callus was easily formed and indirect regeneration of plants was possible. The lowest ZEN concentration (1.5 micrroM) induced ovary swelling in 42.8/100 pollinated florets and embryo growth in 3.3 out of 100 pollinated florets. The embryos were regular in shape, and almost half of them germinated in vitro while callus induction from them failed. The concentration of 3 microM ZEN had an intermediate effect. The type of response of the various wheat genotypes was similar, while frequencies were different, with cv. Izolda being the most responsive. The results show that ZEN has some of the properties of an auxin analogue, while other effects of its action are unique. It can be used in the maize pollination system of doubled haploid production to replace auxin analogues when indirect regeneration of plants via callus tissue is planned.

Dose-Response Relationship, Drug↗

Temporal and sexual variation of leaf-produced pollinator-attracting odours in the dwarf palm.

Information on intra-specific variation in pollinator-attracting floral traits provides clues to selective pressures imposed by pollinators. However, these traits also reflect constraints related to floral phenology or morphology. The specific weevil pollinator Derelomus chamaeropsis of the dioecious Mediterranean dwarf palm Chamaerops humilis is attracted by volatile compounds that leaves, and not flowers, release during anthesis. Production of these olfactory cues is thus probably not constrained by any other floral function. This provides the opportunity to study variation of a "floral" trait that is not produced by a floral organ. We studied volatile compounds emitted by leaves of 12 individual C. humilis over the whole flowering season. The quantity of volatile compounds emitted by leaves reached a maximum when plants required pollinator visits. The relationship between odour emission and floral phenology was slightly different between male and female plants, probably reflecting differences in the exact time at which females and males benefit from pollinator visitation. Male plants produced higher quantities of volatile compounds than females. Odour composition was highly variable among individuals but did not differ between male and female plants. In this system, female C. humilis are pollinated by deceit and pollinators should be selected to avoid visiting them. The absence of sexual difference in blend composition may thus prevent pollinators from discriminating between male and female plants.

Analysis of Variance↗

Floral scent emission and pollinator attraction in two species of Gymnadenia (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.

Adaptation, Physiological↗

Higher pollinator effectiveness by specialist than generalist flower-visitors of unspecialized Knautia arvensis (Dipsacaceae).

A critical issue in pollination ecology is the evolution of generalist pollination systems, and under which conditions floral specializations evolve from these. The gynodioecious herb Knautia arvensis (Dipsacaceae) exhibits a generalized pollination system, but is visited by both generalist and specialist flower-visitors. The present study tested pollinator effectiveness and pollinator importance of the pollen specialist solitary bee Andrena hattorfiana (Andrenidae) vs. the generalist flower-visitors to K. arvensis on the island of Oland, SE Sweden. Females of the specialist bee removed more pollen per inflorescence-visit than the major groups of generalist visitors such as bumblebees and flies. They also deposited more pollen per inflorescence-visit than any of the generalist visitor groups. The females have a preference for pollen-presenting vs. stigma-presenting inflorescences, a pattern shared with most of the generalist flower-visitors. Females of the specialist exert such a strong preference that they, despite their great pollinator effectiveness, make modest contribution to pollen transfer in K. arvensis. The females of A. hattorfiana accounted for 14.2% of the overall visits and 5.8% of the total pollination, the rest being performed by generalist visitors and males of A. hattorfiana. This study shows that pollinator effectiveness of a specialist can be superior while generalist flower-visitors select floral characters towards generalization through their greater contribution to overall pollen flow.

Animals↗

Proteome comparison following self- and across-pollination in self-incompatible apricot (Prunus armeniaca L.).

The study compared the protein differences between self- and across-pollinated self-incompatible (SI) apricots by two-dimensional gel electrophoresis and liquid chromatography-electrospray ion trap tandem mass spectrometry, the results showed that nine protein spots were expressed in self-pollinated pistil and only one was expressed in cross-pollinated pistils. Sixteen and three protein spots were up- and down-regulated in cross-pollinated pistils, respectively, compared with self-pollinated pistils. Seven protein spots were identified unambiguously by SEQUEST in NCBI protein database: Actin-12, enolase, MYB transcription-factor-like protein, heat-shock protein 70 were upregulated in cross-pollinated pistils compared with self-pollinated pistils; and actin-7, actin-8 and fructose bisphosphate aldolase-like protein were detected only in self-pollinated pistils.

Amino Acid Sequence↗

The smallest of all worlds: pollination networks.

A pollination network may be either 2-mode, describing trophic and reproductive interactions between communities of flowering plants and pollinator species within a well-defined habitat, or 1-mode, describing interactions between either plants or pollinators. In a 1-mode pollinator network, two pollinator species are linked to each other if they both visit the same plant species, and vice versa for plants. Properties of 2-mode networks and their derived 1-mode networks are highly correlated and so are properties of 1-mode pollinator and 1-mode plant networks. Most network properties are scale-dependent, i.e. they are dependent upon network size. Pollination networks have the strongest small-world properties of any networks yet studied, i.e. all species are close to each other (short average path length) and species are highly clustered. Species in pollination networks are much more densely linked than species in traditional food webs, i.e. they have a higher density of links, a shorter distance between species, and species are more clustered.

Animals↗

Flower visitors and pollination in the Oriental (Indomalayan) Region.

Current knowledge of flower visitors and pollination in the Oriental Region is summarised. Much less is known about pollination than seed dispersal and the coverage of habitats and taxa in the region is very uneven. The available evidence suggests that pollination in lowland forests is dominated by highly social bees (mainly Trigona and Apis species), with beetles probably the next most important group, followed by other bees and flies. In comparison with the better-studied Neotropics, large solitary bees, moths, Lepidoptera and vertebrates are relatively less important. These differences are greatest in the canopy of the lowland dipterocarp forests of Southeast Asia, where they probably reflect the unique temporal pattern of floral resource availability resulting from 'general flowering' at supra-annual intervals. Apis bees (but not Trigona species) are also important in most montane, subtropical and non-forest habitats. Apart from the figs (Ficus spp.), there are few well-documented examples of plant species visited by a single potential pollinator and most plant-pollinator relationships in the region appear to be relatively generalised. The small sizes of most pollinators and the absence of direct human exploitation probably make pollination mutualisms less vulnerable to failure as a result of human impacts than dispersal mutualisms, but more subtle impacts, as a result of altered gene flows, are likely to be widespread. On current evidence, pollination systems in the Oriental Region do not require any specific conservation action, but this review reinforces arguments for making the preservation (or restoration) of habitat connectivity the major focus of Oriental conservation.

Animals↗

Non-Euglossine bees also function as pollinators of Sinningia species (Gesneriaceae) in southeastern brazil.

Pollination by male and female Euglossini bees, euglossophily, was suggested for a number of neotropical Gesneriaceae species. Information on bee species other than Euglossini as pollinators of neotropical members of this family is limited, and in the tribe Sinningieae data about bee pollination are still lacking. Here, we report on floral biology and bee pollination of four Sinningia species: S. schiffneri, S. eumorpha, S. villosa, and Sinningia "canastrensis". The flower features, such as corolla size, shape, and colour, are very different among the four species, but all conform to the melittophilous syndrome. The average nectar volume and sugar amount is low in S. schiffneri, S. eumorpha, and Sinningia "canastrensis", when compared to that of S. villosa, but low nectar amounts is a general feature of Sinningia species. The main pollinators of the four species are: small Tapinotaspidini (Trigonopedia ferruginea) of S. schiffneri, large Bombini (Bombus morio) and large Centridini (Epicharis morio) of S. eumorpha, large Euglossini (Eulaema cingulata and Eufriesea surinamensis) of S. villosa, and large Euglossini (Eufriesea violascens) and Megachilini (Megachile sp.) of Sinningia "canastrensis". Out of the four species, only S. villosa is exclusively Euglossini-pollinated. The marked differences in flower features and nectar production of these Sinningia species may reflect their pollination by distinct groups of bees. These results strengthen the idea of multiple origins for the pollination systems involving bees within this genus, which is highly supported by molecular phylogenetic analyses.

Animals↗

Pollinator preference and the evolution of floral traits in monkeyflowers (Mimulus).

A paradigm of evolutionary biology is that adaptation and reproductive isolation are caused by a nearly infinite number of mutations of individually small effect. Here, we test this hypothesis by investigating the genetic basis of pollinator discrimination in two closely related species of monkeyflowers that differ in their major pollinators. This system provides a unique opportunity to investigate the genetic architecture of adaptation and speciation because floral traits that confer pollinator specificity also contribute to premating reproductive isolation. We asked: (i) What floral traits cause pollinator discrimination among plant species? and (ii) What is the genetic basis of these traits? We examined these questions by using data obtained from a large-scale field experiment where genetic markers were employed to determine the genetic basis of pollinator visitation. Observations of F2 hybrids produced by crossing bee-pollinated Mimulus lewisii with hummingbird-pollinated Mimulus cardinalis revealed that bees preferred large flowers low in anthocyanin and carotenoid pigments, whereas hummingbirds favored nectar-rich flowers high in anthocyanins. An allele that increases petal carotenoid concentration reduced bee visitation by 80%, whereas an allele that increases nectar production doubled hummingbird visitation. These results suggest that genes of large effect on pollinator preference have contributed to floral evolution and premating reproductive isolation in these monkeyflowers. This work contributes to growing evidence that adaptation and reproductive isolation may often involve major genes.

Journal Article↗

Pollination-induced ethylene promotes the early phase of pollen tube growth in Petunia inflata.

In Petunia inflata, a species with gametophytic self-incompatibility, pollination triggers two phases of ethylene production by the pistil, the first of which peaks 3 hours after pollination with compatible or incompatible pollen. To investigate the physiological significance of the first phase of ethylene production, pollinated flowers were treated with 2,5-norbornadiene (NBD), an inhibitor of ethylene action. Treatment with NBD reduced pollen tube growth in a dose-dependent manner during the first six hours after pollination; however, pollen tube growth was insensitive to NBD if the treatment was applied 6 hours or more after pollination. Simultaneous application of exogenous ethylene substantially offset the inhibitory effects of NBD in flowers pollinated for 4 hours. Another inhibitor of ethylene action, 1-methylcyclopropene (1-MCP), also produced a strong inhibition of pollen tube growth during the first six hours of pollination. The experiments with 1-MCP pretreatment indicate that pistil tissues are the primary target of the pollination-induced ethylene.

Culture Techniques↗