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Anther cap retention prevents self-pollination by elaterid beetles in the South African orchid Eulophia foliosa.

BACKGROUND AND AIMS: Pollination by insects that spend long periods visiting many flowers on a plant may impose a higher risk of facilitated self-pollination. Orchids and asclepiads are particularly at risk as their pollen is packaged as pollinia and so can be deposited on self-stigmas en masse. Many orchids and asclepiads have adaptations to limit self-deposition of pollinia, including gradual reconfiguration of pollinaria following removal. Here an unusual mechanism--anther cap retention--that appears to prevent self-pollination in the South African orchid Eulophia foliosa is examined. METHODS: Visits to inflorescences in the field were observed and pollinators collected. Visitation rates to transplanted inflorescences were compared between a site where putative pollinators were abundant and a site where they were rare. Anther cap retention times were determined for removed pollinaria and atmospheric vapour pressure deficit was recorded concurrently. Anther cap anatomy was examined using light microscopy. KEY RESULTS: Eulophia foliosa is pollinated almost exclusively by Cardiophorus obliquemaculatus (Elateridae) beetles, which remain on the deceptive inflorescences for on average 301 s (n = 18). The anther cap that covers the pollinarium is retained for an average of 512 s (n = 24) after pollinarium removal by beetles. In all populations measured, anther cap dimensions are greater than those of the stigmatic cavity, thus precluding the deposition of self-pollinia until after the anther cap has dropped. An anatomical investigation of this mechanism suggests that differential water loss from regions of the anther cap results in opening of the anther cap flaps. This is supported by observations that as atmospheric vapour pressure deficits increased, the duration of anther cap retention was reduced. CONCLUSIONS: Flowers of E. foliosa are specialized for pollination by elaterid beetles. Retention of anther caps for a period exceeding average visit times by beetles to inflorescences appears to prevent facilitated self-pollination in E. foliosa effectively.

Animals↗

Behavior and pollination efficiency of Nannotrigona perilampoides (Hymenoptera: Meliponini) on greenhouse tomatoes (Lycopersicon esculentum) in subtropical México.

The acclimation, foraging behavior, and pollination efficiency of stingless bees of the species Nannotrigona perilampoides Cresson were evaluated in tomato (Lycopersicon esculentum Mill.) plants cultivated in two greenhouses. The greenhouses were divided into three areas of 16 m2, and one of the following treatments was used for pollination: stingless bees (SB), mechanical vibration (MV), and no pollination (NP). Observations were conducted once a week from 0800 to 1600 hours during 2 mo. The acclimation of the bees to the greenhouses was estimated by the number of bees that did not return to the hive (lost bees) and by comparing the population of the colonies (brood and adults). The foraging activity of the bees across the day was evaluated by comparing the number of foragers per hour. The influence of environmental variables on the foraging activity was also analyzed. The pollination efficiency was compared among treatments through the percentage of fruit set, weight of individual fruit, kilograms of fruit produced per square meter, and the number of seed per fruit. The bees started foraging on the flowers approximately 7 d after the colonies were introduced to the greenhouse. There was a decline in the population of the colonies across the experiment, but colonies did not die out. Correlations of environmental variables with the foraging activity of the bees showed that none of them had a significant influence on pollen foraging. However, water collection was positively correlated with the temperature and negatively correlated with the humidity inside the greenhouse. The estimation of the pollination efficiency per treatment showed that there were significant differences in fruit set in SB (83 +/- 4.2) and MV (78.5 +/- 6.4) compared with NP (52.6 +/- 7.6). However, the average weight of the fruit was similar for the three treatments (65 g). There were significant differences for seed number in SB (200 +/- 15.3) and MV (232 +/- 21.4) compared with NP (120 +/- 16.6). The productivity in kilograms of fruit per square meter was higher in SB (5.72 +/- 0.61) and MV (5.66 +/- 0.58 kg) compared with NP (3.34 +/- 0.72). The number of seed was positively correlated with the weight of the fruit. We conclude that the use of Nannotrigona testaceicornis Rondani, for pollinating greenhouse tomatoes in tropical climates, could be an alternative to the use of highly defensive African-derived Apis mellifera or non-native bumble bees (Bombus spp.). However, more research is needed to evaluate the cost/benefit on large-scale greenhouse pollination using N. perilampoides Cresson against other bee species and pollination methods.

Agriculture↗

Effects of Inhibitors of Protein Serine/Threonine Phosphatases on Pollination in Brassica.

We have examined the effect of the protein phosphatase inhibitors okadaic acid and microcystin on pollen-pistil interactions in Brassica. Inhibitor-treated flowers or floral buds were pollinated with untreated pollen and examined for pollen tube growth by fluorescence microscopy. Our results show that type 1 or type 2A serine/threonine phosphatases play a crucial role in the pollination responses of Brassica. We observed two distinct effects of protein phosphatase inhibitors on pollination: (a) the inhibition of pollen tube growth during cross-pollination in flowers, and (b) the break-down of self-incompatibility or promotion of pollen tube growth during self-pollination in flower buds just prior to anthesis. Thus, treatment of flower pistils with protein phosphatase inhibitors resulted in the inhibition of pollen tube growth at the surface of the papillar cells of the stigma in crosses between different self-incompatible Brassica oleracea strains, in an interspecific cross between B. oleracea and Brassica campestris, and in self-pollinations of a self-fertile Brassica napus cultivar. With four different self-incompatibility genotypes, treatment of mature flowers with protein phosphatase inhibitors had no effect on self-pollination response. In contrast, treatment of flower buds just prior to the anthesis stage allowed self-pollen tube invasion of papillar cells. However, the magnitude of this effect was genotype dependent, being most pronounced in the S22 genotype. The data support the conclusion that pollinations in Brassica are controlled in part by the presence of phosphorylated proteins in the papillar cells of the stigma, and that the quantity of these proteins or their levels of phosphorylation changes during stigma development.

Journal Article↗

Development and Pollination Regulated Accumulation and Glycosylation of a Stylar Transmitting Tissue-Specific Proline-Rich Protein.

The extracellular matrix of stylar transmitting tissues of many angiosperms is enriched in secretory materials that are believed to be important for interactions with pollen tubes. We have previously characterized two related cDNAs (TTS-1 and TTS-2) for stylar transmitting tissue-specific proline-rich proteins (TTS proteins) from Nicotiana tabacum. We show here that TTS proteins are highly glycosylated proteins with apparent molecular masses ranging between 50 and 100 kD. Results from chemical and enzymatic deglycosylation suggest that TTS proteins have N-linked glycosyl groups, and the extensive glycosylation most probably has resulted from modifications at the proline residues. TTS proteins are localized to the intercellular regions between neighboring transmitting tissue cells, the space in which pollen tubes elongate as they migrate from the stigma toward the ovary. TTS mRNA and protein levels are regulated during pistil development and by pollination. The levels of TTS mRNAs and proteins increase with flower development and reach the maximal levels as flowers approach anthesis. These maximal levels are maintained in the styles for at least 3 to 4 days after pollination, during which time pollen tubes elongate and reach the ovary. Spatially, TTS mRNAs and proteins accumulate first in the stigmatic end of young styles, and their levels progressively increase toward the basal end as pistils mature. Pollination stimulates the levels of TTS mRNAs and proteins in hand-pollinated young styles, which normally accumulate relatively low levels of these TTS gene products. Pollination also qualitatively affects TTS mRNAs and proteins. In pollinated styles, TTS mRNAs are shorter than those in unpollinated styles and underglycosylated TTS protein species begin to accumulate. The elaborate regulatory mechanisms governing TTS mRNAs and proteins during development and by pollination strongly suggest that these proteins may play a functional role in the process of pollination.

Journal Article↗

Pollinator preferences for Nicotiana alata, N. forgetiana, and their F1 hybrids.

The role of pollinators in plant speciation and maintenance of species boundaries is dubious because most plant species are visited by several types of pollinators, and most pollinator species visit several species of plants. We investigated pollinator preferences and their efficacy as ethological isolation mechanisms between two interfertile species, Nicotiana alata and N. forgetiana and their F1 hybrids. Hawkmoths pollinate N. alata, while primarily hummingbirds and occasionally small hawkmoths visit N. forgetiana. F1 hybrids are easily produced in the greenhouse and although the species grow in similar habitats, hybrids have not been found in nature. In Rio Grande do Sul, Brazil, near where both species are found, experimental plots were studied containing both species, and both species plus F1 hybrids. In the mixed-species plots, hawkmoths showed a strong preference for N. alata. Hummingbirds were less common and only visited N. forgetiana. Hybrid seed was produced but plants made significantly fewer hybrid offspring than predicted by the frequency of interspecific pollinator movements. Nicotiana forgetiana was the seed parent of 97% of the F1 offspring, suggesting an asymmetry in pollen delivery or postpollination processes. In plots containing F1 hybrids plus both parental species, hawkmoths preferred N. alata and undervisited the other two phenotypes, except that in the third plot they visited hybrids in proportion to the hybrid frequency. Hummingbirds strongly preferred N. forgetiana in all plots but also visited F1 hybrids in proportion to their frequency in the third plot. Overall, F1 hybrids were well pollinated and were frequently visited immediately before or after one of the parental species. Thus hybrids could facilitate gene flow between the parental species. We conclude that pollinator discrimination among species is strong but is an imperfect isolation mechanism, especially if hybrids are present.

Animals↗

Relating geographical variation in pollination types to environmental and spatial factors using novel statistical methods.

The relative frequencies of functional traits of plant species show notable spatial variation, which is often related to environmental factors. Pollination type (insect-, wind- or self-pollination) is a critical trait for plant reproduction and provision of ecosystem services. Here, we mapped the distribution of pollination types across Germany by combining databases on plant distribution and plant pollination types. Applying a new method, we modelled the composition of pollination types using a set of 12 environmental variables as predictors within a Bayesian framework which allows for the analysis of compositional data in the presence of spatial autocorrelation. A clear biogeographical pattern in the distribution of pollination types was revealed which was adequately captured by our model. The most striking relationship was a relative increase in insect-pollination and a corresponding decrease of selfing with increasing altitude. Further important factors were wind speed, geology and land use. We present a powerful tool to analyse the distribution patterns of plant functional types such as pollination types and their relationship with environmental parameters in a spatially explicit framework.

Demography↗

New evidence for the Darwinian hypothesis of heterostyly: breeding systems and pollinators in Narcissus sect. Apodanthi.

Here we analysed the role played by breeding systems and pollinators in the evolution of heterostyly by testing whether evolution towards heterostyly is associated with style polymorphism and changes in pollinator proficiency or breeding system variation (Darwinian hypothesis). We studied pollinators, pollen-transfer efficiency, and incompatibility systems in all seven species of Narcissus sect. Apodanthi for which we also obtained chloroplast DNA (cpDNA) sequences from three spacers to infer phylogenetic relationships. Five species are self-incompatible and within-morph cross-compatible. Heterostylous (Narcissus albimarginatus) and style-dimorphic (Narcissus cuatrecasasii) species that have a high degree of reciprocity in stigma and anther height are primarily pollinated by solitary bees. The style-monomorphic species (Narcissus watieri) and the style-dimorphic species with the least stigma-anther reciprocity (Narcissus rupicola) are both self-compatible and pollinated by butterflies, moths and hover flies. Phylogenetic reconstruction of character transitions indicates that the shift from style dimorphism to distyly is associated with a shift to bee pollination. Pollination by lepidopterans and flies is associated with stable style dimorphism and monomorphism. Evolution and maintenance of style polymorphisms in this group of species are independent of incompatibility systems. Taken together, our results strongly support the pollinator-based model for evolution of heterostyly and style length polymorphisms in general.

Animals↗

POLLINATION REGULATION OF FLOWER DEVELOPMENT.

Pollination regulates a syndrome of developmental responses that contributes to successful sexual reproduction in higher plants. Pollination-regulated developmental events collectively prepare the flower for fertilization and embryogenesis while bringing about the loss of floral organs that have completed their function in pollen dispersal and reception. Components of this process include changes in flower pigmentation, senescence and abscission of floral organs, growth and development of the ovary, and, in certain cases, pollination also triggers ovule and female gametophyte development in anticipation of fertilization. Pollination-regulated development is initiated by the primary pollination event at the stigma surface, but because developmental processes occur in distal floral organs, the activity of interorgan signals that amplify and transmit the primary pollination signal to floral organs is implicated. Interorgan signaling and signal amplification involves the regulation of ethylene biosynthetic gene expression and interorgan transport of hormones and their precursors. The coordination of pollination- regulated flower development including gametophyte, embryo, and ovary development; pollination signaling; the molecular regulation of ethylene biosynthesis; and interorgan communication are presented.

Journal Article↗

Bumble bee (Hymenoptera: Apidae) activity and pollination levels in commercial tomato greenhouses.

Commercial greenhouse studies were conducted to assess levels of pollination of tomato (Lycopersicon esculentum Mill.) flowers in relation to bumble bee (Bombus impatiens Cresson) colony activity and colony densities. For the assessment of pollination levels of tomato flowers, five categories were defined based on bruising levels caused by bumble bee pollination. Colony activity was measured as bee trips per ha/d using electric powered photodiode monitors inserted into the hive entrance. Levels of pollination were positively correlated with bee activity levels, up to a mean of approximately 400 pollen grains per stigma per day, after which greater activity did not result in further increases in daily pollination levels. Densities of colonies in the commercial greenhouses studied ranged from 7.6 to 19.8 colonies per hectare with a mean of 11.6 +/- 0.9. We found that an average activity of 2,000 bee trips per hectare per day was more than adequate to ensure sufficient pollination, and that this level of activity could be achieved with 7-15 colonies per hectare, depending on greenhouse conditions. Greenhouses requiring >15 colonies per hectare to achieve this level of pollination may be able to increase bee activity through alteration of greenhouse conditions. Across 50-m rows of tomato plants, levels of pollination decreased with increasing distance from bee colonies, suggesting that colonies should be evenly distributed throughout the greenhouses.

Animals↗

Differential ovule development following self- and cross-pollination: the basis of self-sterility in Narcissus triandrus (Amaryllidaceae).

Self-pollination results in significantly lower seed set than cross-pollination in tristylous Narcissus triandrus. We investigated structural and functional aspects of pollen-pistil interactions and ovule-seed development following cross- and self-pollination to assess the timing and mechanism of self-sterility. Ovule development within an ovary was asynchronous at anthesis. There were no significant differences in pollen tube behavior following cross- vs. self-pollination during the first 6 d of growth, regardless of style morph type. Double fertilization was significantly higher following cross- vs. self-pollination. Aborted embryo development was not detected following either pollination type up to seed maturity. Prior to pollen tube entry, a significantly greater number of ovules ceased to develop following self- vs. cross-pollination. These results indicate that self-sterility in N. triandrus operates prezygotically but does not involve differential pollen tube growth typical of many self-incompatibility (SI) systems. Instead, low seed set following self-pollination is caused by a reduction in ovule availability resulting from embryo sac degeneration. We hypothesize that this is due to the absence of a required stimulus for normal ovule development. If this is correct, current concepts of SI may need to be broadened to include a wider range of pollen-pistil interactions.

Journal Article↗

Pollen dispersal in Yucca filamentosa (Agavaceae): the paradox of self-pollination behavior by Tegeticula yuccasella (Prodoxidae).

We investigated pollen dispersal in an obligate pollination mutualism between Yucca filamentosa and Tegeticula yuccasella. Yucca moths are the only documented pollinator of yuccas, and moth larvae feed solely on developing yucca seeds. The quality of pollination by a female moth affects larval survival because flowers receiving small amounts of pollen or self-pollen have a high abscission probability, and larvae die in abscised flowers. We tested the prediction that yucca moths primarily perform outcross pollinations by using fluorescent dye to track pollen dispersal in five populations of Y. filamentosa. Dye transfers within plants were common in all populations (mean ± 1 SE, 55 ± 3.0%), indicating that moths frequently deposit self-pollen. Distance of dye transfers ranged from 0 to 50 m, and the mean number of flowering plants between the pollen donor and recipient was 5 (median = 0), suggesting that most pollen was transferred among near neighbors. A multilocus genetic estimate of outcrossing based on seedlings matured from open-pollinated fruits at one site was 94 ± 6% (mean ± 1 SD). We discuss why moths frequently deposit self-pollen to the detriment of their offspring and compare the yucca-yucca moth interaction with other obligate pollinator mutualisms in which neither pollinator nor plant benefit from self-pollination.

Journal Article↗

Bird-pollination of three Durio species (Bombacaceae) in a tropical rainforest in Sarawak, Malaysia.

Pollination ecology of three Durio species, D. grandiflorus, D. oblongus, and D. kutejensis (Bombacaceae), was studied in a lowland dipterocarp forest in Sarawak, Malaysia, during a peak flowering period when at least 305 species of plants bloomed in 1996. Durio has been reported to be pollinated by bats in Peninsular Malaysia. However, my observations of flower visitors and pollination experiments indicated that two species, D. grandiflorus and D. oblongus, were pollinated by spiderhunters (Nectariniidae) and that the other species, D. kutejensis, was pollinated by giant honey bees and bats as well as birds. Hand-pollination experiments showed that all three species were obligate outbreeders. A resource limitation in fruit production was suggested. The former two species were visited only by spiderhunters, and the bagged flowers that were opened for animal visitors only at night bore no fruit, while those that were opened only during the day bore fruits, at comparable fruiting ratios to open pollination. Durio kutejensis was observed to be visited by giant honey bees, birds, and bats at different times of day, and three series of bagged experiments that exposed the flowers to animal visitors at different times of day bore fruits at a comparable ratio to open-pollination.

Journal Article↗

Fly-pollinated Pleurothallis (Orchidaceae) species have high genetic variability: evidence from isozyme markers.

We conducted an isozyme study in 22 populations of five Pleurothallis (Orchidaceae) species (12 loci in nine enzymatic systems). The genetic variability in all populations is surprisingly high (P = 58-83%, A = 2.1-3.8, H(e) = 0.25-0.43) in spite of the fact that the five species are pollinated by small flies whose behavior enables self-pollination. We suggest that self-incompatibility, inbreeding depression, and mechanical barriers that prevent self-pollination in these species are responsible for the maintainance of the high genetic variability. These traits are uncommon in Orchidaceae, but have been observed in these and some other species pollinated by flies or other pollinators with behavior that facilitates self-pollination. The genetic similarity among conspecific populations is also high for species with very short-range flying pollinators. Only one population of P. teres presented values of genetic similarity lower than usually observed in allopatric conspecific populations. Morphology, however, does not support its segregation as a new taxon. All species can be recognized by their enzymatic patterns, and the results agree with recently proposed taxonomic realignments. Conversely, the supposed affinities among these species based on floral morphology are not supported, and we hypothesize that it may be due to convergence in species with similar pollinators.

Journal Article↗

Phylogenetic relationships of fig wasps pollinating functionally dioecious Ficus based on mitochondrial DNA sequences and morphology.

The obligate mutualism between pollinating fig wasps in the family Agaonidae (Hymenoptera: Chalcidoidea) and Ficus species (Moraceae) is often regarded as an example of co-evolution but little is known about the history of the interaction, and understanding the origin of functionally dioecious fig pollination has been especially difficult. The phylogenetic relationships of fig wasps pollinating functionally dioecious Ficus were inferred from mitochondrial cytochrome oxidase gene sequences (mtDNA) and morphology. Separate and combined analyses indicated that the pollinators of functionally dioecious figs are not monophyletic. However, pollinator relationships were generally congruent with host phylogeny and support a revised classification of Ficus. Ancestral changes in pollinator ovipositor length also correlated with changes in fig breeding systems. In particular, the relative elongation of the ovipositor was associated with the repeated loss of functionally dioecious pollination. The concerted evolution of interacting morphologies may bias estimates of phylogeny based on female head characters, but homoplasy is not so strong in other morphological traits. The lesser phylogenetic utility of morphology than of mtDNA is not due to rampant convergence in morphology but rather to the greater number of potentially informative characters in DNA sequence data; patterns of nucleotide substitution also limit the utility of mtDNA findings. Nonetheless, inferring the ancestral associations of fig pollinators from the best-supported phylogeny provided strong evidence of host conservatism in this highly specialized mutualism.

Animals↗

[Mechanisms that limit pollinator range in Ericaceae].

Studies were conducted in 2001-2003 at Valdai National Park (Novgorod region) and at the Zvenigorod biological station of Moscow State University. The morphology of flowers, flowering dynamics and composition of insect visiting flowers of Ericaceae species: Andromeda polifolia, Chamaedaphne calyculata, Ledum palustre, Oxycoccus palustris, Vaccinium myrtillus, V. uliginosum, and V. vitis-idaea L. were studied. Some species of insects visiting flowers were excluded from the list of pollinators on the basis of observation on their behavior. L. palustre was visited mainly by flies where as other investigated species were visited mainly by bumblebees. In some cases bumblebees were the only visitors of the investigated plants. Mechanisms that protect flowers from flies and short-tongued solitary bees visits and ensure a best pollination by bumblebees are various among different species of Ericaceae. Efficiency of nectary protection also differs among different plant species and is defined by particularities of their habitats and flowering phenology. As far as all species of this family during the flowering are dominants in typical habitats, a competition for the pollination with species of other families in most cases is megligible. Flowering periods of V. vitis-idaea and V. myrtillus in forest ecosystems overlapped weakly. Moreover, V. myrtillus is pollinated mainly by bumblebee queens where as pollinators of V. vitis-idaea are bumblebee workers, solitary bees and horse flies. The other investigated plant species inhabit only oligotrophic peat bogs. Thery are pollinated by bumblebees but periods of flowering are not overlapped and consequently follow one after another. L. palustre and V. uliginosum flower simultaneosly but they are pollinated by different pollinators.

Animals↗

On mutualists and exploiters: plant-insect coevolution in pollinating seed-parasite systems.

We investigate the coevolution of time of flowering and time of pollinator emergence in an obligate association between a plant and an insect that both pollinates and parasitizes flowers. Numerical analysis shows that the system in general evolves towards a time of flowering different from the time favoured by the abiotic environment. The equilibrium towards which the system evolves is a local fitness maximum (an ESS) with respect to mutational variation in flowering time but, for the insect, it can be a local fitness minimum at which selection on mutational variation in the time of insect emergence is disruptive. A consequence of evolutionary convergence to a fitness minimum is that pollinators having an earlier phenology can coexist with pollinators having a later phenology. Since late emerging insects are more likely to encounter and oviposit within previously pollinated flowers, their effect on the plant is more exploitative, leading them to function as cheaters within the system. Thus, in the long term, pollinators and exploiters are likely to be found in stable coexistence in pollinating seed-parasite systems.

Animals↗

A weevil pollinating the Canary Islands date palm: between parasitism and mutualism.

Palm pollination systems are highly diverse, including by wind and by several different groups of insects. Many palm species are associated with more or less specific pollinating weevils that are also floral herbivores. For many such palms, the importance of these "palm flower weevils" as pollinators has not been examined. Here we describe a new ex situ method of demonstrating insect pollination when pollinator exclusion is not possible. We show that Neoderelomus piriformis beetles carry pollen and deposit it on the stigma of Phoenix canariensis. Up until now, pollination systems in Phoenix have been unclear, despite the economic importance of the date palm P. dactylifera. We demonstrate here that small weevils that visit inflorescences and often inconspicuously hide there could be efficient pollinators.

Animals↗

Morning floral heat as a reward to the pollinators of the Oncocyclus irises.

Relationships between flowering plants and their pollinators are usually affected by the amount of reward, mainly pollen or nectar, offered to pollinators by flowers, with these amounts usually positively correlated with floral display. The large Oncocyclus iris flowers, despite being the largest flowers in the East Mediterranean flora, are nectarless and have hidden pollen. No pollinators visit the flowers during daytime, and these flowers are pollinated only by night-sheltering solitary male bees. These iris flowers are partially or fully dark-colored, suggesting that they gather heat by absorbing solar radiation. Here we test the hypothesis that the dark-colored flowers of the Oncocyclus irises offer heat reward to their male solitary bee pollinators. Floral temperature was higher by 2.5 degrees C than ambient air after sunrise. Solitary male bees emerged earlier after sheltering in Oncocyclus flowers than from other experimental shelter types. Pollination tunnels facing east towards the rising sun hosted more male bees than other aspects. We suggest that floral heat reward can explain the evolution of dark floral colors in Oncocyclus irises, mediated by the pollinators' behavior.

Adaptation, Physiological↗