From near extinction to diversification by means of a shift in pollination mechanism in the gymnosperm relict Ephedra (Ephedraceae,Gnetales)

Pollination in gymnosperms is usually accomplished by means of wind, but some groups are insect‐pollinated. We show that wind and insect pollination occur in the morphologically uniform genus Ephedra (Gnetales). Based on field experiments over several years, we demonstrate distinct differences between two Ephedra species that grow in sympatry in Greece in pollen dispersal and clump formation, insect visitations and embryo formation when insects are denied access to cones. Ephedra distachya, nested in the core clade of Ephedra, is anemophilous, which is probably the prevailing state in Ephedra. Ephedra foeminea, sister to the remaining species of the genus, is entomophilous and pollinated by a range of diurnal and nocturnal insects. The generalist entomophilous system of E. foeminea, with distinct but infrequent insect visitations, is in many respects similar to that reported for Gnetum and Welwitschia and appears ancestral in Gnetales. The Ephedra lineage is well documented already from the Early Cretaceous, but the diversity declined dramatically during the Late Cretaceous, possibly to near extinction around the Cretaceous–Palaeogene boundary. The clade imbalance between insect‐ and wind‐pollinated lineages is larger than expected by chance and the shift in pollination mode may explain why Ephedra escaped extinction and began to diversify again.     

Aerodynamics of Ephedra trifurca

Computer simulations are used to predict the behavior of pollen grains with different physical properties within the acceleration field created around the ovules of the gymnosperm Ephedra trifurca. A modelling procedure is given that (1) calculates the number of pollen grains captured by an ovule's pollination-droplet and (2) gives a correlation between pollination efficiency and the physical properties (= mass, size) of different types of pollen. Based on this procedure, the number of Ephedra pollen grains captured by micropyles can be less than the number captured from other species. However, the mass and size of Ephedra pollen grains appear to coincide with those predicted to yield a local maximum of pollination efficiency, i.e. slightly larger or smaller values of either mass or size would decrease the probability of capture. In addition, the properties of Ephedra pollen grains operate synergistically in the aerodynamic environment around ovules and are focused to collide with pollination-droplets. By analogy, the properties of Ephedra pollen coincide with those predicted for a localized adaptive peak. The physical properties of pollen grain types other than E. trifurca that can maximize pollen capture are not generally represented in the aerobiology of Ephedra during the pollination season. Therefore, the phenology of pollen release, community taxonomic-composition, and the physics of particle capture play collectively important roles in the reproductive success of Ephedra trifurca.     

POLLEN ENERGETICS AND POLLINATION MODES

The caloric content of pollen for several different species was determined by using a semimicro oxygen bomb calorimeter. The species studied were divided into subsets for statistical comparisons. The subsets considered were wind-pollinated dicots, monocots and gymnosperms; insect-pollinated dicots; and taxonomic subclasses of the wind-pollinated angiosperms (Asteridae, Hamamelidae, Caryophyllidae and Commelinidae). The results indicated a statistically significant caloric content difference (0.05 or higher) between wind-pollinated dicots vs. monocots and gymnosperms. There was no statistical difference between wind-pollinated dicots and insect-pollinated dicots. All subclasses were significantly different from one another. A statistically significant difference was found between two tribes in the Asteraceae which have shifted from insect to wind pollination, providing additional evidence for an independent evolution of the two. The need for future research in this area is discussed.     

Transition from wind pollination to insect pollination in sedges: experimental evidence and functional traits

Transitions from wind pollination to insect pollination were pivotal to the radiation of land plants, yet only a handful are known and the trait shifts required are poorly understood. We tested the hypothesis that a transition to insect pollination took place in the ancestrally wind-pollinated sedges (Cyperaceae) and that floral traits modified during this transition have functional significance. We paired putatively insect-pollinated Cyperus obtusiflorus and Cyperus sphaerocephalus with related, co-flowering, co-occurring wind-pollinated species, and compared pairs in terms of pollination mode and functional roles of floral traits. Experimentally excluding insects reduced seed set by 56-89% in putatively insect-pollinated species but not in intermingled wind-pollinated species. The pollen of putatively insect-pollinated species was less motile in a wind tunnel than that of wind-pollinated species. Bees, beetles and flies preferred inflorescences, and color-matched white or yellow models, of putatively insect-pollinated species over inflorescences, or color-matched brown models, of wind-pollinated species. Floral scents of putatively insect-pollinated species were chemically consistent with those of other insect-pollinated plants, and attracted pollinators; wind-pollinated species were unscented. These results show that a transition from wind pollination to insect pollination occurred in sedges and shed new light on the function of traits involved in this important transition.     

Wind of change: new insights on the ecology and evolution of pollination and mating in wind-pollinated plants

Background

The rich literature that characterizes the field of pollination biology has focused largely on animal-pollinated plants. At least 10 % of angiosperms are wind pollinated, and this mode of pollination has evolved on multiple occasions among unrelated lineages, and hence this discrepancy in research interest is surprising. Here, the evolution and functional ecology of pollination and mating in wind-pollinated plants are discussed, a theoretical framework for modelling the selection of wind pollination is outlined, and pollen capture and the occurrence of pollen limitation in diverse wind-pollinated herbs are investigated experimentally.

Scope and Conclusions

Wind pollination may commonly evolve to provide reproductive assurance when pollinators are scarce. Evidence is presented that pollen limitation in wind-pollinated plants may not be as common as it is in animal-pollinated species. The studies of pollen capture in wind-pollinated herbs demonstrate that pollen transfer efficiency is not substantially lower than in animal-pollinated plants as is often assumed. These findings challenge the explanation that the evolution of few ovules in wind-pollinated flowers is associated with low pollen loads. Floral and inflorescence architecture is crucial to pollination and mating because of the aerodynamics of wind pollination. Evidence is provided for the importance of plant height, floral position, and stamen and stigma characteristics in promoting effective pollen dispersal and capture. Finally, it is proposed that geitonogamous selfing may alleviate pollen limitation in many wind-pollinated plants with unisexual flowers.Key words: Wind pollination, reproductive assurance, pollen limitation, geitonogamy, sex allocation, inflorescence architecture, mating systems     

Phylogenetic relationships in gnetophyta deduced fromrbcL gene sequences

Part of the large subunit of the ribulose-1,5-bisphosphate carboxylase gene (rbcL) was sequenced (1333 base pairs) from three species of gymnosperms:Ephedra sinica Gnetum parvifolium, Welwitschia mirabilis. Phylogenetic trees inferred from the neighbor joining, Wagner parsimony and maximum likelihood methods showed thatGnetum andWelwitschia were more closely related to each other than either is toEphedra within Gnetophyta, and the result supports previous cladistical analysis of morphological data.     

The pollination seesaw of Rhynchospora cephalotes (L.) Vahl (Cyperaceae): Influence of plant location on the role of wind and insects as pollen vectors

Ambophily (wind and insect pollination) has been reported for some genera of the typically wind-pollinated family Cyperaceae, including the genus Rhynchospora. The significance of wind and insect pollination can vary, depending on local microclimatic conditions. Rhynchospora cephalotes is an ambophilous species that can grow under different environmental conditions, either along forest edges or inside forest fragments. This study, therefore, tests the hypothesis that (a) there is greater contribution by wind than insects to fruit set of the individuals at the forest edge and (b) there is greater contribution by insects than wind to fruit set of the individuals inside the forest. Field work was carried out in a fragment of Atlantic Forest in northeastern Brazil. We quantified the number of visits by insects, wind velocity and the reproductive success (fruit set) of R. cephalotes provided by wind and insects through exclusion experiments performed at the edge of and inside the forest. We observed a greater number of visits per day by pollinating bees to individuals inside the forest (36.83 ± 7.46) than to individuals at the edge (16.66 ± 6.53). The wind speed was significantly higher at the edge (1.71 ± 0.46 m/s) than inside the forest (0.97 ± 0.18 m/s). Bees and wind are both pollen vectors of R. cephalotes, but bees were the pollen vector that contributed most to fruit set (63.3%) for individuals inside the forest, whereas wind was the primary vector for individuals at the forest edge (76.6%). This seesaw in importance of each pollen vector in the two different environments guarantees high fruit set in R. cephalotes under different microclimatic conditions.     

An enigmatic Ephedra-like fossil lacking micropylar tube from the Lower Cretaceous Yixian Formation of Liaoning,China

Angiosperms and gymnosperms are two well-separated groups in seed plants according to the current understanding. The huge gap between these two groups constitutes a serious threat against the Darwinism, which expects a continuous transitional series between them. The Lower Cretaceous Yixian Formation of Liaoning, China is famous for its megafossil angiosperms, including some early angiosperms and putative gnetalean plants. Here we document another Ephedra-like fossil plant, Pseudoephedra n. gen. n. sp., from the Yixian Formation on the basis of light microscopic (LM) and scanning electron microscopic (SEM) observations. Although its general morphology demonstrates a great resemblance to Ephedra, the expected micropylar tube characteristic of Ephedra is missing in Pseudoephedra. Instead a solid projection is seen on the top of the female parts. Such a puzzling character combination makes Pseudoephedra perplexing in seed plant phylogeny. If put in Ephedraceae (Gnetales), Pseudoephedra would destroy the only synapomorphy (micropylar tube) of the BEG clade. If put in angiosperms, Pseudoephedra would bridge the formerly huge gap between gymnosperms and angiosperms. Apparently, further investigation is needed to clarify the uncertain position of Pseudoephedra.     

Relative importance of diurnal and nocturnal pollinators for reproduction in the early spring flowering shrub Stachyurus praecox (Stachyuraceae)

Generalized pollination systems may be favored in early spring flowering plants, as during this period pollinator activity is unpredictable. Many previous studies have concentrated on the importance of diurnal visitors in pollination, and consequently, information on the contribution of nocturnal visitors to pollination in early spring is limited. This study was conducted to evaluate the relative importance of diurnal and nocturnal pollinators in the early spring flowering dioecious shrub Stachyurus praecox (Stachyuraceae), in two temperate forests in central Japan. Visitors to the female and male flowers were observed during day and night, and their relative contributions to seed set were compared. The pollinator observations revealed that the diurnal and nocturnal insects visited both male and female flowers, and that the main flower visitors were diurnal small bees and flies as well as nocturnal settling moths. The diurnal and nocturnal flower visitors also acted as pollinators, as the pollen grains of S. praecox were attached to the insects collected from the female flowers. Pollination experiments demonstrated that the contributions of diurnal pollinators to the seed set were higher than those of the nocturnal pollinators. The results of this study indicate that S. praecox has a generalized pollination system, comprising both diurnal insects and nocturnal settling moths. Although the roles of diurnal insects are more important in the pollination of S. praecox, nocturnal settling moths may have a complementary role in early spring.     

Influence of environmental variation on the pollination of the ambophilous sedge <Emphasis Type="Italic">Rhynchospora ciliata</Emphasis> (Cyperaceae)

In typically anemophilous families, there are species, such as Rhynchospora ciliata (Cyperaceae), in which both wind and insects have been registered as pollen vectors (ambophily). However, the contribution of each vector to pollination of ambophilous species can change under different environmental conditions. Therefore, this study aimed to determine the primary pollination system, abiotic or biotic, of R. ciliata, comparing wet and dry seasons. We tested the following hypotheses: (1) in dry months, wind is the main pollen vector of R. ciliata; (2) in wet months, insects will contribute more to the reproductive success of R. ciliata and compensate for low fruit formation by the abiotic vector. Our hypothesis was not confirmed. We found that higher wind speed can favor pollination by wind in R. ciliata even in wetter months. Because insects were less susceptible to environmental variation, they contributed more in dry months when wind speed was low, thus compensating for lower fruit set by wind. High rainfall and humidity can result in faster settling of pollen grains suspended in the air; however, rainfall does not prevent pollination by wind. Since individuals of R. ciliata are located near the edge of the forest, grouped quite close together, and as the inflorescence is located at the apex of the floral scape, wind can act as pollen vector and the inhibiting effects of rainfall can be minimized. The contribution of each pollen vector is important to the reproductive success of R. ciliata and the effectiveness of wind varies with local environmental conditions.     

NEAR-VESSELLESSNESS IN EPHEDRA AND ITS SIGNIFICANCE

Wood of two specimens of Ephedra from high alpine localities is nearly vesselless: E. gerardiana from 4,750 m in Tibet, and E. rupestris from 4,300 m in Argentina. Ephedra gerardiana from lower elevations in the Himalayas has more vessels, although even there they are not abundant. Instances in other species of Ephedra in which extensive areas of latewood or even entire growth rings are vesselless are cited; these tend to occur in high deserts with climates somewhat less extreme than those of the alpine localities. Near-vessellessness is interpreted as an adaptive condition in which the proportion of tracheids, conductively safer than vessel elements, reaches a maximum. Near-vessellessness in Ephedra wood is probably reversible within limits, and should be likened to production of latewood to the exclusion of earlywood. Comparisons are offered to dicotyledons in which abundance of vessels is lowered with respect to ecological extremes.     

Ecological aspects of the rise of angiosperms: a challenge to the reproductive superiority hypotheses

The rise to dominance of the angiosperms is frequently considered to be directly or indirectly due to co-evolutionary mutualisms with vertebrate seed dispersers and insect pollinators ('reproductive co-evolution hypothesis'). The decline of the gymnosperms is considered to be due to the inefficiency of wind pollination in maintaining heterozygosity in angiosperm-enriched communities or the limited scope for speciation of wind-pollinated taxa. Contrasting hypotheses for the rise of the angiosperms focus on either environmental change or superior reproductive and vegetative innovations leading to faster growth rates.
We suggest that plant-animal interactions have been overemphasized and competitive interactions largely overlooked, as determinants of the Cretaceous revolution in plant history.     

Pollination of Richea dracophylla (Ericaceae) and the role of the nocturnal ant Notoncus hickmani (Formicinae)

Our observation of large numbers of ants Notoncus hickmani (Formicinae) on the flowers of a Tasmanian endemic shrub Richea dracophyllum (Ericacea) during the night in winter prompted us to carry out a more extensive study of pollination of R. dracophyllum at two sites. Terminal panicles bear inflorescences of 300+ flowers whose petals are fused into calyptrae covering the reproductive organs, and the calyptrae must be removed before pollination can occur. To exclude birds, we placed mesh bags over some inflorescences. Inflorescences with exclusion bags had a high proportion of retained calyptra and the numbers of viable seeds were greatly reduced. We found R. dracophyllum to have a very long flowering period (more than 6 months) and a range of floral visitors, including honey-eaters (Meliphagidae) and flying insects. At one site we observed large numbers (up to 100) of N. hickmani on inflorescences between dusk and midnight during winter at air temperatures down to 2.5°C. They fed on nectar from flowers that had lost their calyptra where they were exposed to pollen from the anthers and scanning electron microscopy showed high numbers of pollen grains being carried on the ants. The feeding behaviour of these ants would facilitate pollination between adjacent flowers (geitonogamy). Smaller numbers of dolichoderine ants (Iridomyrmex bicknelli?) were observed during the daytime in spring but mainly fed under partially detached calyptrae where they had little exposure to pollen. We suggest that R. dracophylla is pollinated by a range of species and functional groups—birds, flying insects and ants, and that the pollinating species will vary during the year and between sites, although insects depend on birds to remove the calyptrae. Ant pollination is rare, nocturnal ant pollination is extremely rare, and pollination by nocturnal cryophilic ants has not been documented previously.     

Singing in the moonlight: dawn song performance of a diurnal bird varies with lunar phase

It is well established that the lunar cycle can affect the behaviour of nocturnal animals, but its potential to have a similar influence on diurnal species has received less research attention. Here, we demonstrate that the dawn song of a cooperative songbird, the white-browed sparrow weaver (Plocepasser mahali), varies with moon phase. When the moon was above the horizon at dawn, males began singing on average 10 min earlier, if there was a full moon compared with a new moon, resulting in a 67% mean increase in performance period and greater total song output. The lack of a difference between full and new moon dawns when the moon was below the horizon suggests that the observed effects were driven by light intensity, rather than driven by other factors associated with moon phase. Effects of the lunar cycle on twilight signalling behaviour have implications for both pure and applied animal communication research.     

Contrasting flowering phenology and species richness in abiotically and biotically pollinated angiosperms

Abstract Biotic pollination is thought to correlate with increased interspecific competition for pollination among plants and a higher speciation rate. In this study we compared patterns of flowering phenology and species richness between abiotically (wind) and biotically pollinated plants, using phylogenetically independent contrasts. We compiled phenological data from eight local seasonal floras, in which we found geographically overlapping sister clades. Of 65 documented origins of wind pollination, we were able to use up to 17 independent contrasts. In contrast to previous studies we found no difference in global species richness between wind- and biotically pollinated sister clades. Regarding phenology, we found wider phenological spread in biotically pollinated clades, earlier flowering onset in wind-pollinated trees, but no difference in duration of flowering between pollination modes. These results corroborate previous views that niche space is more constrained for wind-pollinated species, and that niche partitioning is less important between wind-pollinated plants compared to plants pollinated by animals.     

Beetle pollination of the fruit-scented cones of the South African cycad Stangeria eriopus

There has been considerable uncertainty about the importance of wind vs. insects in cycad pollination, but recent studies in several cycad genera have indicated that these are pollinated primarily, if not exclusively, by insects. Stangeria represents an isolated southern African cycad lineage previously thought to be wind-pollinated. Unlike in most other cycads, there is no evidence of cone thermogenesis in Stangeria. We found that the scent of both male and female Stangeria cones mimics that of fermented fruit, the main volatiles being esters of acetic acid, ketones, and aldehydes. We found a large variety of insect visitors on the cones, the most common ones being sap and rove beetles (Coleoptera: Nitidulidae, Staphylinidae) and fruit flies (Diptera: Drosophilidae). Of these, only sap beetles (Nitidulidae) were able to effect pollination under experimental conditions. Because sap beetles are also pollinators of Cycas and members of several ancient angiosperm families, their role in the pollination of Stangeria adds interesting details to the role this group of insects has played in the history of plant-pollinator interactions.     

A high frequency of allopolyploid speciation in the gymnospermous genus Ephedra and its possible association with some biological and ecological features

The origin and evolution of polyploids have been studied extensively in angiosperms and ferns but very rarely in gymnosperms. With the exception of three species of conifers, all natural polyploid species of gymnosperms belong to Ephedra, in which more than half of the species show polyploid cytotypes. Here, we investigated the origin and evolution of polyploids of Ephedra distributed in the Qinghai–Tibetan Plateau (QTP) and neighbouring areas. Flow cytometry (FCM) was used to measure the ploidy levels of the sampled species that are represented by multiple individuals from different populations, and then, two single‐copy nuclear genes (LFY and DDB2) and two chloroplast DNA fragments were used to unravel the possible origins and maternal donors of the polyploids. The results indicate that the studied polyploid species are allopolyploids, and suggest that allotetraploidy is a dominant mode of speciation in Ephedra. The high percentage of polyploids in the genus could be related to some of its biological attributes such as vegetative propagation, a relatively high rate of unreduced gamete formation, and a small genome size relative to most other gymnosperms. Significant ecological divergences between allotetraploids and their putative progenitors were detected by PCAs and anova and Tukey's tests, with the exception of E. saxatilis. The overlap of geographical distributions and ecological niches of some diploid species could have provided opportunities for interspecific hybridization and allopolyploid speciation.     

THE AERODYNAMICS OF POLLEN CAPTURE IN TWO SYMPATRIC EPHEDRA SPECIES

Wind-tunnel analyses of the behavior of airborne pollen around ovules of two Ephedra species (E. trifurca and E. nevadensis) indicate that at certain airflow speeds (0.5 m/sec and 1.0 m/sec) each species is capable of biasing pollination in favor of conspecific pollen. A computer procedure was designed to evaluate the physical basis for this aerodynamic discrimination. This procedure indicates that differences in size and density confer significantly different inertial properties to the two pollen species. Operating within the specific aerodynamic environments generated around ovules from each species, these differences are sufficient to account for the biases observed in the probability of pollination. Within natural populations, there exists significant variation in pollen size (and possibly in density). Accordingly, it is possible that, under certain ambient wind conditions, ovules from each species can select subsets of the entire airborne population of Ephedra pollen.     

Population Structure and Reproductive Biology of Saururus cernuus L. (Saururaceae)

Abstract The population structure and reproductive biology of Saururus cernuus (Lizard's Tail; Saururaceae; Piperales), is documented in five sites in southern Louisiana (Mississippi Delta). The species is common throughout the southeastern United States in marshes, along streams, edges of lakes, and in the understory of moist forests. The clonal species exhibits sexual and vegetative reproduction. Wind and insects both play important roles in pollination. Pollen may be borne by insects directly. Alternatively, the pollen may be borne by wind after its release is triggered from pendulous floral spikes (the “Lizard's Tail”) by either wind or insect landings (insect-mediated wind pollination). The plants are self-incompatible and seed set results from a combination of pollination modes with wind pollination being the primary mode (rare in the Magnoliidae). Inflorescence and floral structure exhibit adaptive features that facilitate the various modes of pollen transfer, viz., numerous, small scented, protogynous flowers with no perianth, ultraviolet patterns produced by stamen filaments, small pollen grains, curved inflorescences, and exerted stamens, etc. Fruit production and seed germination are documented and plant growth and densities are compared in sunny versus forest sites.     

Insects and plants in the pollination ecology of the boreal zone

Pollination systems in the boreal zone range from generalist to specialist, both entomologically and botanically. The relative importance of wind pollination, insect pollination, sexual separation between and within plants, and between flowers, hermaphroditism of flowers, and various breeding systems are related to plant growth form and habitat. The diversity and specializations of anthophilous insects parallel those in other biogeographic zones, but seem less developed. We suggest that this reflects the combined effects of evolutionary youth, severity of climate, restriction of symbiont ranges within those of their hosts, and the naturally frequent perturbations by fire or insect outbreaks in the zone, requiring faunal and floral vagility and constraining specialization in mutualism. Modern perturbations by logging and pesticides seem to be well buffered because of the relative openness of the ecosystem (compared to others), although damage has been documented. Insect pollination is as much a keystone process in the boreal forest as elsewhere, despite the immediate counter-impression given by the dominance of wind-pollinated conifers. Nevertheless, there are few studies, botanical or entomologicalin situ. The boreal system offers important opportunities in general and applied research in pollination ecology and synecology generally.