Background and Aims
Until recently, there was no consensus regarding the phylogenetic relationships of the Neotropical orchid genera Scuticaria Lindl. and Dichaea Lindl. However, recent evidence derived from both gross morphological and molecular studies supports the inclusion of Scuticaria and Dichaea in sub-tribes Maxillariinae and Zygopetalinae, respectively. The present paper describes the labellar micromorphology of both genera and seeks to establish whether labellar characters support the assignment of Scuticaria and Dichaea to these sub-tribes.
The labella of four species of Scuticaria and 14 species of Dichaea were examined using light microscopy and scanning electron microscopy, and their micromorphology was compared with that of representative species of Maxillariinae sensu lato and Zygopetalinae (Huntleya clade).
Key Results and Conclusions
In most specimens of Scuticaria examined, the papillose labella bear uniseriate, multicellular, unbranched trichomes. However, in S. steelii (Lindl.) Lindl., branched hairs may also be present and some trichomes may fragment and form pseudopollen. Multicellular, leaf-like scales were also present in one species of Scuticaria. Similar, unbranched hairs are present in certain species of Maxillaria Ruiz & Pav. (Maxillariinae sensu stricto) and Chaubardia Rchb.f. (Huntleya clade). As yet, moniliform, pseudopollen-forming hairs have not been observed for Zygopetalinae, but their presence in Scuticaria steelii, Maxillaria and Heterotaxis Lindl. supports the placing of Scuticaria in Maxillariinae. As other genera are sampled, the presence of branched hairs, hitherto unknown for Maxillariinae sensu lato, may prove to be a useful character in taxonomy and phylogenetic studies. Euglossophily occurs in Dichaea, as well as Chondrorhyncha Lindl. and Pescatorea Rchb.f. (Huntleya clade), and all three genera tend to lack distinctive labellar features. Instead, lip micromorphology is relatively simple and glabrous or papillose. However, two of the Dichaea species examined bear unicellular, labellar trichomes very similar to those found in Bifrenaria Lindl. (pollinated by both euglossine bees and Bombus spp.), and this feature may have arisen by convergence in response to similar pollination pressures.
Bifrenaria; Bifrenaria clade; Chaubardia; Chondrorhyncha; Dichaea; Dichaeinae; Heterotaxis; Huntleya clade; Huntleyinae; labellum; Maxillaria; Maxillariinae; papillae; Pescatorea; scales; Scuticaria; trichomes; Zygopetalinae
• Background and Aims The two closely related subtribes Bifrenariinae Dressler and Maxillariinae Benth. are easily distinguished on morphological grounds. Recently, however, molecular techniques have supported the inclusion of Bifrenariinae within a more broadly defined Maxillariinae. The present paper describes the diverse labellar micromorphology found amongst representatives of Bifrenariinae (Bifrenaria Lindl., Rudolfiella Hoehne, Teuscheria Garay and Xylobium Lindl.) and compares it with that found in Maxillaria Pabst & Dungs and Mormolyca Fenzl (Maxillariinae).
• Methods The labella of 35 specimens representing 22 species of Bifrenariinae were examined by means of light microscopy and scanning electron microscopy and their micromorphology compared with that of Maxillaria sensu stricto and Mormolyca spp. The labellar epidermis of representatives of Bifrenaria, Xylobium and Mormolyca was tested for protein, starch and lipids in order to ascertain whether this tissue is involved in the rewarding of pollinators.
• Key Results and Conclusions The labella of Bifrenaria spp. and Mormolyca spp. are densely pubescent but those of Xylobium, Teuscheria and Rudolfiella are generally papillose. However, whereas the trichomes of Bifrenaria and Mormolyca are unicellular, those found in the other three genera are multicellular. Hitherto, no unicellular trichomes have been described for Maxillaria, although the labella of a number of species secrete a viscid substance or bear moniliform, pseudopollen-producing hairs. Moniliform hairs and secretory material also occur in certain species of Xylobium and Teuscheria and these genera, together with Maxillaria, are thought to be pollinated by stingless bees (Meliponini). Differences in the labellar micromorphology of Bifrenaria and Mormolyca are perhaps related to Euglossine- and/ or bumble bee-mediated pollination and pseudocopulation, respectively. Although Xylobium and Teuscheria share a number of labellar features with Maxillaria sensu stricto, this does not necessarily reflect taxonomic relationships but may be indicative of convergence in response to similar pollinator pressures.
Bifrenaria; Bifrenariinae; Maxillaria; Maxillariinae; Meliponini; papillae; pollination; pseudopollen; Rudolfiella; Teuscheria; trichomes; Xylobium
Background and Aims
The orchid genus Dichaea, with over 100 species found throughout the neotropics, is easily recognized by distichous leaves on long stems without pseudobulbs and flowers with infrastigmatic ligules. The genus has previously been divided into four sections based primarily on presence of ovary bristles and a foliar abscission layer. The aim of this work is to use DNA sequence data to estimate phylogenetic relationships within Dichaea and map the distribution of major morphological characters that have been used to delimit subgenera/sections.
Sequence data for the nuclear ribosomal internal transcribed spacers and plastid matK, trnL intron, trnL-F spacer and ycf1 for 67 ingroup and seven outgroup operational taxonomic units were used to estimate phylogenetic relationships within Dichaea. Taxa from each of the four sections were sampled, with the greatest representation from section Dichaea, the most diverse and taxonomically puzzling group.
Molecular data and morphology support monophyly of Dichaea. Results indicate that section Dichaeopsis is polyphyletic and based on symplesiomorphies, including deciduous leaves and smooth ovaries that are widespread in Zygopetalinae. There are at least three well-supported clades within section Dichaeopsis. Section Pseudodichaea is monophyletic and defined by setose ovaries and leaves with an abscission layer. Sections Dichaea and Dichaeastrum are monophyletic and defined by pendent habit and persistent leaves. Section Dichaeastrum, distinguished from section Dichaea primarily by a glabrous ovary, is potentially polyphyletic.
The leaf abscission layer was lost once, occurring only in the derived sections Dichaea and Dichaeastrum. The setose fruit is a more homoplasious character with several losses and gains within the genus. We propose an informal division of the genus based upon five well-supported clades.
Dichaea; matK; nrITS; Orchidaceae; trnL intron; trnL-F spacer; ycf1; Zygopetalinae
Background and Aims
Gross vegetative and floral morphology, as well as modern molecular techniques, indicate that Cryptocentrum Benth. and Sepalosaccus Schltr. are related to Maxillaria Ruiz & Pav. However, they differ from Maxillaria in their possession of floral spurs and, in this respect, are atypical of Maxillariinae. The labellar micromorphology of Maxillaria, unlike that of the other two genera, has been extensively studied. In the present report, the labellar micromorphology of Cryptocentrum and Sepalosaccus is compared with that of Maxillaria and, for the first time, the micromorphology of the floral spur as found in Maxillariinae is described.
Labella and dissected floral spurs of Cryptocentrum and Sepalosaccus were examined using light microscopy (LM) and scanning electron microscopy (SEM).
In each case, the labellum consists of a papillose mid-lobe (epichile), a cymbiform region (hypochile) and, proximally, a spur, which is pronounced in Cryptocentrum but short and blunt in Sepalosaccus. The inner epidermal surface of the spur of Cryptocentrum is glabrous or pubescent, and the bicellular hairs, where present, are unlike any hitherto described for Maxillariinae. Similar but unicellular hairs also occur in the floral spur of Sepalosaccus, whereas the glabrous epidermis lining the spur of C. peruvianum contains putative nectar pores.
The labellar micromorphology of Cryptocentrum and Sepalosaccus generally resembles that of Maxillaria. The floral spur of Cryptocentrum displays two types of organization in that the epidermal lining may be glabrous (possibly with nectar pores) or pubescent. This may have taxonomic significance and perhaps reflects physiological differences relating to nectar secretion. The trichomes found within the spurs of Cryptocentrum and Sepalosaccus more closely resemble the hairs of certain unrelated, nectariferous orchid taxa than those found in the largely nectarless genus Maxillaria, and this further supports the case for parallelism.
Labellum; Maxillariinae; micromorphology; nectar pore; nectary; spur; trichome
• Background and Aims Pollen characters have been widely used in defining evolutionary trends in orchids. In recent years, information on pollination biology and phylogenetic patterns within Orchidinae has become available. Hence, the aim of the presented work is to re-evaluate exine micromorphology of Orchidinae in light of recent phylogenetic studies and to test whether pollen micromorphology strictly depends on phylogenetic relationships among species or whether it is influenced by the marked differences in pollination ecology also reported among closely related species.
• Methods Pollen sculpturing of 45 species of Orchidinae and related taxa was investigated using scanning electron microscopy. To cover potential intraspecific variation, several accessions of the same species were examined.
• Key Results Orchidinae show remarkable variation in exine sculpturing, with a different level of variation within species groups. In some genera, such as Serapias (rugulate) and Ophrys (psilate to verrucate), intrageneric uniformity corresponds well to a common pollination strategy and close relationships among species. However, little exine variability (psilate–scabrate and scabrate–rugulate) was also found in the genus Anacamptis in spite of striking differences in floral architecture and pollination strategies. A larger variety of exine conditions was found in genera Dactylorhiza (psilate, psilate–scabrate and reticulate) and Orchis s.s. (psilate, reticulate, perforate–rugulate and baculate) where no unequivocal correspondence can be found to either phylogenetic patterns or pollination strategies.
• Conclusions Changes in pollen characteristics do not consistently reflect shifts in pollination strategy. A unique trend of exine evolution within Orchidinae is difficult to trace. However, the clades comprising Anacamptis, Neotinea, Ophrys and Serapias show psilate to rugulate or scabrate pollen, while that of the clade comprising Chamorchis, Dactylorhiza, Gymnadenia, Orchis s.s., Platanthera, Pseudorchis and Traunsteinera ranges from psilate to reticulate. Comparison of the data with exine micromorphology from members of the tribe Orchidieae and related tribes suggests a possible general trend from reticulate to psilate.
SEM; exine; phylogeny; pollination biology; pollen; orchids
Background and Aims
Most neotropical Melastomataceae have bee-pollinated flowers with poricidal anthers. However, nectar rewards are known to be produced in about 80 species in eight genera from four different tribes. These nectar-producing species are pollinated by both vertebrates and invertebrates.
The floral morphology and anatomy of 14 species was studied in six genera of nectar-producing Melastomataceae (Blakea, Brachyotum, Charianthus, Huilaea, Meriania and Miconia). Anatomical methods included scanning electron microscopy, and serial sections of paraffin-embedded flowers.
All vertebrate-pollinated melastome flowers have petals that do not open completely at anthesis, thus forming a pseudo-tubular corolla, while closely related species that are bee pollinated have rotate or reflexed corollas. In most species, nectar secretion is related to stomatal or epidermal nectaries and not filament slits as previously reported. Moreover, the nectar is probably supplied by large vascular bundles near the release area. Blakea and Huilaea have nectary stomata located upon the dorsal anther connective appendages. Brachyotum also has nectary stomata on the anther connectives, but these are distributed lengthwise along most of the connective. Meriania may release nectar through the anther connective, but has additional nectary stomata on the inner walls of the hypanthium. Miconia has nectary stomata on the ovary apex. Charianthus nectaries were not found, but there is circumstantial evidence that nectar release occurs through the epidermis at the apex of the ovary and the lower portions of the inner wall of the hypanthium.
Nectar release in Melastomataceae is apparently related to nectary stomata and not filament slits. The presence of nectary stomata on stamens and on ovary apices in different lineages suggests that the acquisition of nectaries is a derived condition. Nectary location also supports a derived condition, because location is strongly consistent within each genus, but differs between genera.
Blakea; Brachyotum; Charianthus; Huilaea; Meriania; Melastomataceae; Miconia; nectaries; nectary stomata; pollination
• Background Seventeen distinct pollination systems are known for genera of sub-Saharan African Iridaceae and recurrent shifts in pollination system have evolved in those with ten or more species. Pollination by long-tongued anthophorine bees foraging for nectar and coincidentally acquiring pollen on some part of their bodies is the inferred ancestral pollination strategy for most genera of the large subfamilies Iridoideae and Crocoideae and may be ancestral for the latter. Derived strategies include pollination by long-proboscid flies, large butterflies, night-flying hovering and settling moths, hopliine beetles and sunbirds. Bee pollination is diverse, with active pollen collection by female bees occurring in several genera, vibratile systems in a few and non-volatile oil as a reward in one species. Long-proboscid fly pollination, which is apparently restricted to southern Africa, includes four separate syndromes using different sets of flies and plant species in different parts of the subcontinent. Small numbers of species use bibionid flies, short-proboscid flies or wasps for their pollination; only about 2 % of species use multiple pollinators and can be described as generalists.
• Scope Using pollination observations for 375 species and based on repeated patterns of floral attractants and rewards, we infer pollination mechanisms for an additional 610 species. Matching pollination system to phylogeny or what is known about species relationships based on shared derived features, we infer repeated shifts in pollination system in some genera, as frequently as one shift for every five or six species of southern African Babiana or Gladiolus. Specialized systems using pollinators of one pollination group, or even a single pollinator species are the rule in the family. Shifts in pollination system are more frequent in genera of Crocoideae that have bilaterally symmetric flowers and a perianth tube, features that promote adaptive radiation by facilitating precise shifts in pollen placement, in conjunction with changes in flower colour, scent and tube length.
• Conclusions Diversity of pollination systems explains in part the huge species diversity of Iridaceae in sub-Saharan Africa, and permits species packing locally. Pollination shifts are, however, seen as playing a secondary role in speciation by promoting reproductive isolation in peripheral, ecologically distinct populations in areas of diverse topography, climate and soils. Pollination of Iridaceae in Eurasia and the New World, where the family is also well represented, is poorly studied but appears less diverse, although pollination by both pollen- and oil-collecting bees is frequent and bird pollination rare.
Floral form; fragrance chemistry; guilds; keystone species; nectar chemistry; Coleoptera; Hymenoptera; Lepidoptera; Nectarinia
Background and Aims
Pollen fates strongly influence mating success in plants but are difficult to quantify. By promoting foraging constancy in pollinators, floral rewards such as nectar may enhance the overall efficiency of pollen transfer. However, this can also lead to high levels of geitonogamy. Pollen fates were studied in Acrolophia cochlearis, a member of a terrestrial epidendroid orchid genus that includes both rewarding and deceptive species.
Pollinator observations were conducted. Pollen transfer efficiency (PTE), the proportion of removed pollinia deposited on stigmas, was measured in a large population at regular intervals throughout the 5-month flowering season. The level of cross-pollination in two populations was estimated from the percentage of seeds with embryos in naturally pollinated fruits.
Acrolophia cochlearis (and a congener A. micrantha) produce minute but concentrated nectar rewards. Observations showed that A. cochlearis is pollinated exclusively by a solitary bee species, Colletes claripes. Although both sexes visited flowers, only males carried pollinaria. Overall levels of pollination and PTE of the rewarding A. cochlearis were much higher than in a deceptive congener, A. capensis. Seeds resulting from self-fertilization had a significantly lower probability of containing viable embryos than did those from cross-fertilization. This dichotomy in fruit quality was used to estimate that cross-pollination occurred in approx. 66 % of A. cochlearis flowers in a large dense population and approx. 10 % in a small sparse population. Traits of A. cochlearis that limit geitonogamy include pollinarium reconfiguration that exceeds the visit time of pollinators and rapid flower senescence following visitation.
Presence of a nectar reward in Acrolophia cochlearis results in high levels of PTE. It is estimated that approx. 33–90 % of fruits in natural populations arise from self-pollination in this species.
Reward; deception; pollen transfer efficiency; pollen tracking; geitonogamy; Acrolophia cochlearis; epidendroid orchid; Cape floral region
Background and Aims
Many orchid flowers have glands called elaiophores and these reward pollinating insects with oil. In contrast to other reward-producing structures such as nectaries, the anatomy of the elaiophore and the process of oil secretion have not been extensively studied. In this paper, elaiophore structure is described for two members of Oncidiinae, Oncidium trulliferum Lindl. and Ornithophora radicans (Rchb.f.) Garay & Pabst.
Elaiophores of both species were examined using light microscopy, scanning electron microscopy and transmission electron microscopy.
Key Results and Conclusions
In flowers of Oncidium trulliferum and Ornithophora radicans, oil is secreted by morphologically distinct elaiophores associated with the labellar callus. However, in O. trulliferum, elaiophores also occur on the lateral lobes of the labellum. In both these species, the epithelial elaiophores are composed of a single layer of palisade-like epidermal cells and a distinct subepithelial layer. Secretory elaiophore cells may contain numerous, starchless plastids, mitochondria and smooth endoplasmic reticulum profiles. In O. trulliferum, the cytoplasm contains myelin-like figures but these are absent from O. radicans. In the former species, cavities occur in the cell wall and these presumably facilitate the passage of oil onto the elaiophore surface. In O. radicans, the accumulation of oil between the outer tangential wall and the cuticle causes the latter to become distended. Since it is probable that the full discharge of oil from the elaiophores of O. radicans occurs only when the cuticle is ruptured by a visiting insect, this may contribute towards pollinator specificity. The structure of the elaiophore in these species resembles both that found in previously investigated species of Oncidiinae and that of certain members of the Malpighiaceae.
Elaiophore; Oncidium trulliferum; Ornithophora radicans; Orchidaceae; oil secretion; pollination
Background and Aims
If stabilizing selection by pollinators is a prerequisite for pollinator-mediated floral evolution, spatiotemporal variation in the pollinator assemblage may confuse the plant–pollinator interaction in a given species. Here, effective pollinators in a living fossil plant Nelumbo nucifera (Nelumbonaceae) were examined to test whether beetles are major pollinators as predicted by its pollination syndrome.
Pollinators of N. nucifera were investigated in 11 wild populations and one cultivated population, and pollination experiments were conducted to examine the pollinating role of two major pollinators (bees and beetles) in three populations.
Lotus flowers are protogynous, bowl shaped and without nectar. The fragrant flowers can be self-heating during anthesis and produce around 1 million pollen grains per flower. It was found that bees and flies were the most frequent flower visitors in wild populations, contributing on average 87·9 and 49·4 % of seed set in Mishan and Lantian, respectively. Beetles were only found in one wild population and in the cultivated population, but the pollinator exclusion experiments showed that beetles were effective pollinators of Asian sacred lotus.
This study indicated that in their pollinating role, beetles, probable pollinators for this thermoregulating plant, had been replaced by some generalist insects in the wild. This finding implies that contemporary pollinators may not reflect the pollination syndrome.
Nelumbo nucifera; beetle pollinated; pollination syndrome; effective pollinator; generalized flower; thermoregulation; Nelumbonaceae
Background and Aims
Previous studies have suggested that velamen characteristics are useful as taxonomic markers in Orchidaceae. Members of tribe Cranichideae have been assigned to two velamen types constructed based on combinations of characters such as the presence of secondary cell-wall thickenings and pores. However, such characters have not been analysed on an individual basis in explicit cladistic analyses.
The micromorphology of roots of 26 species of Cranichideae was examined through scanning electron microscopy and light microscopy, scoring the variation and distribution of four characters: number of velamen cell layers, velamen cell-wall thickenings, presence and type of tilosomes, and supraendodermal spaces. The last three characters were analysed cladistically in combination with DNA sequence data of plastid trnK/matK and nuclear ribosomal internal transcribed spacer (ITS) regions and optimized on the resulting phylogenetic tree.
Thickenings of velamen cell walls group Prescottiinae with Spiranthinae, whereas tilosomes, documented here for the first time in Cranichideae, provide an unambiguous synapomorphy for subtribe Spiranthinae. Supraendodermal spaces occur mostly in species dwelling in seasonally dry habitats and appear to have evolved three times.
Three of the four structural characters assessed are phylogenetically informative, marking monophyletic groups recovered in the combined molecular–morphological analysis. This study highlights the need for conducting character-based structural studies to overcome analytical shortcomings of the typological approach.
Cranichideae; Cranichidinae; ITS; Orchidaceae; phylogeny; Prescottiinae; trnK/matK; root anatomy; Spiranthinae; tilosomes; velamen
Background and Aims
Hummingbird flower mites feed and reproduce in flowers of host plants pollinated by hummingbirds, and use the nostrils and bill of the hummingbird to move from plant to plant. These mites compete with the pollinator for the nectar produced by flowers. An investigation was made of the relationship between the pattern of nectar production and the effects of hummingbird flower mites in the flowers of two sympatric species of Heliconia (Heliconiaceae).
Nectar production was sampled by carrying out two experiments: 2-hour intervals and accumulated nectar. Flowers with and without mites were used in both experiments.
Exclusion of mites increased nectar production, especially in accumulated daily production (a maximum of 49 % more nectar). Both Heliconia species had the same pattern of nectar production: a high concentration in the morning, which was progressively reduced as the day passed. This pattern of nectar production coincides with the behaviour of the pollinator, which makes more frequent visits in the morning, as observed in a previous study.
The results suggest that the impact of mites on nectar availability of Heliconia is more important with regard to total volume of nectar producted irrespective of flower longevity. A high variation among individuals in nectar produced in the populations was also observed. Hummingbird flower mites strongly affect availability of nectar for hummingbirds.
Brazilian Atlantic Forest; hummingbird flower mites; nectar production; Ascidae; nectarivory; Heliconia spathocircinata; Heliconia laneana var. flava
Declines in pollinator colonies represent a worldwide concern. The widespread use of agricultural pesticides is recognized as a potential cause of these declines. Previous studies have examined the effects of neonicotinoid insecticides such as imidacloprid on pollinator colonies, but these investigations have mainly focused on adult honey bees. Native stingless bees (Hymenoptera: Apidae: Meliponinae) are key pollinators in neotropical areas and are threatened with extinction due to deforestation and pesticide use. Few studies have directly investigated the effects of pesticides on these pollinators. Furthermore, the existing impact studies did not address the issue of larval ingestion of contaminated pollen and nectar, which could potentially have dire consequences for the colony. Here, we assessed the effects of imidacloprid ingestion by stingless bee larvae on their survival, development, neuromorphology and adult walking behavior. Increasing doses of imidacloprid were added to the diet provided to individual worker larvae of the stingless bee Melipona quadrifasciata anthidioides throughout their development. Survival rates above 50% were only observed at insecticide doses lower than 0.0056 µg active ingredient (a.i.)/bee. No sublethal effect on body mass or developmental time was observed in the surviving insects, but the pesticide treatment negatively affected the development of mushroom bodies in the brain and impaired the walking behavior of newly emerged adult workers. Therefore, stingless bee larvae are particularly susceptible to imidacloprid, as it caused both high mortality and sublethal effects that impaired brain development and compromised mobility at the young adult stage. These findings demonstrate the lethal effects of imidacloprid on native stingless bees and provide evidence of novel serious sublethal effects that may compromise colony survival. The ecological and economic importance of neotropical stingless bees as pollinators, their susceptibility to insecticides and the vulnerability of their larvae to insecticide exposure emphasize the importance of studying these species.
Background and Aims
Spatial variation in pollinator composition and abundance is a well-recognized phenomenon. However, a weakness of many studies claiming specificity of plant–pollinator interactions is that they are often restricted to a single locality. The aim of the present study was to investigate pollinator effectiveness of the different flower visitors to the terrestrial orchid Eulophia alta at three different localities and to analyse whether differences in pollinator abundance and composition effect this plant's reproductive success.
Natural pollination was observed in vivo, and manipulative experiments were used to study the pollination biology and breeding system of E. alta at three sites near Manaus, Brazil. To gain a better understanding of the underlying mechanisms of pollinator attraction, nectar composition and secretion patterns were also studied, floral scent composition was analysed and a bioassay was conducted.
Flower visitors, pollinator composition, pollinia transfer efficiency of particular pollinator species and natural fruit set differed among the investigated populations of E. alta. Flowers were self-compatible, partially autogamous and effectively pollinated by five bee species (four Centris species and Xylocopa muscaria). Visiting insects appeared to imbibe small amounts of hexose-rich nectar. Nectar sugar content was highest on the third day after flower opening. Floral fragrance analyses revealed 42 compounds, of which monoterpenes and benzenoids predominated. A bioassay using floral parts revealed that only floral tissue from the labellum chamber and labellum tip was attractive to flower visitors.
The data suggest that observed differences in reproductive success in the three populations cannot be explained by absolute abundance of pollinators alone. Due to behavioural patterns such as disturbance of effective pollinators on flowers by male Centris varia bees defending territory, pollinia transfer efficiencies of particular pollinator species also vary between study sites and result in differing reproductive success.
Eulophia alta; Orchidaceae; floral biology; floral volatiles; GC-MS; nectar composition; pollinator performance; reproductive success
Background and Aims
Abolboda (Xyridaceae) belongs to the Poales, a predominantly wind-pollinated order whose phylogeny has been widely studied in recent years. The reproductive biology of Abolboda pulchella and A. poarchon was studied to determine the main pollination system of these species, providing the first experimental data on reproduction in the Xyridaceae.
A field study was conducted, including observations on the morphology and biology of the flowers, insect visits and pollinator behaviour. Experimental pollination treatments were performed to assess agamospermy, spontaneous self-pollination and self-compatibility. Pollination success was determined by pollen tube growth, and reproductive success was assessed by fruit- and seed-set.
Abolboda pulchella and A. poarchon were pollinated by Apidae, Megachilidae and Halictidae bees. The floral resources were pollen and nectar that was produced by stylar appendages, an uncommom nectary type for monocotyledons. The species were self-compatible, and pollen tube growth from self-pollen was similar to that of cross-pollen. However, herkogamy prevented spontaneous selfing, rendering the plants dependent on the pollinator's activity. There was no production of seeds by agamospermy.
Melittophily is the main pollination system of these two Abolboda species. Nectar production was first recorded here for Xyridaceae, and along with self-compatibility, herkogamy and bee pollination, is an informative characteristic that can be used in future phylogenetic analyses of the family as well as Poales.
Abolboda pulchella; Abolboda poarchon; bee pollination; herkogamy; nectar; Poales; reproduction; self-compatibility; stylar appendages; Xyridaceae
Tropical South America is rich in different groups of pollinators, but the biotic and abiotic factors determining the geographical distribution of their species richness are poorly understood. We analyzed the species richness of three groups of pollinators (bees and wasps, butterflies, hummingbirds) in six tropical forests in the Bolivian lowlands along a gradient of climatic seasonality and precipitation ranging from 410 mm to 6250 mm. At each site, we sampled the three pollinator groups and their food plants twice for 16 days in both the dry and rainy seasons. The richness of the pollinator groups was related to climatic factors by linear regressions. Differences in species numbers between pollinator groups were analyzed by Wilcoxon tests for matched pairs and the proportion in species numbers between pollinator groups by correlation analyses. Species richness of hummingbirds was most closely correlated to the continuous availability of food, that of bees and wasps to the number of food plant species and flowers, and that of butterflies to air temperature. Only the species number of butterflies differed significantly between seasons. We were not able to find shifts in the proportion of species numbers of the different groups of pollinators along the study gradient. Thus, we conclude that the diversity of pollinator guilds is determined by group-specific factors and that the constant proportions in species numbers of the different pollinator groups constitute a general pattern.
It has been suggested that the absence of floral rewards in many orchid species causes pollinators to probe fewer flowers on a plant, and thus reduces geitonogamy, i.e. self-pollination between flowers, which may result in inbreeding depression and reduced pollen export. We examined the effects of nectar addition on pollinator visitation and pollen transfer by tracking the fate of colour-labelled pollen in Anacamptis morio, a non-rewarding orchid species pollinated primarily by queen bumble-bees. Addition of nectar to spurs of A. morio significantly increased the number of flowers probed by bumble-bees, the time spent on an inflorescence, pollinarium removal and the proportion of removed pollen involved in self-pollination through geitonogamy, but did not affect pollen carryover (the fraction of a pollinarium carried over from one flower to the next). Only visits that exceeded 18 s resulted in geitonogamy, as this is the time taken for removed pollinaria to bend into a position to strike the stigma. A mutation for nectar production in A. morio would result in an initial 3.8-fold increase in pollinarium removal per visit, but also increase geitonogamous self-pollination from less than 10% of pollen depositions to ca. 40%. Greater efficiency of pollen export will favour deceptive plants when pollinators are relatively common and most pollinaria are removed from flowers or when inbreeding depression is severe. These findings provide empirical support both for Darwin's contention that pollinarium bending is an anti-selfing mechanism in orchids and for the idea that floral deception serves to maximize the efficiency of pollen export.
Pollinator specificity has traditionally been considered the main reproductive isolation mechanism in orchids. Among Mediterranean orchids, however, many species attract and deceive pollinators by mimicking nectar-rewarding plants. To test the extent to which deceptive orchid species share pollinators, we collected and identified hemipollinaria-carrying insects, and used ribosomal sequences to identify the orchid species from which hemipollinaria were removed. We found that social and solitary bees, and also flies, carried hemipollinaria belonging to nine orchid species with different degrees of specialization. In particular, Anacamptis morio, Dactylorhiza romana and Orchis mascula used a large set of pollinator species, whereas others such as Orchis quadripunctata seemed to be pollinated by one pollinator species only. Out of the insects with hemipollinaria, 19% were found to carry hemipollinaria from more than one orchid species, indicating that sympatric food-deceptive orchids can share pollinators. This sharing was apparent even among orchid sister-species, thus revealing an effective overlap in pollinator sets among closely related species. These results suggest varying degrees of pollinator specificity in these orchids, and indicate that pollinator specificity cannot always act as the main isolation mechanism in food-deceptive temperate orchids.
bees; hemipollinaria; nuclear rDNA internal transcribed spacer; pollination; orchids; reproductive isolation
The interaction between flowers and insect pollinators is an important aspect of the reproductive mechanisms of many plant species. Several laboratory and field studies indicate that raising flower temperature above ambient can be an advantage in attracting pollinators. Here we demonstrate that this preference for warmer flowers is, in fact, context-dependent. Using an Australian native bee as a model, we demonstrate for the first time a significant shift in behaviour when the ambient temperature reaches 34°C, at which point bees prefer ambient temperature nectar over warmer nectar. We then use thermal imaging techniques to show warmer nectar maintains the flight temperature of bees during the period of rest on flowers at lower ambient temperatures but the behavioural switch is associated with the body temperature rising above that maintained during flight. These findings suggest that flower-pollinator interactions are dependent upon ambient temperature and may therefore alter in different thermal environments.
Background and Aims
Pollen-collecting bees are among the most important pollinators globally, but are also the most common pollen thieves and can significantly reduce plant reproduction. The pollination efficiency of pollen collectors depends on the frequency of their visits to female(-phase) flowers, contact with stigmas and deposition of pollen of sufficient quantity and quality to fertilize ovules. Here we investigate the relative importance of these components, and the hypothesis that floral and inflorescence characteristics mediate the pollination role of pollen collection by bees.
For ten Aloe species that differ extensively in floral and inflorescence traits, we experimentally excluded potential bird pollinators to quantify the contributions of insect visitors to pollen removal, pollen deposition and seed production. We measured corolla width and depth to determine nectar accessibility, and the phenology of anther dehiscence and stigma receptivity to quantify herkogamy and dichogamy. Further, we compiled all published bird-exclusion studies of aloes, and compared insect pollination success with floral morphology.
Species varied from exclusively insect pollinated, to exclusively bird pollinated but subject to extensive pollen theft by insects. Nectar inaccessibility and strong dichogamy inhibited pollination by pollen-collecting bees by discouraging visits to female-phase (i.e. pollenless) flowers. For species with large inflorescences of pollen-rich flowers, pollen collectors successfully deposited pollen, but of such low quality (probably self-pollen) that they made almost no contribution to seed set. Indeed, considering all published bird-exclusion studies (17 species in total), insect pollination efficiency varied significantly with floral shape.
Species-specific floral and inflorescence characteristics, especially nectar accessibility and dichogamy, control the efficiency of pollen-collecting bees as pollinators of aloes.
Pollen theft; pollination efficiency; dichogamy; floral morphology; Aloe; Alooideae; Xanthorrhoeaceae; Asphodeloideae
The potential for reduced pollination ecosystem service due to global declines of bees and other pollinators is cause for considerable concern. Habitat degradation, destruction and fragmentation due to agricultural intensification have historically been the main causes of this pollinator decline. However, despite increasing and accelerating levels of global urbanization, very little research has investigated the effects of urbanization on pollinator assemblages. We assessed changes in the diversity, abundance and species composition of bee and hoverfly pollinator assemblages in urban, suburban, and rural sites across a UK city.
Bees and hoverflies were trapped and netted at 24 sites of similar habitat character (churchyards and cemeteries) that varied in position along a gradient of urbanization. Local habitat quality (altitude, shelter from wind, diversity and abundance of flowers), and the broader-scale degree of urbanization (e.g. percentage of built landscape and gardens within 100 m, 250 m, 500 m, 1 km, and 2.5 km of the site) were assessed for each study site. The diversity and abundance of pollinators were both significantly negatively associated with higher levels of urbanization. Assemblage composition changed along the urbanization gradient with some species positively associated with urban and suburban land-use, but more species negatively so. Pollinator assemblages were positively affected by good site habitat quality, in particular the availability of flowering plants.
Our results show that urban areas can support diverse pollinator assemblages, but that this capacity is strongly affected by local habitat quality. Nonetheless, in both urban and suburban areas of the city the assemblages had fewer individuals and lower diversity than similar rural habitats. The unique development histories of different urban areas, and the difficulty of assessing mobile pollinator assemblages in just part of their range, mean that complementary studies in different cities and urban habitats are required to discover if these findings are more widely applicable.
Background and Aims
The genus Melocactus comprises 36 species of globose cacti with the most derived traits in the Cereeae tribe. It is the proper study system to examine what are the most derived reproductive strategies within that tribe. This study aims to characterize the reproductive biology and to estimate the mating system parameters of two Andean melocacti, Melocactus schatzlii and M. andinus.
The reproductive attributes of the two species were described, including floral morphology, anthesis patterns, floral rewards, floral visitors and visitation patterns. Levels of self-compatibility and autonomous self-pollination were estimated by hand-pollination experiments. Mating system estimates were obtained by conducting progeny array analyses using isozymes.
The flowers of the two species present the typical hummingbird-pollination syndrome. Despite their morphological resemblance, the two species differ in flower size, pollen and ovule production and anthesis pattern. Their main pollinator agents are hummingbirds, four species in M. schatzlii and one species in M. andinus. Both cacti are self-compatible and capable of self-pollination without the aid of pollen vectors. Population-level outcrossing rate was higher for M. schatzlii (tm=0·9) than for M. andinus (tm=0·4). At the family level, outcrossing rates for most mothers of M. schatzlii were higher (tm>0·8) than for M. andinus (tm<0·5).
Although the two cacti are capable of selfing, M. schatzlii is a predominantly outcrossing species, while M. andinus behaves as a mixed-mating cactus. Hummingbirds are the only pollinators responsible for outcrossing and gene flow events in these species. In their absence, both melocacti set seeds by selfing. Based on its low population size, restricted distribution in Venezuela, low rates of floral visits, and high levels of inbreeding, M. andinus is considered to be an endangered species deserving further study to define its conservation status.
Andes; Cactaceae; hummingbird; isozymes; mating system; Melocactus andinus; Melocactus schatzlii; reproductive biology
Background and Aims
Increasing evidence challenges the conventional perception that orchids are the most distinct example of floral diversification due to floral or prezygotic isolation. Regarding the relationship between co-flowering plants, rewarding and non-rewarding orchids in particular, few studies have investigated whether non-rewarding plants affect the pollination success of rewarding plants. Here, floral isolation and mutual effects between the rewarding orchid Galearis diantha and the non-rewarding orchid Ponerorchis chusua were investigated.
Flowering phenological traits were monitored by noting the opening and wilting dates of the chosen individual plants. The pollinator pool and pollinator behaviour were assessed from field observations. Key morphological traits of the flowers and pollinators were measured directly in the field. Pollinator limitation and interspecific compatibility were evaluated by hand-pollination experiments. Fruit set was surveyed in monospecific and heterospecific plots.
The species had overlapping peak flowering periods. Pollinators of both species displayed a certain degree of constancy in visiting each species, but they also visited other flowers before landing on the focal orchids. A substantial difference in spur size between the species resulted in the deposition of pollen on different regions of the body of the shared pollinator. Hand-pollination experiments revealed that fruit set was strongly pollinator-limited in both species. No significant difference in fruit set was found between monospecific plots and heterospecific plots.
A combination of mechanical isolation and incomplete ethological isolation eliminates the possibility of pollen transfer between the species. These results do not support either the facilitation or competition hypothesis regarding the effect of nearby rewarding flowers on non-rewarding plants. The absence of a significant effect of non-rewarding P. chusua on rewarding G. diantha can be ascribed to low levels of overlap between the pollinator pools of two species.
Galearis diantha; Ponerorchis chusua; rewarding and non-rewarding; mechanical isolation; ethological isolation; pollinator limitation; fruit set
Background and Aims
In the sexually deceptive Ophrys genus, species isolation is generally considered ethological and occurs via different, specific pollinators, but there are cases in which Ophrys species can share a common pollinator and differ in pollen placement on the body of the insect. In that condition, species are expected to be reproductively isolated through a pre-mating mechanical barrier. Here, the relative contribution of pre- vs. post-mating barriers to gene flow among two Ophrys species that share a common pollinator and can occur in sympatry is studied.
A natural hybrid zone on Sardinia between O. iricolor and O. incubacea, sharing Andrena morio as pollinator, was investigated by analysing floral traits involved in pollinator attraction as odour extracts both for non-active and active compounds and for labellum morphology. The genetic architecture of the hybrid zone was also estimated with amplified fragment length polymorphism (AFLP) markers, and pollination fitness and seed set of both parental species and their hybrids in the sympatric zone were estimated by controlled crosses.
Although hybrids were intermediate between parental species in labellum morphology and non-active odour compounds, both parental species and hybrids produced a similar odour bouquet for active compounds. However, hybrids produced significantly lower fruit and seed set than parental species, and the genetic architecture of the hybrid zone suggests that they were mostly first-generation hybrids.
The two parental species hybridize in sympatry as a consequence of pollinator overlap and weak mechanical isolation, but post-zygotic barriers reduce hybrid frequency and fitness, and prevent extensive introgression. These results highlight a significant contribution of late post-mating barriers, such as chromosomal divergence, for maintaining reproductive isolation, in an orchid group for which pre-mating barriers are often considered predominant.
AFLP markers; floral scent variation; hybrid zone; hybrid fitness; Ophrys iricolor; Ophrys incubacea; reproductive isolation; sexual deception
Background and Aims
The study of specialized interactions between species is crucial to our understanding of processes in evolutionary ecology due to their profound effect on life cycles and diversification. Obligate pollination by a single wasp species is rare in Orchidaceae except in species with sexually deceptive flowers that are pollinated exclusively by male insects. The object of this study was to document pollination of the food-deceptive flowers of Coelogyne fimbriata, a species pollinated exclusively by female wasps.
Field observations and experiments were conducted in two populations of C. fimbriata. Floral phenology was recorded, and functional floral architecture was measured. Insect visitors to flowers were observed from 2005 to 2007. Bioassay experiments were conducted to check whether the floral odour attracted pollinators. Natural (insect-mediated) rates of pollinarium removal, pollinium deposition on stigmas, and fruit set were recorded. To determine the importance of cross-pollination, the breeding system was assessed via controlled, hand-pollination experiments.
Two populations of C. fimbriata with fragrant, nectarless flowers are pollinated by females of the same Vespula species (Vespidae, Hymenoptera). Experiments on wasps show that they crawl towards the source of the odour. The flowering period appears to coincide with an annual peak in Vespula colony expansion when additional workers forage for carbohydrates. Rates of pollinarium removal (0·069–0·918) and pollinium deposition on stigmas (0·025–0·695) are extremely variable. However, fruit set in C. fimbriata is always low (0·014–0·069) and appears to be based on self-incompatibility coupled with intraclonal (geitonogamous) deposition of pollinia.
Coelogyne fimbriata and Steveniella satyrioides are now the only orchid species known to have food-deceptive flowers that are pollinated exclusively by eusocial, worker wasps. In C. fimbriata, floral odour appears to be the major attractant. Sub-populations may go through flowering seasons when pollinators are abundant or infrequent, but fruit set always remains low because the obligate pollinator does not often appear to transfer pollinaria between intercompatible genets.
Coelogyne fimbriata; Vespula wasps; food deception; floral odour; pollinarium removal; pollinium deposition; self-incompatibility