The uncommon cavitated secretory trichomes in Bauhinia s.s. (Fabaceae): the same roles in different organs

Cavitated secretory trichomes are characterized by a short or absent stalk that is connected to a secretory hollow head. They are rare structures in angiosperms; in Fabaceae, they have been recorded in only seven genera, including Bauhinia s.s. Because B. curvula and B. rufa exhibit glands that are responsible for attracting pollinators to flowers, this study aimed to test whether the cavitated secretory trichomes present in the flowers of these species have an attraction function. As leaf trichomes are commonly related to plant defence, comparative analyses of the morphology, ontogeny, ultrastructure and chemical profile of the secretory trichomes present in flowers and leaves were conducted. It was found that cavitated secretory trichomes are similar in their external morphology and development, regardless of the organ or species analysed. However, interspecific differences were found in the secretion process and chemical profile of the exudate. The differences found in the cavitated secretory trichomes between species indicate that they secrete distinct compounds, whereas the similarities found in these structures between vegetative and reproductive organs indicate that the cavitated trichomes have equivalent ecological functions within a species, probably in plant defence during organ development. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2016, 180 , 104–122.     

Are deception-pollinated species more variable than those offering a reward?

In most pollination systems, animals transfer pollen among plants of a given species. Pollinator visitations do not come without cost, so plants usually offer a reward. However, the flowers of some plant species, mostly orchids, lack rewards and deceive animals into visiting their flowers. Deceptive species are thought to have high levels of variation in traits associated with advertisement and pollinator attraction, which have been attributed to genetic drift, or disruptive selection due to pollinator behavior. Rewarding species are assumed to have less variation due to stabilizing selection. We compared variability in floral morphology and fragrance composition between deceptive and rewarding species. Because both suites of traits are often linked with floral advertisement and pollinator attraction, we expected variation to be greater in species with deceptive pollination systems than in those offering rewards. We obtained floral morphology metrics for 20 deceptive species and 41 rewarding species native or naturalized in Puerto Rico, Venezuela, and Ecuador. Floral fragrances were sampled from eight deceptive species and four rewarding species. We found that the amplitude of variation in floral morphology and fragrance composition covaries significantly. Comparison of coefficients of variation for morphology indicated that, overall, deceptive species show significantly higher variation than rewarding species, and this pattern was also found among just orchids or just nonorchids. There were no statistical differences in morphological variation between orchids and nonorchids within a functional pollination group. Fragrance variation, measured by Jaccard distance, tended to be greater for deceptive species than for rewarding species. Although overlap in measures of variation occurs between the two groups, the data support the hypothesis that populations of deception-pollinated species are more variable than rewarding species in traits associated with pollinator attraction.     

What do floral anatomy and micromorphology tell us about Neotropical Bulbophyllum section Didactyle (Orchidaceae: Bulbophyllinae)?

Bulbophyllum section Didactyle comprises seven species, but distinction between these is often problematic. These species are pollinated by milichiid flies and air currents move the hinged labellum and press the pollinator against the gynostemium. The labellum structure is considered to be homogeneous and conservative for the genus. Therefore, the floral anatomy and micromorphology of B. section Didactyle were studied in order to identify characters useful for distinguishing the species. All species have sunken glandular trichomes on the abaxial surface of the sepals (possible osmophores) and a trilobed labellum, clothed with trichomes, with a secretory cavity in the callus that is bound by scale‐like papillae. Of the c. 100 characters assessed, 25 varied between species, and each pair of species differed by at least four character states, mainly occurring on the labellum. The data presented allow for a distinction to be made between species and corroborates their grouping in B. section Didactyle, as proposed previously. The presence of osmophores and a nectary on the labellum is confirmed, although their structure is more diverse than anticipated. Moreover, structural differences between B. weddellii and the core of the section might be the result of the odour‐mediated attraction of pollinators rather than flower morphology and thus phylogeny. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 438–452.     

The diversity of elaborate petals in Isopyreae (Ranunculaceae): a special focus on nectary structure

Elaborate petals are highly diverse in morphology, structure, and epidermal differentiation and play a key role in attracting pollinators. There have been few studies on the elaborate structure of petals in the tribe Isopyreae (Ranunculaceae). Seven genera in Isopyreae (Aquilegia, Semiaquilegia, Urophysa, Isopyrum, Paraquilegia, Dichocarpum, and Leptopyrum) have petals that vary in morphology, and two genera (Enemion and Thalictrum) have no petals. The petals of nine species belonged to 7 genera in the tribe were studied to reveal their nectary structure, epidermal micromorphology and ancestral traits. The petal nectaries of Isopyreae examined in this study were located at the tip of spurs (Aquilegia yabeana and A. rockii), or the bottom of shallow sacs (Semiaquilegia adoxoides, Urophysa henryi, Isopyrum manshuricum, and Paraquilegia microphylla), a cup-shaped structure (Dichocarpum fargesii) and a bilabiate structure (Leptopyrum fumarioides). The petal nectary of eight species in Isopyreae (except A. ecalcarata) was composed of secretory epidermis, nectary parenchyma, and vascular tissues, and some sieve tubes reached the secretory parenchyma cells. Among the eight species with nectaries examined in the present study, A. yabeana had the most developed nectaries, with 10–15 layers of secretory parenchyma cells. The epidermal cells of mature petals of the nine species were divided into 11 types. Among these 11 types, there were two types of secretory cells and two types of trichomes. Aquilegia yabeana and A. rockii had the highest number of cell types (eight types), and I. manshuricum and L. fumarioides had the lowest number of cell types (three types). Aquilegia ecalcarata had no secretory cells, and the papillose conical polygonal secretory cells of D. fargesii were different from those of the other seven species with nectaries. Trichomes were found only in Aquilegia, Semiaquilegia, Urophysa, and Paraquilegia. The ancestral mode of nectar presentation in Isopyreae was petals with hidden nectar (70.58%). The different modes of nectar presentation in petals may reflect adaptations to different pollinators in Isopyreae.

     

Evolutionary relationships among pollinators and repeated pollinator sharing in sexually deceptive orchids

The mechanism of pollinator attraction is predicted to strongly influence both plant diversification and the extent of pollinator sharing between species. Sexually deceptive orchids rely on mimicry of species‐specific sex pheromones to attract their insect pollinators. Given that sex pheromones tend to be conserved among related species, we predicted that in sexually deceptive orchids, (i) pollinator sharing is rare, (ii) closely related orchids use closely related pollinators and (iii) there is strong bias in the wasp lineages exploited by orchids. We focused on species that are pollinated by sexual deception of thynnine wasps in the distantly related genera Caladenia and Drakaea, including new field observations for 45 species of Caladenia. Specialization was extreme with most orchids using a single pollinator species. Unexpectedly, seven cases of pollinator sharing were found, including two between Caladenia and Drakaea, which exhibit strikingly different floral morphology. Phylogenetic analysis of pollinators using four nuclear sequence loci demonstrated that although orchids within major clades primarily use closely related pollinator species, up to 17% of orchids within these clades are pollinated by a member of a phylogenetically distant wasp genus. Further, compared to the total diversity of thynnine wasps within the study region, orchids show a strong bias towards exploiting certain genera. Although these patterns may arise through conservatism in the chemical classes used in sex pheromones, apparent switches between wasp clades suggest unexpected flexibility in floral semiochemical production. Alternatively, wasp sex pheromones within lineages may exhibit greater chemical diversity than currently appreciated.     

Floral biology and histochemical analysis of Vanilla edwallii Hoehne (Orchidaceae: Vanilloideae): an orchid pollinated by Epicharis (Apidae: Centridini)

The genus Vanilla is the most diverse in Vanilloideae, with ca 90 species distributed among tropical regions. Despite their economic importance, studies on pollination of Vanilla are very scarce and data on pollinators of species endemic to Brazil are lacking. Based on fieldwork and laboratory investigations, the floral biology of V. edwallii was studied. The pollinators and pollination process were recorded at the Serra do Japi reserve, state of São Paulo, southeastern Brazil, and the presence of floral reward was also investigated. Vanilla edwallii blooms in summer. The lateral inflorescences produce up to four pale green flowers. The white labellum is united to the base of the column forming a mentum. In the studied population V. edwallii is pollinated by Epicharis (Hoplepicharis) affinis, where the males exhibit a territorial behavior, defending flowers from other possible flower visitors. The pollen is deposited on the scutellum of bees when they abandon the flower. The mentum region is dry, suggesting no nectar production. The only secretory structures are osmophores dispersed on the inner surface of the lip responsible for production of a sweet fragrance, which together with color and morphology of flowers is related to bee attraction. The labellum is rich in mucilaginous cells, while the mucilaginous substance is retained inside the cells. The histochemical analysis also detected the presence of phenolic compounds and starch concentrated mainly at the adaxial surface of the lip and around the vascular bundles.     

POLLINATION OF DALECHAMPIA MAGNOLIIFOLIA (EUPHORBIACEAE) BY MALE EUGLOSSINE BEES

Flowers of Dalechampia magnoliifolia in eastern Peru were pollinated primarily by male Eulaema meriana and E. cingulata, which collected fragrance from the secretory gland borne in the staminate cymule. The fragrance contains carvone oxide, benzyl acetate, limonene, α-pinene, myrcine, β-pinene/camphene, and carvone. Trigona cf. pallens was the most abundant visitor, but was primarily a pollen thief. The rate of contact with the stigmas by Trigona, and hence the bee's efficacy as a pollinator, was highly variable and was determined by variation in the separation of anthers and stigmas. Dalechampia magnoliifolia co-occurred with a congeneric species, D. cissifolia, but did not share pollinators with it.     

How to be an attractive male: floral dimorphism and attractiveness to pollinators in a dioecious plant

Background  

Sexual selection theory predicts that males are limited in their reproductive success by access to mates, whereas females are more limited by resources. In animal-pollinated plants, attraction of pollinators and successful pollination is crucial for reproductive success. In dioecious plant species, males should thus be selected to increase their attractiveness to pollinators by investing more than females in floral traits that enhance pollinator visitation. We tested the prediction of higher attractiveness of male flowers in the dioecious, moth-pollinated herb Silene latifolia, by investigating floral signals (floral display and fragrance) and conducting behavioral experiments with the pollinator-moth, Hadena bicruris.     

Scent glands in legume flowers

Scent glands, or osmophores, are predominantly floral secretory structures that secrete volatile substances during anthesis, and therefore act in interactions with pollinators. The Leguminosae family, despite being the third largest angiosperm family, with a wide geographical distribution and diversity of habits, morphology and pollinators, has been ignored with respect to these glands. Thus, we localised and characterised the sites of fragrance production and release in flowers of legumes, in which scent plays an important role in pollination, and also tested whether there are relationships between the structure of the scent gland and the pollinator habit: diurnal or nocturnal. Flowers in pre‐anthesis and anthesis of 12 legume species were collected and analysed using immersion in neutral red, olfactory tests and anatomical studies (light and scanning electron microscopy). The main production site of floral scent is the perianth, especially the petals. The scent glands are distributed in a restricted way in Caesalpinia pulcherrima, Anadenanthera peregrina, Inga edulis and Parkia pendula, constituting mesophilic osmophores, and in a diffuse way in Bauhinia rufa, Hymenaea courbaril, Erythrostemon gilliesii, Poincianella pluviosa, Pterodon pubescens, Platycyamus regnellii, Mucuna urens and Tipuana tipu. The glands are comprised of cells of the epidermis and mesophyll that secrete mainly terpenes, nitrogen compounds and phenols. Relationships between the presence of osmophores and type of anthesis (diurnal and nocturnal) and the pollinator were not found. Our data on scent glands in Leguminosae are original and detail the type of diffuse release, which has been very poorly studied.     

RETRACTED ARTICLE: The AtCCX1 transporter mediates salinity tolerance in both <Emphasis Type="Italic">Arabidopsis</Emphasis> and yeast

The only species in the genus Passiflora (Passifloraceae) known to produce resin glands is P. foetida. These glands are secretory trichomes mainly present on the floral bracts and leaf stipules. The secretion produced by these glands has received attention recently due to the presence of substances with pharmacological properties. Attempts to apply in vitro cell culture methods for the large scale production of highly valuable metabolites has been rather limited due to the fact that these compounds are produced by highly differentiated secretory cells in trichomes which are seldom obtained or because differentiation is inhibited by in vitro conditions. Here we describe the in vitro plant regeneration of P. foetida obtained via organogenesis, using mature zygotic embryos as explants. Differentiated plantlets and, more important, the de novo differentiation of secretory trichomes in vitro could be observed in less than 30 days. There was a clear effect of the concentration of 2,4-dichlorophenoxyacetic acid in the culture media on the regeneration of plants and on the differentiation of glandular trichomes. Our results should be useful for the micropropagation of P. foetida, as well as for studies of the process of secretory trichome differentiation and the implemention of biotechnological methodologies for in vitro mass production of passifloricin and/or other substances present in the P. foetida resin.     

Trichome micromorphology in Turkish species of Ziziphora (Lamiaceae)

Ziziphora L. is represented by 5 species and 2 subspecies in the flora of Turkey: Z. clinopodioides, Z. capitata, Z. persica, Z. tenuior, Z. taurica subsp. taurica, Z. taurica subsp. cleonioides. It is difficult to distinguish between some Ziziphora taxa because of their morphological similarities. In this study, the leaf and calyx trichomes of Ziziphora taxa in Turkey were studied in order to assess anatomical variations that may serve as distinguishing characters. Their micromorphological features were surveyed by scanning electron microscopy (SEM) and light microscopy (LM). Trichomes on leaves and calyx can be divided into two general types: non‐glandular trichomes and glandular (secretory) trichomes. The non‐ glandular trichomes are simple, acicular or curved with cuticular micropapillae. They usually consist of one or more additional cells. The glandular trichomes are divided into two types: peltate and capitate and Ziziphora taxa can easily be distinguished by presence/absence, density and types of glandular trichomes on leaves and calyx. The peltate trichomes consist of 12 or 18 secretory head cells in a single disc; four or six central cells surrounded by eight or twelve peripheral ones. Peltate trichomes are absent on the adaxial leaf surface of Z. capitata and Z. persica. Two types of capitate trichomes are present in Ziziphora. The capitate trichomes are only absent on the calyx surface of Z. persica. In addition, the trichome micromorphology provides some support for separating the two subspecies of Z. taurica. In conclusion, Ziziphora taxa can easily be distinguished by cell number, cell shape presence/absence and density of the glandular trichomes on leaves and calyx.     

Pollinator attracting odour signals in sexually deceptive orchids of the Ophrys fusca group

We investigated patterns of volatiles of several allopatric and sympatric species of the Ophrys fusca group and one species of the O. mammosa/sphegodes group pollinated by either Andrena nigroaenea or A. flavipes, using electrophysiology (gas chromatography coupled with electroantennography; GC-EAD) and chemical analyses. We found 52 GC-EAD active compounds, mainly saturated and unsaturated hydrocarbons with chain lengths of 21 to 31, aldehydes, an ester, and an acid. Based on the relative proportions of all GC-EAD active compounds, the investigated species were compared using various statistical methods (ANOVA, principle component analyses, discriminant function analyses and cluster analyses). Our results show that Ophrys species with the same pollinator – independent of their phylogenetic relationship–use the same volatiles for pollinator attraction. Differences between the species mainly involve different quantitative patterns of volatiles. Our results are in congruence with previous studies that showed different odour bouquets to be responsible for the specific attraction of different pollinators and that alkanes and alkenes are most important for pollinator attraction.     

Flower morphology evolution in Jaborosa (Solanaceae): shape and size variation associated with contrasting pollination modes

• Flower morphology is considered an important factor in species diversification because it may influence the efficiency of pollination in different ways (e.g. attraction and mechanical fit with different groups of pollinators). In the present study, we quantified the variation in flower morphology (i.e. shape and size) of the diverse South American genus Jaborosa Juss. (Solanaceae) in relation to contrasting pollination modes: rewarding pollination either by moths or by generalist small insects versus brood-site deceptive pollination by saprophilous flies.
• We examined variations of flower morphology in frontal (pollinator attraction) and sagittal (functional fit with pollinators) views in 12 Jaborosa species using geometric morphometric methods and comparative approaches to infer whether flower shape evolution, not attributable to flower allometry or phylogenetic relationship, is associated with shifts in pollination modes.
• We found remarkable variation in flower morphology among both Jaborosa species and pollination modes, largely in sagittal view. Evolutionary trends in shape of fly-pollinated flowers were mainly attributable to changes in developmental trajectories. Variation in flower architecture facilitated differential pollen placement – on the proboscis of moths, and either on the back or ventral region of saprophilous flies – promoting diversification of the genus.
• Diversification of shape, independent of size, in most of the studied Jaborosa species would indicate adaptation to contrasting pollination modes.

     

Development and ultrastructure of glandular trichomes in<Emphasis Type="Italic">pelargonium x fragrans</Emphasis> ‘mabel grey’ (geraniaceae)

The glandular trichomes of leaves fromPelargonium xfragrans ‘Mabel Grey’ (Geraniaceae) were examined by light, scanning, and transmission electron microscopy. These trichomes had unicellular globular heads and stalks of different lengths and features. Two types were classified: Type I, with an elongated, large head and a short (100 μm), cylindrical stalk that was more apparent on the adaxial surface; and Type II, with a spherical, small head and a long (300μm), conical stalk that was more pronounced on the abaxial surface. The ultrastructure of secretory cells from both types was distinguished by a well-developed endoplasmic reticulum, mitochondria, plastids, dictyosomes, and numerous vacuoles that likely were involved in the storage and transport of lipophilic substances. Plasmodesmata were frequent on the walls of the secretory and stalked cells. Here, we discuss the implication of structural differentiation in these trichomes.     

Plant reproductive strategies vary under low and high pollinator densities

Long‐term variation in the population density of honey bees Apis mellifera across landscapes has been shown to correlate with variation in the floral traits of plant populations in these landscapes, suggesting that variations in pollinator population density and foraging rates can drive floral trait evolution of their host plants. However, it remained to be determined whether this variation in plant traits is associated with adaptive variation in plant reproductive strategies under conditions of high and low pollinator densities. Here we conducted a reciprocal transplant experiment to examine how this variation in floral traits, under conditions of either high and low pollinator density, impacted seed production in the Tibetan lotus Saussurea nigrescens. In 2014 and 2015, we recorded the floral traits, pollinator visitation rates, and seed production of S. nigrescens populations grown in both home sites and foreign sites, where sites varied in honey bee population density. Our results demonstrated that the floral traits reflected those of their original population, regardless of their current location. However, seed production varied with both population origin and transplant site. Seed number was positively correlated with flower abundance in the pollinator‐rich sites, but with nectar production in the pollinator‐poor sites. Pollinator visitation rate was also positively correlated with flower number at pollinator‐rich sites, and with nectar volume at pollinator‐poor sites. Overall, the local genotype had higher seed production than nonlocal genotypes in home sites. However, when pollen is hand‐supplemented, plants from pollinator‐rich populations had higher seed production than plants from pollinator‐poor populations, regardless of whether they were transplanted to pollinator‐rich or ‐poor sites. These results suggest that the plant genotypic differences primarily drive variation in pollinator attraction, and this ultimately drives variation in seed: ovule ratio. Thus, our results suggest that flowering plant species use different reproductive strategies to respond to high or low pollinator densities.     

Genic rather than genome‐wide differences between sexually deceptive Ophrys orchids with different pollinators

High pollinator specificity and the potential for simple genetic changes to affect pollinator attraction make sexually deceptive orchids an ideal system for the study of ecological speciation, in which change of flower odour is likely important. This study surveys reproductive barriers and differences in floral phenotypes in a group of four closely related, coflowering sympatric Ophrys species and uses a genotyping‐by‐sequencing (GBS) approach to obtain information on the proportion of the genome that is differentiated between species. Ophrys species were found to effectively lack postpollination barriers, but are strongly isolated by their different pollinators (floral isolation) and, to a smaller extent, by shifts in flowering time (temporal isolation). Although flower morphology and perhaps labellum coloration may contribute to floral isolation, reproductive barriers may largely be due to differences in flower odour chemistry. GBS revealed shared polymorphism throughout the Ophrys genome, with very little population structure between species. Genome scans for F_ST outliers identified few markers that are highly differentiated between species and repeatable in several populations. These genome scans also revealed highly differentiated polymorphisms in genes with putative involvement in floral odour production, including a previously identified candidate gene thought to be involved in the biosynthesis of pseudo‐pheromones by the orchid flowers. Taken together, these data suggest that ecological speciation associated with different pollinators in sexually deceptive orchids has a genic rather than a genomic basis, placing these species at an early phase of genomic divergence within the ‘speciation continuum’.     

Does competition with wind‐pollinated species alter Echium plantagineum's attractiveness to a common pollinator Bombus terrestris?

1. In insect‐pollinated plants, pollinator attraction is influenced by flowers (e.g. number, size) and their associated rewards (e.g. pollen, nectar). These traits can depend on plant interactions. Indeed, below‐ground competition between plants can lead to a decrease in flower or reward production in insect‐pollinated species. 2. Wind‐pollinated plants, in particular, which are almost never studied in plant–pollinator networks, can alter insect‐pollinated plants' attractiveness through competition for nutrients. The response of pollinators to such changes has never been investigated. 3. A pot experiment was carried out in which an insect‐pollinated species, Echium plantagineum, was grown in binary mixture with three wind‐pollinated species selected to exert a panel of competitive interactions. Below‐ground competition was controlled using dividers limiting interspecific root competition. Floral traits of E. plantagineum (i.e. flower production, floral display size, flower size and nectar production) were measured. For each species mixture, the visits (i.e. first visit, number of visits, 10‐min sequences) of Bombus terrestris individuals released in a flight cage containing two pots were followed, one with and one without below‐ground competition. 4. Below‐ground competition significantly affected nectar's sucrose concentration but did not influence flower and nectar production. Likewise, pollinator visits were not influenced by below‐ground competition. Competitor identity significantly influenced flower and reward production of E. plantagineum, with a decrease in the presence of the most competitive wind‐pollinated species. A tendency for faster flower visitation events was also detected in the presence of the least competitive competitor. This study raises new questions regarding the influence of wind‐pollinated plants on plant–pollinator interactions.     

Structural investigation of the glandular trichomes of <Emphasis Type="Italic">Salvia argentea</Emphasis>

The morphology, anatomy and distribution of glandular trichomes on the aerial organs of Salvia argentea L. has been investigated. Two morphologically distinct types of glandular trichomes were determined. Capitate glandular trichomes forming a base 1–7 celled, a stalk 1–5 celled or no stalk and a head uni- or bicellular had various types. In capitate trichomes, the neck cell that has an important role especially for xeroformic plants, acting to prevent the backflow of secreted substance through the apoplast has been distinctively observed in the investigated species. The capitate trichomes were present abundantly on all aerial organs of S. argentea. Peltate glandular trichomes had a large secretory head forming 1–5, 8 central and 8–10, 12, 14 peripheral cells. Peltate trichomes are present on all aerial organs, except petiole, being the most abundant on calyx and corolla. Results were shown by tables and photographs.     

Ontogeny resolves gland classification in two caesalpinoid legumes

Robust glandular appendages are reported in legumes of the Caesalpinieae tribe. Most studies only attempt to describe the external morphology of these structures, without providing a distinction between glandular trichomes and emergences. This study employed ontogeny to resolve the terminology of these structures present in flowers of two tropical woody legumes of Caesalpinieae, Erythrostemon gilliesii and Poincianella pluviosa, through surface, anatomical and ultrastructural analyses. Flowers of both species exhibit branched and non-branched glandular trichomes since these structures originate from a single protodermal cell. Non-branched glandular trichomes occur on the inflorescence axis, pedicel, sepals and ovary; in P. pluviosa, they also occur in the unguicle of wings and standard, filaments, anthers and style. This type of trichome shows a non-secretory multiseriate stalk and a secretory multicellular head. Branched glandular trichomes, with similar morphology but exhibiting non-secretory branches, occur in the inflorescence axis, pedicel and sepals; in P. pluviosa, they also occur in the unguicle of wings. During the secretory phase, the trichome head cells have large nuclei, cytoplasm rich in vacuoles, oleoplasts, mitochondria, rough endoplasmic reticulum and free ribosomes. The content is released in the intercellular spaces of the head in a merocrinous mechanism and reaches the surface through cuticle rupture. We emphasized the importance of ontogenetic studies to clarify the terminology of secretory structures. This type of study should be performed in other caesalpinoids so that such robust glandular appendages can be correctly interpreted and used with phylogenetic value in the group.