Nectar trichome structure of aquatic bladderworts from the section <Emphasis Type="Italic">Utricularia</Emphasis> (Lentibulariaceae) with observation of flower visitors and pollinators

In Utricularia, the flower spur is a nectary and in this organ, nectar is produced and stored. This study aimed to examine the structure of the nectary trichomes in four Utricularia species (Utricularia vulgaris L., U. australis R.Br., U. bremii Heer and U. foliosa L.) from the generic section Utricularia. We have investigated whether species with different spur morphology had similar spur anatomy and nectary trichome structure. In Utricularia flowers, nectar is produced by spur capitate trichomes (sessile or stalked). Our results showed that regardless of the various spur morphology, trichomes have similar architecture and ultrastructure. Head cells of these trichomes are transfer cells with an eccrine nectar secretion. Examined species differed in the micromorphology of papillae in spurs. The fly Eristalis tenax was found to be a pollinator of U. vulgaris. Small Halictidae bees seem to be pollinators of U. foliosa.     

Flower palate ultrastructure of the carnivorous plant <Emphasis Type="Italic">Genlisea hispidula</Emphasis> Stapf with remarks on the structure and function of the palate in the subgenus <Emphasis Type="Italic">Genlisea</Emphasis> (Lentibulariaceae)

In the genus Genlisea as well as in its sister genus Utricularia, the palate probably plays a key role in providing the colour, mechanical and olfactory stimuli to attract insect pollinators and to guide them to the generative structures and the nectary spur. However, information about the micro-morphology of the palate of Genlisea is scarce. This study aims to examine the structure of the palate in Genlisea hispidula in detail as well as the palate from other five species from the subgenus Genlisea. In particular, its aim is to ascertain whether these palates function as an area for the osmophores in the flower or whether they produce nectar. We showed that the palate in all of the species that were examined was the glandular type and that it had capitate, glandular trichomes, which had a similar general architecture across the species that were examined. No nectar secretion was observed on the palates. The ultrastructure of the palate trichomes showed that the palate glandular trichomes most probably function as scent glands that produce an olfactory stimulus for flower pollinators.     

Bee pollination and evidence of substitutive nectary in <Emphasis Type="Italic">Anadenanthera colubrina</Emphasis> (Leguminosae-Mimosoideae)

Anadenanthera colubrina (Vell.) Brenan (Leguminosae-Mimosoideae) is a widely-distributed tree in seasonally dry tropical forests of South America that was classified previously as lacking nectaries. However, some studies have stated that its flowers produce nectar, while others analyzed the composition of unifloral honey produced from A. colubrina flowers, raising the question about nectar production in the species. We studied the pollination and reproductive biology of A. colubrina var. cebil (Griseb.) Altschul in a natural population in the Caatinga, northeastern Brazil. Reproductive phenology, sexual system, floral biology, resource, and pollinators were investigated. We analyzed the breeding system through controlled pollinations for addressing its dependence on pollen vectors for reproduction. Anadenanthera colubrina flowered in the dry season, flower heads are heteromorphic, with staminate flowers at the base and perfect flowers at the apex of the inflorescence, characterizing andromonoecy. Anthesis is diurnal. We observed small drops of nectar at the apex of the petals of some flowers per inflorescence. Together with observations on flower visitor behavior and histochemical tests, we propose that A. colubrina produces floral nectar at the apex of the corolla, characterizing a substitutive nectary (sensu Vogel). This is the first record of substitutive nectary in the Mimosoideae and the first record of andromonoecy in the genus. Bees were the main pollinators (higher frequency), although other insects such as wasps, butterflies, and small beetles were also observed collecting nectar and/or pollen. The species is self-incompatible, thus depending on insect pollen vectors, mainly bees, for reproduction.     

Micromorphological and histochemical attributes of flowers and floral reward in <Emphasis Type="Italic">Linaria vulgaris</Emphasis> (Plantaginaceae)

The self-incompatible flowers of Linaria vulgaris have developed a range of mechanisms for attraction of insect visitors/pollinators and deterrence of ineffective pollinators and herbivores. These adaptive traits include the flower size and symmetry, the presence of a spur as a “secondary nectar presenter,” olfactory (secondary metabolites) and sensual (scent, flower color, nectar guide—contrasting palate) signals, and floral rewards, i.e. pollen and nectar. Histochemical tests revealed that the floral glandular trichomes produced essential oils and flavonoids, and pollen grains contained flavonoids, terpenoids, and steroids, which play a role of olfactory attractants/repellents. The nectary gland is disc-shaped and located at the base of the ovary. Nectar is secreted through numerous modified stomata. Nectar secretion began in the bud stage and lasted to the end of anthesis. The amount of produced nectar depended on the flower age and ranged from 0.21 to 3.95 mg/flower (mean?=?1.51 mg). The concentration of sugars in the nectar reached up to 57.0%. Both the nectar amount and sugar concentration demonstrated a significant year and population effect. Pollen production was variable between the years of the study. On average, a single flower of L. vulgaris produced 0.31 mg of pollen. The spectrum of insect visitors in the flowers of L. vulgaris differed significantly between populations. In the urban site, Bombus terrestris and Apis mellifera were the most common visitors, while a considerable number of visits of wasps and syrphid flies were noted in the rural site.     

Comparative reproductive biology reveals two distinct pollination strategies in Neotropical twig-epiphyte orchids

Members of Oncidiinae are widely known for their interactions with oil-collecting bees that explore lipophilic secretions on flowers. They may also be pollinated through food deception and the offering of nectar. Although data on breeding systems are available for many Oncidiinae orchids, little is known about the reproductive strategies in Rodriguezia, a neotropical genus of ca. 55 species. In this paper, we explore the reproductive biology of two species of Rodriguezia with distinctive morphologies: R. decora and R. lanceolata. Floral features, spectral reflectance, pollinators and pollination mechanisms, and breeding systems were studied. Both species are scentless and produce nectar as a reward. Floral nectar is secreted by a gland at the base of the labellum and stored into the sepaline spur. Rodriguezia decora reflects mainly in the blue and red regions of the light spectrum, while R. lanceolata reflects in the red region. Rodriguezia decora is exclusively visited and pollinated by butterflies, while Trochilidae hummingbirds are the pollinators of R. lanceolata. Pollinaria attach to the upper third of the proboscis of butterflies (R. decora), and to the bill of hummingbirds (R. lanceolata), during the collection of nectar from the spur. Both Rodriguezia species are self-sterile. Flower features and floral reflectance support the occurrence of psychophily in R. decora and ornithophily in R. lanceolata.     

Glandular trichomes of <Emphasis Type="Italic">Tussilago Farfara</Emphasis> (Senecioneae,Asteraceae)

Main conclusion

The glandular trichomes are developed on the aerial organs of Tussilago farfara ; they produce phenols and terpenoids. Smooth endoplasmic reticulum and leucoplasts are the main organelles of the trichome secretory cells. The aim of this study was to characterise the morphology, anatomy, histochemistry and ultrastructure of the trichomes in Tussilago farfara as well as to identify composition of the secretory products. Structure of trichomes located on the peduncles, bracts, phyllaries, and leaves were studied by light and electron microscopy. The capitate glandular trichomes consist of a multicellular head and a biseriate long stalk. Histochemical tests and fluorescence microscopy reveal phenols and terpenoids in the head cells. During secretory stage, the head cells contain smooth and rough endoplasmic reticulum, Golgi apparatus, diversiform leucoplasts with opaque contents in lamellae, chloroplasts, mitochondria, and microbodies. In the capitate glandular trichomes of T. farfara subcuticular cavity is absent, unlike glandular trichomes in other Asteraceae species. For the first time, content of metabolites in the different vegetative and reproductive organs as well as in the isolated capitate glandular trichomes was identified by GC–MS. Forty-five compounds, including organic acids, sugars, polyols, phenolics, and terpenoids were identified. It appeared that metabolite content in the methanol extracts from peduncles, bracts and phyllaries is biochemically analogous, and similar to the metabolites from leaves, in which photosynthesis happens. At the same time, the metabolites from trichome extracts essentially differ and refer to the above-mentioned secondary substances. The study has shown that the practical value of the aerial organs of coltsfoot is provided with flavonoids produced in the capitate glandular trichomes.

     

Dimorphism in trichome production of <Emphasis Type="Italic">Persicaria lapathifolia</Emphasis> var. <Emphasis Type="Italic">lapathifolia</Emphasis> and Its multiple effects on a leaf beetle

Polymorphisms in plants are main factors that determine the diversity of associated animal communities and their population dynamics. Typically, Persicaria lapathifolia var. lapathifolia (Polygonaceae) has no trichomes on leaf surfaces (glabrous type), but a hairy type does sometimes occur. Based on a cultivation experiment, the presence or absence of trichomes is clarified to be under genetic control. To reveal the defensive function of trichomes against herbivores, laboratory experiments were conducted using a major herbivore, Galerucella grisescens (Coleoptera: Chrysomelidae). In both choice and no-choice feeding tests, the glabrous type was significantly more consumed by G. grisescens adults, while the hairy type was not consumed. In the hairy leaf treatment, larval duration tended to become longer, the adult body weight became significantly lower, and adults laid significantly more eggs than in the glabrous leaf treatment. Hairy leaves contained significantly more total phenolics and condensed tannins than glabrous leaves, suggesting that the hairy type allocates more resources for physical and chemical defence. Because no significant differences in leaf consumption were detected in the feeding experiment using powdered host leaves, G. grisescens seems to have adapted to the chemical defences of P. lapathifolia var. lapathifolia. These results clearly indicate that leaf trichomes of P. lapathifolia var. lapathifolia effectively act as a physical defence against G. grisescens.     

Floral nectar production and carbohydrate composition and the structure of receptacular nectaries in the invasive plant <Emphasis Type="Italic">Bunias orientalis</Emphasis> L. (Brassicaceae)

The data relating to the nectaries and nectar secretion in invasive Brassicacean taxa are scarce. In the present paper, the nectar production and nectar carbohydrate composition as well as the morphology, anatomy and ultrastructure of the floral nectaries in Bunias orientalis were investigated. Nectary glands were examined using light, fluorescence, scanning electron and transmission electron microscopy. The quantities of nectar produced by flowers and total sugar mass in nectar were relatively low. Total nectar carbohydrate production per 10 flowers averaged 0.3 mg. Nectar contained exclusively glucose (G) and fructose (F) with overall G/F ratio greater than 1. The flowers of B. orientalis have four nectaries placed at the base of the ovary. The nectarium is intermediate between two nectary types: the lateral and median nectary type (lateral and median glands stay separated) and the annular nectary type (both nectaries are united into one). Both pairs of glands represent photosynthetic type and consist of epidermis and glandular tissue. However, they differ in their shape, size, secretory activity, dimensions of epidermal and parenchyma cells, thickness of secretory parenchyma, phloem supply, presence of modified stomata and cuticle ornamentation. The cells of nectaries contain dense cytoplasm, plastids with starch grains and numerous mitochondria. Companion cells of phloem lack cell wall ingrowths. The ultrastructure of secretory cells indicates an eccrine mechanism of secretion. Nectar is exuded throughout modified stomata.     

Nectaries and male-biased nectar production in protandrous flowers of a perennial umbellifer <Emphasis Type="Italic">Angelica sylvestris</Emphasis> L. (Apiaceae)

Nectar is the most common floral pollinator reward. In dichogamous species, floral nectar production rates can differ between sexual phases. We studied the structure of nectaries located on the stylopodium and nectar production in protandrous umbellifer Angelica sylvestris. Our study species produced nectar in both floral sexual phases. Nectar sugar concentration was low (on average 22 ± 11 %, mean ± SD) and the nectar hexose rich and composed of sucrose, glucose, fructose and a small amount of amino acids, including β-alanine, a non-protein amino acid. Although nectar composition and sugar concentration varied little between floral sexual phases, nectar production showed a threefold reduction during the stigma receptive period. This is in contrast to other studies of Apiaceae that have reported female-biased nectar production, but in the direction predicted by plant sexual selection theory, suggesting that in pollen-unlimited species, floral rewards mainly enhance male reproductive success. The structure of the nectary was similar at the two sexual stages investigated, and composed of a secretory epidermis and several layers of nectariferous and subsecretory parenchyma. The nectary cells were small, had large nuclei, numerous small vacuoles and dense, intensely staining cytoplasm with abundant endoplasmic reticulum, mitochondria and secretory vesicles. They contained abundant resin-like material that may potentially act as defence against microbes. Starch was rarely observed in the nectary cells, occurring predominantly at the female stage and mainly in guard and parenchyma cells in close proximity to stomata, and in subsecretory parenchyma. The main route of nectar release in A. sylvestris seems to be via modified stomata.     

<Emphasis Type="Italic">Virola sebifera</Emphasis> Aubl. (Myristicaceae) leaf chemical composition and implications on leaf-cutter ant foraging choice

Leaf-cutter ant plant material choice is essential for colony maintenance and growth. Plant material is used as a substrate for cultivating symbiotic fungus, and the ants’ preference for particular leaves, tends to be determined by vegetal age-related physicochemical factors. The plant species Virola sebifera Aubl. (Myristicaceae), for example, shows a large number of leaf surface trichomes. Although non-glandular, V. sebifera trichomes may gradually retain an increasing amount of chemical compounds over the lifetime of the leaf. Thus, the present study aims to investigate the role of plant chemical compounds on Atta sexdens rubropilosa preference for V. sebifera leaves of different ages. For this purpose, the chemical composition of trichomes on young and senescent leaves was analyzed, and ants’ preference tested. The chemical compositions differ between V. sebifera young and senescent leaves, with triacontane (C30) predominance in young leaves and tetratriacontane (C34) predominance in senescent leaves. Ants’ preference choice was tested by randomly offering leaves of different ages to A. sexdens rubropilosa workers, following six different treatments: (1) young leaf fragments; (2) young leaf fragments with few trichomes removed; (3) loose trichomes from young leaves; (4) senescent leaf fragments; (5) senescent leaf fragments with few trichomes removed, and (6) loose trichomes from senescent leaves. Ants’ preference was observed for young leaves fragments with a few trichomes removed and also for young leaves loose trichomes. Ants’ preference might be due to specific volatile compounds (GLV) preset in V. sebifera young leaves. Results suggest occurrence of ants’ selectivity resulting from changes on trichomes chemical composition between V. sebifera leaves different age stages.     

Multiple invasions in urbanized landscapes: interactions between the invasive garden ant <Emphasis Type="Italic">Lasius neglectus</Emphasis> and Japanese knotweeds (<Emphasis Type="Italic">Fallopia</Emphasis> spp.)

Urbanized landscapes are the theater of multiple simultaneous biological invasions likely to affect spread dynamics when co-occurring introduced species interact with each other. Interactions between widespread invaders call for particular attention because they are likely to be common and because non-additive outcomes of such associations might induce negative consequences (e.g., enhanced population growth increasing impacts or resistance to control). We explored the invasions of two widespread invasive taxa: the Japanese knotweed species complex Fallopia spp. and the invasive garden ant Lasius neglectus, in the urban area of Lyon (France). First, we investigated landscape habitat preferences as well as co-occurrence rates of the two species. We showed that Fallopia spp. and L. neglectus had broadly overlapping environmental preferences (measured by seven landscape variables), but their landscape co-occurrence pattern was random, indicating independent spread and non-obligatory association. Second, as Fallopia spp. produce extra-floral nectar, we estimated the amount of nectar L. neglectus used under field conditions without ant competitors. We estimated that L. neglectus collected 150–321 kg of nectar in the month of April (when nectar production is peaking) in a 1162 m² knotweed patch, an amount likely to boost ant population growth. Finally, at six patches of Fallopia spp. surveyed, herbivory levels were low (1–6% loss of leaf surface area) but no relationship between ant abundance (native and invasive) and loss of leaf surface was found. Co-occurrences of Fallopia spp. and L. neglectus are likely to become more common as both taxa colonize landscapes, which could favor the spread and invasion success of the invasive ant.     

Nectar palatability can selectively filter bird and insect visitors to coral tree flowers

Secondary compounds in nectar may play a decisive role in determining the spectrum of floral visitors on plants. Flowers of the African coral tree Erythrina caffra are visited mainly by generalist passerine nectarivores, such as weavers and bulbuls. As the nectar of this species tastes very bitter to humans, it was hypothesized that secondary compounds may repel sunbirds and honeybees which are common in the same habitats yet seldom consume the nectar. We conducted choice tests using fresh nectar and both sucrose and hexose (glucose/fructose) solutions of the same concentration as the nectar. White-bellied Sunbirds (Cinnyris talatala) were repelled by nectar of both E. caffra and a related species Erythrina lysistemon, but Dark-capped Bulbuls (Pycnonotus tricolor) did not discriminate between the Erythrina nectar and control sugar solution in terms of amounts consumed. Honeybees (Apis mellifera scutellata) probed exposed droplets of E. caffra nectar and a control sugar solution at the same rate, suggesting that there is no volatile deterrent, but they immediately withdrew their proboscis far more often from the droplets of Erythrina nectar than they did from the sugar solution, suggesting that they find Erythrina nectar distasteful. These results contribute to a growing awareness that non-sugar components of nectar can play important functional roles in plant pollination systems.     

Influence of Trichomes in Strawberry Cultivars on the Feeding Behavior of <Emphasis Type="Italic">Chaetosiphon fragaefolii</Emphasis> (Cockerell) (Hemiptera: Aphididae)

Chaetosiphon fragaefolii (Cockerell) (Hemiptera: Aphididae) is the predominant aphid in strawberry (Fragaria × ananassa Duchesne) production systems in Brazil. This pest species directly damages the plants and is also responsible for spreading viruses. Further, C. fragaefolii often renders strawberry cultivation unviable, because of its high reproductive rate, as well as the large number of individuals generated through parthenogenesis. The present study aimed to (1) evaluate the feeding behavior of C. fragaefolii in four strawberry cultivars (Albion, Aromas, Camarosa, and San Andreas) and (2) identify the resistance factors associated with the number and type of trichomes in the cultivars, and also its effect on the feeding behavior of C. fragaefolii, using the electrical penetration graph (EPG) technique. The results revealed an intrinsic relationship between the number of trichomes on the cultivar and feeding behavior of C. fragaefolii. A higher number of trichomes, both tector and glandular, was observed in Albion compared to that of other cultivars, resulting in a longer no probing (Np) period per insect, and a longer Np phase. A relatively short phloem phase and ingestion time of the phloem sieve elements were also observed in Albion. These results suggest that the trichomes act as a physical barrier creating difficulties for C. fragaefolii to feed, thereby altering its feeding behavior in the four cultivars studied.     

Waterbird response indicates floodplain wetland restoration

Filamentous cyanobacteria disturb food collection in Daphnia by mechanical interference with the filtering apparatus by the long trichomes. The intensity of this interference depends on the water temperature and the Daphnia body size. However, Daphnia are capable of breaking down the filaments, therefore improving the palatability of the cyanobacteria. The main objective of this study was to test whether the shortening of cyanobacterial filaments and the ensuing clearance rate of Daphnia would increase at higher temperatures to a greater degree in small-bodied Daphnia species than in large-bodied one. Laboratory feeding experiments were conducted in order to measure variation in the length of Cylindrospermopsis raciborskii trichomes and to calculate clearance rate. The filament length and the cyanobacteria clearance rate by Daphnia were calculated following their exposure to grazing by large-bodied D. pulicaria and small-bodied D. longispina in 20, 24, and 28°C. Rising temperature did not affect the intensity of breakage of C. raciborskii trichomes by D. pulicaria and caused decrease in clearance rate of this species, whereas for D. longispina, the temperature increase enhanced both filament breakage and clearance rate. We suggest that these temperature-related changes may affect relative competitive performance of Daphnia species in the presence of cyanobacteria.     

Anatomy,pollen, and chromosomes of <Emphasis Type="Italic">Adenoa</Emphasis> (Turneraceae), a monotypic genus endemic to Cuba

The monotypic genus Adenoa is endemic to Cuba. Its name alludes to the presence of minute glands on the petal margin, identified in the present study as lachrymiform colleters. Here we describe the morphological, anatomical, palynological, and chromosome features that characterize Adenoa cubensis. The indumentum of Adenoa consists only of stellate trichomes. Unlike many species of the new world genera Piriqueta and Turnera, Adenoa lacks glandular hairs and extrafloral nectaries. Adenoa, Piriqueta, and Turnera share the presence of standard, sessile, and lachrymiform colleters. The leaves of Adenoa have xeromorphic features, which include entire, revolute blade margins, an adaxial hypodermis, and stomata restricted to the abaxial surface. The chromosome number is 2n?=?14, which is probably the ancestral number of the family. Adenoa chromosomes are similar in size to those of Turnera, and are larger than those of Piriqueta. Using the available data, we discuss relationships among the new world genera of Turneraceae.     

Oviposition by herbivorous insects induces changes in optical and mechanical properties of <Emphasis Type="Italic">Prunus avium</Emphasis> leaves

This study of animal–plant interaction focused on the impact of oviposition by an insect on the leaves of Prunus avium (cherries). We examined whether the oviposition by Caliroa cerasi affects leaf mechanical and spectral traits in P. avium. Three cultivars of P. avium were studied. Infested leaves had from 1 to 18 eggs and exhibited higher leaf dry mass per area (LMA) than leaves without eggs. Leaf dry weight and LMA were positively correlated with egg number per leaf. Infested leaves tended to have higher number of trichomes. Leaf thickness and material and structural resistance tended to increase in infested leaves. The reflectance across all wavelengths (500–700 nm) in leaves with larger number of eggs was higher compared to leaves without eggs. Photosynthetic performance was reduced and oxidative activity was increased in leaves with eggs. Extrafloral nectaries increased with increasing the number of eggs per leaf and thus play an important role in defense against herbivores by providing nectar rewards that attract their depredators. These responses to oviposition may be beneficial for the plants in terms of resistance to feeding larvae.