Nectar Research Articles

Chemical markers of a rare honey from the traditional spice plant Amomum tsao–ko Crevost et Lemarié, via integrated GC–MS and LC-MS approaches

The precious medicinal plant, Amomum tsao-ko Crevost et Lemarié, is the nectariferous plant from which the rare Amomum tsao-ko Crevost et Lemarié honey (ATH) is produced. Presently, chemical markers for authentication of this honey are not available due to the lack of data on its chemical composition. Here, we analyzed the volatile components and their odor activity values (OAVs), which revealed that the unique aroma was mildly flowery and fruity, accompanied by subtle sweet and fresh undertones. Since non-volatile chemicals are more reliable markers for routine authentication, we used a metabolomic approach combined with NMR-based identification to find and confirm a suitable compound to unambiguously distinguish ATH from other honeys. Isorhamnetin 3-O-neohesperidoside ranged from 3.62 to 9.38 mg/kg in ATH and was absent in the other tested honeys. In sum, the study uncovered unique chemical characteristics of ATH that will be helpful to control its quality.

Phenology and foraging bias contribute to sex-specific foraging patterns in the rare declining butterfly Argynnis idalia idalia.

Variation in pollinator foraging behavior can influence pollination effectiveness, community diversity, and plant-pollinator network structure. Although effects of interspecific variation have been widely documented, studies of intraspecific variation in pollinator foraging are relatively rare. Sex-specific differences in resource use are a strong potential source of intraspecific variation, especially in species where the phenology of males and females differ. Differences may arise from encountering different flowering communities, sex-specific traits, nutritional requirements, or a combination of these factors. We evaluated sex-specific foraging patterns in the eastern regal fritillary butterfly (Argynnis idalia idalia), leveraging a 21-year floral visitation dataset. Because A. i. idalia is protandrous, we determined whether foraging differences were due to divergent phenology by comparing visitation patterns between the entire season with restricted periods of male-female overlap. We quantified nectar carbohydrate and amino acid contents of the most visited plant species and compared those visited more frequently by males versus females. We demonstrate significant differences in visitation patterns between male and female A. i. idalia over two decades. Females visit a greater diversity of species, while dissimilarity in foraging patterns between sexes is persistent and comparable to differences between species. While differences are diminished or absent in some years during periods of male-female overlap, remaining signatures of foraging dissimilarity during implicate mechanisms other than phenology. Nectar of plants visited more by females had greater concentrations of total carbohydrates, glucose, and fructose and individual amino acids than male-associated plants. Further work can test whether nutritional differences are a cause of visitation patterns or consequence, reflecting seasonal shifts in the nutritional landscape encountered by male and female A. i. idalia. We highlight the importance of considering sex-specific foraging patterns when studying interaction networks, and in making conservation management decisions for this at-risk butterfly and other species exhibiting strong intraspecific variation.

A synthetic lure for Anopheles gambiae (Diptera: Culicidae) based on the attractive plant Parthenium hysterophorus.

Sugar is the sole diet for male mosquitoes and a complementary meal for females. Searching for natural sources of sugar is mediated by semiochemicals. Floral nectars, extra floral nectaries, damaged tissues of plants and rotten fruits are the most common sources of sugar in nature. I provide laboratory evidence of the high attraction of Parthenium hysterophorus L., a weed that grows in tropical climates, to Anopheles gambiae Giles. This study has tried to identify the chemicals which might be involved in the chemical attraction of A. gambiae to this plant. Using quantitative GC-MS analysis, α-pinene, camphene, 1-octen-3-ol, β-pinene, cis-β-ocimene, bornyl acetate, α-caryophyllene, hexadecanoic acid, and α-linolenic acid were identified as the main constituents of P. hysterophorus volatiles. Successive olfactory assays helped a better understanding of the more attractive chemicals of P. hysterophorus to A. gambiae which was the basis for testing a possible synthetic blend. Olfactory experiments proved this synthetic blend to be as attractive as Parthenium intact plants for A. gambiae. A minimal blend, consisting of only α-pinene, camphene, and cis-β-ocimene, was also produced and laboratory experiments indicated its relative attraction for A. gambiae. This blend can be tested in the attractive toxic sugar bait stations for sampling, surveillance, or control programs of mosquitoes in tropical Africa, where A. gambiae sensu stricto transfer malaria among residents.

Effects of diesel exhaust particles on the health and survival of the buff-tailed bumblebee Bombus terrestris after acute and chronic oral exposure

The diesel-powered transportation sector is a major producer of environmental pollution in the form of micro- and nanoscale diesel exhaust particles (DEP). Pollinators, such as wild bees, may inhale DEP or ingest it orally through plant nectar. However, if these insects are adversely affected by DEP is largely unknown. To investigate potential health threats of DEP to pollinators, we exposed individuals of Bombus terrestris to different concentrations of DEP. We analysed the polycyclic aromatic hydrocarbons (PAH) content of DEP since these are known to elicit adverse effects on invertebrates. We investigated the dose-dependent effects of those well-characterized DEP on survival and fat body content, as a proxy for the insects’ health condition, in acute and chronic oral exposure experiments. Acute oral exposure to DEP showed no dose-dependent effects on survival or fat body content of B. terrestris. However, we could show dose-dependent effects after chronic oral exposure with high doses of DEP where significantly increased mortality was observed. Further, there was no dose-dependent effect of DEP on the fat body content after exposure. Our results give insights into how the accumulation of high concentrations of DEP e.g., near heavily trafficked sites, can influence insect pollinators’ health and survival.

Floral nectar reabsorption and a sugar concentration gradient in two long-spurred Habenaria species (Orchidaceae)

BackgroundFloral nectar is the most common reward flowers offered to pollinators. The quality and quantity of nectar produced by a plant species provide a key to understanding its interactions with pollinators and predicting rates of reproductive success. However, nectar secretion is a dynamic process with a production period accompanied or followed by reabsorption and reabsorption remains an understudied topic. In this study, we compared nectar volume and sugar concentration in the flowers of two long-spurred orchid species, Habenaria limprichtii and H. davidii (Orchidaceae). We also compared sugar concentration gradients within their spurs and rates of reabsorption of water and sugars.ResultsBoth species produced diluted nectar with sugar concentrations from 17 to 24%. Analyses of nectar production dynamics showed that as flowers of both species wilted almost all sugar was reabsorbed while the original water was retained in their spurs. We established a nectar sugar concentration gradient for both species, with differences in sugar concentrations at their spur’s terminus and at their spur’s entrance (sinus). Sugar concentration gradient levels were 1.1% in H. limprichtii and 2.8% in H. davidii, both decreasing as flowers aged.ConclusionWe provided evidence for the reabsorption of sugars but not water occurred in wilted flowers of both Habenaria species. Their sugar concentration gradients vanished as flowers aged suggesting a slow process of sugar diffusion from the nectary at the spur’s terminus where the nectar gland is located. The processes of nectar secretion/reabsorption in conjunction with the dilution and hydration of sugar rewards for moth pollinators warrant further study.

Tripartite interactions comprising yeast-endobacteria systems in the gut of vector mosquitoes.

It is shown that bacteria use yeast as a niche for survival in stressful conditions, therefore yeasts may act as temporary or permanent bacterial reservoirs. Endobacteria colonise the fungal vacuole of various osmotolerant yeasts which survive and multiply in sugar-rich sources such as plant nectars. Nectar-associated yeasts are present even in the digestive system of insects and often establish mutualistic symbioses with both hosts. Research on insect microbial symbioses is increasing but bacterial-fungal interactions are yet unexplored. Here, we have focused on the endobacteria of Wickerhamomyces anomalus (formerly Pichia anomala and Candida pelliculosa), an osmotolerant yeast associated with sugar sources and the insect gut. Symbiotic strains of W. anomalus influence larval development and contribute digestive processes in adults, in addition to exerting wide antimicrobial properties for host defence in diverse insects including mosquitoes. Antiplasmodial effects of W. anomalus have been shown in the gut of the female malaria vector mosquito Anopheles stephensi. This discovery highlights the potential of utilizing yeast as a promising tool for symbiotic control of mosquito-borne diseases. In the present study, we have carried out a large Next Generation Sequencing (NGS) metagenomics analysis including W. anomalus strains associated with vector mosquitoes Anopheles, Aedes and Culex, which has highlighted wide and heterogeneous EB communities in yeast. Furthermore, we have disclosed a Matryoshka-like association in the gut of A stephensi that comprises different EB in the strain of W. anomalus WaF17.12. Our investigations started with the localization of fast-moving bacteria-like bodies within the yeast vacuole of WaF17.12. Additional microscopy analyses have validated the presence of alive intravacuolar bacteria and 16S rDNA libraries from WaF17.12 have identified a few bacterial targets. Some of these EB have been isolated and tested for lytic properties and capability to re-infect the yeast cell. Moreover, a selective competence to enter yeast cell has been shown comparing different bacteria. We suggested possible tripartite interactions among EB, W. anomalus and the host, opening new knowledge on the vector biology.

Gut microbiome helps honeybee (Apis mellifera) resist the stress of toxic nectar plant (Bidens pilosa) exposure: Evidence for survival and immunity.

Honeybee (Apis mellifera) ingestion of toxic nectar plants can threaten their health and survival. However, little is known about how to help honeybees mitigate the effects of toxic nectar plant poisoning. We exposed honeybees to different concentrations of Bidens pilosa flower extracts and found that B. pilosa exposure significantly reduced honeybee survival in a dose-dependent manner. By measuring changes in detoxification and antioxidant enzymes and the gut microbiome, we found that superoxide dismutase, glutathione-S-transferase and carboxylesterase activities were significantly activated with increasing concentrations of B. pilosa and that different concentrations of B. pilosa exposure changed the structure of the honeybee gut microbiome, causing a significant reduction in the abundance of Bartonella (p < 0.001) and an increase in Lactobacillus. Importantly, by using Germ-Free bees, we found that colonization by the gut microbes Bartonella apis and Apilactobacillus kunkeei (original classification as Lactobacillus kunkeei) significantly increased the resistance of honeybees to B. pilosa and significantly upregulated bee-associated immune genes. These results suggest that honeybee detoxification systems possess a level of resistance to the toxic nectar plant B. pilosa and that the gut microbes B. apis and A. kunkeei may augment resistance to B. pilosa stress by improving host immunity.

Soil salinization disrupts plant-plant signaling effects on extra-floral nectar induction in wild cotton.

Plant-plant interactions via volatile organic compounds (VOCs) have received much attention, but how abiotic stresses affect these interactions is poorly understood. We tested the effect of VOCs exposure from damaged conspecifics on the production of extra-floral nectar (EFN) in wild cotton plants (Gossypium hirsutum), a coastal species in northern Yucatan (Mexico), and whether soil salinization affected these responses. We placed plants in mesh cages, and within each cage assigned plants as emitters or receivers. We exposed emitters to either ambient or augmented soil salinity to simulate a salinity shock, and within each group subjected half of the emitters to no damage or artificial leaf damage with caterpillar regurgitant. Damage increased the emission of sesquiterpenes and aromatic compounds under ambient but not under augmented salinity. Correspondingly, exposure to VOCs from damaged emitters had effect on receiver EFN induction, but this effect was contingent on salinization. Receivers produced more EFN in response to damage after being exposed to VOCs from damaged emitters when the latter were grown under ambient salinity, but not when they were subjected to salinization. These results suggest complex effects of abiotic factors on VOC-mediated plant interactions.

1064. Agapetes grandiflora Hook.f.: Ericaceae

SummaryA specimen of Agapetes from Myanmar is described and illustrated as Agapetes grandiflora Hook.f., as a result of comparison with A. burmanica W.E.Evans, A. macrantha (Hook.) Hook.f. and A. variegata (Roxb.) D.Don ex G.Don. This species has extrafloral nectaries on the margins of young leaves and at the apex of calyx lobes.

An overview of Selected Economically Important Genera of Family Caesalpinioideae

In this chapter, economically important plant species belonging to the Caesalpinioideae family are discussed. The plant family Caesalpinioideae is a subfamily belonging to Leguminosae. The plant species of Caesalpinioideae are trees or shrubs with alternate, compound leaves. They are provided with extra-floral nectaries or glands on the rachis. The diagnostic feature of the family is the posterior petal being overlapped by the lateral petals. Since the family is of great importance, this chapter provides detailed information on the uses, folk medicine, chemistry, distribution, cultivation, harvesting, yields and economics of selected economically important genera of the family.

Aedes albopictus microbiome derives from environmental sources and partitions across distinct host tissues.

The mosquito microbiome consists of a consortium of interacting microorganisms that reside on and within culicid hosts. Mosquitoes acquire most of their microbial diversity from the environment over their life cycle. Once present within the mosquito host, the microbes colonize distinct tissues, and these symbiotic relationships are maintained by immune-related mechanisms, environmental filtering, and trait selection. The processes that govern how environmental microbes assemble across the tissues within mosquitoes remain poorly resolved. We use ecological network analyses to examine how environmental bacteria assemble to form bacteriomes among Aedes albopictus host tissues. Mosquitoes, water, soil, and plant nectar were collected from 20 sites in the Mānoa Valley, Oahu. DNA was extracted and associated bacteriomes were inventoried using Earth Microbiome Project protocols. We find that the bacteriomes of A. albopictus tissues were compositional taxonomic subsets of environmental bacteriomes and suggest that the environmental microbiome serves as a source pool that supports mosquito microbiome diversity. Within the mosquito, the microbiomes of the crop, midgut, Malpighian tubules, and ovaries differed in composition. This microbial diversity partitioned among host tissues formed two specialized modules: one in the crop and midgut, and another in the Malpighian tubules and ovaries. The specialized modules may form based on microbe niche preferences and/or selection of mosquito tissues for specific microbes that aid unique biological functions of the tissue types. A strong niche-driven assembly of tissue-specific microbiotas from the environmental species pool suggests that each tissue has specialized associations with microbes, which derive from host-mediated microbe selection.

Choice of forage plants by the giant honey bee Apis laboriosa Smith in a captive condition outside their native geographic range

The giant honey bee, Apis laboriosa, is native to the central and eastern Himalayas, Hengduan, Arakan, Annamite and adjoining mountain ranges of Southeast Asia. The present work documented forage plants of A. laboriosa in a captive condition outside its natural geographic range for 09 months and tested their choice based on the abundance of the forager bees on flowers. The study was conducted by introducing a combless colony of A. laboriosa in a climate control greenhouse. A total of 68 plant species of 34 families were offered, out of which the foragers accepted the floral rewards from 28 species of 20 plant families. The foragers collected pollens and nectar from 10 and 18 plant species, respectively. A maximum number of foragers visited Mesua ferrea for pollen (16.94 ± 1.25 foragers/flower/hour) and Indigofera sp. for nectar (9.60 ± 1.24 foragers/flower/hour). Tukey’s Multiple Comparison test showed significant variation in the abundance of foragers on some plants. Analysis of variance showed no significant difference in the number of foragers at different time intervals of a day except for the nectar plants Cucumis sativus and Indigofera sp. The study also showed that the choice of the foragers was not associated with floral colours and plant family, but possibly dependent on some common cues of the forage plants resulting from convergent evolution. Foraging from plants of low altitudes also indicated that they can forage from plants outside their native geographic range.

Caffeine Consumption Helps Honey Bees Fight a Bacterial Pathogen.

Caffeine has long been used as a stimulant by humans. Although this secondary metabolite is produced by some plants as a mechanism of defense against herbivores, beneficial or detrimental effects of such consumption are usually associated with dose. The Western honey bee, Apis mellifera, can also be exposed to caffeine when foraging at Coffea and Citrus plants, and low doses as are found in the nectar of these plants seem to boost memory learning and ameliorate parasite infection in bees. In this study, we investigated the effects of caffeine consumption on the gut microbiota of honey bees and on susceptibility to bacterial infection. We performed in vivo experiments in which honey bees, deprived of or colonized with their native microbiota, were exposed to nectar-relevant concentrations of caffeine for a week, then challenged with the bacterial pathogen Serratia marcescens. We found that caffeine consumption did not impact the gut microbiota or survival rates of honey bees. Moreover, microbiota-colonized bees exposed to caffeine were more resistant to infection and exhibited increased survival rates compared to microbiota-colonized or microbiota-deprived bees only exposed to the pathogen. Our findings point to an additional benefit of caffeine consumption in honey bee health by protecting against bacterial infections. IMPORTANCE The consumption of caffeine is a remarkable feature of the human diet. Common drinks, such as coffee and tea, contain caffeine as a stimulant. Interestingly, honey bees also seem to like caffeine. They are usually attracted to the low concentrations of caffeine found in nectar and pollen of Coffea plants, and consumption improves learning and memory retention, as well as protects against viruses and fungal parasites. In this study, we expanded these findings by demonstrating that caffeine can improve survival rates of honey bees infected with Serratia marcescens, a bacterial pathogen known to cause sepsis in animals. However, this beneficial effect was only observed when bees were colonized with their native gut microbiota, and caffeine seemed not to directly affect the gut microbiota or survival rates of bees. Our findings suggest a potential synergism between caffeine and gut microbial communities in protection against bacterial pathogens.

Ecological risks of pesticide pollution of beekeeping products

The article is focused on the actuality of the ecological risks researching of pesticide pollution of agricultural soils, which are the feed and raw material base for beekeeping. There is mentioned the importance of permanent control over the content of dangerous toxicants, in particular chlororganic pesticides, in the bee honey and other beekeeping products in the article. The results of researching the content of residual amounts of persistent chlororganic pesticides hexachlorocyclohexane (HCH) and dichlorodiphenyltrichlormethylmethane (DDT) in samples of the sunflower honey which are taken from the bee farm of three administrative districts of Cherkasy region are presented. Significant pollution of the sunflower honey from the bee farm which is placed in the village of Novoselytsia by residual amounts of pesticides is detected, the concentration of the sum of metabolites and isomers DDT exceeding the maximum allowable standards in 1.3 to 2.3 times. It should be noted that the determined concentrations of the above mentioned pesticide in the soil (from 6.41 to 23.53 mcg/kg for the MRL is equal 100 mcg/kg) cannot be a source of significant pollution of bee honey. It is suggested that the reason for the xenobiotic entering to bee products through the biological cycle (soil – honey plant – honey) is the presence in the range of honey collection area of anonactive compound of toxic chemicals, which is a powerful source of toxicants, including pesticides into environmental objects. It is detected the direct correlation dependance between the content of pesticides in honey and their concentration in the soil (in the range from r=0.61 for the village of Stebne to r=0.98 for villages of Novoselytsia and Chervone). The importance of assessment of ecological risks of environmental pollution by pesticide residues was proved, since even a small concentration of a toxic substance in soil, water, air, nectar or pollen of honey plants often leads to massive damage and death of bees. It is noted that honey can be used as a suitable bioindicator for assessing the ecological risks of environmental pollution by toxic substances, including chlororganic pesticides.

Cowpea extrafloral nectar has potential to provide ecosystem services lost in agricultural intensification and support native parasitoids that suppress the wheat stem sawfly.

The native parasitoids Bracon cephi (Gahan) and B. lissogaster Muesebeck (Hymenoptera: Braconidae) reduce populations of Cephus cinctus Norton (Hymenoptera: Cephidae), a native grassland species, and major wheat pest on the Northern Great Plains of North America. Non-host feeding adults of these braconids increase longevity, egg load, and egg volume when provisioned carbohydrate-rich diets. Nutrition from nectar can enhance the success of natural enemies in pest management programs. Cowpea, Vigna unguiculata (L.) Walpers, is a potential cover-crop that could add resilient features to the landscape and has extrafloral nectaries (EFN), easy-access nectar sources for beneficial insects. If more cowpea was grown on the Northern Great Plains, would B. cephi and B. lissogaster benefit from foraging on putatively beneficial EFN? We investigated cowpea inflorescence stalk extrafloral nectars (IS-EFN) and leaf stipel extrafloral nectars (LS-EFN) as potential food sources for these parasitoids. Females were caged on EFN sources on living cowpea plants to assess longevity. Egg load and volume were measured at 2, 5, and 10 days after placement. Bracon cephi survived 10 days on water, 38 days on IS-EFN; B. lissogaster 6 days on water, 28 days on IS-EFN. Bracon lissogaster maintained a constant egg load and volume across treatments while B. cephi produced 2.1-fold more eggs that were 1.6-fold larger on IS-EFN. Y-tube olfactometry indicated adult females were attracted to airstreams containing cowpea volatiles. These results demonstrate that non-native, warm-season cowpea benefits these native parasitoids and may improve conservation biocontrol of C. cinctus.

Dear neighbor: Trees with extrafloral nectaries facilitate defense and growth of adjacent undefended trees.

Plant diversity can increase productivity. One mechanism behind this biodiversity effect is facilitation, which is when one species increases the performance of another species. Plants with extrafloral nectaries (EFNs) establish defense mutualisms with ants. However, whether EFN plants facilitate defense of neighboring non-EFN plants is unknown. Synthesizing data on ants, herbivores, leaf damage, and defense traits from a forest biodiversity experiment, we show that trees growing adjacent to EFN trees had higher ant biomass and species richness and lower caterpillar biomass than conspecific controls without EFN-bearing neighbors. Concurrently, the composition of defense traits in non-EFN trees changed. Thus, when non-EFN trees benefit from lower herbivore loads as a result of ants spilling over from EFN tree neighbors, this may allow relatively reduced resource allocation to defense in the former, potentially explaining the higher growth of those trees. Via this mutualist-mediated facilitation, promoting EFN trees in tropical reforestation could foster carbon capture and multiple other ecosystem functions.

On Calliandra and Afrocalliandra (Fabaceae-Caesalpinioideae)

The history of the genus Calliandra is reviewed, with particular reference to the two African species C. gilbertii and C. redacta. These species have been segregated as Afrocalliandra, based on proposed differences from a neotropical Calliandra. The stated differences are acalymmate, 7-celled (versus calymmate, 8-celled) polyads, presence (versus absence) of extrafloral nectaries, and presence (versus absence) of spines or thorns. A scrutiny of these claims shows that neither number of cells in the polyads, nor extrafloral nectaries, nor armature can be used to separate the two African species from the rest of the genus. The only remaining difference is the acalymmate polyads, a plesiomorphic state among mimosoids. On the contrary, a circumscription of Calliandra that includes also the two African species is, apart from being robustly supported by molecular data, also very strongly supported morphologically by 8-celled asymmetrical polyads with tail cells provided with viscin bodies (which are unique among legumes), by expanded stigmas with a wide area of polyad receptivity (which are unique among mimosoids), and by sagittate, petiolate and persistent cotyledons. A return to a circumscription of Calliandra that includes also the two African species is therefore proposed. Calliandra is subdivided into subg. Calliandra, with ca. 140 species in the Neotropics, and subg. Afrocalliandra, comb. et stat. nov., with C. gilbertii in Somalia and Kenya, and C. redacta in South Africa.

Nectar guides and floral nectary in Lamium album L. subsp. album: structure and histochemistry in light, fluorescence, and electron microscopy

Lamium album is a native common plant growing in Eurasia. It is used in medicine and cosmetics and is highly valued in apiculture. The aim of the study was to investigate the structure of the floral nectary in three stages of flower development. Additionally, histochemical studies of the nectary and nectar guides present on the lower corolla lobe were carried out. No detailed analyses of nectary tissues in this species have been conducted to date. The present analyses were performed with the use of light, fluorescence, and scanning electron microscopy. The nectary gland in the flowers of Lamium album subsp. album formed an incomplete ring at the ovary base. The nectarostomata were arranged in clusters only in the adaxial epidermis of the anterior part of the nectary. During the secretory activity of the nectary (1st day of flowering), numerous small vacuoles and cells with large lobulate nuclei with surrounding plastid clusters were observed in the epidermis and glandular parenchyma cells. The vascular bundles contained xylem and phloem elements. Corolla wilting (3rd day of flowering) was accompanied by destructive changes in the nectary parenchyma, leading to the formation of empty spaces and appearance of cell remnants on the nectary surface. The histochemical analyses revealed the presence of starch and phenolic compounds as well as acidic and neutral lipids, which are characteristic of essential oils, in the nectary tissues. The nectar guides were composed of large yellow papillae containing phenolic compounds and acidic and neutral lipids, which were also present in glandular trichomes and abaxial parenchyma cells. The present study has demonstrated that the scent of Lamium album subsp. album flowers is produced with the involvement of essential oils contained in adaxial and abaxial epidermis cells, glandular trichomes, and nectary tissues.

Plant species with larger extrafloral nectaries produce better quality nectar when needed and interact with the best ant partners.

Few studies have explored the phenotypic plasticity of nectar production on plant attractiveness to ants. Here, we investigate the role of extrafloral nectary (EFN) size on the productivity of extrafloral nectar in three sympatric legume species. We hypothesized that plant species with larger EFNs (i) have higher induced nectar secretion after herbivory events, and (ii) are more likely to interact with more protective (i.e. dominant) ant partners. We target 90 plants of three Chamaecrista species in the field. We estimated EFN size and conducted field experiments to evaluate any differences in nectar traits before and after leaf damage to investigate the phenotypic plasticity of nectar production across species. We conducted multiple censuses of ant species feeding on EFNs over time. Plant species increased nectar descriptors after leaf damage, but in different ways. Supporting our hypothesis, C. duckeana, with the largest EFN size, increased all nectar descriptors, with most intense post-herbivory-induced response, taking its place as the most attractive to ants, including dominant species. EFN size variation was an excellent indicator of nectar productivity across species. The higher control over reward production in plants with larger sized EFNs reflects an induction mechanism under damage that reduces costs and increases the potential benefits of indirect biotic defences.

Elevated ozone and carbon dioxide affects the composition of volatile organic compounds emitted by Vicia faba (L.) and visitation by European orchard bee (Osmia cornuta).

Recent studies link increased ozone (O3) and carbon dioxide (CO2) levels to alteration of plant performance and plant-herbivore interactions, but their interactive effects on plant-pollinator interactions are little understood. Extra floral nectaries (EFNs) are essential organs used by some plants for stimulating defense against herbivory and for the attraction of insect pollinators, e.g., bees. The factors driving the interactions between bees and plants regarding the visitation of bees to EFNs are poorly understood, especially in the face of global change driven by greenhouse gases. Here, we experimentally tested whether elevated levels of O3 and CO2 individually and interactively alter the emission of Volatile Organic Compound (VOC) profiles in the field bean plant (Vicia faba, L., Fabaceae), EFN nectar production and EFN visitation by the European orchard bee (Osmia cornuta, Latreille, Megachilidae). Our results showed that O3 alone had significant negative effects on the blends of VOCs emitted while the treatment with elevated CO2 alone did not differ from the control. Furthermore, as with O3 alone, the mixture of O3 and CO2 also had a significant difference in the VOCs' profile. O3 exposure was also linked to reduced nectar volume and had a negative impact on EFN visitation by bees. Increased CO2 level, on the other hand, had a positive impact on bee visits. Our results add to the knowledge of the interactive effects of O3 and CO2 on plant volatiles emitted by Vicia faba and bee responses. As greenhouse gas levels continue to rise globally, it is important to take these findings into consideration to better prepare for changes in plant-insect interactions.