Sugar and amino acid composition of ant‐attended nectar and honeydew sources from an Australian rainforest

Abstract Ants (Hymenoptera: Formicidae) consume a broad spectrum of liquid food sources including nectar and honeydew, which play a key role in their diet especially in tropical forests. This study compares carbohydrates and amino acids from a representative spectrum of liquid sources used by ants in the canopy and understorey of a tropical rainforest in northern Queensland, Australia. Eighteen floral nectars, 16 extrafloral nectars, two wound sap and four homopteran honeydew sources were analysed using high performance liquid chromatography. Wounds comprised flower abscission scars on Normanbya normanbyi L. H. Bailey and bitemarks on Cardwellia sublimis F. Muell. where ants were actively involved in wounding. Discriminant analysis was performed to model differences between food sources in sugar and amino acid concentration and composition. All characteristics varied significantly among plant species. Honeydew contained a broader spectrum of sugars (including melezitose, raffinose, melibiose, lactose and maltose) than nectar (sucrose, glucose, fructose), but certain extrafloral nectars had similar amino acid profiles and, like honeydew sources, were often monopolized by ants. Most common amino acids across the sources were proline, alanine and threonine among 17 α‐amino acids identified. Interspecific variability concealed characteristic differences in sugar and amino acid parameters between nectar, honeydew and wound sap across all plants, but these types differed significantly when found on the same plant. Among all sources studied, only a few flower nectars were naturally not consumed by ants and they were significantly less attended than sugar controls in feeding trials. These nectars did not differ in sugars and amino acids from ant‐attended flower nectars, suggesting the activity of repellents. Apart from these exceptions, variability in amino acids and carbohydrates is proposed to play a key role in ant preferences and nutrition.     

Preference of cabbage white butterflies and honey bees for nectar that contains amino acids

Summary Amino acids occur in most floral nectars but their role in pollinator attraction is relatively unstudied. Nectars of butterfly-pollinated flower tend to have higher concentrations of amino acids than do flowers pollinated by bees and many other animals, suggesting that amino acids are important attractants of butterflies to flowers. In order to determine whether amino acids are important in attracting butterflies and bees, we tested the preference of cabbage white butterflies (Pieris rapae) and honey bees (Apis mellifera) by allowing them to feed from artificial flowers containing sugar-only or sugar-amino acid mimics ofLantana camara nectar. Honey bees and female cabbage white butterflies consumed more sugar-amino acid nectar than sugar-only nectar. In addition, female cabbage white butterflies visited artificial flowers containing sugar-amino acid nectars more frequently than flowers containing sugar-only nectars; honey bees spent more time consuming the sugar-amino acid nectar. Male cabbage white butterflies did not discriminate between the two nectars. These results support the hypothesis that the amino acids of nectar contribute to pollinator attraction and/or feeding.     

FACULTATIVE MUTUALISM BETWEEN WEEDY VETCHES BEARING EXTRAFLORAL NECTARIES AND WEEDY ANTS IN CALIFORNIA

Vetches (Vicia spp.) were studied in the San Francisco Bay Area of California in the spring of 1978. The stipular nectaries of the vetches are visited by the Argentine ant, Iridomyrmex humilis Mayr. The nectaries were removed to exclude ants in controlled experiments to determine if these ants protect the vetches from herbivores or seed predators. Plants with excised nectaries suffered substantially greater damage to their foliage than control plants, indicating that ants protect the foliage. There was no indication that ants protect the vetches from seed predators, but fruit set was substantially lower in plants with excised nectaries. Analysis of sugar and amino acid composition of extrafloral nectar served as a basis for feeding tests with Argentine ants by using artificial nectar solutions. Ants preferred sucrose and glucose solutions over fructose. They showed no preference for any one sugar mixture over another, nor did they exhibit differential recruitment to artificial nectar solutions containing only sugars or sugars and amino acids.     

THE EXTRAFLORAL NECTARIES OF IPOMOEA CARNEA (CONVOLVULACEAE)

Ipomoea carnea (Convolvulaceae) possesses two types of extrafloral nectaries, located on the petiole and on the pedicel. These secrete a complex nectar containing sugars and amino acids. The insects attracted to the extrafloral nectaries are predominantly ants and they are relatively abundant throughout the year. A number of incidents of plant defense as a result of the presence of extrafloral nectary visitors at the extrafloral nectaries of I. carnea were observed and are consistent with the ant-guard theory of the function of extrafloral nectaries.     

Detection of amino acids in artificial nectars by two tropical ants,Leptothorax and Monomorium

Summary Many ants forage at extrafloral nectar on plants and provide the plant with some measure of protection from herbivory. These nectars contain sugars, amino acids and, often, other compounds. The role of amino acids in attracting ants to extrafloral nectars was studied by baiting with Karo-syrup-based solutions. Control (without amino acids) and experimental (with amino acids) solutions were placed in second growth forest in Trinidad, W.I. The number of ants visiting the solutions was counted at fiveminute intervals for 45 min. In tests of solutions with only one amino acid, both Leptothorax sp. and Monomorium sp. visited solutions with alanine, arginine, serine, cysteine, methionine or aspartic acid more frequently than sugaronly controls. Monomorium preferred control solutions to tyrosine solutions; Leptothorax preferred control solutions to histidine solutions. Leptothorax did not discriminate between control and tyrosine solutions; Monomorium did not discriminate between control and histidine solutions. However, in six of eight tests of combinations of amino acids, ants visited control solutions more frequently than experimental solutions. These results suggest that ants can act as selective agents, favoring plants with particular amino acids in their nectars.     

Facultative Ant-Plant Interactions: Nectar Sugar Preferences of Introduced Pest Ant Species in South Florida1

We observed nectar use by native and exotic ant species in nature, garden, and urban situations, and found ants utilizing floral and extrafloral nectar of a variety of flowering plant species. We collected 31 plant nectars (29 floral, 2 extrafloral) and used them in feeding preference tests against standard solutions of sugars (20 percent fructose, glucose, and sucrose, and their mixture), 10 trials for each nectar-ant comparison. We compared time-to-discovery and total ant visits to each droplet using ANOVA, and found that both trial and solution contributed significantly to the variation in most experiments. Seven of the floral nectars tested were significantly more attractive to certain ant species than the sugar solutions. Not only do ants use floral nectar, but it appears that some floral nectars contain compounds that are especially attractive to ants.     

Herbivory-induced extrafloral nectar increases native and invasive ant worker survival

Ascertaining the costs and benefits of mutualistic interactions is important for predicting their stability and effect on community dynamics. Despite widespread designation of the interaction between ants and extrafloral nectaries (EFNs) as a mutualism and over 100 years of studies on ant benefits to plants, the benefits to ants have never been experimentally quantified. The success of invasive ants is thought to be linked to the availability of carbohydrate-rich resources, though reports of invasive ant visits to EFNs are mixed. In two laboratory experiments, we compared worker survival of one native (Iridomyrmex chasei) and two invasive ant species (Linepithema humile and Pheidole megacephala) exposed to herbivorized or non-herbivorized EFN-bearing plants (Acacia saligna) or positive and negative controls. We found that non-herbivorized plants did not produce any measurable extrafloral nectar, and ants with access to non-herbivorized plants had the same survival as ants with access to an artificial plant and water (unfed ants). Ants given herbivorized plants had 7–11 times greater worker survival relative to unfed ants, but there were no differences in survival between native and invasive ants exposed to herbivorized plants. Our results reveal that ants cannot induce A. saligna extrafloral nectar production, but workers of both native and invasive ant species can benefit from extrafloral nectar as much as they benefit from sucrose.     

THE FUNCTION OF EXTRAFLORAL NECTARIES IN OPUNTIA ACANTHOCARPA (CACTACEAE)

Opuntia acanthocarpa (Cactaceae) possesses extrafloral nectaries embedded in the areoles of new reproductive and vegetative growth. The nectar secreted by these glands attracts ants and is a nutritional food source. Members of one attracted ant species, Crematogaster opuntiae (Myrmicinae), are aggressive and efficient defenders of the plants against cactus-feeding insects. The results of our study are consistent with the ant-guard hypothesis for the role of extrafloral nectaries in O. acanthocarpa. Additionally, individuals of O. acanthocarpa are well protected in comparison with those of O. phaeacantha. The latter generally possess ephemeral extrafloral nectaries and consistently maintain fewer ants.     

Ant visitation of extrafloral nectaries of Passiflora: the effects of nectary attributes and ant behavior on patterns in facultative ant-plant mutualisms

Extrafloral nectary (EFN) plants are widespread and can be quite species-rich in some communities. Thus, ants that utilize extrafloral nectar may have the opportunity to discriminate among a wide variety of nectar sources, resulting in variation in the ant attention EFN plants receive. In this study, we compare ant visitation rates of three Passiflora species that coexist in an early successional neotropical forest. These three vine species (Passiflora auriculata, P. biflora, and P. oerstedii) differ in their extrafloral nectary structure and placement, and thus may attract different numbers or species of ants. Through censuses of ants tending extrafloral nectaries, we found that P. auriculata received significantly higher numbers of ant visitors than P. oerstedii, but did not differ significantly from P. biflora in its attractiveness to ants. We also found that termite worker baits (simulating herbivores) placed on P. auriculata and P. biflora were discovered by ants significantly more quickly than baits placed on P. oerstedii. In both ant visitation censuses and in termite bait trials, we found no significant associations between Passiflora species and the species of ant visitors. We also performed experimental manipulations of several characteristics of P. auriculata, which resulted in changes in levels of ant visitation. When petiolar nectaries of P. auriculata were experimentally blocked, visitation by the common ant Ectatomma ruidum declined, even though nectaries on the leaf surfaces were still functional. Connections with other vegetation also had an effect on ant visitation. Though experimental creation of connections between growing P. auriculata shoots and other vegetation did not enhance ant visitation, eliminating connections resulted in a significant decline in the number of ant visitors. The results of this study suggest factors that may contribute to variation in ant visitation of extrafloral nectary plants. In addition, this study demonstrates that extrafloral nectary plants co-occurring in a habitat and available to the same ants may differ in patterns of visitation by ants and perhaps in the quality of protection from herbivores that they receive.     

Relationships between the ant Brachymyrmex obscurior (Hymenoptera, Formicidae) and Acacia pennatula (Fabaceae)

Summary. In central Mexico, the ant Brachymyrmex obscurior Forel feeds on nectar produced by extrafloral nectaries of Acacia pennatula (Schlecht. & Cham.) Benth. However, no studies have determined whether the ants visitation is related to plant nectar availability and whether ants protect A. pennatula from herbivory. The objectives of this 2-yr study (2000–2001) were to assess whether seasonal changes in ant visitation coincide with extrafloral nectar productivity in A. pennatula and to determine whether ants protect the plant. At the end of the dry season (April–June) B. obscurior was the only ant species on A. pennatula and extrafloral nectar production is limited to this period. Exclusion experiments, performed at the end of the dry season showed that A. pennatula did not receive a protective benefit when visited by ants. Branches with ants and branches where ants are excluded had similar numbers of the nonmyrmecophile leafhopper Sibovia sp. which was the only herbivore observed under natural conditions.Received 24 March 2004; revised 4 September 2004; accepted 8 September 2004.     

Argentine ants displace floral arthropods in a biodiversity hotspot

Argentine ant (Linepithema humile (Mayr)) invasions are often associated with the displacement of ground‐dwelling arthropods. Argentine ant invasions can also exert other effects on the community through interactions with plants and their associated arthropods. For example, carbohydrate resources (e.g. floral or extrafloral nectar) may influence foraging behaviour and interactions among ants and other arthropods. In South Africa's Cape Floristic Region, Argentine ants and some native ant species are attracted to the floral nectar of Leucospermum conocarpodendron Rourke (Proteaceae), a native tree that also has extrafloral nectaries (EFNs). Despite having relatively low abundance in pitfall traps, Argentine ants visited inflorescences more frequently and in higher abundance than the most frequently observed native ants, Camponotus spp., though neither native nor Argentine ant floral foraging was influenced by the EFNs. Non‐metric multidimensional scaling revealed significant dissimilarity in arthropod communities on inflorescences with Argentine ants compared to inflorescences with native or no ants, with Coleoptera, Diptera, Hymenoptera, Arachnida, Orthoptera, and Blattaria all being underrepresented in inflorescences with Argentine ants compared to ant‐excluded inflorescences. Native honeybees (Apis mellifera capensis Eschscholtz) spent 75% less time foraging on inflorescences with Argentine ants than on inflorescences without ants. Neither Argentine ant nor native ant visits to inflorescences had a detectable effect on seed set of Le. conocarpodendron. However, a pollen supplementation experiment revealed that like many other proteas, Le. conocarpodendron is not pollen‐limited. Flower predation was negatively associated with increased ant visit frequency to the inflorescences, but did not differ among inflorescences visited by native and Argentine ants. Displacement of arthropods appears to be a consistent consequence of Argentine ant invasions. The displacement of floral arthropods by Argentine ants may have far‐reaching consequences for this biodiversity hotspot and other regions that are rich in insect‐pollinated plants.     

Extranuptial nectaries in flowers: ants increase the reproductive success of the ant‐plant Miconia tococa (Melastomataceae)

• Although the production of extranuptial nectar is a common strategy of indirect defence against herbivores among tropical plants, the presence of extranuptial nectaries in reproductive structures is rare, especially in ant‐plants. This is because the presence of ants in reproductive organs can generate conflicts between the partners, as ants can inhibit the activity of pollinators or even castrate their host plants. Here we evaluate the hypothesis that the ant‐plant Miconia tococa produces nectar in its petals which attracts ants and affects fruit set.
• Floral buds were analysed using anatomical and histochemical techniques. The frequency and behaviour of floral visitors were recorded in field observations. Finally, an ant exclusion experiment was conducted to evaluate the effect of ant presence on fruit production.
• The petals of M. tococa have a secretory epidermis that produces sugary compounds. Nectar production occurred during the floral bud stage and attracted 17 species of non‐obligate ants (i.e. have a facultative association with ant‐plants). Ants foraged only on floral buds, and thus did not affect the activity of pollinators in the neighbouring open flowers. The presence of ants in the inflorescences increased fruit production by 15%.
• To our knowledge, the production of extranuptial nectar in the reproductive structures of a myrmecophyte is very rare, with few records in the literature. Although studies show conflicts between the partners in the ant–plant interaction, ants that forage on M. tococa floral buds protect the plant against floral herbivores without affecting bee pollination.

     

Influence of light,dark, temperature and drought on metabolite and ion composition in nectar and nectaries of an epiphytic bromeliad species (Aechmea fasciata)

• Research into the influence of stress factors, such as drought, different temperatures and/or varied light conditions, on plants due to climate changes is becoming increasingly important. Epiphytes, like many species of the Bromeliaceae, are particularly affected by this, but little is known about impacts on nectar composition and nectary metabolism.
• We investigated the influence of drought, different temperatures and light–dark regimes on nectar and nectaries of the epiphytic bromeliad species, Aechmea fasciata, and also the influence of drought with the terrestrial bromeliad, Billbergia nutans. The content of sugars, amino acids and ions in nectar and nectaries was analysed using HPLC. In addition, the starch content and the activities of different invertases in nectaries were determined.
• Compositions of nectar and nectaries were hardly influenced, neither by light nor dark, nor by different temperatures. In contrast, drought revealed changes in nectar volumes and nectar sugar compositions in the epiphytic bromeliad as well as in the terrestrial bromeliad. In both species, the sucrose‐to‐hexose ratio in nectar decreased considerably during the drought period. These changes in nectar sugar composition do not correlate with changes in the nectaries. The total sugar, amino acid and ion concentrations remained constant in nectar as well as in nectaries during the drought period.
• Changes in nectar composition or in the production of floral pollinator rewards are likely to affect plant–pollinator interactions. It remains questionable how far the adaptations of the bromeliads to drought and diverse light or temperature conditions are still sufficient.

     

Nectar quality affects ant aggressiveness and biotic defense provided to plants

Ant–plant mutualisms are useful models for investigating how plant traits mediate interspecific interactions. As plant‐derived resources are essential components of ant diets, plants that offer more nutritious food to ants should be better defended in return, as a result of more aggressive behavior toward natural enemies. We tested this hypothesis in a field experiment by adding artificial nectaries to individuals of the species Vochysia elliptica (Vochysiaceae). Ants were offered one of four liquid foods of different nutritional quality: amino acids, sugar, sugar + amino acids, and water (control). We used live termites (Nasutitermes coxipoensis) as herbivore competitors and observed ant behavior toward them. In 88 hr of observations, we recorded 1,009 interactions with artificial nectaries involving 1,923 individual ants of 26 species. We recorded 381 encounters between ants and termites, of which 38% led to attack. Sixty‐one percent of these attacks led to termite exclusion from the plants. Recruitment and patrolling were highest when ants fed upon nectaries providing sugar + amino acids, the most nutritious food. This increase in recruitment and patrolling led to higher encounter rates between ants and termites, more frequent attacks, and faster and more complete termite removal. Our results are consistent with the hypothesis that plant biotic defense is mediated by resource quality. We highlight the importance of qualitative differences in nectar composition for the outcome of ant–plant interactions. Abstract in Portuguese is available with online material.     

The role of alanine synthesis and nitrate-induced nitric oxide production during hypoxia stress in Cucurbita pepo nectaries

Floral nectar is a sugary solution produced by nectaries to attract and reward pollinators. Nectar metabolites, such as sugars, are synthesized within the nectary during secretion from both pre-stored and direct phloem-derived precursors. In addition to sugars, nectars contain nitrogenous compounds such as amino acids; however, little is known about the role(s) of nitrogen (N) compounds in nectary function. In this study, we investigated N metabolism in Cucurbita pepo (squash) floral nectaries in order to understand how various N-containing compounds are produced and determine the role of N metabolism in nectar secretion. The expression and activity of key enzymes involved in primary N assimilation, including nitrate reductase (NR) and alanine aminotransferase (AlaAT), were induced during secretion in C. pepo nectaries. Alanine (Ala) accumulated to about 35% of total amino acids in nectaries and nectar during peak secretion; however, alteration of vascular nitrate supply had no impact on Ala accumulation during secretion, suggesting that nectar(y) amino acids are produced by precursors other than nitrate. In addition, nitric oxide (NO) is produced from nitrate and nitrite, at least partially by NR, in nectaries and nectar. Hypoxia-related processes are induced in nectaries during secretion, including lactic acid and ethanolic fermentation. Finally, treatments that alter nitrate supply affect levels of hypoxic metabolites, nectar volume and nectar sugar composition. The induction of N metabolism in C. pepo nectaries thus plays an important role in the synthesis and secretion of nectar sugar.     

Induced Floral and Extrafloral Nectar Production Affect Ant‐pollinator Interactions and Plant Fitness

Thousands of plant species throughout tropical and temperate zones secrete extrafloral nectar to attract ants, whose presence provides an indirect defense against herbivores. Extrafloral nectaries are located close to flowers and may modify competition between ants and pollinators. Here, we used Lima bean (Phaseolus lunatus L.) to study the plants interaction between ants and flower visitors and its consequences for plant fitness. To test these objectives, we carried out two field experiments in which we manipulated the presence of ants and nectar production via induction with jasmonic acid (JA). We then measured floral and extrafloral nectar production, the number of patrolling ants and flower visitors as well as specific plant fitness traits. Lima bean plants under JA induction produced more nectar in both extrafloral nectaries and flowers, attracted more ants and produced more flowers and seeds than non‐induced plants. Despite an increase in floral nectar in JA plants, application of this hormone had no significant effects on flower visitor attraction. Finally, ant presence did not result in a decrease in the number of visits, but our results suggest that ants could negatively affect pollination efficiency. In particular, JA‐induced plants without ants produced a greater number of seeds compared with the JA‐treated plants with ants.     

Variation in the use of orchid extrafloral nectar by ants

Summary The relation between ant-plant specificity and the use of host plants as a resource was investigated in the facultative, myrmecophytic orchid, Caularthron bilamellatum (Rchg.f.) Schult. Using stable isotopes, we determined the portion of the ants' diets derived from host plants. We documented that six ant species inhabiting the orchid: (1) derived nutritional benefit from host orchids, and (2) had species-specific levels of extrafloral nectar use. Proportionate contribution of extrafloral nectar to ant diets ranged from 11 to 48%. These results demonstrate extreme interspecific differences in the nutritional benefits received by ants from host orchids. Interspecific differences in nutritional benefits from orchid nectar may be affected by colony size, nutritional needs, behavioral ecology of the ants, and the abundance of alternate food sources.     

Why do ants shift their foraging from extrafloral nectar to aphid honeydew?

When aphids parasitize plants with extrafloral nectaries (EFNs) and aphid colony size is small, ants frequently use EFNs but hardly tend aphids. However, as the aphid colony size increases, ants stop using EFNs and strengthen their associations with aphids. Although the shift in ant behavior is important for determining the dynamics of the ant–plant–aphid interaction, it is not known why this shift occurs. Here, we test two hypotheses to explain the mechanism responsible for this behavioral shift: (1) Extrafloral nectar secretion changes in response to aphid herbivory, or (2) plants do not change extrafloral nectar secretion, but the total reward to ants from aphids will exceed that from EFNs above a certain aphid colony size. To judge which mechanism is plausible, we investigated secretion patterns of extrafloral nectar produced by plants with and without aphids, compared the amount of sugar supplied by EFNs and aphids, and examined whether extrafloral nectar or honeydew was more attractive to ants. Our results show that there was no inducible extrafloral secretion in response to aphid herbivory, but the sugar concentration in extrafloral nectar was higher than in honeydew, and more ant workers were attracted to an artificial extrafloral nectar solution than to an artificial aphid honeydew solution. These results indicate that extrafloral nectar is a more attractive reward than aphid honeydew per unit volume. However, even an aphid colony containing only two individuals can supply a greater reward to ants than EFNs. This suggests that the ant behavioral shift may be explained by the second hypothesis.     

Extrafloral‐nectaries and interspecific aggressiveness regulate day/night turnover of ant species foraging for nectar on Bionia coriacea

Plants bearing extrafloral nectaries (EFNs) vary the secretion of nectar between day and night, which creates turnover in the composition of interacting ant species. Daily variation in the composition of ant species foraging on vegetation is commonly observed, but its mechanisms are poorly understood. We evaluated the daily variation in nectar availability and interspecific aggressiveness between ants as possible regulatory mechanisms of the turnover in ant–plant interactions. We hypothesized that (i) plants would interact with more ant species during periods of higher secretion of nectar and that (ii) aggressive ant species would compete for nectar, creating a daily turnover of species collecting nectar. We tested this hypothesis by measuring the production of nectar during the day and night and by experimentally removing EFNs of Bionia coriacea (=Camptosema coriaceum) (Nees & Mart.) Benth. (Fabaceae: Faboideae) plants in a Brazilian savanna (Cerrado). We then compared the abundance and composition of ant species between those treatments and during the day. Our results indicate that more ant workers forage on plants during the day, when nectar was sugary, while more ant species forage at night, when aggressiveness between ant species was lower. We also detected a day/night turnover in ant species composition. Ant species foraging for nectar during the day were not the same at night, and this turnover did not occur on plants without EFNs. Both dominant ant species, diurnal Camponotus crassus (Hymenoptera: Formicidae) and nocturnal Camponotus rufipes (Hymenoptera: Formicidae), were the most aggressive species, attacking other ants in their specific periods of forage while also being very aggressive toward each other. However, this aggressiveness did not occur in the absence of nectar, which allowed non‐aggressive nocturnal ant species to forage only during the daytime, disrupting the turnover. We conclude that extrafloral‐nectar presence and interspecific aggressiveness between ants, along with other environmental factors, are important mechanisms creating turnovers in ants foraging on plants.