Aphid–ant mutualism: how honeydew sugars influence the behaviour of ant scouts

Honeydew is the keystone on which ant–aphid mutualism is built. The present study investigates how each sugar identified in Aphis fabae Scopoli honeydew acts upon the feeding and the laying of a recruitment trail by scouts of the aphid‐tending ant Lasius niger Linnaeus, and thus may enhance collective exploitation by the ant mutualists. The feeding preferences shown by L. niger for honeydew sugars are: melezitose = sucrose = raffinose > glucose = fructose > maltose = trehalose = melibiose = xylose. Although feeding is a prerequisite to the launching of trail recruitment, the reverse is not necessarily true: not all ingested sugar solutions elicit a trail‐laying behaviour among fed scouts. Trail mark laying is only triggered by raffinose, sucrose or melezitose, with the latter sugar being specific to honeydew. By comparing gustatory and recruitment responses of ant foragers to sugar food sources, the present study clarifies the role of honeydew composition both as a source of energy and as a mediator in ant–aphid interactions. Lasius niger feeding preferences can be related to the physiological suitability of each sugar (i.e. their detection by gustatory receptors as well as their ability to be digested and converted into energy). Regarding recruitment, the aphid‐synthesized oligosaccharide (melezitose) could be used by ant scouts as a cue indicative of a long‐lasting productive resource that is worthy of collective exploitation and defence against competitors or aphid predators.     

Honeydew amino acids in relation to sugars and their role in the establishment of ant-attendance hierarchy in eight species of aphids feeding on tansy (Tanacetum vulgare)

Abstract. The ratio of the concentration of honeydew total amino acids to total sugars in the honeydew of eight species of aphids, all feeding on tansy, Tanacetum vulgare (L.), was determined and correlated with honeydew production and ant‐attendance. The honeydew of the five ant‐attended aphid species [Metopeurum fuscoviride (Stroyan), Trama troglodytes (v. Hayd), Aphis vandergooti (Börner), Brachycardus cardui (L.), Aphis fabae (Scopoli)] was rich in total amino acids, ranging from 12.9 to 20.8 nmol µL^?1 compared with the unattended aphid Macrosiphoniella tanacetaria (Kalt.) with only 3 nmol µL^?1. Asparagine, glutamine, glutamic acid and serine (all nonessential amino acids) were the predominant amino acids in the honeydew of all species. The total concentration of amino acids in the phloem sap of tansy was much higher (78.7 nmol µL^?1) then in the honeydew samples, and the predominant amino acids were glutamate (34.3%) and threonine (17.7%). A somewhat unexpected result was the finding that those aphid species with the highest total amino acid concentration in the honeydew always had the highest concentration of sugars. The lowest amino acid–sugar combined value was 104–28.8 nmol µL^?1 in the non ant‐attended species M. tanacetaria, and the highest value was an average of 270–89.9 nmol µL^?1 for the three most intensely attended aphid species M. fuscoviride, A. vandergooti and T. troglodytes. There is no evidence that any single amino acid or group of amino acids in the honeydew acted as an attractant for ant‐attendance in these eight aphid species. The richness of the honeydew (rate of secretion × total concentration of sugars), along with the presence of the attractant sugar melezitose, comprised the critical factors determining the extent of ant‐attendance of the aphids feeding on T. vulgare. The high total amino acid concentration in sugar‐rich honeydews can be explained by the high flow‐through of nutrients in aphids that are particularly well attended by ants.     

Age-specific patterns in honeydew production and honeydew composition in the aphid Metopeurum fuscoviride: implications for ant-attendance

The intensity of the mutualistic relationship between aphids and ants depends mainly on the composition and amount of honeydew. We used the model system Tanacetum vulgare-Metopeurum fuscoviride to study age-related differences in honeydew production and composition and its effect on the mutualism between M. fuscoviride and the ant Lasius niger. First and second instar larvae of M. fuscoviride produced only half of the amount of honeydew as older larvae or adults. There were, however, no differences between age classes in the total honeydew sugar concentration, which averaged approx. 80 μg sugar/μl honeydew. Honeydew sugar composition also did not differ between age classes, and melezitose was the dominant sugar (59% in all classes). The amino acid concentration, by contrast, increased significantly with aphid age, reaching 22.6 nmol per μl honeydew in adult M. fuscoviride. This increase was mainly caused by asparagine and glutamine, while there were no differences in the concentrations of the five other regularly detected amino acids and cystine, respectively. The intensity of ant-attendance was significantly lower in colonies of first and second instar larvae than in colonies of older age classes. Ant-attendance correlated with the amount of honeydew produced, and not with the total amino acid concentration.     

The effect of host plants on genotype variability in fitness and honeydew composition of Aphis fabae

Aphid species can be polyphagous, feeding on multiple host plants across genera. As host plant species can have large variation in their phloem composition, this can affect aphid fitness and honeydew composition. Previous research showed significant intraspecific genotype variation in the composition of the honeydew carbohydrates of the black bean aphid Aphis fabae, with the ant attractant trisaccharide melezitose showing especially large variation across different genotypes. In this study, we test if variation in melezitose and carbohydrate composition of aphid honeydew could be linked to the adaptation of specific aphid genotypes to particular host plants. To this end, 4 high and 5 low melezitose secreting genotypes of the black bean aphid Aphis fabae were reared on 4 common host plants: broad bean, goosefoot, beet, and poppy. The carbohydrate composition, and in particular melezitose secretion, showed important aphid genotype and host plant interactions, with some genotypes being high melezitose secreting on 1 host plant but not on another. However, the interaction effects were not paralleled in the fitness measurements, even though there were significant differences in the average fitness across the different host plants. On the whole, this study demonstrates that aphid honeydew composition is influenced by complex herbivore–plant interactions. We discuss the relevance of these findings in the context of ant–aphid mutualisms and adaptive specialization in aphids.     

Wedged between bottom-up and top-down processes: aphids on tansy

Abstract. 1. Many species of aphids exploit a single host‐plant species and have to cope with changing environmental conditions. They often vary greatly in abundance even when feeding on the same host. In a field experiment, the bottom‐up (plant quality/patch type frequency) and top‐down (ant attendance/predation) effects on the abundance of four species of aphids feeding on tansy (Tanacetum vulgare) were tested using a full factorial design. In addition, a model was used to examine these patch characteristics for their relative effects on the population dynamics and abundance of different aphid species. 2. Aphid numbers changed significantly depending on the quality of the host plant and the presence/absence of attending ants. The obligate myrmecophile, Metopeurum fuscoviride, was abundant on high‐quality plants, while on poor quality plants or on plants without attending ants these aphids did not survive until the end of the experiment. The facultative myrmecophiles, Aphis fabae and Brachycaudus cardui, and the unattended aphid species, Macrosiphoniella tanacetaria, all reached similar peak population densities, but M. tanacetaria did best in poor quality patches. 3. Natural enemies reduced aphid numbers, but those species feeding on high‐quality plants survived longer than those on poor‐quality plants, which existed only for a short period of time, especially when associated with ants. Losses due to migration of winged morphs and mortality caused by parasitoids were insignificant. 4. Varying the frequency of different patch types in a model indicates that different degrees of associations with ants are favoured in different environments. If the proportion of high‐quality patches in a habitat is large, obligate myrmecophiles do best. On increasing the number of poor‐quality patches, unattended species become more abundant. 5. The results suggest that, in spite of large species specific differences in growth rates, degree of myrmecophily or life cycle features, the temporal and spatial variability in top‐down and bottom‐up forces differentially affects aphid species and allows the simultaneous exploitation of a shared host‐plant species.     

The cost of ant attendance and melezitose secretion in the black bean aphid Aphis fabae

1. The aphid Aphis fabae (Scopoli) is facultatively tended by Lasius niger (Linnaeus) ants. Previously, we found that A. fabae colonies can be made up of several clones, and that clones display significant differences in the composition of their honeydew sugars, especially in the amount of the ant attractant sugar melezitose that they produce. 2. These clonal differences could greatly impact the strength of the mutualistic interaction with ants as well as the aphids' fitness. 3. Hence, the aim of this study was to compare the fitness of different A. fabae clones that differed in their melezitose secretion, and whether or not they were tended by ants. 4. Individual fitness indices, colony growth, and alate production of single‐clone aphid colonies were analysed. 5. The results demonstrate that the fitness consequences of ant attendance critically depend on an interaction between levels of melezitose production. In particular, we show that high‐melezitose secreting clones produce fewer alates and hence might have a lower dispersal ability in the presence of ants. 6. Furthermore, these data confirm previous evidence that ant attendance is costly and results in the production of fewer apterae.     

The Foraging Behavior of <Emphasis Type="Italic">Lysiphlebus fabarum</Emphasis> (Marshall), a Thelytokous Parasitoid of the Black Bean Aphid in Iran

A series of laboratory experiments examined the foraging behavior of a thelytokous strain of Lysiphlebus fabarum (Marshall), a strongly proovigenic parasitoid of Aphis fabae Scopoli, in Iran. Females use chemical camoflage to forage undisturbed in ant-tended aphid colonies and solicit honeydew from aphids in the manner of ants. Rates of oviposition are very low (∼ 1.2 eggs / h) despite many aphid encounters and persistent ovipositor probing which appears to prime aphids for subsequent honeydew solicitation. Starved females spent 3.6 times longer in host patches (leaf disks with 15 2nd–3rd instar A. fabae) than did females sated on honey, and 40% of this time was spent soliciting honeydew. Five d-old females spent longer in patches than did one d-old females, and parasitized three times as many aphids. A 24 h pre-trial foraging experience did not reduce mean egg load significantly compared to a one h experience, but was sufficient to reduce patch residence time and number of aphids parasitized. Wasps reared under short day conditions (L:D = 10:14) were more pessimistic foragers (remained longer in patches and parasitized more aphids) than females raised under long days (L:D = 16:8). Wasps that encountered aphids previously parasitized by conspecifics began to superparasitize and remained longer in patches than females that encountered only unparasitized aphids. Encounters with other females had no effect on foraging behavior, possibly because cuticular camoflage interferes with conspecific recognition. The exceptionally low oviposition rate of this wasp may reflect a life history in which individual fitness has evolved to be strongly dependent on continued ant attendance throughout the period of progeny development.     

Responses of the ant Lasius niger to various compounds perceived as sweet in humans: a structure-activity relationship study

A behavioural study on the ant Lasius niger was performed by observing its feeding responses to 85 compounds presented in a two-choice situation (tested compound versus water control or sucrose solution). Among these compounds, only 21 were phagostimulating: six monosaccharides (D-glucose, 6-deoxy-D-glucose, L-galactose, L-fucose, D-fructose, L-sorbose), four derivatives of D-glucose (methyl alpha-D-glucoside, D-gluconolactone and 6-chloro- and 6-fluoro-deoxy-D-glucose), five disaccharides (sucrose, maltose, palatinose, turanose and isomaltose), one polyol glycoside (maltitol), three trisaccharides (melezitose, raffinose and maltotriose) and two polyols (sorbitol and L-iditol). None of the 16 non-carbohydrate non-polyol compounds tested, although perceived as sweet in humans, was found to be active in ants. The molar order of effectiveness of the major naturally occuring compounds (melezitose > sucrose = raffinose > D-glucose > D-fructose = maltose = sorbitol) is basically different from the molar order of their sweetness potency in humans (sucrose > D-fructose > melezitose > maltose > D-glucose = raffinose = sorbitol). On a molar basis melezitose is in L. niger about twice as effective as sucrose or raffinose, while D-glucose and D-fructose are three and four times less effective, respectively, than sucrose or raffinose. From a structure-activity relationship study it was inferred that the active monosaccharides and polyols should interact with the ant receptor through only one type of receptor, through the same binding pocket and the same binding residues, via a six-point interaction. The high effectiveness of melezitose in L. niger mirrors the feeding habits of these ants, which attend homopterans and are heavy feeders on their honeydew, which is very rich in this carbohydrate.     

Large interclone differences in melezitose secretion in the facultatively ant-tended black bean aphid Aphis fabae

Many aphids are known to engage in a trophic mutualism with ants, whereby the aphids secrete sugary-rich honeydew which is collected by the ants for food, and the ants, in exchange, protect the aphids against natural enemies. Previous results, however, suggest that the production of some of the honeydew sugars, such as the ant-attractant trisaccharide melezitose, may induce an indirect cost to the aphids. This led us to believe that large differences in the nature of the secreted honeydew might exist, due to some clones capitalizing more or less on their mutualistic interaction with ants, or due to some “cheater” clones foregoing the production of particular sugars, instead taking advantage of the ant-attracting effect of other non sugar-deficient clones, co-occurring on the same plant. Here we present data on clonal variation in the composition of honeydew of the black bean aphid Aphis fabae which confirm this prediction. In particular, our results show that there was large interclone variation in the amount of glucose, melezitose and total sugar produced. The variation in the production of melezitose, however, showed particularly large differences, with 54% (7 out of 13) of the clones screened being virtually deficient for the production of this sugar, irrespective of whether the aphid colonies were ant-tended or not. The consequences of this finding in the context of the evolution and maintenance of the ant–aphid mutualism, as well as the adaptive benefits of oligosaccharide synthesis in aphids and other insects are discussed.     

The influence of facultative endosymbionts on honeydew carbohydrate and amino acid composition of the black bean aphid Aphis fabae

The facultative endosymbionts Hamiltonella defensa and Regiella insecticola are commonly found in aphids. They are linked with various ecological benefits but generally occur at low prevalence, which indicates a possible harbouring cost. Little is known about how the presence of facultative endosymbionts is reflected in honeydew composition. Honeydew is the key mediator of the mutualism between aphids and their tending ants. The present study examines whether endosymbionts have an influence on aphid honeydew quality by comparing the amino acid and carbohydrate concentrations between infected and uninfected aphids. To this end, two genetic lines of the aphid Aphis fabae Scopoli are experimentally infected with different strains of Hamiltonella and Regiella. Infected aphids are shown to have reduced concentrations of amino acids in the honeydew compared with uninfected aphids. However, the presence of endosymbionts has no effect on the absolute amount of carbohydrates produced. Nevertheless, interclonal variation in honeydew composition between aphid genotypes is observed for both carbohydrate and amino acid production. These results imply that the nutritional value of honeydew depends on aphid genotype, as well as on the presence of secondary bacterial endosymbionts, which suggests that there is a physiological cost of harbouring endosymbionts and which could also impact aphid attractiveness to tending ants.     

Effect of willow cultivar and plant age on the melezitose content of giant willow aphid (Tuberolachnus salignus) honeydew

1. The giant willow aphid Tuberolachnus salignus is an invasive pest in New Zealand, attacking over 50 species and hybrids of willow. The aphids produce copious amounts of honeydew, which is used by other insects as a food source.
2. When foraged by honeybees, T. salignus honeydew causes honey to crystallize in the comb and affects bee health; these effects are associated with the elevated melezitose content in the honeydew. The impact of host plant-related factors on T. salignus honeydew melezitose content remains unknown.
3. This study investigated the effect of willow cultivar and plant age on the melezitose content (and that of other sugars) of T. salignus honeydew. To do so, we conducted high-performance liquid chromatography analyses of honeydew samples from 13 willow clones collected in the same season (autumn) from 1- and 2-year old plants under field conditions.
4. Melezitose was the most abundant of the measured sugars in most samples, but its content did not vary significantly with willow cultivar or plant age. By contrast, sucrose was significantly affected by both factors. Fructose and glucose were significantly impacted by willow plant age and cultivar, respectively. A significant cultivar*age interaction was observed for all sugars.
5. We recommend the selection of resistant willow cultivars and further research on potential biocontrol agents to lessen melezitose-related problems in apiculture industries.

     

Multiple Cues for Winged Morph Production in an Aphid Metacommunity

Environmental factors can lead individuals down different developmental pathways giving rise to distinct phenotypes (phenotypic plasticity). The production of winged or unwinged morphs in aphids is an example of two alternative developmental pathways. Dispersal is paramount in aphids that often have a metapopulation structure, where local subpopulations frequently go extinct, such as the specialized aphids on tansy (Tanacetum vulgare). We conducted various experiments to further understand the cues involved in the production of winged dispersal morphs by the two dominant species of the tansy aphid metacommunity, Metopeurum fuscoviride and Macrosiphoniella tanacetaria. We found that the ant-tended M. fuscoviride produced winged individuals predominantly at the beginning of the season while the untended M. tanacetaria produced winged individuals throughout the season. Winged mothers of both species produced winged offspring, although in both species winged offspring were mainly produced by unwinged females. Crowding and the presence of predators, effects already known to influence wing production in other aphid species, increased the percentage of winged offspring in M. tanacetaria, but not in M. fuscoviride. We find there are also other factors (i.e. temporal effects) inducing the production of winged offspring for natural aphid populations. Our results show that the responses of each aphid species are due to multiple wing induction cues.     

Herbivory damage does not indirectly influence the composition or excretion of aphid honeydew

Recent research has shown that extrafloral nectar secretion by plants, which also attracts ants, is a defense inducible by herbivory damage. Our research addresses the question of whether plants can manipulate the honeydew secretions of homopterans as an inducible defense in response to herbivory in the same manner as extrafloral nectaries. We investigated changes in honeydew composition and excretion rate by the facultatively ant-attended aphid Chaitophorus saliniger Shinji (Homoptera: Aphididae) depending on the presence or absence of herbivory by caterpillars (Clostera anastomosis L.; Lepidoptera: Notodontidae) on their host plant, the willow Salix gilgiana Seemen. Our results found no evidence to suggest that herbivory damage to the aphids host plant causes significant changes in aphid population growth, honeydew droplet volume, volume of honeydew excreted per aphid per hour, or composition of three abundant sugars in aphid honeydew as a result of herbivory damage to the aphids host plant, suggesting that, in this particular system, the plant is not manipulating the aphids honeydew output for its own benefit.     

Effects of aphid honeydew sugars on the longevity and fecundity of the aphidophagous gall midge Aphidoletes aphidimyza

The predatory gall midge Aphidoletes aphidimyza, which is used for augmentative biological control of aphids in greenhouses, uses aphid honeydew as an energy source. Currently, Rhopalosiphum padi on barley plants and Melanaphis sacchari on sorghum plants are used as the banker plant systems for A. aphidimyza to control Aphis gossypii on eggplant in Japan. We analyzed the sugar components of the honeydew of these three species by HPLC. The major components of honeydew were sucrose, fructose, and melezitose for A. gossypii; glucose and fructose for R. padi; and glucose, fructose, and melezitose for M. sacchari. Maltose and trehalose were minor components of the honeydew of these species. We investigated the influence of sugars, including three artificial aphid honeydews, six sugar components, and distilled water, on the longevity of unmated A. aphidimyza. Both females and males attained the greatest longevity on sucrose and artificial honeydew of A. gossypii. Mean longevities of both females and males were shortest when they were provided only with water. We conducted another experiment in which a mated female was released in a cage with an eggplant seedling infested with A. gossypii and fed with sucrose or only water. Females fed with sucrose lived significantly longer and had higher lifetime fecundity than those fed only water.     

Influence of aphid honeydew on the foraging behaviour of Hippodamia convergens larvae

1. Environmental cues associated with prey are known to increase predator foraging efficiency. Ladybird larvae are major predators of aphids. The sugary excretion of aphids (honeydew) has been proposed to serve as a prey‐associated cue for ladybird larvae. 2. Ladybird larvae are regularly found on the ground moving between plants or after falling off plants. The use of prey‐associated cues would be particularly beneficial for ladybird larvae on the ground in that such cues would help them to decide which plants to climb because aphids are patchily distributed within as well as amongst plants and, as a result, many plants are either not infested with aphids or do not host an aphid species of high nutritional value for ladybird larvae. 3. Laboratory experiments with larvae of Hippodamia convergens Guérin‐Méneville (Coleoptera: Coccinellidae) were carried out to explore whether honeydew accumulated on the ground is used as a foraging cue. The study also investigated whether, if honeydew is a foraging cue, larvae show differential responses to honeydew of high‐quality prey Acyrthosiphon pisum Harris compared with that of low‐quality prey Aphis fabae Scopoli (both: Homoptera: Aphididae). 4. Hippodamia convergens larvae stayed longer in areas containing honeydew but did not engage in longer bouts of searching. Furthermore, larvae did not distinguish between honeydew from high‐ and low‐quality aphid prey.     

Bacteria may contribute to distant species recognition in ant–aphid mutualistic relationships

Mutualistic interactions between ant and aphid species have been the subject of considerable historical and contemporary investigations, the primary benefits being cleaning and protection for the aphids and carbohydrate‐rich honeydew for the ants. Questions remained, however, as to the volatile semiochemical factor influencing this relationship. A recent study highlighted the role of bacterial honeydew volatile compounds in ant attraction. Here, ant's ability to distantly discriminate 2 aphid species was investigated based on bacterial honeydew semiochemicals emissions using a two‐way olfactometer. Both the mutualistic aphid Aphis fabae L. and the nonmyrmecophilous aphid Acyrthosiphon pisum Harris were found to be attractive for the ant Lasius niger L. The level of attraction was similar in both assays (control vs. one of the aphid species). However, when given a choice between these 2 aphid species, ants showed a significant preference for Aphis fabae. Honeydew volatiles, mostly from bacterial origins, are known to be a key element in ant attraction. Using the same olfactometry protocol, the relative attractiveness of volatiles emitted by honeydews collected from each aphid species and by bacteria isolated from each honeydew was investigated. Again, ants significantly preferred volatiles released by Aphis fabae honeydew and bacteria. This information suggests that microbial honeydew volatiles enable ants to distantly discriminate aphid species. These results strengthen the interest of studying the occurrence and potential impact of microorganisms in insect symbioses.     

Influence of climatic and nutritional factors on yeast population dynamics in the phyllosphere of wheat

The role of saprophytic phyllosphere yeasts in removing aphid honeydew and other nutrients from wheat leaves was evaluated in growth cabinet experiments at different temperatures and relative humidities. Population densities of both pink and white yeasts (Sporobolomyces roseus and Cryptococcus laurentii, respectively) increased between 12 and 24°C, if nutrients were supplied. White yeast numbers increased rapidly at a constant vapor pressure deficit (VPD) of 0.10 kPa and alternating VPDs of 0.10 and 0.61 kPa (each 12 hours per day) but decreased at a constant VPD of 0.61 kPa. In growth cabinet experiments with aphids on wheat plants, the amount of aphid honeydew on the leaves was lower when yeast population densities were high. Addition of amino acids to leaves with honeydew had no effect on yeast population density or the rate of honeydew consumption. This indicated that low concentrations of amino acids in aphid honeydew are not a limiting factor for honeydew consumption by the yeasts. The naturally occurring saprophytes efficiently removed fructose, sucrose, and melezitose from the phyllosphere of field-grown wheat plants.     

Effects of honeydew sugar composition on the longevity of Aphidius ervi

Feeding on sugar‐rich foods such as nectar and honeydew is important for survival of many adult parasitoids. Especially in agricultural systems, honeydew is often the most prevalent carbohydrate source. However, relative to plant nectar, honeydew may be relatively unsuitable, as a result of an unfavourable sugar composition or the presence of secondary plant compounds. We studied survival of the aphid parasitoid Aphidius ervi Haliday (Hymenoptera: Braconidae) on honeydew collected from various aphid species feeding on potato (Solanum tuberosum L., cv. Desiree) (Solanaceae), wheat (Triticum aestivum L., cv. Bobwhite) (Poaceae), or artificial diet, as well as the sugar composition of the different honeydews. Honeydews from the tested aphid species on potato, wheat, or artificial diet were found to be relatively suitable food sources for adult A. ervi, although not always as suitable as a 2 M sucrose solution. There were differences in honeydew sugar composition among the different aphid species on the various host plants. Multivariate statistics showed that the factor ‘aphid species’ had a significant influence on the sugar composition of the honeydew, explaining 27% of the variation in the potato system and 89% in the wheat system. When exploring the relationship between carbohydrate composition of the honeydews from aphids on potato and wheat plants, and their nutritional value for A. ervi, data revealed that differences in parasitoid longevity can to some extent be explained by carbohydrate composition. Furthermore, our results confirm that sucrose and its hexose components glucose and fructose are very suitable carbohydrate sources for hymenopteran parasitoids and show that parasitoid survival on an equimolar solution of the two monosaccharides glucose and fructose does not exceed performance on the disaccharide sucrose.     

Oligosaccharide synthesis regulates gut osmolality in the ant-attended aphid Metopeurum fuscoviride but not in the unattended aphid Macrosiphoniella tanacetaria

Abstract:  An artificial phloem sap (APS) for Metopeurum fuscoviride and Macrosiphoniella tanacetaria , based on analysis of their host plant, Tanacetum vulgare , phloem sap, contained 308  μ g/ μ l (900  μ mol/ μ l) sucrose (and no other sugars) and a mixture of 14 amino acids with a total concentration of 82.5 nmol/ μ l. There was no significant difference in the total amino acid concentration of the honeydew of adult M. fuscoviride fed on the host plant and aphids fed on APS. Incubation of isolated guts in APS indicated no role of gut bacteria or gut enzymes on the amino acid pattern in the gut. The sugar composition of the honeydew of the ant-attended M. fuscoviride indicated a rapid digestion of sucrose into glucose and fructose, and the simultaneous synthesis of considerable amounts of melezitose and some trehalose. The sugar composition of the honeydew of the unattended M. tanacetaria in contrast showed only traces of trehalose and melezitose, but up to 20% erlose in plant-fed aphids. Incubation of isolated guts of M. fuscoviride in APS demonstrated a steady high rate of melezitose synthesis by gut enzymes over an 8-h period. Incubation of isolated guts of M. tanacetaria on the other hand demonstrated only a moderate rate of erlose synthesis and no detectable melezitose or trehalose. Melezitose in the aphid M. fuscoviride is a signal sugar for ants (ecological function), indicating the presence of abundant sugar rich honeydew [ Woodring et al. (2004) Physiol. Entomol. , vol. 29, pp. 311–319]. It was estimated that melezitose reduces the gut osmolality of M. fuscoviride to approximately 25–35% of what it would be without the synthetases (physiological function). M. tanacetaria on the other hand produces very little honeydew, is not attended by ants, and thus there is little need to synthesize large amounts of oligosaccharides to attract ants or for osmoregulation.     

Interactions between ants attendingAphis fabae ssp.cirsiiacanthoidis on thistles and foraging parasitoid wasps

We describe the behavioral interactions between honeydew-collecting workers of the ants Lasius nigerand Myrmica ruginodisand females of three species of aphidiid wasps (Lysiphlebus cardui, Lysiphlebus testaceipes, Trioxys angelicae)foraging for their aphid host, Aphis fabaessp. cirsiiacanthoidis,on thistles. Using field and laboratory experiments, we show that the ant-parasitoid interactions are species specific. Workers of both ant species generally attacked and killed females of T. angelicae,but they ignored those of L. cardui.This pattern was not altered when we anesthetized the wasps slightly with carbon dioxide to reduce their mobility. Prior contacts between L. carduiand either conspecific L. nigerfrom a different nest or workers of a different ant species (M. ruginodis)did not influence L. niger'snonaggressive behavior. The number of aphids parasitized by L. testaceipeswas significantly reduced in aphid colonies attended by L. niger,although this parasitoid was rarely attacked by ants. In encounters between these species of ants and wasps, ant aggression is consistent with differences in wasp behavior. We suggest that, in addition, chemical cues located in the cuticula may enable L. carduito avoid detection by honeydew-collecting ants.