Phloem amino acids and the host plant range of the polyphagous aphid, Aphis fabae

This study investigated the relationship between the essential amino acid requirement of the aphid Aphis fabae Scop. and the phloem sap amino acid composition of its host plants. The dietary amino acid requirement of A. fabae varied between clones. One or more of the eight clones of A. fabae tested displayed depressed larval survival, larval growth rate, or r_m on diets lacking histidine, methionine, threonine, and valine, but none of the other five essential amino acids. The required amino acids corresponded closely to the essential amino acids that varied in relative concentrations among 16 plant species tested: histidine, threonine, tryptophan, and valine. It is suggested that the interclonal variation in the dietary requirements of an aphid species may contribute to the intraspecific variation in plant utilisation patterns. The phloem sap amino acid composition and sucrose : amino acid ratio did not differ consistently between host plant species of A. fabae and non‐host species, indicating that phloem amino acid composition is not an important factor in determining the host plant range of this aphid species.     

How nutritionally imbalanced is phloem sap for aphids?

Aphids harbour intracellular symbionts (Buchnera) that provide their host with amino acids present in low amounts in their diet, phloem sap. To find out the extent to which aphids depend on their symbionts for synthesis of individual essential amino acids, we have evaluated how well phloem sap amino acid composition matches the aphids' needs. Amino acid compositions of the ingested phloem sap were compared to amino acids in aphid body proteins and also to available information about optimal diet composition for other plant feeding insects. Phloem sap data from severed stylets of two aphid species, Rhopalosiphum padi (L.) (Homoptera: Aphididae) feeding on wheat, and Uroleucon sonchi (L.) (Homoptera: Aphididae) feeding on Sonchus oleraceus (L.), together with published information on phloem sap compositions from other plant species were used.Phloem sap has in general only around 20% essential amino acids, with a range from 15–48%. Aphid body proteins and optimal diets for two other plant feeding insects have around 50%. The phloem sap of early flowering S. oleraceus ingested by U. sonchi contained 48%, which seems to be exceptional. Aphids feeding on different plants appear to be very differently dependent on their symbionts for their overall essential amino acid synthesis, due to the large variation in proportion of essential amino acids in phloem sap from different plants.The profile of the essential amino acids in phloem sap from different plant species corresponds rather well to profiles of both aphid body proteins and optimal diets determined for plant feeding insects. However, methionine and leucine in phloem sap are in general low in these comparisons, suggesting a higher dependence on the symbiont for synthesis of these amino acids. Concentrations of several essential amino acids in phloem from different plant species seem to vary together, suggesting that levels of symbiont provisioning of different amino acids are adjusted in parallel.     

Nutritional enhancement of host plants by aphids - a comparison of three aphid species on grasses

Three aphid species were compared with respect to ability of enhancing the nutritional quality of their host plants. Rhopalosiphum padi, which does not induce macroscopic changes in its host plants, was compared with Schizaphis graminum and Diuraphis noxia, both of which induce distinctive types of chlorotic lesions. Phloem sap samples were collected from severed stylets of feeding aphids and from exudates of cut leaves of plants uninfested or infested with each aphid species. Samples were analyzed for concentrations of individual amino acids.Compared to R. padi, S. graminum ingested phloem sap with a two-fold higher concentration of amino acids and a much higher proportion of essential amino acids. Similar differences between these two aphid species were observed on both wheat and barley. For each aphid species, the absolute concentrations of amino acids and the relative proportions of essential amino acids were similar between the two host plants. Effects of D. noxia were similar to those of S. graminum, though less dramatic. Exudates from leaves infested with each aphid species showed relative concentrations of individual amino acids that were similar to those in the corresponding stylet exudates. Based on comparison of stylet exudates and cut leaf exudates from infested and uninfested plants, R. padi seems to have little effect on amino acid composition of phloem. Changes in the phloem induced by both S. graminum and D. noxia appear to be systemic, affecting at least the whole leaf they are feeding on. The changes observed for D. noxia and for S. graminum are likely to be nutritionally advantageous for the aphids and are expected to affect the aphids' dependence on nutritional supplementation by intracellular symbionts (Buchnera).     

Patterns in aphid honeydew production parallel diurnal shifts in phloem sap composition

Variation in phloem sap composition is important in determining aphid performance and is known to occur at both diurnal timescales and in response to plant age. For field grown potato plants, Solanum tuberosum L. (Solanaceae), we determined diurnal variation in components of phloem sap, measured by ethylene diamine tetra‐acetate exudation, and tested for impacts of plant age. The effects of plant age and diurnal cycles on honeydew production by Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer) (both Hemiptera: Aphididae) were also quantified. Both the ratio of sucrose to amino acids and the composition of amino acids in phloem sap varied significantly with time of day. Dietary essential amino acids contributed a smaller proportion of amino acids in the phloem sap of older plants and during early phases of the diurnal cycle. The only significant effect on aphid honeydew production was of the diurnal cycle for Ma. euphorbiae, although increased honeydew production during the day when compared with the production at night, was consistent across the two species. In contrast with studies carried out at seasonal scales, we found limited evidence for variation in phloem sap composition in response to plant age, consistent with our results for honeydew production. These data highlight the need for improved understanding of how seasonal and diurnal physiology of plants influence performance in phloem sap feeding insects.     

Seasonal changes in the free amino acids of oat and barley phloem sap in relation to plant growth stage and growth of Rhopalosiphum padi

The concentrations and composition of free amino acids in phloem sap from two cultivars of oats and barley, both susceptible to the aphid Rhopalosiphum padi, were determined by means of high performance liquid chromatography. Sap was collected from excised aphid stylets at three developmental stages (seedlings, tillering plants and plants undergoing stem elongation) from plants given or not given fertiliser and grown outdoors. In connection, the growth of individual R. padi nymphs was estimated at the same phenological stages on plants grown in the greenhouse. The content of free amino acids was consistently higher in seedlings than in plants at the early tillering stage. Only in seedlings did the addition of fertiliser increase amino acid levels. Barley phloem sap contained more free amino acids than that of oats when fertiliser was added and at later developmental stages. Phloem sap of oats and barley showed similar patterns in their composition of free amino acids at the seedling stage, but as the plants grew older the patterns became increasingly different. Plants given fertiliser had higher amounts of dicarboxylic amino acids (glutamic and aspartic acid) than unfertilised plants. The concentrations of γ-amino butyric acid, glycine, histidine, and methionine were very low in all treatments. The relative growth rates of R. padi nymphs were low when amino acid content was low and vice versa. The results are discussed in relation to host plant suitability and plant resistance mechanisms.     

Free amino acid content of phloem sap from Brassica plants in relation to performance of Lipaphis erysimi (Hemiptera: Aphididae)

Cultivars of Brassica juncea, B. napus and B. campestris, differing in host-suitability to the mustard aphid (Lipaphis erysimi pseudobrassicae Kalt.), were analysed for their phloem sap content of free amino acids. Sap was collected from excised mustard aphid stylets and analysed by means of high performance liquid chromatography. Aspartic and glutamic acid were, together with glutamine, the dominating amino acids. All plants had very little glycine, γ-aminobutyric acid (GABA), methionine and tryptophan in their sap. This amino acid pattern closely resembled that of cereals. The total concentration and the individual composition of amino acids were not found to be related to the level of aphid resistance previously found in the investigated Brassica species. However, one B. campestris cultivar, yellow sarson YSB-9, on which the aphids were shown to grow more slowly, had less amino acid content than other B. campestris varieties. The slow population growth of aphids on B. juncea, compared to that on B. campestris, is probably due to other than nutritional factors.     

Role of phloem sap quality and exudation characteristics on performance of pea aphid grown on lucerne genotypes

The phloem sap of two lucerne (Medicago sativa L.) genotypes inducing differences in pea aphid,Acyrthosiphon pisum Harris (Homoptera: Aphididae) performances (weight, fecundity, survival), was collected by stylectomy (radio frequency microcautery). Sugars and amino acids were assayed. No significant difference in their concentrations could be established between genotypes. It is thus unlikely that a substantial part of the varietal resistance of these lucerne genotypes to pea aphid could be related to the content in sugars or amino acids. The same conclusion was drawn for the sugar/amino acid ratio. The amino acid profiles were globally similar. They nevertheless allowed a discrimination between the two genotypes on a few amino acids (alanine, leucine, isoleucine, arginine, ornithine) only. Moreover, the degree of resistance can be related neither to GABA concentration, nor to that of sulfur amino acids, because these constituants were either absent, or in trace amounts, whatever the lucerne genotype taken into account. Stylectomy parameters revealed some interesting differences in the sap exudation of the two genotypes. Average volume of exudation by stylet and average duration of exudation were respective 2.8 and 2.3 times higher in the susceptible genotype. These results suggest a reduced flow of phloem sap, after puncturing of the sieve-tubes by the aphid stylets on the resistant genotype; this could result in a greater difficulty in phloem sap ingestion by aphids settled on the resistant genotype and thus in suboptimal performances.     

Free amino Acid composition of leaf exudates and Phloem sap : a comparative study in oats and barley

Comparisons were made between the free amino acid composition in leaf exudates and that in pure phloem sap, using twin samples taken from a single leaf of two oat (Avena sativa L.) and three barley (Hordeum vulgare L.) varieties. Leaf exudate was collected in a 5 mm EDTA-solution (pH 7.0) from cut leaf blades and phloem sap was obtained through excised aphid (Rhopalosiphum padi L.) stylets. Fluorescent derivatives of amino acids were obtained using 9-fluorenylmethyl chloroformate and were separated by means of high performance liquid chromatography. The total concentration of free amino acids varied considerably in the exudate samples. There was no correlation between the total amino acid content in the exudate samples and that of the corresponding phloem sap samples, but the amino acid composition of the corresponding samples was highly correlated (median R²-value 0.848). There was only limited between-plant variation in phloem sap amino acid composition. Nevertheless, in comparisons involving all samples, many of the amino acids showed significant correlations between their relative amounts in exudate and phloem sap. The results presented here indicate that the exudate technique holds great promise as an interesting alternative to the laborious and time-consuming stylet-cutting technique of obtaining samples for comparative studies of phloem sap.     

Sources of variation in dietary requirements in an obligate nutritional symbiosis

The nutritional symbiosis between aphids and their obligate symbiont, Buchnera aphidicola, is often characterized as a highly functional partnership in which the symbiont provides the host with essential nutrients. Despite this, some aphid lineages exhibit dietary requirements for nutrients typically synthesized by Buchnera, suggesting that some aspect of the symbiosis is disrupted. To examine this phenomenon in the pea aphid, Acyrthosiphon pisum, populations were assayed using defined artificial diet to determine dietary requirements for essential amino acids (EAAs). Six clones exhibiting dependence on EAAs in their diet were investigated further. In one aphid clone, a mutation in a Buchnera amino acid biosynthesis gene could account for the clone''s requirement for dietary arginine. Analysis of aphid F₁ hybrids allowed separation of effects of the host and symbiont genomes, and revealed that both affect the requirement for dietary EAAs in the clones tested. Amino acid requirements were minimally affected by secondary symbiont infection. Our results indicate that variation among pea aphids in dependence on dietary amino acids can result from Buchnera mutation as well as variation in the host genotype.     

How aphids decide what is good for them: experiments to test aphid feeding behaviour on Tanacetum vulgare (L.) using different nitrogen regimes

Leaf-chewing herbivores select food with a protein/carbohydrate ratio of 0.8–1.5, whereas phloem sap, which aphids feed on, has a ratio of ~0.1. Enhanced N fertilization increases the amino acid concentration in phloem sap and elevates the N/C ratio. The study examines: (1) whether aphids select between plants of different N nutrition, (2) whether feeding time correlates with the amino acid composition of phloem sap, and (3) at which stage of probing aphids identify the quality of the plant. Uroleucon tanaceti (Mordvilko) and Macrosiphoniella tanacetaria (Kaltenbach), specialist aphids feeding on tansy (Tanacetum vulgare L.), were reared on this host plant grown essentially hydroponically (in Vermiculite) in the greenhouse on 1, 3, 6, or 12 mM NH₄NO₃. One and 3 mM NH₄NO₃ corresponds to the situation found in natural tansy stands. Aphid stylet penetration was monitored by electrical penetration graphs whilst phloem sap was sampled by stylectomy. Both aphid species settled 2–3 times more frequently on plants fertilized with 6 or 12 mM NH₄NO₃. The phloem sap of these plants contained up to threefold higher amino acid concentrations, without a change in the proportion of essential amino acids. No time differences were observed before stylet penetration of plant tissue. After the first symplast contact, most aphids penetrated further, except M. tanacetaria on low-N plants, where 50% withdrew the stylet after the first probing. The duration of phloem feeding was 2–3 times longer in N-rich plants and the time spent in individual sieve tubes was up to tenfold longer. Aphids identified the nutritional quality of the host plant mainly by the amino acid concentration of phloem sap, not by leaf surface cues nor the proportion of essential amino acids. However, U. tanaceti infestation increased the percentage of methionine plus tryptophan in phloem tenfold, thus manipulating the plants nutritional quality, and causing premature leaf senescence.     

Amino acid budgets in three aphid species using the same host plant

Abstract. Nutrient provisioning in aphids depends both on the composition of ingested phloem sap and on the biosynthetic capabilities of the aphid and its intracellular symbionts. Amino acid budgets for three aphid species, Rhopalosiphum padi (L.), Schizaphis graminum (Rondani) and Diuraphis noxia (Mordvilko), were compared on a single host plant species, wheat Triticum aestivum L. Ingestion of amino acids from phloem, elimination of amino acids in honeydew, and the content of amino acids in aphids tissue were measured. From these values, ingestion rates were estimated and compared to honeydew and to estimated composition of aphid proteins. Ingestion rate was lowest in D. noxia due to low growth rate and low honeydew production; intermediate in S. graminum due to higher growth rate and intermediate honeydew production; and highest in R. padi , which had the highest rates for both variables. Both D. noxia and S. graminum induced increases in the amino acid content of ingested phloem. These changes in phloem content, combined with differences in ingestion rates, resulted in large differences among aphids in estimated rates of ingestion of individual amino acids. In honeydew, most essential amino acids were found in low amounts compared with the amounts ingested, especially for methionine and lysine. A few amino acids (arginine, cystine, histidine and tryptophan) were more abundant in honeydew of some aphids, suggesting oversupply. Aphid species differed in the composition of free amino acids in tissue but showed very similar composition in protein, implying similar requirements among the aphids. In R. padi and D. noxia , most essential amino acids were ingested in amounts insufficient for growth, implying dependence on symbiont provisioning. In S. graminum , most amino acids were ingested in amounts apparently sufficient for growth.     

Effects of host plant drought stress on the performance of the bird cherry-oat aphid, Rhopalosiphum padi (L.): a mechanistic analysis

Abstract.  1. The growth (increase in height and leaf number) of four grass species was reduced by a −0.5 MPa drought stress, but the performance of an associated herbivore, Rhopalosiphum padi (L.), was not affected consistently. The intrinsic rate of increase of R. padi was reduced by drought stress on three grass species, including Dactylis glomerata (L.), but was unaffected on Arrhenatherum elatius (L.). Therefore, there is no general relationship in the effect of plant drought on an insect herbivore, even among closely related host plant species.
2. Drought stress increased the quality of plant phloem sap, as indicated by increased sieve element osmotic pressure and essential amino acid concentrations. Thus, diet quality could not account for the reduced performance of R. padi under drought stress. The concentration of essential amino acids in the phloem of well-watered A. elatius was, however, lower than that of well-watered D. glomerata , correlating with the decreased performance of aphids on well-watered A. elatius .
3. There were no differences in aphid feeding duration between watering treatments or plant species but sap ingestion rates were reduced significantly under drought stress.
4. Using the measure of dietary amino acid concentrations and the estimate of sap ingestion, the essential amino acid flux through aphids was calculated. Compared with the flux through aphids feeding on well-watered D. glomerata , there was a reduction in aphids feeding on drought-stressed D. glomerata and drought-stressed A. elatius due to lower sap ingestion rates. The flux through aphids on well-watered A. elatius was also reduced due to low phloem essential amino acid concentrations. Thus, the performance of an aphid is correlated with the availability and accessibility of essential amino acids.     

Plant penetration by pea aphids (Acyrthosiphon pisum) of different plant range

Plant penetration by the stylets of six clones of the pea aphid, Acyrthosiphon pisum, on Vicia faba (acceptable to all clones) and Pisum sativum (acceptable to 3/6 clones) was investigated by the DC electrical penetration graph technique. In a 10 h recording period, 93% of 144 aphids exhibited sustained feeding on phloem sap. Significant interclonal differences were observed for the incidence of potential drops (indicative of brief punctures of plant cells) and the duration of waveform E1 (insect salivation into a sieve element). In addition, the total duration of the sieve element phase and the duration of completed bouts of sustained feeding differed between the two test plants, in a fashion varying between clones. However, these differences could not be related to the acceptability of plants to the different aphid clones. The duration of the stylet pathway phase preceding the first sustained feeding on phloem sap did not vary significantly with either aphid clone or plant. It is concluded that the resistance of P. sativum to certain A. pisum clones does not arise from factors impeding either stylet penetration through the plant tissues or the maintenance of feeding from the sieve elements. It is proposed that host plant affiliation of A. pisum may be mediated primarily by specific olfactory or gustatory cues, before the aphid initiates stylet penetration of the plant.     

Impact of plant nutrients on the relationship between a herbivorous insect and its symbiotic bacteria

The interactions between herbivorous insects and their symbiotic micro-organisms can be influenced by the plant species on which the insects are reared, but the underlying mechanisms are not understood. Here, we identify plant nutrients, specifically amino acids, as a candidate factor affecting the impact of symbiotic bacteria on the performance of the phloem-feeding aphid Aphis fabae. Aphis fabae grew more slowly on the labiate plant Lamium purpureum than on an alternative host plant Vicia faba, and the negative effect of L. purpureum on aphid growth was consistently exacerbated by the bacterial secondary symbionts Regiella insecticola and Hamiltonella defensa, which attained high densities in L. purpureum-reared aphids. The amino acid content of the phloem sap of L. purpureum was very low; and A. fabae on chemically defined diets of low amino acid content also grew slowly and had elevated secondary symbiont densities. It is suggested that the phloem nutrient profile of L. purpureum promotes deleterious traits in the secondary symbionts and disturbs insect controls over bacterial abundance.     

Phloem sap intricacy and interplay with aphid feeding

Aphididae feed upon the plant sieve elements (SE), where they ingest sugars, nitrogen compounds and other nutrients. For ingestion, aphid stylets penetrate SE, and because of the high hydrostatic pressure in SE, phloem sap exudes out into the stylets. Severing stylets to sample phloem exudates (i.e. stylectomy) has been used extensively for the study of phloem contents. Alternative sampling techniques are spontaneous exudation upon wounding that only works in a few plant species, and the popular EDTA-facilitated exudation technique. These approaches have allowed fundamental advances on the understanding of phloem sap composition and sieve tube physiology, which are surveyed in this review. A more complete picture of metabolites, ions, proteins and RNAs present in phloem sap is now available, which has provided large evidence for the phloem role as a signalling network in addition to its primary role in partitioning of photo-assimilates. Thus, phloem sap sampling methods can have remarkable applications to analyse plant nutrition, physiology and defence responses. Since aphid behaviour is suspected to be affected by phloem sap quality, attempts to manipulate phloem sap content were recently undertaken based on deregulation in mutant plants of genes controlling amino acid or sugar content of phloem sap. This opens up new strategies to control aphid settlement on a plant host.     

High variation in host adaptation among clones of the pea aphid,Acyrthosiphon pisum on peas,Pisum sativum

The performance of one clone of the pea aphid,Acyrthosiphon pisum (Harris), was assessed on 37 different cultivars and species ofPisum L. In addition, random samples of 36 pea aphid clones collected on alfalfa and clover were tested on a selection of fivePisum sativum L. cultivars. Aphid performance was evaluated in terms of the mean relative growth rate (MRGR) during the first five days of life or other life history variables. The MRGR of the first-mentioned pea aphid clone differed little between cultivars. No significant differences in MRGR were found between wild and cultivatedPisum species or between modern and oldP. sativum cultivars. There was considerable variation in host adaptation among the 36 pea aphid clones within each sampled field. The pea aphid clones showed no consistent pattern in performance on four of the five pea cultivars i.e. there was a significant pea aphid genotype —pea genotype interaction. On one of the cultivars all clones performed well. Pea aphid clones collected from red clover generally performed relatively poorly on pea cultivars, in contrast to the pea aphid clones collected on alfalfa. There was no difference in performance between the two pea aphid colour forms tested. Possible reasons for the high variation and the observed adaptation patterns are discussed. The fact that all clones were collected in two adjacent fields indicates thatA. pisum shows high local intraspecific variability in terms of host adaptation.     

Phloem-sap feeding by animals: problems and solutions

The incidence of phloem sap feeding by animals appears paradoxical. Although phloem sap is nutrient-rich compared with many other plant products and generally lacking in toxins and feeding deterrents, it is consumed as the dominant or sole diet by a very restricted range of animals, exclusively insects of the order Hemiptera. These insects display two sets of adaptations. First, linked to the high ratio of non-essential:essential amino acids in phloem sap, these insects contain symbiotic micro-organisms which provide them with essential amino acids. For example, bacteria of the genus Buchnera contribute up to 90% of the essential amino acids required by the pea aphid Acyrthosiphon pisum feeding on Vicia faba. Second, the insect tolerance of the very high sugar content and osmotic pressure of phloem sap is promoted by their possession in the gut of sucrase-transglucosidase activity, which transforms excess ingested sugar into long-chain oligosaccharides voided via honeydew. Various other animals consume phloem sap by proxy, through feeding on the honeydew of phloem-feeding hemipterans. Honeydew is physiologically less extreme than phloem sap, with a higher essential:non-essential amino acid ratio and lower osmotic pressure. Even so, ant species strongly dependent on honeydew as food may benefit from nutrients derived from their symbiotic bacteria Blochmannia.     

Changes in the amino acid composition and protein modification in phloem sap of rice

Pure phloem sap was collected from insects feeding on rice (Oryza sativa L.) leaves by a laser technique similar to the aphid stylet technique. Rapid circulation of nitrogen in the sieve tubes was demonstrated directly using ¹⁵N as a tracer. Application to the roots of the metabolic inhibitors of amino acids, aminooxyacetate and methioninesulfoximine, changed the amino acid composition in the sieve tubes. Feeding methionine to leaf tips resulted in its bulk transfer into the sieve tubes. In vitro experiments confirmed the existence of protein kinases in the pure rice phloem sap. The phosphorylation status of the sieve tube sap proteins was affected by the light regime. The possibility that changes in chemical composition or protein modification such as phosphorylation in the sieve tubes might affect plant growth are discussed.Analysis of pure phloem sap collected from rice plants by insect laser technique has shown dynamic changes in the chemical composition and the quality of proteins in the sap.     

Barley yellow dwarf virus, wheat, and Sitobion avenae: a case of trilateral interactions

We analysed interactions in the system of two Barley Yellow Dwarf Virus (BYDV) strains (MAV and PAV), and wheat (cv. Tinos) as host plant for the virus, and the cereal aphid Sitobion avenae (F.) as vector, in particular whether or not infection by the virus might alter host plant suitability in favour of vector development. By measuring the amino acid and sugar content in the phloem sap of infected and non‐infected wheat plants we found a significant reduction in the concentration of the total amount of amino acids on BYDV‐infected plants. Qualitative and quantitative analysis of honeydew and honeydew excretion indicated a lower efficiency of phloem sap utilisation by S. avenae on infected plants. In addition, S. avenae excreted less honeydew on infected plants. Both BYDV strains significantly affected aphid development by a reduction in the intrinsic rate of natural increase. Hence, infection by the virus reduced the host suitability in terms of aphid population growth potential on BYDV‐infected plants. However, more alate morphs developed on virus‐infected plants. These findings are discussed in relation to the population dynamics of S. avenae, and, as a consequence, the spread of BYDV.     

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.