Prey and Non-prey Arthropods Sharing a Host Plant: Effects on Induced Volatile Emission and Predator Attraction

It is well established that plants infested with a single herbivore species can attract specific natural enemies through the emission of herbivore-induced volatiles. However, it is less clear what happens when plants are simultaneously attacked by more than one species. We analyzed volatile emissions of lima bean and cucumber plants upon multi-species herbivory by spider mites (Tetranychus urticae) and caterpillars (Spodoptera exigua) in comparison to single-species herbivory. Upon herbivory by single or multiple species, lima bean and cucumber plants emitted volatile blends that comprised mostly the same compounds. To detect additive, synergistic, or antagonistic effects, we compared the multi-species herbivory volatile blend with the sum of the volatile blends induced by each of the herbivore species feeding alone. In lima bean, the majority of compounds were more strongly induced by multi-species herbivory than expected based on the sum of volatile emissions by each of the herbivores separately, potentially caused by synergistic effects. In contrast, in cucumber, two compounds were suppressed by multi-species herbivory, suggesting the potential for antagonistic effects. We also studied the behavioral responses of the predatory mite Phytoseiulus persimilis, a specialized natural enemy of spider mites. Olfactometer experiments showed that P. persimilis preferred volatiles induced by multi-species herbivory to volatiles induced by S. exigua alone or by prey mites alone. We conclude that both lima bean and cucumber plants effectively attract predatory mites upon multi-species herbivory, but the underlying mechanisms appear different between these species.     

Systemically Induced Plant Volatiles Emitted at the Time of “Danger”

Feeding by Pieris brassicae caterpillars on the lower leaves of Brussels sprouts (Brassica oleracea var. gemmifera) plants triggers the release of volatiles from upper leaves. The volatiles are attractive for a natural antagonist of the herbivore, the parasitoid Cotesia glomerata. Parasitoids are attracted only if additional damage is inflicted on the systemically induced upper leaves and only after at least three days of herbivore feeding on the lower leaves. Upon termination of caterpillar feeding, the systemic signal is emitted for a maximum of one more day. Systemic induction did not occur at low levels of herbivore infestation. Systemically induced leaves emitted green leaf volatiles, cyclic monoterpenoids, and sesquiterpenes. GC-MS profiles of systemically induced and herbivore-infested leaves did not differ for most compounds, although herbivore infested plants did emit higher amounts of green leaf volatiles. Emission of systemically induced volatiles in Brussels sprouts might function as an induced defense that is activated only when needed, i.e., at the time of caterpillar attack. This way, plants may adopt a flexible management of inducible defensive resources to minimize costs of defense and to maximize fitness in response to unpredictable herbivore attack.     

Molecular,Biochemical, and Organismal Analyses of Tomato Plants Simultaneously Attacked by Herbivores from Two Feeding Guilds

Previous work identified aphids and caterpillars as having distinct effects on plant responses to herbivory. We sought to decipher these interactions across different levels of biological organization, i.e., molecular, biochemical, and organismal, with tomato plants either damaged by one 3rd-instar beet armyworm caterpillar (Spodoptera exigua), damaged by 40 adult potato aphids (Macrosiphum euphorbiae), simultaneous damaged by both herbivores, or left undamaged (controls). After placing insects on plants, plants were transferred to a growth chamber for 5 d to induce a systemic response. Subsequently, individual leaflets from non-damaged parts of plants were excised and used for gene expression analysis (microarrays and quantitative real-time PCR), C/N analysis, total protein analysis, proteinase inhibitor (PI) analysis, and for performance assays. At the molecular level, caterpillars up-regulated 56 and down-regulated 29 genes systemically, while aphids up-regulated 93 and down-regulated 146 genes, compared to controls. Although aphids induced more genes than caterpillars, the magnitude of caterpillar-induced gene accumulation, particularly for those associated with plant defenses, was often greater. In dual-damaged plants, aphids suppressed 27% of the genes regulated by caterpillars, while caterpillars suppressed 66% of the genes regulated by aphids. At the biochemical level, caterpillars induced three-fold higher PI activity compared to controls, while aphids had no effects on PIs either alone or when paired with caterpillars. Aphid feeding alone reduced the foliar C/N ratio, but not when caterpillars also fed on the plants. Aphid and caterpillar feeding alone had no effect on the amount of protein in systemic leaves; however, both herbivores feeding on the plant reduced the amount of protein compared to aphid-damaged plants. At the organismal level, S. exigua neonate performance was negatively affected by prior caterpillar feeding, regardless of whether aphids were present or absent. This study highlights areas of concordance and disjunction between molecular, biochemical, and organismal measures of induced plant resistance when plants are attacked by multiple herbivores. In general, our data produced consistent results when considering each herbivore separately but not when considering them together.     

Induction of Direct and Indirect Plant Responses by Jasmonic Acid, Low Spider Mite Densities, or a Combination of Jasmonic Acid Treatment and Spider Mite Infestation

Jasmonic acid (JA) and the octadecanoid pathway are involved in both induced direct and induced indirect plant responses. In this study, the herbivorous mite, Tetranychus urticae, and its predator, Phytoseiulus persimilis, were given a choice between Lima bean plants induced by JA or spider mites and uninduced control plants. Infestation densities resulting in the induction of predator attractants were much lower than thus far assumed, i.e., predatory mites were significantly attracted to plants that were infested for 2 days with only one or four spider mites per plant. Phytoseiulus persimilis showed a density-dependent response to volatiles from plants that were infested with different numbers of spider mites. Similarly, treating plants with increasing concentrations of JA also led to increased attraction of P. persimilis. Moreover, the duration of spider mite infestation was positively correlated with the proportion of predators that were attracted to mite-infested plants. A pretreatment of the plants with JA followed by a spider mite infestation enhanced the attraction of P. persimilis to plant volatiles compared to attraction to volatiles from plants that were only infested with spider mites and did not receive a pretreatment with JA. The herbivore, T. urticae preferred leaf tissue that previously had been infested with conspecifics to uninfested leaf tissue. In the case of choice tests with JA-induced and control leaf tissue, spider mites slightly preferred control leaf tissue. When spider mites were given a choice between leaf discs induced by JA and leaf discs damaged by spider mite feeding, they preferred the latter. The presence of herbivore induced chemicals and/or spider mite products enhanced settlement of the mites, whereas treatment with JA seemed to impede settlement.     

Polymorphism in Plant Defense Against Herbivory: Constitutive and Induced Resistance in Cucumis sativus

Theory predicts that plant resistance to herbivores is determined by both genetic and environmentally induced components. In this study, we demonstrate that the phenotypic expression of plant resistance to spider mite herbivory in Cucumis sativus is determined by genetic and environmental factors and that there is an interaction between these factors. Previous feeding by spider mites induced systemic resistance to subsequent attack over several spatial scales within plants, reducing the population growth of mites compared to that on control plants. Effects of induction were effective locally over the short term, but resulted in local increased susceptibility to spider mite attack after several days. However, this local induced susceptibility on the damaged leaf was associated with induced resistance on newer leaves. Induced resistance was correlated with increases in cucurbitacin content of leaves, but was not associated with changes in the density of leaf trichomes. Induced resistance to herbivory was not detected in plants of a genotype lacking constitutive expression of cucurbitacins, which were in general highly susceptibile to mite attack. Allocation trade-offs between growth and defense are often invoked to explain the maintenance of variation in the levels of plant resistance. Contrary to current thinking, neither constitutive nor herbivore-induced plant resistance were associated with reductions in plant allocation to root and shoot growth. However, plants that had high levels of induced resistance to spider mites were the most susceptible to attack by a specialist beetle. Such ecological trade-offs between resistance to generalist herbivores and susceptibility to specialist herbivores may be important in the maintenance of variation of plant resistance traits. In summary, C. sativus exhibits strong genetic variation for constitutive and induced resistance to spider mites, and this variation in resistance is associated with ecological trade-offs.     

Brassica Plant Responses to Mild Herbivore Stress Elicited by Two Specialist Insects from Different Feeding Guilds

Compensation growth and chemical defense are two components of plant defense strategy against herbivores. In this study, compensation growth and the response of primary and secondary metabolites were investigated in Brassica rapa plants subjected to infestation by two herbivores from contrasting feeding guilds, the phloem-feeding aphid Brevicoryne brassicae and the leaf-feeding caterpillar Pieris brassicae. These specialist herbivores were used at two different densities and allowed to feed for seven days on a young caged leaf. Changes in growth rates were assessed for total leaf area and bulb mass, whereas changes in primary and secondary metabolites were evaluated in young and mature leaves, roots, and bulbs. Mild stress by caterpillars on young plants enhanced mean bulb mass and elicited a contrasting regulation of aliphatic and indolic glucosinolates in the leaves. In contrast, mild stress by aphids enhanced leaf growth and increased glucosinolate concentrations in the bulb, the most important storage organ of B. rapa. A similar mild stress by either herbivore to older plants did not alter plant growth parameters or concentrations of the metabolites analyzed. In conclusion, Brassica plant growth was either maintained or enhanced under mild herbivore stress, and defense patterns differed strongly in response to herbivore type and plant development stage. These results have implications for the understanding of plasticity in plant defenses against herbivores and for the management of Brassica rapa in agroecosystems.     

Volatile Stimuli Related to Feeding Activity of Nonprey Caterpillars, Spodoptera exigua, Affect Olfactory Response of the Predatory Mite Phytoseiulus persimilis

The effect of volatiles related to feeding activity of nonprey caterpillars, Spodoptera exigua, on the olfactory response of the predatory mites Phytoseiulus persimilis was examined in a Y-tube olfactometer. At a low caterpillar density (20 caterpillars on 10 Lima bean leaves), the predators were significantly more attracted to volatiles from infested leaves on which the caterpillars and their products were present or from infested leaves from which the caterpillars and their products had been removed when compared to volatiles from uninfested leaves. The predators, however, significantly avoided odors from 20 caterpillars and their products (mainly feces) removed from bean leaves. In contrast, at a higher caterpillar density (100 caterpillars on 10 Lima bean leaves), the predators avoided volatiles from caterpillar-infested bean leaves. Volatiles from infested leaves from which the caterpillars and their products had been removed were not preferred over volatiles from uninfested leaves. Volatiles from feces collected from 100 caterpillars were strongly avoided by the predators, while the behavior of the predatory mites was not affected by volatiles from 100 caterpillars removed from a plant. The data show that carnivorous arthropods may avoid nonprofitable herbivores. This avoidance seems to result from an interference of volatiles from herbivore products with the attraction to herbivore-induced plant volatiles.     

Induction of Plant Volatiles by Herbivores with Different Feeding Habits and the Effects of Induced Defenses on Host-Plant Selection by Thrips

Induced plant responses to attack by chewing insects have been intensively studied, but little is known about plant responses to nonchewing insects or to attack by multiple herbivores with different feeding habits. We examined volatile emissions by tobacco, Nicotiana tabacum, in response to feeding by the piercing–sucking insect western flower thrips (WFT), Frankliniella occidentalis, the chewing herbivore Heliothis virescens, and both herbivores simultaneously. In addition, we examined the effects of herbivore-induced plant defenses on host-plant selection by WFT. Plants responded to thrips feeding by consistently releasing five compounds. Simultaneous feeding by WFT and H. virescens elicited the same 11 compounds emitted in response to caterpillar feeding alone; however, two compounds, α-humulene and caryophyllene oxide, were produced in greater amounts in response to simultaneous herbivory. In choice tests, thrips consistently preferred uninduced plants over all other treatments and preferred plants damaged by caterpillars and those treated with caterpillar saliva over those treated with caterpillar regurgitant. The results are consistent with a previous finding that caterpillar regurgitant induces the release of significantly more volatile nicotine than plants damaged by caterpillars or plants treated with caterpillar saliva. A repellent effect of nicotine on WFT was confirmed by encircling unwounded plants with septa releasing volatile nicotine. Our results provide the first direct evidence that thrips feeding induces volatile responses and indicates that simultaneous herbivory by insects with different feeding habits can alter volatile emissions. In addition, the findings demonstrate that induced plant responses influence host-plant selection by WFT and suggest that the induction of volatile nicotine may play a role in this process.     

Folivory Affects Composition of Nectar,Floral Odor and Modifies Pollinator Behavior

Herbivory induces changes in plants that influence the associated insect community. The present study addresses the potential trade-off between plant phytochemical responses to insect herbivory and interactions with pollinators. We used a multidisciplinary approach and have combined field and greenhouse experiments to investigate effects of herbivory in plant volatile emission, nectar production, and pollinator behavior, when Pieris brassicae caterpillars were allowed to feed only on the leaves of Brassica nigra plants. Interestingly, volatile emission by flowers changed upon feeding by herbivores on the leaves, whereas, remarkably, volatile emission by leaves did not significantly differ between infested and non-infested flowering plants. The frequency of flower visits by pollinators was generally not influenced by herbivory, but the duration of visits by honeybees and butterflies was negatively affected by herbivore damage to leaves. Shorter duration of pollinator visits could be beneficial for a plant, because it sustains pollen transfer between flowers while reducing nectar consumption per visit. Thus, no trade-off between herbivore-induced plant responses and pollination was evident. The effects of herbivore-induced plant responses on pollinator behavior underpin the importance of including ecological factors, such as herbivore infestation, in studies of the ecology of plant pollination.     

Herbivore-Induced Extrafloral Nectar Production in Lima Bean Plants Enhanced by Previous Exposure to Volatiles from Infested Conspecifics

In response to herbivory by spider mites (Tetranychus urticae), lima bean plants produced significantly greater quantities of extrafloral nectar (EFN) than intact conspecific plants. Moreover, EFN amounts of infested plants depended on exposure to odor of infested neighbor plants. Two d after spider mite infestation, a test plant produced more EFN when exposed prior to infestation to volatiles from infested neighbor plants than when exposed to volatiles from uninfested conspecific plants. However, this effect was only detectable 2 d after spider mite infestation and vanished 4 d after infestation. These results suggest that EFN production is enhanced during the earlier stages of damage by T. urticae in response to previous exposure to volatiles from infested neighbor plants.     

Identification of Volatiles That Are Used in Discrimination Between Plants Infested with Prey or Nonprey Herbivores by a Predatory Mite

Carnivorous arthropods can use herbivore-induced plant volatiles to locate their herbivorous prey. In the field, carnivores are confronted with information from plants infested with herbivores that may differ in their suitability as prey. Discrimination by the predatory mite Phytoseiulus persimilis between volatiles from lima bean plants infested with the prey herbivore Tetranychus urticae, or plants infested with the nonprey caterpillar Spodoptera exigua, depends on spider mite density. In this article, we analyzed the chemical composition of the volatile blends from T. urticae-infested lima bean plants at different densities of spider mites, and from S. exigua-infested plants. Based on the behavioral preferences of P. persimilis and the volatile profiles, we selected compounds that potentially enable the mite to discriminate between T. urticae-induced and S. exigua-induced volatiles. Subsequently, we demonstrated in Y-tube olfactometer assays that the relatively large amounts of methyl salicylate and (3E, 7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene emitted by T. urticae-infested bean plants compared to S. exigua-infested plants enable the predators to discriminate. Our data show that specific compounds from complex herbivore-induced volatile blends can play an important role in the selective foraging behavior of natural enemies of herbivorous arthropods.     

Herbivory induces systemic production of plant volatiles that attract predators of the herbivore: Extraction of endogenous elicitor

It was previously shown that in response to infestation by spider mites (Tetranychus urticae), lima bean plants produce a volatile herbivoreinduced synomone that attracts phytoseiid mites (Phytoseiulus persimilis) that are predators of the spider mites. The production of predator-attracting infochemicals was established to occur systemically throughout the spider mitein-fested plant. Here we describe the extraction of a water-soluble endogenous elicitor from spider mite-infested lima bean leaves. This elicitor was shown to be transported out of infested leaves and was collected in water in which the petiole of the infested leaf was placed. When the petioles of uninfested lima bean leaves were placed in water in which infested leaves had been present for the previous seven days, these uninfested lima bean leaves became highly attractive to predatory mites in an olfactometer when an appropriate control of uninfested lima bean leaves was offered as alternative. The strength of this effect was dependent on the number of spider mites infesting the elicitor-producing leaves. Higher numbers of spider mites resulted in an elicitor solution with a stronger effect. In addition, spider mite density was important. The elicitor obtained from one leaf with 50 spider mites had a stronger effect on the attractiveness of uninfested leaves than the elicitor obtained from three leaves with 17 spider mites each. This suggests that the stress intensity imposed on a plant is an important determinant of the elicitor quantity. While the elicitor has a strong effect on the attractiveness of uninfested leaves, spider mite-infested leaves are still much more attractive to predatory mites than elicitor-exposed leaves. The data are discussed in the context of systemic effects in plant defense and the biosynthesis of herbivore-induced terpenoids in plants.     

Specificity of Systemically Released Cotton Volatiles as Attractants for Specialist and Generalist Parasitic Wasps

Cotton plants under herbivore attack release volatile semiochemicals that attract natural enemies of the herbivores to the damaged plant. The volatiles released in response to herbivory are not only released from the damaged leaves but from the entire cotton plant. We found that cotton plants that released myrcene, (Z)-3-hexenyl acetate, (E)--ocimene, linalool, (E)-4,8-dimethyl-1,3,7-nonatriene, (E)--farnesene, and (E, E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene systemically from undamaged leaves of caterpillar damaged plants were attractive to the generalist parasitoid Cotesia marginiventris and the specialist parasitoid Microplitis croceipes. Plants from which the caterpillar damaged leaves were removed and that released those compounds systemically were significantly preferred over undamaged control plants in two-choice experiments in a flight tunnel. Artificially damaged cotton plants that released green leafy volatiles and constitutive terpenoids were less attractive for M. croceipes and C. marginiventris. Only C. marginiventris preferred artificially damaged plants over undamaged control plants, whereas M. croceipes showed no preference. The apparent lack of specificity of systemically released compounds in response to different herbivores feeding on the lower leaves is discussed.     

Plant–herbivore–carnivore Interactions in Cotton, <Emphasis Type="Italic">Gossypium hirsutum</Emphasis>: Linking Belowground and Aboveground

Most studies on plant–herbivore interactions focus on either root or shoot herbivory in isolation, but above- and belowground herbivores may interact on a shared host plant. Cotton (Gossypium spp.) produces gossypol and a variety of other gossypol-like terpenoids that exhibit toxicity to a wide range of herbivores and pathogens. Cotton plants also can emit herbivore-induced volatile compounds at the site of damage and systemically on all tissues above the site of damage. As these volatile compounds attract natural enemy species of the herbivore, they are thought to represent an indirect plant defense. Our study quantified gossypol and gossypol-like compounds in cotton plants with foliage feeding (Heliocoverpa zea), root feeding (Meloidogyne incognita), or their combination. Cotton plants with these treatments were studied also with respect to induced local and systemic volatile production and the attraction of the parasitic wasp Microplitis croceipes to those plants. We also evaluated whether foliage or root feeding affected foliar nitrogen levels in cotton. After 48 hr of leaf feeding and 5 wk of root feeding, local and systemic induction of volatiles (known to attract parasitoids such as M. croceipes) occurred with herbivore damage to leaves, and it increased in levels when root herbivory was added. Nevertheless, M. croceipes were equally attracted to plants with both leaf and root damage and leaf damage only. In contrast to previous studies in cotton, production of gossypol and gossypol-like compounds was not induced in leaf and root tissue following foliage or root herbivory, or their combination. We conclude that root feeding by M. incognita has little influence on direct and indirect defenses of Gossypium hirsutum against insect herbivory.     

Defense Suppression through Interplant Communication Depends on the Attacking Herbivore Species

In response to herbivory, plants emit volatile compounds that play important roles in plant defense. Herbivore-induced plant volatiles (HIPVs) can deter herbivores, recruit natural enemies, and warn other plants of possible herbivore attack. Following HIPV detection, neighboring plants often respond by enhancing their anti-herbivore defenses, but a recent study found that herbivores can manipulate HIPV-interplant communication for their own benefit and suppress defenses in neighboring plants. Herbivores induce species-specific blends of HIPVs and how these different blends affect the specificity of plant defense responses remains unclear. Here we assessed how HIPVs from zucchini plants (Cucurbita pepo) challenged with different herbivore species affect resistance in neighboring plants. Volatile “emitter” plants were damaged by one of three herbivore species: saltmarsh caterpillars (Estigmene acrea), squash bugs (Anasa tristis), or striped cucumber beetles (Acalymma vittatum), or were left as undamaged controls. Neighboring “receiver” plants were exposed to HIPVs or control volatiles and then challenged by the associated herbivore species. As measures of plant resistance, we quantified herbivore feeding damage and defense-related phytohormones in receivers. We found that the three herbivore species induced different HIPV blends from squash plants. HIPVs induced by saltmarsh caterpillars suppressed defenses in receivers, leading to greater herbivory and lower defense induction compared to controls. In contrast, HIPVs induced by cucumber beetles and squash bugs did not affect plant resistance to subsequent herbivory in receivers. Our study shows that herbivore species identity affects volatile-mediated interplant communication in zucchini, revealing a new example of herbivore defense suppression through volatile cues.

     

Feeding Experience Affects the Behavioral Response of Polyphagous Gypsy Moth Caterpillars to Herbivore-induced Poplar Volatiles

Plant volatiles influence host selection of herbivorous insects. Since volatiles often vary in space and time, herbivores (especially polyphagous ones) may be able to use these compounds as cues to track variation in host plant quality based on their innate abilities and previous experience. We investigated the behavioral response of naïve (fed on artificial diet) and experienced (fed on poplar) gypsy moth (Lymantria dispar) caterpillars, a polyphagous species, towards constitutive and herbivore-induced black poplar (Populus nigra) volatiles at different stages of herbivore attack. In Y-tube olfactometer assays, both naïve and experienced caterpillars were attracted to constitutive volatiles and volatiles released after short-term herbivory (up to 6 hr). Naïve caterpillars also were attracted to volatiles released after longer-term herbivory (24–30 hr), but experienced caterpillars preferred the odor of undamaged foliage. A multivariate statistical analysis comparing the volatile emission of undamaged plants vs. plants after short and longer-term herbivory, suggested various compounds as being responsible for distinguishing between the odors of these plants. Ten compounds were selected for individual testing of caterpillar behavioral responses in a four-arm olfactometer. Naïve caterpillars spent more time in arms containing (Z)-3-hexenol and (Z)-3-hexenyl acetate than in solvent permeated arms, while avoiding benzyl cyanide and salicyl aldehyde. Experienced caterpillars avoided benzyl cyanide and preferred (Z)-3-hexenyl acetate and the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) over solvent. Only responses to DMNT were significantly different when comparing experienced and naïve caterpillars. The results show that gypsy moth caterpillars display an innate behavioral response towards constitutive and herbivore-induced plant volatiles, but also that larval behavior is plastic and can be modulated by previous feeding experience.     

Does Aphid Infestation Interfere with Indirect Plant Defense against Lepidopteran Caterpillars in Wild Cabbage?

Attraction of parasitoids to plant volatiles induced by multiple herbivory depends on the specific combinations of attacking herbivore species, especially when their feeding modes activate different defense signalling pathways as has been reported for phloem feeding aphids and tissue feeding caterpillars. We studied the effects of pre-infestation with non-host aphids (Brevicoryne brassicae) for two different time periods on the ability of two parasitoid species to discriminate between volatiles emitted by plants infested by host caterpillars alone and those emitted by plants infested with host caterpillars plus aphids. Using plants originating from three chemically distinct wild cabbage (Brassica oleracea) populations, Diadegma semiclausum switched preference for dually infested plants to preference for plants infested with Plutella xylostella hosts alone when the duration of pre-aphid infestation doubled from 7 to 14 days. Microplitis mediator, a parasitoid of Mamestra brassicae caterpillars, preferred dually-infested plants irrespective of aphid-infestation duration. Separation of the volatile blends emitted by plants infested with hosts plus aphids or with hosts only was poor, based on multivariate statistics. However, emission rates of individual compounds were often reduced in plants infested with aphids plus hosts compared to those emitted by plants infested with hosts alone. This effect depended on host caterpillar species and plant population and was little affected by aphid infestation duration. Thus, the interactive effect of aphids and hosts on plant volatile production and parasitoid attraction can be dynamic and parasitoid specific. The characteristics of the multi-component volatile blends that determine parasitoid attraction are too complex to be deduced from simple correlative statistical analyses.     

Effects of Root Herbivory by Nematodes on the Performance and Preference of a Leaf-Infesting Generalist Aphid Depend on Nitrate Fertilization

The performance and behavior of herbivores is strongly affected by the quality of their host plants, which is determined by various environmental conditions. We investigated the performance and preference of the polyphagous shoot-infesting aphid Myzus persicae on the host-plant Arabidopsis thaliana in a two-factorial design in which nitrate fertilization was varied by 33 %, and the root-infesting cyst-nematode Heterodera schachtii was present or absent. Aphid performance was influenced by these abiotic and biotic factors in an interactive way. Nematode presence decreased aphid performance when nitrate levels were low, whereas nematode infestation did not influence aphid performance under higher nitrate fertilization. Aphids followed the “mother knows best” principle when given a choice, settling preferentially on those plants on which they performed best. Hence, they preferred nematode-free over nematode-infested plants in the low fertilization treatment but host choice was not affected by nematodes under higher nitrate fertilization. The amino acid composition of the phloem exudates was significantly influenced by fertilization but also by the interaction of the two treatments. Various glucosinolates in the leaves, which provide an estimate of phloem glucosinolates, were not affected by the individual treatments but by the combination of fertilization and herbivory. These changes in primary and secondary metabolites may be decisive for the herbivore responses. Our data demonstrate that abiotic and biotic factors can interactively affect herbivores, adding a layer of complexity to plant-mediated herbivore interactions.