Root damage to apple plants by cockchafer larvae induces a change in volatile signals below‐ and above‐ground

Volatile organic compounds (VOCs) mediate communication between plants and insects. Plants under insect herbivore attack release VOCs either at the site of attack or systemically, indicating within‐plant communication. Some of these VOCs, which may be induced only upon herbivore attack, recruit parasitoids and predatory insects to feed on the attacking insects. Moreover, some plants are able to ‘eavesdrop’ on herbivore‐induced plant volatiles (HIPVs) to prime themselves against impending attack; such eavesdropping exemplifies plant–plant communication. In apple orchards, the beetle Melolontha melolontha L. (Coleoptera: Scarabaeidae) is an important insect pest whose larvae live and feed on roots for about 4 years. In this study, we investigated whether the feeding activity of M. melolontha larvae (1) alters the volatile profile of apple roots, (2) induces the release of HIPVs systemically in the leaves, and (3) whether infested plants communicate to neighbouring non‐infested conspecifics through HIPVs. To answer these questions, we collected constitutive VOCs from intact M9 roots as well as M. melolontha larvae‐damaged roots using a newly designed ‘rhizobox’, to collect root‐released volatiles in situ, without damaging the plant root system. We also collected VOCs from the leaf‐bearing shoots of M9 whose roots were under attack by M. melolontha larvae and from shoots of neighbouring non‐infested conspecifics. Gas chromatography‐mass spectrometry analysis showed that feeding activity of M. melolontha larvae induces the release of specific HIPVs; for instance, camphor was found in the roots only after larvae caused root damage. Melolontha melolontha also induced the systemic release of methyl salicylate and (E,E)‐α‐farnesene from the leaf‐bearing shoots. Methyl salicylate and (E,E)‐α‐farnesene were also released by the shoots of non‐infested neighbouring conspecifics. These phenomena indicate the induction of specific VOCs below‐ and above‐ground upon M. melolontha larvae feeding on apple roots as well as plant–plant communication in apple plants.     

Plant‐mediated interactions among above‐ground and below‐ground life stages of a root‐feeding weevil

1. Interactions among herbivores mediated by plant responses to herbivore injury may have large impacts on herbivore population densities. Responses may persist for weeks after injury and may affect not only the initial (inducing) herbivore, but also herbivores that are spatially or temporally separated from the initial attacker. 2. In many plant–insect interactions, multiple life stages of the insect may be associated with the same plant, and these various stages may interact indirectly with one another via induced responses. The rice water weevil (RWW), Lissorhoptrus oryzophilus, a serious global pest of rice, is one such insect. A series of experiments were performed with root‐feeding larvae and leaf‐feeding adults of the RWW using three conventional rice varieties. 3. The first objective of this study was to test whether RWW adult feeding on rice leaves resulted in altered oviposition by subsequent adults. The hypothesis for the first objective was that RWW adult feeding would decrease plant suitability, resulting in reduced oviposition by subsequent adults. 4. The second objective was to test whether injury by RWW larvae to rice roots resulted in altered oviposition by subsequent adults. The hypothesis for the second objective was that below‐ground RWW larval feeding would decrease plant suitability of rice to above‐ground RWW adults, resulting in decreased oviposition. 5. Results provided inconsistent support for the first hypothesis, indicating that responses differed among combinations of variety and injury level. Conversely, consistent support for the second hypothesis was found, indicating that larval feeding on roots decreased suitability of rice plants for oviposition.     

Instar-specific sensitivity of specialist Manduca sexta larvae to induced defences in their host plant Nicotiana attenuata

1. The time delay associated with the activation of induced defences is thought to be a liability for this type of defence because it allows herbivores to remove biomass before the defence is fully induced. When defences are costly and plants grow with competitors, however, it may be more advantageous not to induce defences too fast and motivate the herbivore to move to the neighbour when it is most voracious. 2. Such a strategy can only work when the costs for the herbivore of moving to a neighbouring plant are smaller than the costs of staying on a fully induced plant. For lepidopteran herbivores, both the sensitivity to induced defences and the costs of moving may vary considerably between instars and this variation may constrain the plant's defensive opportunities. 3. This study was designed to examine whether the cost of moving, mimicked by a starvation period of 8 h, was larger than the cost of staying on a fully induced plant for each larval instar of the specialist Manduca sexta feeding on induced and control tissues of Nicotiana attenuata. 4. For first‐ and second‐instar larvae, the costs of moving were larger than the costs of staying on a fully induced plant. In contrast, feeding on induced plant material retarded development in third‐instar larvae more than did starvation, indicating that in this instar the costs of leaving are smaller than the costs of staying on an induced plant. More than 98% of the lifetime leaf mass consumed by a M. sexta larva is consumed during the fourth and fifth instars, and during these instars larval development was not affected by either induced defences or starvation. Thus the third instar, the stage just before larvae cause the majority of damage, represents a window of sensitivity to induced defences during which larvae can be motivated to change plants. 5. These results suggest that N. attenuata plants, which commonly compete with conspecifics in nature, have the opportunity to manipulate the behaviour of the specialist herbivore M. sexta to minimise the fitness effects of inducing defences when these defences are most costly, i.e. when plants grow under intraspecific competition.     

Nucleopolyhedrovirus infection enhances plant defences by increasing plant volatile diversity

Plant–herbivore–entomopathogen tri-trophic interactions and biodiversity are relatively understudied topics in ecology. Particularly, the effects of entomopathogens on herbivore-induced plant volatiles and plant volatile diversity on the defensive function of plants have not been studied in detail. We used soybean (Glycine max), beet armyworm larvae (Spodoptera exigua), and nucleopolyhedrovirus (NPV) as a tri-trophic system to determine whether NPV infection can promote the emission and diversity of volatiles from plants. We also investigated whether NPV infection affects the attraction of Microplitis pallidipes, an important endoparasitoid of larval S. exigua. Uninfested soybean plants released 7 detectable volatile compounds while plants fed upon by healthy and NPV-infected S. exigua larvae released 12 and 15 volatiles, respectively. Female parasitoids were more attracted to the volatiles from plants that were fed upon by NPV-infected larvae than healthy larvae, and more attracted to the volatiles from plants that were fed upon by healthy larvae than no larvae. The selective responses of parasitoids to plant odours increased as plant volatile diversity increased. Our study suggests that the NPV infection facilitates the release of plant volatiles and enhances the defensive function of plants by increasing plant volatile diversity which in turn attracts more parasitoids. Also, this work reveals that plants might accrue two indirect benefits from NPV infection, cessation of herbivore feeding and more parasitisation.     

Metabolomics reveals herbivore‐induced metabolites of resistance and susceptibility in maize leaves and roots

Plants respond to herbivory by reprogramming their metabolism. Most research in this context has focused on locally induced compounds that function as toxins or feeding deterrents. We developed an ultra‐high‐pressure liquid chromatography time‐of‐flight mass spectrometry (UHPLC‐TOF‐MS)‐based metabolomics approach to evaluate local and systemic herbivore‐induced changes in maize leaves, sap, roots and root exudates without any prior assumptions about their function. Thirty‐two differentially regulated compounds were identified from Spodoptera littoralis‐infested maize seedlings and isolated for structure assignment by microflow nuclear magnetic resonance (CapNMR). Nine compounds were quantified by a high throughput direct nano‐infusion tandem mass spectrometry/mass spectrometry (MS/MS) method. Leaf infestation led to a marked local increase of 1,3‐benzoxazin‐4‐ones, phospholipids, N‐hydroxycinnamoyltyramines, azealic acid and tryptophan. Only few changes were found in the root metabolome, but 1,3‐benzoxazin‐4‐ones increased in the vascular sap and root exudates. The role of N‐hydroxycinnamoyltyramines in plant–herbivore interactions is unknown, and we therefore tested the effect of the dominating p‐coumaroyltyramine on S. littoralis. Unexpectedly, p‐coumaroyltyramine was metabolized by the larvae and increased larval growth, possibly by providing additional nitrogen to the insect. Taken together, this study illustrates that herbivore attack leads to the induction of metabolites that can have contrasting effects on herbivore resistance in the leaves and roots.     

The role of herbivore‐ and plant‐related experiences in intraspecific host preference of a relatively specialized parasitoid

Parasitoids use odor cues from infested plants and herbivore hosts to locate their hosts. Specialist parasitoids of generalist herbivores are predicted to rely more on herbivorederived cues than plant-derived cues. Microplitis croceipes (Cresson)(Hymenoptera: Braconidae) is a relatively specialized larval endoparasitoid of Heliothis virescens (F.)(Lepidoptera: Noctuidae), which is a generalist herbivore on several crops including cotton and soybean. Using M. croceipes/H. virescens as a model system, we tested the following predictions about specialist parasitoids of generalist herbivores:(i) naive parasitoids will show innate responses to herbivore-emitted kairomones, regardless of host plant identity and (ii) herbivore-related experience will have a greater influence on intraspecific oviposition preference than plant-related experience. Inexperienced (naive) female M. croceipes did not discriminate between cotton-fed and soybean-fed H. virescens in oviposition choice tests, supporting our first prediction. Oviposition experience alone with either host group influenced subsequent oviposition preference while experience with infested plants alone did not elicit preference in M. croceipes, supporting our second prediction. Furthermore, associative learning of oviposition with host-damaged plants facilitated host location. I terestingly, naive parasitoids attacked more soybeathan cotton-fed host larvae in two-choice tests when a background of host-infested cotton odor was supplied, and vice versa. This suggests that plant volatiles may have created an olfactory contrast effect. We discussed ecological significance of the results and concluded that both plant- and herbivore-related experiences play important role in parasitoid host foraging.     

Within‐plant distribution of 1,4‐benzoxazin‐3‐ones contributes to herbivore niche differentiation in maize

Plant defences vary in space and time, which may translate into specific herbivore‐foraging patterns and feeding niche differentiation. To date, little is known about the effect of secondary metabolite patterning on within‐plant herbivore foraging. We investigated how variation in the major maize secondary metabolites, 1,4‐benzoxazin‐3‐one derivatives (BXDs), affects the foraging behaviour of two leaf‐chewing herbivores. BXD levels varied substantially within plants. Older leaves had higher levels of constitutive BXDs while younger leaves were consistently more inducible. These differences were observed independently of plant age, even though the concentrations of most BXDs declined markedly in older plants. Larvae of the well‐adapted maize pest Spodoptera frugiperda preferred and grew better on young inducible leaves irrespective of plant age, while larvae of the generalist Spodoptera littoralis preferred and tended to grow better on old leaves. In BXD‐free mutants, the differences in herbivore weight gain between old and young leaves were absent for both species, and leaf preferences of S. frugiperda were attenuated. In contrast, S. littoralis foraging patterns were not affected. In summary, our study shows that plant secondary metabolites differentially affect performance and foraging of adapted and non‐adapted herbivores and thereby likely contribute to feeding niche differentiation.     

Herbivores and plant defences affect selection on plant reproductive traits more strongly than pollinators

Pollinators and herbivores can both affect the evolutionary diversification of plant reproductive traits. However, plant defences frequently alter antagonistic and mutualistic interactions, and therefore, variation in plant defences may alter patterns of herbivore‐ and pollinator‐mediated selection on plant traits. We tested this hypothesis by conducting a common garden field experiment using 50 clonal genotypes of white clover (Trifolium repens) that varied in a Mendelian‐inherited chemical antiherbivore defence—the production of hydrogen cyanide (HCN). To evaluate whether plant defences alter herbivore‐ and/or pollinator‐mediated selection, we factorially crossed chemical defence (25 cyanogenic and 25 acyanogenic genotypes), herbivore damage (herbivore suppression) and pollination (hand pollination). We found that herbivores weakened selection for increased inflorescence production, suggesting that large displays are costly in the presence of herbivores. In addition, herbivores weakened selection on flower size but only among acyanogenic plants, suggesting that plant defences reduce the strength of herbivore‐mediated selection. Pollinators did not independently affect selection on any trait, although pollinators weakened selection for later flowering among cyanogenic plants. Overall, cyanogenic plant defences consistently increased the strength of positive directional selection on reproductive traits. Herbivores and pollinators both strengthened and weakened the strength of selection on reproductive traits, although herbivores imposed ~2.7× stronger selection than pollinators across all traits. Contrary to the view that pollinators are the most important agents of selection on reproductive traits, our data show that selection on reproductive traits is driven primarily by variation in herbivory and plant defences in this system.     

Plant-mediated interactions between the rice water weevil and fall armyworm in rice

Greenhouse studies were conducted to investigate plant-mediated interactions between an above-ground and a below-ground herbivore when sharing a common host plant, rice (Oryza sativa L). Two common pests of rice were used: the rice water weevil (RWW), Lissorhoptrus oryzophilus Kuschel, as the root herbivore, and the fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) as the foliage-feeding herbivore. Rice water weevil larval performance was assessed by measuring larval density and average weight in response to different levels of defoliation by FAW larvae. The reciprocal experiment was done to evaluate FAW performance (growth rate) in response to RWW feeding. Severe defoliation by FAW decreased RWW densities by 32% and reduced larval weights by 48% compared to larvae on roots of non-defoliated plants. Effects in the converse experiments were not as strong. FAW growth rates were reduced 9–37% when feeding on rice leaves from plants damaged by RWW compared to larvae feed leaves from the no damage treatment. These reciprocal negative effects show that RWW and FAW are potential competitors when sharing a rice plant. Because RWW and FAW did not interact directly, competition was plant-mediated.     

The jasmonate pathway alters herbivore feeding behaviour: consequences for plant defences

The jasmonate pathway is a highly conserved defensive cascade in plants that regulates the induction of resistance against herbivores; however, its role in herbivore feeding behaviour remains unknown. We used a mutant tomato plant (def‐1) deficient in the production of jasmonate‐related defensive proteins to test the hypothesis that genotypes with a reduced ability to induce resistance have a higher and more concentrated pattern of herbivore damage. Wild‐type and def‐1 plants received either damage by Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) caterpillars or no damage. After treatment, we tested for systemic responses by allowing a free roaming S. exigua caterpillar to feed on the undamaged portions of plants. Weight‐gain and leaf consumption of S. exigua were highest on def‐1 plants, regardless of prior herbivore damage. Def‐1 plants also had fewer numbers of leaves and leaflets eaten, and fewer feeding holes, which was indicative of a more concentrated distribution of damage on mutant compared to wild‐type plants. Following these results, we mimicked the amount and distribution of feeding damage that wild‐type or jasmonate‐deficient plants would receive on wild‐type plants to test whether changes in feeding behaviour may feedback to influence the expression of induced resistance. We mimicked the distribution of damage in wild‐type and jasmonate‐deficient plants by allowing caterpillars to feed on either one (leaf 1 or 2) or two leaves (leaf 1 and 2). Increased herbivore damage resulted in higher proteinase inhibitor (PI) activity, a jasmonate‐regulated defensive protein, and lower S. exigua performance on wild‐type but not jasmonate‐deficient plants. Compared to undamaged plants, a concentrated pattern of herbivore damage increased systemic resistance; these induced responses were greater on leaflets with stronger vascular connections to the damaged leaf. A more dispersed pattern of caterpillar damage altered the expression of induced responses, but the outcome depended on the specific pattern of damage. When leaf 1 was damaged and then leaf 2, the undamaged (third) leaf (which is more strongly connected to leaf 1 than 2) expressed reduced the PI activity compared to plants receiving concentrated damage to leaf 1; whereas in plants where leaf 2 was first damaged and then leaf 1, there were no differences in PI activity in leaf 3 compared to plants receiving concentrated damage to leaf 2. Thus, induction of the jasmonate pathway may not only determine the amount and distribution of feeding damage by herbivores, but this may feedback to affect the subsequent expression of plant defence.     

Aboveground phytochemical responses to belowground herbivory in poplar trees and the consequence for leaf herbivore preference

Belowground (BG) herbivory can influence aboveground (AG) herbivore performance and food preference via changes in plant chemistry. Most evidence for this phenomenon derives from studies in herbaceous plants but studies in woody plants are scarce. Here we investigated whether and how BG herbivory on black poplar (Populus nigra) trees by Melolontha melolontha larvae influences the feeding preference of Lymantria dispar (gypsy moth) caterpillars. In a food choice assay, caterpillars preferred to feed on leaves from trees that had experienced attack by BG herbivores. Therefore, we investigated the effect of BG herbivory on the phytochemical composition of P. nigra trees alone and in combination with AG feeding by L. dispar caterpillars. BG herbivory did not increase systemic AG tree defences like volatile organic compounds, protease inhibitors and salicinoids. Jasmonates and salicylic acid were also not induced by BG herbivory in leaves but abscisic acid concentrations drastically increased together with proline and few other amino acids. Leaf coating experiments with amino acids suggest that proline might be responsible for the caterpillar feeding preference via presumptive phagostimulatory properties. This study shows that BG herbivory in poplar can modify the feeding preference of AG herbivores via phytochemical changes as a consequence of root‐to‐shoot signaling.     

Asymmetric effects of a leaf‐chewing herbivore on aphid population growth

1. Plant responses to herbivory are often specific to the feeding guild of the attacking herbivore. These phytochemical responses to herbivore damage can affect herbivore performance and activity. Comprehensive studies on the ecological consequences of multi‐herbivore plant interactions are key to understanding plant–herbivore community dynamics. 2. This study examined how feeding damage by co‐occurring herbivores from separate feeding guilds, Myzus persicae (Sulzer), a sucking herbivore, and Leptinotarsa decemlineata (Say), a chewing herbivore, alter plant chemistry and indirectly affect herbivore performance. Performance was measured when each insect fed on plants individually, sequentially, or simultaneously in laboratory and field experiments. Phytohormone and glycoalkaloid content were measured for each feeding sequence to evaluate plant responses to herbivory by each guild. Mid‐season and end‐of‐season tuber yield were evaluated in the field study. 3. Damage by L. decemlineata negatively impacted M. persicae performance in both laboratory and field settings. Damage by M. persicae did not affect L. decemlineata performance in laboratory assays. However, L. decemlineata performance was positively affected by M. persicae herbivory in the field, but this effect was temporary. Although phytohormones and plant defences varied across treatments, they provide little resolution on interaction outcomes. 4. These results confirm that the presence of multiple feeding guilds on a single plant can affect these chewing and sucking herbivores differentially, but given the variability in our phytochemical analyses compared with other studies, the mechanism remains unclear. The study's findings show that aphids are negatively affected by chewing herbivores across systems, while aphids temporarily affected beetles positively.     

Novel evidence for systemic induction of silicon defences in cucumber following attack by a global insect herbivore

1. Silicon (Si) is a beneficial nutrient that has been reported to ameliorate many abiotic and biotic stresses in plants, including insect herbivory. Insect herbivory has been shown to induce Si defences in plants, although the magnitude and nature of induction remain largely ambiguous. In particular, it is unclear whether herbivore induction of Si defences is confined to attacked tissues (local) or occurs elsewhere in the plant (systemic). 2. We grew cucumber, Cucumis sativus L. plants (var. Burpless F1 and Beit Alpha), an intermediate Si accumulator, hydroponically under Si-supplemented or Si-free conditions and measured the level of Si induction caused by a polyphagous chewing insect, the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). We also examined the impacts of Si on insect performance by conducting in vitro feeding assays on excised leaves (ex situ) and intact leaves on plants (in situ). 3. Herbivory significantly increased Si accumulation both locally in attacked leaves (21% increase in Beit Alpha and 17% in Burpless F1) and systemically in non-attacked leaves (19% increase in Beit Alpha and 10% in Burpless F1). Si supplementation significantly increased % foliar Si and C:N ratio, while significantly decreasing larval relative consumption (RC) and relative growth rate (RGR) in the in situ assays. In ex situ assays, however, Si only reduced larval RGR when fed on Beit Alpha plants. 4. Our results confirm that Si-based defences can also operate in moderate Si-accumulating plants and, for the first time, that insect herbivory induces systemic Si accumulation equivalently between plant varieties.     

Maintenance of narrow diet breadth in the monarch butterfly caterpillar: response to various plant species and chemicals

In order to better understand the maintenance of a fairly narrow diet breadth in monarch butterfly larvae, Danaus plexippus L. (Lepidoptera: Nymphalidae: Danainae), we measured feeding preference and survival on host and non-host plant species, and sensitivity to host and non-host plant chemicals. For the plant species tested, a hierarchy of feeding preferences was observed; only plants from the Asclepiadaceae were more or equally preferred to Asclepias curassavica, the common control. The feeding preferences among plant species within the Asclepiadaceae are similar to published mean cardenolide concentrations. However, since cardenolide data were not collected from individual plants tested, definitive conclusions regarding cardenolide concentrations and plant acceptability cannot be made. Although several non-Asclepiadaceae were eaten in small quantities, all were less preferred to A. curassavica. Additionally, these non-Asclepiadaceae do not support continued feeding, development, and survival of first and fifth-instar larvae. Preference for a host versus a non-host (A. curassavica versus Vinca rosea) increased for A. curassavica reared larvae as compared to diet-reared larvae suggesting plasticity in larval food preferences. Furthermore, host species were significantly preferred over non-host plant species in bioassays using a host plant or sucrose as a common control. Larval responses to pure chemicals were examined in order to determine if host and non-host chemicals stimulate or deter feeding in monarch larvae. We found that larvae were stimulated to feed by some ubiquitous plant chemicals, such as sucrose, inositol, and rutin. In contrast, several non-host plant chemicals deterred feeding: caffeine, apocynin, gossypol, tomatine, atropine, quercitrin, and sinigrin. Additionally the cardenolides digitoxin and ouabain, which are not in milkweed plants, were neutral in their influence on feeding. Another non-milkweed cardenolide, cymarin, significantly deterred feeding. Extracts of A. curassavica leaves were tested in bioassays to determine which components of the leaf stimulate feeding. Both an ethanol extract of whole leaves and a hexane leaf-surface extract are phagostimulatory, suggesting the involvement of both polar and non-polar plant compounds. These data suggest that the host range of D. plexippus larvae is maintained by both feeding stimulatory and deterrent chemicals in host and non-host plants.     

More than herbivory: levels of silica‐based defences in grasses vary with plant species,genotype and location

Silica defences in grasses have recently been suggested to be a potential driver of vole population dynamics. However, the ability of grasses to induce silica in response to herbivory has not been tested in northern ecosystems where small rodents are important herbivores. We conducted a large‐scale field experiment in subarctic tundra using three river catchments differing in herbivore densities, and examined the effects of small rodent and/or reindeer exclusion on leaf silica levels in five grass species (Avenella flexuosa, Anthoxanthum nipponicum, Calamagrostis phragmitoides, Deschampsia cespitosa and Phleum alpinum). We also conducted a greenhouse experiment using three of these species (A. flexuosa, A. nipponicum and D. cespitosa) and Festuca ovina to determine whether intraspecific genotypic variation affects baseline silica concentrations and the capacity to induce silica in response to simulated grazing. Baseline leaf silica concentrations and silica induction varied with plant species in both experiments, with catchment in the field experiment and with genotype in the greenhouse experiment. These findings show that the allocation to silica defences in grasses is highly variable, and suggest that the combined effects of grazing pressure, plant species and intraspecific genotypic differences are likely to determine the circumstances under which silica induction may be an optimal defence strategy. A better understanding of the interplay between grazing and other factors influencing silica induction is necessary to interpret the role of silica in plant–herbivore interactions.     

Herbivore specific induction of silica-based plant defences

Induced plant responses to herbivory have major impacts on herbivore feeding behaviour, performance and population dynamics. These effects are well established for chemical defences, but induction of physical defences remains far less studied. However, for many plants, it is physical defences that play the major role in regulating the levels of herbivore damage sustained. We provide evidence that, in grasses, induction of physical defences is both specific to herbivore feeding, as opposed to mechanical damage, and may be dependant on the amount of damage imposed. Furthermore, we show that the magnitude of the induction response is sufficient to deter further damage and affect herbivore performance. We compared silica induction in two grass species in response to vertebrate and invertebrate damage, and to mechanical defoliation. Induction was assessed at two levels of damage over 16 months. Foliar silica content did not increase in response to mechanical defoliation, but damage by either voles or locusts resulted in increases in silica content of over 400%. This increase deterred feeding by both voles and locusts. Silica induction in grasses due to repeated damage events over a prolonged period suggests a possible role for silica defence in the cyclical population fluctuations observed in many grass-feeding herbivores.     

Methyl jasmonate does not induce changes in Eucalyptus grandis leaves that alter the effect of constitutive defences on larvae of a specialist herbivore

The up-regulation of secondary metabolic pathways following herbivore attack and the subsequent reduction in herbivore performance have been identified in numerous woody plant species. Eucalypts constitutively express many secondary metabolites in the leaves, including terpenes and formylated phloroglucinol compounds (FPCs). We used clonal ramets from six clones of Eucalyptus grandis and two clones of E. grandis x camaldulensis to determine if methyl jasmonate (MeJA) treatment could induce changes in the foliar concentrations of either of these groups of compounds. We also used bioassays to determine if any changes in the performance of larvae of Paropsis atomaria, a chrysomelid leaf beetle, could be detected in treated ramets versus the untreated controls, thus indicating whether MeJA induced the up-regulation of defences other than terpenes or FPCs. We found no significant effects of MeJA treatment on either the foliar concentrations of terpenes and FPCs or on herbivore performance. We did, however, detect dramatic differences in larval performance between Eucalyptus clones, thereby demonstrating large variations in the levels of constitutive defence. Larval feeding on clones resistant to P. atomaria resulted in high first instar mortality and disruption of normal gregarious feeding behaviour in surviving larvae. Histological examination of larvae feeding on a resistant clone revealed damage to the midgut consistent with the action of a toxin. These findings concur with mounting evidence that most evergreen perennial plants lack foliar-induced defences and suggest that constitutively expressed secondary metabolites other than those commonly examined in studies of interactions between insect herbivores and Eucalyptus may be important in plant defence.