Phratora Vulgatissima Research Articles

Poplar protease inhibitor expression differs in an herbivore specific manner

BackgroundProtease inhibitors are defense proteins widely distributed in the plant kingdom. By reducing the activity of digestive enzymes in insect guts, they reduce the availability of nutrients and thus impair the growth and development of the attacking herbivore. One well-characterized class of protease inhibitors are Kunitz-type trypsin inhibitors (KTIs), which have been described in various plant species, including Populus spp. Long-lived woody perennials like poplar trees encounter a huge diversity of herbivores, but the specificity of tree defenses towards different herbivore species is hardly studied. We therefore aimed to investigate the induction of KTIs in black poplar (P. nigra) leaves upon herbivory by three different chewing herbivores, Lymantria dispar and Amata mogadorensis caterpillars, and Phratora vulgatissima beetles.ResultsWe identified and generated full-length cDNA sequences of 17 KTIs that are upregulated upon herbivory in black poplar leaves, and analyzed the expression patterns of the eight most up-regulated KTIs via qRT-PCR. We found that beetles elicited higher transcriptional induction of KTIs than caterpillars, and that both caterpillar species induced similar KTI expression levels. Furthermore, KTI expression strongly correlated with the trypsin-inhibiting activity in the herbivore-damaged leaves, but was not dependent on damage severity, i.e. leaf area loss, for most of the genes.ConclusionsWe conclude that the induction of KTIs in black poplar is controlled at the transcriptional level in a threshold-based manner and is strongly influenced by the species identity of the herbivore. However, the underlying molecular mechanisms and ecological consequences of these patterns remain to be investigated.

PhenModel: A temperature-dependent phenology/voltinism model for a herbivorous insect incorporating facultative diapause and budburst

A comprehensive phenology/voltinism model was developed for Phratora vulgatissima, an important pest of bioenergy crops. The model, phenModel, was developed based on development times of different life cycle stages (eggs, larvae, pupae, pre-oviposition, oviposition, sexual maturation) obtained from constant temperature laboratory experiments. As part of this study, a number of linear and non-linear models which describe the temperature-dependent development rate (inverse of development time) for each of the different life cycle stages were fitted. Based on the criteria of model parsimony and model fit, the non-linear Lactin-2 model was chosen as the optimum model to describe temperature-driven development in P. vulgatissima. To account for the variation in development times between individuals, an important but often ignored aspect in phenology models, a number of stochastic models (2- and 3- parameter Weibull and logistic models) were evaluated, based on the assumption that normalised development times conform to a similar shaped ('same shape') distribution. Novel aspects of the phenology model include the incorporation of a biologically relevant biofix, based on a budburst model for Salix viminalis, and a photoperiod threshold to induce facultative diapause. The model, which is written in R for accessibility, requires inputs of daily minimum and maximum temperature and site latitude and produces outputs describing the timing of completion of developmental stages for specified proportions of the population. It was evaluated against available field data and found to largely reproduce the observations providing a measure of its potential utility. A key component of the model allows for a sensitivity analysis of the model parameters. The model is structured so that it can easily be adapted for other leaf-feeding beetles which display a facultative reproductive diapause cued by photoperiod, and where the onset of oviposition is dependent on budburst, assuming relevant life cycle stage parameters are available.

Specificity of Herbivore Defense Responses in a Woody Plant, Black Poplar (Populus nigra)

The specificity of woody plant defense responses to different attacking herbivores is poorly known. We investigated the responses of black poplar (Populus nigra) to leaf feeding by three lepidopteran species (Lymantria dispar, Laothoe populi and Amata mogadorensis) and two leaf beetle species (Phratora vulgatissima and Chrysomela populi). Of the direct defenses monitored, increases in trypsin protease inhibitor activity and the salicinoid salicin were triggered by herbivore damage, but this was not herbivore-specific. Moreover, the majority of leaf salicinoid content was present constitutively and not induced by herbivory. On the other hand, volatile emission profiles did vary among herbivore species, especially between coleopterans and lepidopterans. Monoterpenes and sesquiterpenes were induced in damaged and adjacent undamaged leaves, while the emission of green leaf volatiles, aromatic and nitrogen-containing compounds (known to attract herbivore enemies) was restricted to damaged leaves. In conclusion, indirect defenses appear to show more specific responses to attacking herbivores than direct defenses in this woody plant.

Plant sex effects on insect herbivores and biological control in a Short Rotation Coppice willow

In the wild, plant sex can affect plant-herbivore interactions and higher trophic levels, including natural enemies of the herbivores. However, the possibility of manipulating plant sex to improve biological control and reduce herbivory in domesticated dioecious crops remains unexplored. The dioecious bioenergy crop, Salix viminalis, is often planted in monoclonal, and thus monosexual, fields. We investigated whether using plant clones of either sex, or mixing plants of both sexes, reduced the performance and abundance of the herbivorous pest insect Phratora vulgatissima and its main natural enemy, Anthocoris nemorum, and whether predation was affected. The herbivore laid more eggs, and the predator survived longer, on female plants in the lab. However, these effects did not translate into differences in predation rates in laboratory experiments or differential insect abundances on plants of either sex or plantation sex composition in the field. Plant genotype did have a significant effect on insect abundances, but this was due to plant traits other than sex. The results indicate that manipulating plant sex will not lead to improved biological control or reduced insect herbivory in S. viminalis energy forestry, but suggest that a focus on plant genotypic differences offers promise for improving management practices.

Costs and benefits of omnivore-mediated plant protection: effects of plant-feeding on Salix growth more detrimental than expected

Predators can decrease herbivore damage to plants, and this is often assumed to be beneficial to plant growth/reproduction without actual quantification. Moreover, previous studies have been biased towards strict carnivores and neglected the role of omnivorous predators in prey-suppression. Here, we examined the costs (reduction in growth) and benefits (increase in growth) of enemy-mediated plant protection via the omnivorous (prey and plant-feeding) Orthotylus marginalis, relative to herbivory by a detrimental insect pest of Salix spp. plantations, the beetle Phratora vulgatissima. In a first experiment, we compared the cost of adult beetle versus omnivore nymph plant-feeding, and assessed the (non-) additive effects of the two types of damage. In a second experiment, we quantified the reduction in plant damage resulting from beetle-egg feeding by omnivorous nymphs and subsequent benefits to plants. We found that plant-feeding by omnivores negatively affected plant growth and this effect was similar to the cost imposed by beetle herbivory. Furthermore, simultaneous damage effects were additive and more detrimental than individual effects. While egg-predation by omnivore nymphs completely prevented beetle damage to plants, there was no difference in plant growth relative to only herbivore-damaged plants and growth was still reduced compared to control plants. Thus, despite herbivore suppression, there was no benefit to plant growth of omnivore-mediated plant protection and the negative effects of omnivore plant-feeding remained. These results are a first for an omnivorous enemy, and provide novel and timely insights on the underlying assumptions of tri-trophic associations and their use for biocontrol of insect pests.

Predator hunting mode and host plant quality shape attack‐abatement patterns of predation risk in an insect herbivore

AbstractGroup formation reduces individual predation risk when the proportion of prey taken per predator encounter declines faster than the increase in group encounter rate (attack‐abatement). Despite attack‐abatement being an important component of group formation ecology, several key aspects have not been empirically studied, that is, interactions with the hunting mode of the predator and how these relationships are modified by local habitat quality. In 79 cage trials, we examined individual egg predation risk in different‐sized egg clutches from the blue willow beetle Phratora vulgatissima for two predators with different hunting modes (consumption of full group [Orthotylus marginalis] vs. part group [Anthocoris nemorum]). Because these predators also take nutrients from plant sap, we could examine how the quality of alternative food sources (high‐ vs. low‐quality host plant sap) influenced attack‐abatement patterns in the presence of different hunting strategies. For the O. marginalis predator, individual egg predation risk was largely independent of group size. For A. nemorum, egg predation risk clearly declined with increasing group size. However, approximately one‐third of the grouping benefit was lost to an increase in group detectability. There were clear differences in attack‐abatement patterns between plants with high‐ vs. low‐quality sap. When O. marginalis was the predator, there was no clear change in attack‐abatement in relation to host plant quality. However, for A. nemorum there was a clear reduction in overall predation risk and a stronger attack‐abatement pattern with increasing group size when plant sap quality increased. This implies that the relative benefits of prey grouping behavior for any species might show diurnal or seasonal changes as other aspects of resource/habitat quality change for the focal predator. Modulation of attack‐abatement by bottom‐up effects such as plant‐based food resources is yet to be incorporated into general theory, despite the ubiquity of omnivorous predators and with omnivory being important for shaping food webs, ecosystem functions, and in biological control. Thus, ongoing refinement of attack‐abatement theory by focusing on bottom‐up vs. top‐down processes could have significant impacts on many important contemporary fields of study.

Direct effects of elevated temperature on a tri-trophic system: Salix, leaf beetles and predatory bugs

The net effect of climatic change on biotic interactions will depend on how each interacting species is individually affected. Elevated temperatures are predicted to have differential effects on species across trophic levels, due to asymmetric sensitivity to temperature changes. In this study, we examined the direct effects of three temperature regimes (16, 20 and 24 °C) that reflect present and, potentially, future climate conditions on the response of Salix spp. plants, an important bioenergy crop, and its most damaging herbivore (Phratora vulgatissima) and an efficient natural enemy (the omnivorous predator Orthotylus marginalis). We found that plant growth, herbivore oviposition and enemy egg-foraging rate correlated positively with temperature. In the event of elevated temperatures following global climatic changes, these species could potentially respond in tandem. Still, the strength of responses varied among species, with herbivore and natural enemy exhibiting a similar and steeper rate of response relative to plants. Additionally, the herbivore’s response was influenced by plant quality with altered oviposition rates depending on whether it was fed the (previously determined) resistant Salix dasyclados or susceptible S. viminalis. This indicates that host plant chemistry has the potential to mediate differential responses to temperature. Together, our results suggest that indirect effects of elevated temperatures, leading to a disruption of trophic associations, may be less likely or less severe in this tri-trophic system.

Weeds within willow short-rotation coppices alter the arthropod community and improve biological control of the blue willow beetle

Outbreaks of the blue willow beetle Phratora vulgatissima (L.) (Coleoptera: Chrysomelidae) threaten the yield of willow plantations that rely on biological control by natural enemies. Here we show that weed presence increases herbivore and predator/parasitoid diversity on willow shoots and causes increased predation on P. vulgatissima eggs compared to shoots within plots without weeds. We argue that, in addition to higher abundance, the community shift in favour of egg consumers and additive predator effects causes the higher predation pressure. This increase (~35 %) was apparent despite more alternative prey. Neither the willow genotype on which the predator–prey interaction occurred nor the clutch size affected egg predation levels. Our results support root’s enemy hypothesis and suggest that intensive weed control might counteract biological control in willow plantations. However, at least during the establishing phase of the plantation, increased biocontrol probably does not compensate for yield losses caused by competing weeds.

How far away is the next basket of eggs? Spatial memory and perceived cues shape aggregation patterns in a leaf beetle.

Gregarious organisms need to handle the trade-off between increasing food competition and the positive effects of group living, and this is particularly important for ovipositing females. We hypothesized that insect females consider how many conspecifics previously visited a host plant. In a no-choice assay, we show that the gregarious blue willow leaf beetle (Phratora vulgatissima) laid the most eggs and the largest clutches on plants where a sequence of few individual females was released, compared to plants where one or many different females were repeatedly released. Therefore, this species is more sensitive to the indirectly perceived number of conspecifics than the directly perceived number of eggs on a plant. We further hypothesized that females adjust their own intra-plant egg clutch distribution to that of conspecifics and discovered a new behavioral component, i.e., the modulation of distances between clutches. Females adjusted these distances in ways indicating the use of spatial memory, because the largest distance increases were observed on plants with their own clutches, compared to plants with clutches from conspecifics. However, adjustment of aggregation level and distance between clutches occurred only on a suitable, and not on an unsuitable, Salix genotype. We conclude that both behaviors should reduce competition between sibling and non-sibling larvae.

Beetle feeding induces a different volatile emission pattern from black poplar foliage than caterpillar herbivory

Herbivore-induced plant volatile emission is often considered to be attacker species-specific, but most experimental evidence comes from short lived herbaceous species. In a recent study we showed that black poplar (Populus nigra) trees emit a complex blend of volatiles from damaged leaves when they are attacked by generalist gypsy moth (Lymantria dispar) caterpillars. Minor nitrogenous volatiles were especially characteristic of this blend. Here we show that attack on P. nigra by a beetle species, Phratora vulgatissima (Coleoptera, Chrysomelidae), led to the emission of the same compounds as already observed after caterpillar herbivory, but with striking quantitative changes in the blend. The consequences for attraction of herbivore enemies are discussed.

Outbreaking herbivore escapes parasitoid by attaining only a small body size

The reason why parasitoids are often unable to control insect herbivores during outbreak years has remained unclear to modern ecologists. Here I show that the blue willow beetle (Phratora vulgatissima) can escape parasitoids (Perilitus brevicollis) by reaching only a small body size during the increased competition which coincides with outbreaks. Different phases of outbreaks were simulated by allowing leaf‐beetle larvae to develop at different densities on willow. The denser the herbivore populations, the smaller body sizes they attained. In the field too, herbivore body size decreased when the level of defoliation increased. The parasitoids were clearly limited by herbivore body size, exhibiting reduced survival in parallel with decreasing herbivore size, with none of them surviving in beetles from larvae reared at the highest density. The results demonstrate how bottom‐up control of the herbivore becomes more important as outbreaks intensify, while top‐down control gradually becomes less important and collapses during the outbreak peak. Because the mechanism limiting P. brevicollis parasitoids involves herbivore body size, which is reduced at high densities, I anticipate that these parasitoids are unable to terminate herbivore outbreaks.

Phenotypic plasticity in a willow leaf beetle depends on host plant species: release and recognition of beetle odors.

Aggregation behavior of herbivorous insects is mediated by a wide range of biotic and abiotic factors. It has been suggested that aggregation behavior of the blue willow leaf beetle Phratora vulgatissima is mediated by both host plant odor and by odor released by the beetles. Previous studies show that the beetles respond to plant odors according to their prior host plant experiences. Here, we analyzed the effect of the host plant species on odor released and perceived by adult P. vulgatissima. The major difference between the odor of beetles feeding on salicin-rich and salicin-poor host plants was the presence of salicylaldehyde in the odor of the former, where both males and females released this compound. Electrophysiological studies showed that the intensity of responses to single components of odor released by beetles was sex specific and dependent on the host plant species with which the beetles were fed. Finally, behavioral studies revealed that males feeding on salicin-rich willows were attracted by salicylaldehyde, whereas females did not respond behaviorally to this compound, despite showing clear antennal responses to it. Finally, the ecological relevance of the influence of a host plant species on the plasticity of beetle odor chemistry, perception, and behavior is discussed.

Plant‐sex‐biased tritrophic interactions on dioecious willow

Plant sex effects on herbivores are well studied, but little is known about these effects on predators and predator–herbivore dynamics. Here we take a holistic approach to study, simultaneously, plant sex effects on herbivore and predator preference and performance, as well as population densities and predation pressure in the field. For dioecious Salix cinerea (grey willow) we found that male plants represented higher host plant quality than females for an omnivorous predator (Anthocoris nemorum, common flower bug), while host plant quality for its herbivorous prey (Phratora vulgatissima, blue willow beetle) was not sex‐biased. The herbivore strongly preferred the host plant sex (female) that was suboptimal for the predator, which in turn followed its prey to female plants, leading to plant‐sex‐biased predation. These results provide new insight into the far‐reaching effects of plant sex on insect communities, and open up novel opportunities for improving biocontrol of the herbivore in Salix short rotation coppice.

Phenotypic plasticity in host plant preference of the willow leaf beetlePhratora vulgatissima: the impact of experience made by adults

Knowledge of the plasticity of Phratora vulgatissima (Colepotera: Chrysomelidae) behavioural responses to cues of its willow host plants is essential for understanding host affiliations of this species and for developing management strategies. We investigated how the experience obtained by adult P. vulgatissima with two different willow species shapes olfactory, feeding and oviposition preferences in the laboratory. The willow species differed in their leaf odours and phenolic glycoside contents. Females that had experienced Salix viminalis (Salicaceae) neither discriminated between odours of S. viminalis and Salix dasyclados, nor between blends of green leaf volatiles (GLV) that mimic quantitatively those released by S. viminalis and S. dasyclados. However, when females had experienced S. dasyclados, they preferred the experienced odour of S. dasyclados and the respective GLV blend to the odour of S. viminalis or its GLV blend. By contrast, regardless of their experience obtained in the adult stage, females preferred S. viminalis over S. dasyclados for feeding and oviposition. Exposure of beetles to odour of stands with various willow species might affect the experience-induced olfactory preference for a single species and thus impair host location success.

Performance of an Herbivorous Leaf Beetle (Phratora vulgatissima) on Salix F2 Hybrids: the Importance of Phenolics

The genotype of the plant determines, through the expression of the phenotype, how well it is suited as food for herbivores. Since hybridization often results in profound genomic alterations with subsequent changes in phenotypic traits, it has the potential to significantly affect plant-herbivore interactions. In this study, we used a population of F2 hybrids that originated from a cross between a Salix viminalis and a Salix dasyclados genotype, which differed in both phenolic content and resistance to the herbivorous leaf beetle Phratora vulgatissima. We screened for plants that showed a great variability in leaf beetle performance (i.e., oviposition and survival). By correlating leaf phenolics to the response of the herbivores, we evaluated the importance of different phenolic compounds for Salix resistance to the targeted insect species. The performance of P. vulgatissima varied among the F2 hybrids, and two patterns of resistance emerged: leaf beetle oviposition was intermediate on the F2 hybrids compared to the parental genotypes, whereas leaf beetle survival demonstrated similarities to one of the parents. The findings indicate that these life history traits are controlled by different resistance mechanisms that are inherited differently in the hybrids. Salicylates and a methylated luteolin derivative seem to play major roles in hybrid resistance to Phratora vulgatissima. Synergistic effects of these compounds, as well as potential threshold concentrations, are plausible. In addition, we found considerable variation in both distributions and concentrations of different phenolics in the F2 hybrids. The phenolic profiles of parental genotypes and F2 hybrids differed significantly (e.g., novel compounds appeared in the hybrids) suggesting genomic alterations with subsequent changes in biosynthetic pathways in the hybrids.

Beetles Do It Differently: Two Stereodivergent Cyclisation Modes in Iridoid‐Producing Leaf‐Beetle Larvae

Larvae of the Chrysomelina species Phaedon cochleariae, Hydrothassa marginella, Phratora vulgatissima, Gastrophysa viridula, Gastrophysa atrocyanea, Gastrophysa cyanea and Gastrophysa polygoni produce the iridoid chrysomelidial (1) to defend themselves against predators. Feeding experiments with a deuterated precursor ([(2)H(5)]8-hydroxygeraniol 9) and in vitro isotope exchange experiments with defensive secretion in (2)H(2)O revealed differences in the cyclisation of the ultimate precursor 8-oxogeranial (8) to 1, between members of the genus Gastrophysa and all other species. In P. cochleariae, H. marginella and P. vulgatissima 1 is most likely produced by a Rauhut-Currier-type cyclisation via a "transoid dienamine", with loss of a single deuterium atom from C(4) of the precursor. In contrast, members of the genus Gastrophysa cyclise 8 via a "cisoid dienamine" intermediate, with exchange of all three deuterium atoms from the methyl group at C(3). To study whether the different cyclisation modes influence the stereochemistry of 1, the absolute configuration of 1 of the larvae was determined by GC-MS on a chiral column. In accordance with literature (J. Meinwald, T. H. Jones, J. Am. Chem. Soc. 1978, 100, 1883 and N. Shimizu, R. Yakumaru, T. Sakata, S. Shimano, Y. Kuwahara, J. Chem. Ecol. 2012, 38, 29), we found (5S,8S)-chrysomelidial (1) in H. marginella and P. vulgatissima, but P. cochleariae and all investigated members of the genus Gastrophysa synthesise (5R,8R)-chrysomelidial (1).

Food Conditioning Affects Expression of Insect Resistance in Diploid Willows (<i>Salix</i> spp.)

The high energy quota and versatility of use make willows (Salix spp.) attractive as bioenergy crops. Insect defoliation constitutes a threat to the profitability of willow growers. Hitherto, the breeding for resistance against the main insect pests has been hampered by the fact that all known resistant willow clones are polyploids, and existing molecular breeding tools work most effectively for diploids. Here, we firstly report diploid willows highly resistant to the main insect defoliator, the leaf beetle (Phratora vulgatissima), offering new opportunities for breeding resistance. Leaf beetles exposed to three resistant clones (two S. purpurea one S. eriocephala) laid three to 27 times fewer eggs than females on a susceptible S. viminalis clone. Secondly, we show that beetles laid significantly more eggs on resistant clones if they were fed the susceptible clone prior to the oviposition monitoring test compared to when they prefed on resistant clones. Nevertheless, the differences observed between resistant and susceptible clones were pronounced in all cases. The food conditioning effect means that small differences in resistance among clones may be undetected.

Constitutive and herbivore-induced systemic volatiles differentially attract an omnivorous biocontrol agent to contrasting Salix clones

While carnivores are known to be attracted to herbivore-induced plant volatiles, little is known about how such volatiles may affect the behaviour of omnivorous predators that may use both plants and herbivores as food. Here, we examine how systemically produced plant volatiles, in response to local herbivore damage, differentially attract a key omnivorous predator, Anthocoris nemorum (Heteroptera: Anthocoridae), to single clones of three species of Salix: S. viminalis, S. dasyclados and S. cinerea. The profiles of the plant volatiles produced were found to vary among Salix clones and between herbivore-damaged and intact plants. Anthocoris nemorum was attracted to the volatiles released from undamaged plants of all three species, but most strongly to a native S. cinerea clone. Plants damaged by the herbivorous leaf beetle Phratora vulgatissima (Coleoptera: Chrysomelidae) were generally more attractive than undamaged plants, with A. nemorum responding to systemic changes in the damaged plants where the experimental design specifically excluded volatiles released from the actual site of damage. When comparing damaged plants, the S. dasyclados clone was more attractive to A. nemorum than the S. viminalis clone-a somewhat surprising result since this Salix clone is considered relatively resistant to P. vulgatissima, and hence offers a limited amount of prey. Our experiments highlight that both constitutive and induced plant volatiles play a role in omnivore attraction, and this emphasizes the importance of considering odours of released volatiles when cropping and breeding Salix for increased resistance to herbivores.

Host‐plant quality adaptively affects the diapause threshold: evidence from leaf beetles in willow plantations

1. Voltinism of herbivorous insects can vary depending on environmental conditions. The leaf beetle Phratora vulgatissima L. is univoltine in Sweden but will sometimes initiate a second generation in short‐rotation coppice (SRC) willow plantations.2. The study investigated whether increased voltinism by P. vulgatissima in plantations can be explained by (i) rapid life‐cycle development allowing two generations, or (ii) postponed diapause induction on coppiced willows.3. In the field, no difference was found in the phenology or development of first‐generation broods between plantations (S. viminalis) and natural willow habitats (S. cinerea). However, the induction of diapause occurred 1–2 weeks later in SRC willow plantations.4. Laboratory experiments indicated no genetic difference in the critical day‐length for diapause induction between beetles originating from plantations and natural habitats. Development time was unaffected by host‐plant quality but critical day‐length was prolonged by almost an hour when the beetles were reared on a non‐preferred willow species (S. phylicifolia). When reared on new leaves from re‐sprouting shoots of recently coppiced willow plants, diapause incidence was significantly less than when the beetles were reared on mature leaves from uncoppiced plants.5. The study suggests that P. vulgatissima has a plastic diapause threshold influenced by host‐plant quality. The use of host‐plant quality as a diapause‐inducing stimulus is likely to be adaptive in cases where food resources are unpredictable, such as when new host‐plant tissue is produced after a disturbance. SRC willows may allow two beetle generations due to longer growing seasons of coppiced plants that grow vigorously.

Estimating direct resistance in willows against a major insect pest, Phratora vulgatissima, by comparing life history traits

AbstractMany Salix species (Salicaceae) have a high energy quotum and efficient nitrogen use, resulting in a high and stable biomass production, making these plants suitable as bioenergy crops. However, Salix coppices are sensitive to herbivory by chrysomelid beetles, such as Phratora vulgatissima L. (Coleoptera: Chrysomelidae), and to maintain high biomass yields over several years, durable plant resistance is necessary. We have developed a reliable and efficient bioassay that can be used as a tool for detection of resistance against P. vulgatissima in Salix. Screening of six clones of one susceptible (Salix viminalis L.) and one resistant (Salix dasyclados Wimm.) willow species, by the response of several life history traits of larval and adult P. vulgatissima, identified egg production as the most reliable trait for resistance. We show that a 2‐week bioassay of oviposition rate is sufficient for a trustworthy estimation of both total egg production and survival of adult leaf beetles. In addition, to elucidate the variation in resistance among the studied clones, leaf concentrations of various phenolic compounds were compared with the key life history traits of the beetles. Phenolic compounds, especially salicylates, have previously been reported as the main factor controlling Salix resistance against P. vulgatissima. Indeed we found higher levels of salicylates in the resistant clones and larval performance was negatively correlated to the concentration of salicylates in the leaves. In addition, we found indications of negative effects of a luteolin derivative and quercetin‐3‐glucoside on leaf beetle performance, but further studies are needed to elucidate the specific roles that the different phenolic compounds play in plant resistance.