Endophytic Beauveria bassiana induces biosynthesis of flavonoids in oilseed rape following both seed inoculation and natural colonization.

BACKGROUND: Cultivation of oilseed rape Brassica napus is pesticide-intensive, and alternative plant protection strategies are needed because both pesticide resistance and legislation narrow the range of effective chemical pesticides. The entomopathogenic fungus Beauveria bassiana is used as a biocontrol agent against various insect pests, but little is known about its endophytic potential and role in plant protection for oilseed rape. First, we studied whether B. bassiana can establish as an endophyte in oilseed rape, following seed inoculation. To evaluate the plant protection potential of endophytic B. bassiana on oilseed rape, we next examined its ability to induce plant metabolite biosynthesis. In another experiment, we tested the effect of seed inoculation on seedling survival in a semi-field experiment. RESULTS: Beauveria bassiana endophytically colonized oilseed rape following seed inoculation, and, in addition, natural colonization was also recorded. Maximum colonization rate was 40%, and generally increased with inoculation time. Seed inoculation did not affect the germination probability or growth of oilseed rape, but B. bassiana inoculated seeds germinated more slowly compared to controls. Endophytic colonization of B. bassiana induced biosynthesis of several flavonoids in oilseed rape leaves under controlled conditions. In the experiment conducted in semi-field conditions, inoculated seedlings had slightly higher mortality compared to control seedlings. CONCLUSION: Beauveria bassiana showed endophytic potential on oilseed rape via both natural colonization and seed inoculation, and it induced the biosynthesis of flavonoids. However, its use as an endophyte for plant protection against pests or pathogens for oilseed rape remains unclear. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Glyphosate-based herbicide use affects individual microbial taxa in strawberry endosphere but not the microbial community composition.

AIMS: In a field study, the effects of treatments of glyphosate-based herbicides (GBHs) in soil, alone and in combination with phosphate fertilizer, were examined on the performance and endophytic microbiota of garden strawberry. METHODS AND RESULTS: The root and leaf endophytic microbiota of garden strawberries grown in GBH-treated and untreated soil, with and without phosphate fertilizer, were analyzed. Next, bioinformatics analysis on the type of 5-enolpyruvylshikimate-3-phosphate synthase enzyme was conducted to assess the potential sensitivity of strawberry-associated bacteria and fungi to glyphosate, and to compare the results with field observations. GBH treatments altered the abundance and/or frequency of several operational taxonomic units (OTUs), especially those of root-associated fungi and bacteria. These changes were partly related to their sensitivity to glyphosate. Still, GBH treatments did not shape the overall community structure of strawberry microbiota or affect plant performance. Phosphate fertilizer increased the abundance of both glyphosate-resistant and glyphosate-sensitive bacterial OTUs, regardless of the GBH treatments. CONCLUSIONS: These findings demonstrate that although the overall community structure of strawberry endophytic microbes is not affected by GBH use, some individual taxa are.

Legacy of agrochemicals in the circular food economy: Glyphosate-based herbicides introduced via manure fertilizer affect the yield and biochemistry of perennial crop plants during the following year.

Conventional agricultural practices favoring the use of glyphosate-based herbicides (GBHs) increase the risk of GBH residues ending up in animal feed, feces, and, eventually, manure. The use of poultry manure as organic fertilizer in the circular food economy increases the unintentional introduction of GBH residues into horticultural and agricultural systems, with reportedly negative effects on the growth and reproduction of crop plants. To understand the potential lasting effects of exposure to GBH residues via organic manure fertilizers, we studied strawberry (Fragaria x vescana) plant performance, yield quantity, biochemistry, folivory, phytochemistry, and soil elemental composition the year after exposure to GBH. Although plants exposed to GBH residues via manure fertilizer were, on average, 23% smaller in the year of exposure, they were able to compensate for their growth during the following growing season. Interestingly, GBH residue exposure in the previous growing season led to a trend in altered plant size preferences of folivores during the following growing season. Furthermore, the plants that had been exposed to GBH residues in the previous growing season produced 20% heavier fruits with an altered composition of phenolic compounds compared to non-exposed plants. Our results indicate that GBHs introduced via manure fertilizer following circular economy practices in one year can have effects on perennial crop plants in the following year, although GBH residues in soil have largely vanished.

Evolutionary Ecology of Plant-Arthropod Interactions in Light of the "Omics" Sciences: A Broad Guide.

Aboveground plant-arthropod interactions are typically complex, involving herbivores, predators, pollinators, and various other guilds that can strongly affect plant fitness, directly or indirectly, and individually, synergistically, or antagonistically. However, little is known about how ongoing natural selection by these interacting guilds shapes the evolution of plants, i.e., how they affect the differential survival and reproduction of genotypes due to differences in phenotypes in an environment. Recent technological advances, including next-generation sequencing, metabolomics, and gene-editing technologies along with traditional experimental approaches (e.g., quantitative genetics experiments), have enabled far more comprehensive exploration of the genes and traits involved in complex ecological interactions. Connecting different levels of biological organization (genes to communities) will enhance the understanding of evolutionary interactions in complex communities, but this requires a multidisciplinary approach. Here, we review traditional and modern methods and concepts, then highlight future avenues for studying the evolution of plant-arthropod interactions (e.g., plant-herbivore-pollinator interactions). Besides promoting a fundamental understanding of plant-associated arthropod communities' genetic background and evolution, such knowledge can also help address many current global environmental challenges.

Quantification of the Potential Impact of Glyphosate-Based Products on Microbiomes.

Glyphosate-based products (GBP) are the most common broad-spectrum herbicides worldwide. The target of glyphosate is the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in the shikimate pathway, which is virtually universal in plants. The inhibition of the enzyme stops the production of three essential amino acids: phenylalanine, tyrosine, and tryptophan. EPSPS is also present in fungi and prokaryotes, such as archaea and bacteria; thus, the use of GBP may have an impact on the microbiome composition of soils, plants, herbivores, and secondary consumers. This article aims to present general guidelines to assess the effect of GBP on microbiomes from field experiments to bioinformatics analyses and provide a few testable hypotheses. Two field experiments are presented to test the GBP on non-target organisms. First, plant-associated microbes from 10 replicated control and GBP treatment plots simulating no-till cropping are sampled and analyzed. In the second experiment, samples from experimental plots fertilized by either poultry manure containing glyphosate residues or non-treated control manure were obtained. Bioinformatics analysis of EPSPS protein sequences is utilized to determine the potential sensitivity of microbes to glyphosate. The first step in estimating the effect of GBP on microbiomes is to determine their potential sensitivity to the target enzyme (EPSPS). Microbial sequences can be obtained either from public repositories or by means of PCR amplification. However, in the majority of field studies, microbiome composition has been determined based on universal DNA markers such as the 16S rRNA and the internal transcribed spacer (ITS). In these cases, sensitivity to glyphosate can only be estimated through a probabilistic analysis of EPSPS sequences using closely related species. The quantification of the potential sensitivity of organisms to glyphosate, based on the EPSPS enzyme, provides a robust approach for further experiments to study target and non-target resistant mechanisms.

Pollinators and herbivores interactively shape selection on strawberry defence and attraction.

Tripartite interactions between plants, herbivores, and pollinators hold fitness consequences for most angiosperms. However, little is known on how plants evolve in response-and in particular what the net selective outcomes are for traits of shared relevance to pollinators and herbivores. In this study, we manipulated herbivory ("presence" and "absence" treatments) and pollination ("open" and "hand pollination" treatments) in a full factorial common-garden experiment with woodland strawberry (Fragaria vesca L.). This design allowed us to quantify the relative importance and interactive effects of herbivore- and pollinator-mediated selection on nine traits related to plant defence and attraction. Our results showed that pollinators imposed stronger selection than herbivores on traits related to both direct and indirect (i.e., tritrophic) defence. However, conflicting selection was imposed on inflorescence density: a trait that appears to be shared by herbivores and pollinators as a host plant signal. However, in all cases, selection imposed by one agent depended largely on the presence or ecological effect of the other, suggesting that dynamic patterns of selection could be a common outcome of these interactions in natural populations. As a whole, our findings highlight the significance of plant-herbivore-pollinator interactions as potential drivers of evolutionary change, and reveal that pollinators likely play an underappreciated role as selective agents on direct and in direct plant defence.

'Resistance Mixtures' Reduce Insect Herbivory in Strawberry (Fragaria vesca) Plantations.

The transition toward more sustainable plant protection with reduced pesticide use is difficult, because there is no "silver bullet" available among nonchemical tools. Integrating several plant protection approaches may thus be needed for efficient pest management. Recently, increasing the genetic diversity of plantations via cultivar mixing has been proposed as a possible method to reduce pest damage. However, previous studies have not addressed either the relative efficiency of exploiting cultivar mixing and intrinsic plant herbivore resistance or the potential utility of combining these approaches to increase cropping security. Here, using a full factorial experiment with 60 woodland strawberry plots, we tested for the relative and combined effect of cultivar mixing and intrinsic plant resistance on herbivore damage and yield. The experiment comprised two levels of diversity ("high" with 10 varieties and "low" with two varieties) and three levels of resistance ("resistant" comprising only varieties intrinsically resistant against strawberry leaf beetle Galerucella tenella; "susceptible" with susceptible varieties only; and "resistance mixtures" with 50:50 mixtures of resistant and susceptible varieties). The experiment was carried out over two growing seasons. Use of resistant varieties either alone or intermixed with susceptible varieties in "resistance mixtures" reduced insect herbivory. Interestingly, resistant varieties not only reduced the mean damage in "resistance mixtures" by themselves being less damaged, but also protected intermixed susceptible varieties via associational resistance. The effect of higher genetic diversity was less evident, reducing herbivory only at the highest level of herbivore damage. In general, herbivory was lowest in plots with high diversity that included at least some resistant varieties and highest in low diversity plots consisting only of susceptible varieties. Despite this, no significant difference in yield (fruit biomass) was found, indicating that strawberry may be relatively tolerant. Our results demonstrate that combined use of high genetic diversity and resistant varieties can help reduce pest damage and provide a useful tool for sustainable food production. "Resistance mixtures" may be particularly useful for sensitive food crops where susceptible varieties are high yielding that could not be completely replaced by resistant ones.

Risk in the circular food economy: Glyphosate-based herbicide residues in manure fertilizers decrease crop yield.

Glyphosate-based herbicides (GBHs) are the most frequently used herbicides globally. They were launched as a safe solution for weed control, but recently, an increasing number of studies have shown the existence of GBH residues and highlighted the associated risks they pose throughout ecosystems. Conventional agricultural practices often include the use of GBHs, and the use of glyphosate-resistant genetically modified crops is largely based on the application of glyphosate, which increases the likelihood of its residues ending up in animal feed. These residues persist throughout the digestive process of production animals and accumulate in their excretion products. The poultry industry, in particular, is rapidly growing, and excreted products are used as plant fertilizers in line with circular food economy practices. We studied the potential effects of unintentional glyphosate contamination on an agronomically important forage grass, meadow fescue (Festuca pratensis) and a horticulturally important strawberry (Fragaria x vescana) using glyphosate residues containing poultry manure as a plant fertilizer in a common garden experiment. Glyphosate in the manure decreased plant growth in both species and vegetative reproduction in F. x vescana. Furthermore, our results indicate that glyphosate residues in organic fertilizers might have indirect effects on sexual reproduction in F. pratensis and herbivory in F. x vescana because they positively correlate with plant size. Our results highlight that glyphosate can be unintentionally spread via organic fertilizer, counteracting its ability to promote plant growth.

A Glyphosate-Based Herbicide in Soil Differentially Affects Hormonal Homeostasis and Performance of Non-target Crop Plants.

Glyphosate is the most widely used herbicide with a yearly increase in global application. Recent studies report glyphosate residues from diverse habitats globally where the effect on non-target plants are still to be explored. Glyphosate disrupts the shikimate pathway which is the basis for several plant metabolites. The central role of phytohormones in regulating plant growth and responses to abiotic and biotic environment has been ignored in studies examining the effects of glyphosate residues on plant performance and trophic interactions. We studied interactive effects of glyphosate-based herbicide (GBH) residues and phosphate fertilizer in soil on the content of main phytohormones, their precursors and metabolites, as well as on plant performance and herbivore damage, in three plant species, oat (Avena sativa), potato (Solanum tuberosum), and strawberry (Fragaria x ananassa). Plant hormonal responses to GBH residues were highly species-specific. Potato responded to GBH soil treatment with an increase in stress-related phytohormones abscisic acid (ABA), indole-3-acetic acid (IAA), and jasmonic acid (JA) but a decrease in cytokinin (CK) ribosides and cytokinin-O-glycosides. GBH residues in combination with phosphate in soil increased aboveground biomass of potato plants and the concentration of the auxin phenylacetic acid (PAA) but decreased phaseic acid and cytokinin ribosides (CKR) and O-glycosides. Chorismate-derived compounds [IAA, PAA and benzoic acid (BzA)] as well as herbivore damage decreased in oat, when growing in GBH-treated soil but concentrations of the cytokinin dihydrozeatin (DZ) and CKR increased. In strawberry plants, phosphate treatment was associated with an elevation of auxin (IAA) and the CK trans-zeatin (tZ), while decreasing concentrations of the auxin PAA and CK DZ was observed in the case of GBH treatment. Our results demonstrate that ubiquitous herbicide residues have multifaceted consequences by modulating the hormonal equilibrium of plants, which can have cascading effects on trophic interactions.

Wild strawberry shows genetic variation in tolerance but not resistance to a generalist herbivore.

Plants' defenses against herbivores usually include both resistance and tolerance mechanisms. Their deployment has predominantly been studied in either single-plant genotypes or multiple genotypes exposed to single herbivores. In natural situations, however, most plants are attacked by multiple herbivores. Therefore, aims of this study were to assess and compare the effects of single and multiple herbivores on plant resistance and tolerance traits, and the consequences for overall plant performance. For this, we exposed multiple genotypes of wild woodland strawberry (Fragaria vesca) to jasmonic acid (JA), to mimic chewing herbivory and induce the plants' defense responses, and then introduced the generalist herbivore Spodoptera littoralis to feed on them. We found that woodland strawberry consistently showed resistance to S. littoralis herbivory, with no significant genetic variation between the genotypes. By contrast, the studied genotypes showed high variation in tolerance, suggesting evolutionary potential in this trait. Prior JA application did not alter these patterns, although it induced an even higher level of resistance in all tested genotypes. The study provides novel information that may be useful for breeders seeking to exploit tolerance and resistance mechanisms to improve strawberry crops' viability and yields, particularly when multiple herbivores pose significant threats.

Plant resistance does not compromise parasitoid-based biocontrol of a strawberry pest.

Plant nutritional  quality can influence interactions between herbivores and their parasitoids. While most previous work has focused on a limited set of secondary plant metabolites, the tri-trophic effects of overall phenotypic resistance have been understudied. Furthermore, the joint effects of secondary and primary metabolites on parasitoids are almost unexplored. In this study, we compared the performance and survival of the parasitoid species Asecodes parviclava Thompson on wild woodland strawberry (Fragaria vesca L.) genotypes showing variation in resistance against the parasitoid's host, the strawberry leaf beetle (Galerucella tenella L.). Additionally, we related the metabolic profiles of these plant genotypes to the tritrophic outcomes in order to identify primary and secondary metabolites involved in regulating plant potential to facilitate parasitism. We found that parasitoid performance was strongly affected by plant genotype, but those differences in plant resistance to the herbivore were not reflected in parasitoid survival. These findings could be explained in particular by a significant link between parasitoid survival and foliar carbohydrate levels, which appeared to be the most important compounds for parasitism success. The fact that plant quality strongly affects parasitism should be further explored and utilized in plant breeding programs for a synergistic application in sustainable pest management.

Capturing genetic variation in crop wild relatives: An evolutionary approach.

Crop wild relatives (CWRs) offer novel genetic resources for crop improvement. To assist in the urgent need to collect and conserve CWR germplasm, we advance here the concept of an "evolutionary" approach. Central to this approach is the predictive use of spatial proxies of evolutionary processes (natural selection, gene flow and genetic drift) to locate and capture genetic variation. As a means to help validate this concept, we screened wild-collected genotypes of woodland strawberry (Fragaria vesca) in a common garden. A quantitative genetic approach was then used to test the ability of two such proxies-mesoclimatic variation (a proxy of natural selection) and landscape isolation and geographic distance between populations (proxies of gene flow potential)-to predict spatial genetic variation in three quantitative traits (plant size, early season flower number and flower frost tolerance). Our results indicated a significant but variable effect of mesoclimatic conditions in structuring genetic variation in the wild, in addition to other undetermined regional scale processes. As a proxy of gene flow potential, landscape isolation was also a likely determinant of observed patterns-as opposed to, and regardless of, geographic distance between populations. We conclude that harnessing proxies of adaptive and nonadaptive evolutionary processes could provide a robust and valuable means to identify genetic variation in CWRs. We thus advocate wider use and development of this approach amongst researchers, breeders and practitioners, to expedite the capture and in situ conservation of genetic resources provided by crop wild relatives.

Genetic drift precluded adaptation of an insect seed predator to a novel host plant in a long-term selection experiment.

Host specialization is considered a primary driver of the enormous diversity of herbivorous insects. Trade-offs in host use are hypothesized to promote this specialization, but they have mostly been studied in generalist herbivores. We conducted a multi-generation selection experiment to examine the adaptation of the specialist seed-feeding bug, Lygaeus equestris, to three novel host plants (Helianthus annuus, Verbascum thapsus and Centaurea phrygia) and to test whether trade-offs promote specialization. During the selection experiment, body size of L. equestris increased more on the novel host plant H. annuus compared to the primary host plant, Vincetoxicum hirundinaria, but this effect was not observed in other fitness related traits. In addition to selection, genetic drift caused variation among the experimental herbivore populations in their ability to exploit the host plants. Microsatellite data indicated that the level of within-population genetic variation decreased and population differentiation increased more in the selection line feeding on H. annuus compared to V. hirundinaria. We found a negative correlation between genetic differentiation and heterozygosity at the end of the experiment, suggesting that differentiation was significantly affected by genetic drift. We did not find fitness trade-offs between L. equestris feeding on the four hosts. Thus, trade-offs do not seem to promote specialization in L. equestris. Our results suggest that this insect herbivore is not likely to adapt to a novel host species in a time-scale of 20 generations despite sufficient genetic variation and that genetic drift disrupted the response to selection.

Folivory has long-term effects on sexual but not on asexual reproduction in woodland strawberry.

Plant fitness is often a result of both sexual and asexual reproductive success and, in perennial plants, over several years. Folivory can affect both modes of reproduction. However, little is known about the effects of folivory on resource allocation to the two modes of reproduction simultaneously and across years. In a 2-year common garden experiment, we examined the effects of different levels of folivory by the strawberry leaf beetle, Galerucella tenella, on current growth, as well as current and future sexual and asexual reproduction (runners) of perennial woodland strawberry, Fragaria vesca. In addition, we measured the chlorophyll content in leaves in the year of experimental damage to determine whether there was increased photosynthetic activity, and, thus, a compensatory response to herbivory. Finally, we tested whether the previous year's folivory, as a result of its effect on plant fitness, affected the level of natural herbivory the plant experienced during the subsequent year. In the year of experimental damage, plants that were exposed to moderate and high levels of folivory (25% and 50% leaf area consumed, respectively) increased their photosynthetic activity compared to control plants. However, only plants exposed to high folivory exhibited negative effects, with a lower probability of flowering compared to control plants, indicating that plants exposed to low or moderate folivory were able to compensate for the damage. Negative effects of folivory were carried over to the subsequent year. Plants that were exposed to moderate folivory (25% leaf area consumed) during first year produced fewer flowers and fruits in the subsequent year. Runner production was consistently unaffected by folivory. The effects of experimental folivory on the level of natural herbivory were mediated via its effects on plant fitness. Our results show that the negative effects of folivory only influence sexual reproduction in woodland strawberry. Furthermore, even though woodland strawberry can tolerate moderate amounts of folivory in the short term, the negative effects on fitness appear later; this highlights the importance of studying the effects of herbivory over consecutive years in perennial plants.

Direct and Pollinator-Mediated Effects of Herbivory on Strawberry and the Potential for Improved Resistance.

The global decline in pollinators has partly been blamed on pesticides, leading some to propose pesticide-free farming as an option to improve pollination. However, herbivores are likely to be more prevalent in pesticide-free environments, requiring knowledge of their effects on pollinators, and alternative crop protection strategies to mitigate any potential pollination reduction. Strawberry leaf beetles (SLB) Galerucella spp. are important strawberry pests in Northern Europe and Russia. Given that SLB attack both leaf and flower tissue, we hypothesized pollinators would discriminate against SLB-damaged strawberry plants (Fragaria vesca, cultivar 'Rügen'), leading to lower pollination success and yield. In addition we screened the most common commercial cultivar 'Rügen' and wild Swedish F. vesca genotypes for SLB resistance to assess the potential for inverse breeding to restore high SLB resistance in cultivated strawberry. Behavioral observations in a controlled experiment revealed that the local pollinator fauna avoided strawberry flowers with SLB-damaged petals. Low pollination, in turn, resulted in smaller more deformed fruits. Furthermore, SLB-damaged flowers produced smaller fruits even when they were hand pollinated, showing herbivore damage also had direct effects on yield, independent of indirect effects on pollination. We found variable resistance in wild woodland strawberry to SLB and more resistant plant genotypes than the cultivar 'Rügen' were identified. Efficient integrated pest management strategies should be employed to mitigate both direct and indirect effects of herbivory for cultivated strawberry, including high intrinsic plant resistance.

Preference for outbred host plants and positive effects of inbreeding on egg survival in a specialist herbivore.

Inbreeding can profoundly affect the interactions of plants with herbivores as well as with the natural enemies of the herbivores. We studied how plant inbreeding affects herbivore oviposition preference, and whether inbreeding of both plants and herbivores alters the probability of predation or parasitism of herbivore eggs. In a laboratory preference test with the specialist herbivore moth Abrostola asclepiadis and inbred and outbred Vincetoxicum hirundinaria plants, we discovered that herbivores preferred to oviposit on outbred plants. A field experiment with inbred and outbred plants that bore inbred or outbred herbivore eggs revealed that the eggs of the outbred herbivores were more likely to be lost by predation, parasitism or plant hypersensitive responses than inbred eggs. This difference did not lead to differences in the realized fecundity as the number of hatched larvae did not differ between inbred and outbred herbivores. Thus, the strength of inbreeding depression in herbivores decreases when their natural enemies are involved. Plant inbreeding did not alter the attraction of natural enemies of the eggs. We conclude that inbreeding can significantly alter the interactions of plants and herbivores at different life-history stages, and that some of these alterations are mediated by the natural enemies of the herbivores.

Plant-species diversity correlates with genetic variation of an oligophagous seed predator.

Several characteristics of habitats of herbivores and their food-plant communities, such as plant-species composition and plant quality, influence population genetics of both herbivores and their host plants. We investigated how different ecological and geographic factors affect genetic variation in and differentiation of 23 populations of the oligophagous seed predator Lygaeus equestris (Heteroptera) in southwestern Finland and in eastern Sweden. We tested whether genetic differentiation of the L. equestris populations was related to the similarity of vegetation, and whether there was more within-population genetic variation in habitats with a high number of plant species or in those with a large population of the primary food plant, Vincetoxicum hirundinaria. We also tested whether genetic differentiation of the populations was related to the geographic distance, and whether location of the populations on islands or on mainland, island size, or population size affected within-population genetic variation. Pairwise FST ranged from 0 to 0.1 indicating low to moderate genetic differentiation of populations. Differentiation increased with geographic distance between the populations, but was not related to the similarity of vegetation between the habitats. Genetic variation within the L. equestris populations did not increase with the population size of the primary food plant. However, the more diverse the plant community the higher was the level of genetic variation within the L. equestris population. Furthermore, the level of genetic variation did not vary significantly between island and mainland populations. The effect of the population size on within-population genetic variation was related to island size. Usually small populations are susceptible to loss of genetic variation, but small L. equestris populations on large islands seemed to maintain a relatively high level of within-population genetic variation. Our findings suggest that, in addition to geographic and species-specific ecological factors, the plant community affects population genetic structure of oligophagous herbivores.

Simultaneous inbreeding modifies inbreeding depression in a plant-herbivore interaction.

Because inbreeding is common in natural populations of plants and their herbivores, herbivore-induced selection on plants, and vice versa, may be significantly modified by inbreeding and inbreeding depression. In a feeding assay with inbred and outbred lines of both the perennial herb, Vincetoxicum hirundinaria, and its specialist herbivore, Abrostola asclepiadis, we discovered that plant inbreeding increased inbreeding depression in herbivore performance in some populations. The effect of inbreeding on plant resistance varied among plant and herbivore populations. The among-population variation is likely to be driven by variation in plant secondary compounds across populations. In addition, inbreeding depression in plant resistance was substantial when herbivores were outbred, but diminished when herbivores were inbred. These findings demonstrate that in plant-herbivore interactions expression of inbreeding depression can depend on the level of inbreeding of the interacting species. Furthermore, our results suggest that when herbivores are inbred, herbivore-induced selection against self-fertilisation in plants may diminish.