Summary: Herbivore‐induced plant volatiles (HIPVs) are widely recognized as an ecologically important defensive response of plants against herbivory. Although the induction of this ‘cry for help’ has been well documented, only a few studies have investigated the inhibition of HIPVs by herbivores and little is known about whether herbivores have evolved mechanisms to inhibit the release of HIPVs. To examine the role of herbivore effectors in modulating HIPVs and stomatal dynamics, we conducted series of experiments combining pharmacological, surgical, genetic (CRISPR‐Cas9) and chemical (GC‐MS analysis) approaches. We show that the salivary enzyme, glucose oxidase (GOX), secreted by the caterpillarHelicoverpa zeaon leaves, causes stomatal closure in tomato (Solanum lycopersicum) within 5 min, and in both tomato and soybean (Glycine max) for at least 48 h. GOX also inhibits the emission of several HIPVs during feeding byH. zea, including (Z)‐3‐hexenol, (Z)‐jasmone and (Z)‐3‐hexenyl acetate, which are important airborne signals in plant defenses. Our findings highlight a potential adaptive strategy where an insect herbivore inhibits plant airborne defenses during feeding by exploiting the association between stomatal dynamics and HIPV emission.

Chemical cues are essential for many ecological interactions. Previous studies of chemically mediated multitrophic interactions have typically focused on responses to cues from plants or herbivores above‐ground. It is increasingly clear, however, that below‐ground cues and those produced by organisms at higher trophic levels also have ecological importance. Prey animals often avoid predator odours to improve survival, and previous research has documented enhanced plant resistance following contact with below‐ground natural enemies, though the ecological basis was unknown. Here, we investigated plant and insect responses to chemical cues from below‐ground natural enemies and explored the ecological significance of these cues for multitrophic interactions. More specifically, we examined the influence of odours emitted by entomopathogenic nematodes (EPNs), a natural enemy of insect herbivores, on the performance and behaviour of their insect prey and the defence responses of nearby plants. Our findings revealed that EPN‐infected insect cadavers emit a characteristic blend of volatile compounds with bioactivity in plants and insects. EPN chemical cues influenced both performance and preference of a specialist herbivore, Colorado potato beetle (CPB, Leptinotarsa decemlineata), feeding on its host plant, potato (Solanum tuberosum). CPB larvae consumed less leaf tissue and gained less mass feeding on plants exposed to EPN cues compared to control plants. Female CPBs laid fewer eggs on plants with EPN cues than on controls, indicating deterrence by EPN cues or EPN‐altered plant defences. Plant defences were enhanced by exposure to live EPNs or EPN chemical cues. Potato plants exposed to EPN infective juveniles induced higher amounts of the defence hormone salicylic acid (SA) and had higher expression of the pathogenisis‐related gene PR‐1(PR4) in foliar tissue. Exposing plants to EPN cues primed induction of SA and jasmonic acid in response to feeding damage by CPB larvae. These findings suggest that herbivores avoid cues from their EPN natural enemies and plants respond to the beneficial nematodes by enhancing systemic defences that reduce herbivore performance. This work has important implications for the chemical ecology of tritrophic interactions as we report that the third trophic level can play direct and indirect roles in plant defence. A plain language summary is available for this article.

There is increasing evidence that plant-associated microorganisms play important roles in shaping interactions between plants and insect herbivores. Studies of both pathogenic and beneficial plant microbes have documented wide-ranging effects on herbivore behavior and performance. Some studies, for example, have reported enhanced insect-repellent traits or reduced performance of herbivores on microbe-associated plants, while others have documented increased herbivore attraction or performance. Insect herbivores frequently rely on plant cues during foraging and oviposition, suggesting that plant-associated microbes affecting these cues can indirectly influence herbivore preference. We review and synthesize recent literature to provide new insights into the ways pathogenic and beneficial plant-associated microbes alter visual, olfactory, and gustatory cues of plants that affect host-plant selection by insect herbivores. We discuss the underlying mechanisms, ecological implications, and future directions for studies of plant-microbial symbionts that indirectly influence herbivore behavior by altering plant traits.

Above‐ground plant tissues produce characteristic blends of volatile compounds in response to insect herbivory. These herbivore‐induced plant volatiles (HIPVs) function in plant defence and mediate foraging decisions by herbivores and their natural enemies. The ecological roles of HIPVs as foraging cues for different trophic levels highlight an important conflict for herbivores that need to locate suitable host plants while avoiding competition and predation. Plant roots also emit HIPVs following herbivory, but our understanding of root‐produced volatiles and their ecological functions in soil environments remains limited. Moreover, recent studies have documented the effects of temporal dynamics of plant volatile production on ecological interactions, but little is known about how root HIPVs change throughout herbivory or the resulting ecological implications from such changes. In this study, we examined the roles of HIPVs from roots of cucumber plants Cucumis sativus as foraging cues for a specialist herbivore, striped cucumber beetle Acalymma vittatum and its natural enemies, entomopathogenic nematodes (EPNs). We predicted HIPVs from A. vittatum‐damaged roots would attract EPNs, while repelling conspecific larvae that avoid competition, induced plant defences and increased risk of predation by EPNs. To capture the temporal dynamics of root HIPVs, we determined how HIPV‐mediated interactions change over time with sustained herbivory. Initially (after 24 hr), A. vittatum herbivory on C. sativus, or mechanical wounding, induced greater production of root volatiles. These root HIPVs recruited EPNs and repelled foraging A. vittatum larvae, although larval performance was not affected by prior damage. Sustained (7 days) herbivory by larvae reduced HIPVs to levels indistinguishable from undamaged control roots while mechanically damaged roots continued to produce higher levels of volatiles. Attenuation of HIPVs impaired indirect defence responses of C. sativus by reducing recruitment of EPNs and deterrence of A. vittatum larvae. These results suggest that root HIPVs function as honest signals that indicate the presence of herbivores, induction of indirect plant defences and increased risk of predation by natural enemies. However, some herbivores may overcome this line of plant defence by attenuating production of HIPVs and thus altering the outcomes of subsequent interactions among plants, herbivores and natural enemies. A free Plain Language Summary can be found within the Supporting Information of this article.

Host-associated differentiation (HAD) refers to cases in which genetically distinct populations of a species (e.g., herbivores or natural enemies) preferentially reproduce or feed on different host species. In agroecosystems, HAD often results in unique strains or biotypes of pest species, each attacking different species of crops. However, HAD is not restricted to pest populations, and may cascade to the third trophic level, affecting host selection by natural enemies, and ultimately leading to HAD within natural enemy species. Natural enemy HAD may affect the outcomes of biological control efforts, whether classical, conservation, or augmentative. Here, we explore the potential effects of pest and natural enemy HAD on biological control in agroecosystems, with emphases on current knowledge gaps and implications of HAD for selection of biological control agents. Additionally, given the importance of semiochemicals in mediating interactions between trophic levels, we emphasize the role of chemical ecology in interactions between pests and natural enemies, and suggest areas of consideration for biological control. Overall, we aim to jump-start a conversation concerning the relevance of HAD in biological control by reviewing currently available information on natural enemy HAD, identifying challenges to incorporating HAD considerations into biological control efforts, and proposing future research directions on natural enemy selection and HAD.

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