Oviposition-induced Plant Volatiles Research Articles

The oviposition of cotton bollworms stimulates the defense against its eggs and larvae in tomato plants.

Herbivorous insects sustain their populations by oviposition. To reduce the damage caused by herbivores, the host plant triggers a defensive response upon detection of egg deposition. However, the specific impact of the egg deposition time of the cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae), on the tomato plant defense remains obscure. This study investigated the effects of tomato plant defenses on cotton bollworm eggs and larvae at different time intervals following egg deposition. The study revealed that tomato plant defense triggered by egg deposition did not directly affect the hatchability of the eggs. Nevertheless, it attracted Trichogramma chilonis 48 h after the egg deposition. Gas chromatography-mass spectrometry analysis of the oviposition-induced plant volatiles (OIPVs) revealed a considerable increase in the amount of α-pinene released by tomato plants 48 h after egg deposition. The olfactory results from Y-tube experiments showed that α-pinene exhibited a substantial attraction towards T. chilonis. In addition, it was found that the defense response induced by egg deposition for 24 and 48 h significantly inhibited the growth and development of the larvae. Metabolomics analysis revealed that the egg deposition of cotton bollworm altered the metabolite composition and caused significant modifications in the metabolic pathways of tomato plants. These findings provide novel insights into pest management by using egg-induced plant defenses to reduce egg hatching, and impede larval growth and development in herbivorous insects. © 2024 Society of Chemical Industry.

Root inoculation with beneficial soil microbes enhances indirect plant defences induced by insect feeding and egg deposition

Abstract Plants can respond to insect egg deposition by emitting oviposition‐induced plant volatiles (OIPVs) recruiting parasitoids. The recruitment of carnivore insects in response to egg deposition is considered an indirect defence strategy that is widespread in the plant kingdom. In recent years, there has been increasing evidence showing that microbial colonization can influence the strength of plant responses to insect herbivory, yet no information is available on how beneficial microbes modulate indirect defences induced by insect egg deposition. In this work, we evaluated the effects of inoculation with the beneficial soil fungus Trichoderma harzianum strain T22 on a tritrophic system consisting of tomato, the southern green stink bug Nezara viridula and its associated egg parasitoid Trissolcus basalis. We used Y‐tube olfactometer assays to evaluate the behavioural responses of the parasitoids to OIPVs emitted by plants colonized with beneficial soil microbes. We also used gas chromatography coupled with mass spectrometry (GC–MS) to investigate whether and how root inoculation with T. harzianum T22 affects the chemical composition of induced plant volatiles. In olfactometer assays, we found that root inoculation with T. harzianum T22 enhanced the attraction of the egg parasitoid towards tomato plants induced by N. viridula feeding and oviposition activities. In particular, the egg parasitoid preferred OIPVs emitted by tomato plants previously inoculated with T. harzianum T22 over OIPVs emitted by non‐inoculated plants. Furthermore, chemical analysis showed that root inoculation with T. harzianum T22 resulted in changes in the composition of OIPVs, which was consistent with the behavioural observations. Among the compounds that strongly contribute to the chemical differences between OIPVs from non‐inoculated and inoculated plants, chemical analysis identified green leaf volatiles ((Z)‐3‐hepten‐1‐ol, (E,E)‐2,4‐hexadienal), along with terpenoids (terpinen 4‐ol, α‐tujene and δ‐elemene). Taken together our results indicate that beneficial soil microbes enhance indirect plant defences induced by feeding and oviposition, broadening our understanding of plant responses to insect eggs. Our results underscore the importance of taking into account the role of microorganisms to fully comprehend the intricate interactions among plants, herbivore eggs and their associated egg parasitoids. Read the free Plain Language Summary for this article on the Journal blog.

Differential effects of plant-beneficial fungi on the attraction of the egg parasitoid Trissolcus basalisin response to Nezara viridula egg deposition.

There is increasing evidence that plant-associated microorganisms play important roles in defending plants against insect herbivores through both direct and indirect mechanisms. While previous research has shown that these microbes can modify the behaviour and performance of insect herbivores and their natural enemies, little is known about their effect on egg parasitoids which utilize oviposition-induced plant volatiles to locate their hosts. In this study, we investigated how root inoculation of sweet pepper (Capsicum annuum) with the plant-beneficial fungi Beauveria bassiana ARSEF 3097 or Trichoderma harzianum T22 influences the olfactory behaviour of the egg parasitoid Trissolcus basalis following egg deposition by its host Nezara viridula. Olfactometer assays showed that inoculation by T. harzianum significantly enhanced the attraction of the egg parasitoid, while B. bassiana had the opposite effect. However, no variation was observed in the chemical composition of plant volatiles. Additionally, fitness-related traits of the parasitoids (wasp body size) were not altered by any of the two fungi, suggesting that fungal inoculation did not indirectly affect host quality. Altogether, our results indicate that plant inoculation with T. harzianum T22 can be used to enhance attraction of egg parasitoids, which could be a promising strategy in manipulating early plant responses against pest species and improving sustainable crop protection. From a more fundamental point of view, our findings highlight the importance of taking into account the role of microorganisms when studying the intricate interactions between plants, herbivores and their associated egg parasitoids.

Insect oviposition in herbaceous plants attracts egg parasitoids despite fungal phytopathogen infection

Egg parasitoids are important natural enemies of several insect pests. The ability to kill the pest before it can inflict damage to the plant makes egg parasitoids ideal candidates for biological control. Several studies have shown that egg parasitoids exploit oviposition-induced plant volatiles (OIPVs) to locate host eggs laid on plant organs. Yet such studies have often overlooked that, in nature, plants frequently suffer concurrent attack by insect herbivores and phytopathogens. These dual attacks can modify the emission of induced plant volatiles, which may potentially interfere with the host location abilities of egg parasitoids. We investigated this research question using the following study organisms: the broad bean Vicia faba, the plant pathogen Stemphylium sp., the southern green stink bug Nezara viridula and its associated egg parasitoid Trissolcus basalis. We showed that T. basalis is able to exploit OPIVs in order to locate N. viridula egg masses even when V. faba plants were previously infected by Stemphylium sp. Chemical analyses indicate that the egg parasitoid ability to exploit OIPVs persists despite significant alterations of the volatile blends emitted by plants suffering multiple biotic stresses. This study highlights the importance of incorporating the complexity of multiple biotic stresses when studying parasitoid foraging behavior, in order to comprehend how to enhance the effectiveness of natural enemies in crop protection.

Unprecedented oviposition tactics avoid plant defences and reduce attack by parasitic wasps.

Female insects oviposit in sites suitable for the development of their offspring. The Oriental armyworm, Mythimna separata is a serious pest of various crops including wheat and prefers to oviposit on withered leaves rather than on fresh plant material, which is surprisingly different from other insects. Studies here showed that this oviposition tactic enables avoidance of wheat defence against eggs and emerged larvae. Intact plants responded to M. separata egg deposition by releasing oviposition-induced plant volatiles including acetophenone, tetradecene and pentadecane after 24 h. Acetophenone was identified as quantitatively accounting for the attraction of the egg parasitoid wasp (Trichogramma chilonis). Leaf jasmonic acid levels significantly increased after M. separata laid eggs, and primed the plant against emerging larvae. In addition, newly emerged M. separata larvae adopted a fast crawling behaviour and starvation tolerance compared with other noctuid larvae, which enhanced the survival of larvae on the withered leaves. The elucidation of this complex and surprising plant-insect interaction provides the first explanation for a herbivore laying eggs on withered leaves to avoid natural enemies and live-plant defence against emerging larvae.

Tea green leafhopper-induced synomone attracts the egg parasitoids, mymarids to suppress the leafhopper.

The tea green leafhopper, Empoasca flavescens is the most important pest of tea plants in China. Mymarid attractants based on herbivore-induced plant volatiles (HIPVs) from leafhopper feeding and oviposition-induced plant volatiles (OIPVs) were formulated and tested as a novel pest control agent against the leafhopper in tea plantations. Results showed that two mymarid species, Stethynium empoascae and Schizophragma parvula, had a reducing effect on leafhopper populations. The HIPVs and OIPVs were identified and bioassayed to screen the key synomones showing strong attraction to the mymarids. They were formulated into different blends, of which Field Attractant 1, comprising linalool, methyl salicylate, (E)-2-hexenal, perillen and α-farnesene at ratio of 1:2:3:58:146 (20 mg/lure), showed the strongest attraction to the mymarids. In field trials with the attractant, the average parasitism rate (60.46 ± 23.71%) of tea leafhoppers by the two mymarids in the attractant-baited area was significantly higher than that (42.85 ± 19.24%) in the CK area. Also, the average leafhopper density (46 ± 30 per 80 tea shoots) in the attractant-baited area was significantly lower than that (110 ± 70 per 80 tea shoots) in the CK area. This study showed that a synthetic blend of key volatiles from HIPVs and OIPVs at an optimal ratio can be formulated into an attractant with the potential to attract and retain wild mymarid populations to suppress leafhopper populations in infested tea plantations, so as to reduce or avoid the spraying of insecticides. © 2023 Society of Chemical Industry.

Attraction of Trichogramma Wasps to Butterfly Oviposition-Induced Plant Volatiles Depends on Brassica Species, Wasp Strain and Leaf Necrosis

Within the Brassicaceae, wild as well as crop species are challenged by specialist herbivores including cabbage white butterflies (Pieris spp.). The wild crucifer Brassica nigra responds to oviposition by Pieris butterflies by the synergistic expression of two egg-killing traits. Genotypes that express a hypersensitive response (HR)-like necrosis (direct egg-killing) also emit oviposition-induced plant volatiles (OIPVs) attracting Trichogramma egg parasitoids (indirect egg-killing). This so-called double defense line can result in high butterfly egg mortalities. It remains unknown whether this strategy is unique to B. nigra or more common in Brassica species. To test this, we examined the response of different Trichogramma evanescens lines to OIPVs emitted by B. nigra and three close relatives (Brassica napus, Brassica rapa, and Brassica oleracea). Furthermore, we evaluated whether HR-like necrosis played a role in the attraction toward plant volatiles. Our results show a specificity in wasp attraction to different plant species. Three out of four plant species attracted a specific T. evanescens strain, including the crops B. rapa and B. napus. Parasitoid attraction was positively affected by presence of HR-like necrosis in one plant species. Our findings imply that, despite being a true generalist in terms of host range, T. evanescens shows intraspecific variation during host searching, which should be taken into account when selecting parasitoid lines for biocontrol of certain crops. Finally, we conclude that also crop plants within the Brassicaceae family possess egg-killing traits and can exert the double-defense line which may enable effective selection of egg-killing defense traits by cabbage breeders.

Olfactory response of Trichogramma pretiosum (Hymenoptera: Trichogrammatidae) to volatiles induced by transgenic maize.

Plants not only respond to herbivorous damage but adjust their defense system after egg deposition by pest insects. Thereby, parasitoids use oviposition-induced plant volatiles to locate their hosts. We investigated the olfactory behavioral responses of Trichogramma pretiosum Riley, 1879 (Hymenoptera: Trichogrammatidae) to volatile blends emitted by maize (Zea mays L.) with singular and stacked events after oviposition by Spodoptera frugiperda Smith, 1797 (Hymenoptera: Trichogrammatidae) moths. Additionally, we examined possible variations in gene expression and on oviposition-induced volatiles. We used a Y-tube olfactometer to test for the wasp responses to volatiles released by maize plants oviposited by S. frugiperda and not-oviposited plants. Using the real-time PCR technique (qRT-PCR), we analyzed the expression of lipoxygenase and three terpene synthases genes, which are enzymes involved in the synthesis of volatile compounds that attract parasitoids of S. frugiperda. Olfactometer tests showed that T. pretiosum is strongly attracted by volatiles from transgenic maize emitted by S. frugiperda oviposition (VTPRO 3, more than 75% individuals were attracted). The relative expression of genes TPS10, LOX e STC was higher in transgenic hybrids than in the conventional (isogenic line) hybrids. The GC-MS analysis revealed that some volatile compounds are released exclusively by transgenic maize. This study provides evidence that transgenic hybrids enhanced chemical cues under oviposition-induction and helped to increase T. pretiosum efficiency in S. frugiperda control. This finding shows that among the evaluated hybrids, genetically modified hybrids can improve the biological control programs, since they potentialize the egg parasitoid foraging, integrating pest management.

Chemical, Physiological and Molecular Responses of Host Plants to Lepidopteran Egg-Laying.

Plant-lepidopteran interactions involve complex processes encompassing molecules and regulators to counteract defense responses they develop against each other. Lepidoptera identify plants for oviposition and exploit them as larval food sources to complete their development. In turn, plants adopt different strategies to overcome and limit herbivorous damages. The insect egg deposition on leaves can already induce a number of defense responses in several plant species. This minireview deals with the main features involved in the interaction between plants and lepidopteran egg-laying, focusing on responses from both insect and plant side. We discuss different aspects of direct and indirect plant responses triggered by lepidopteran oviposition. In particular, we focus our attention on the mechanisms underlying egg-induced plant defenses that can i) directly damage the eggs such as localized hypersensitive response (HR)-like necrosis, neoplasm formation, production of ovicidal compounds and ii) indirect defenses, such as production of oviposition-induced plant volatiles (OIPVs) used to attract natural enemies (parasitoids) able to kill the eggs or hatching larvae. We provide an overview of chemical, physiological, and molecular egg-mediated plant responses induced by both specialist and generalist lepidopteran species, also dealing with effectors, elicitors, and chemical signals involved in the process. Egg-associated microorganisms are also discussed, although little is known about this third partner participating in plant-lepidopteran interactions.

An Egg Parasitoid Efficiently Exploits Cues From a Coevolved Host But Not Those From a Novel Host.

Egg parasitoids have evolved adaptations to exploit host-associated cues, especially oviposition-induced plant volatiles and odors of gravid females, when foraging for hosts. The entire host selection process is critical for successful parasitism and relevant in defining host specificity of parasitoids. We hypothesized that naïve egg parasitoid females reared on their coevolved host are able to exploit cues related to the coevolved host but not those from a novel host. We used the egg parasitoid Trissolcus japonicus, its coevolved host Halyomorpha halys, and the non-coevolved host Podisus maculiventris to evaluate this hypothesis. H. halys, a polyphagous pest native from Eastern Asia, has invaded North America and Europe, resulting in serious damage to crops. T. japonicus is the most effective egg parasitoid of H. halys in its native area and thus considered a major candidate for biological control. This parasitoid was detected in North America and Europe as a result of accidental introductions. Laboratory host range of T. japonicus includes P. maculiventris, an American predatory stink bug used as a biological control agent of several pests. Using T. japonicus reared on its natural host H. halys, we tested in a Y-tube olfactometer the responses of naïve parasitoid females to volatiles from tomato plants with a deposited egg mass and feeding punctures of either H. halys or P. maculiventris. Additionally, using two different olfactometer set-ups, we tested T. japonicus responses to volatiles emitted by eggs and mature males and females of H. halys or P. maculiventris. Tomato plants subjected to oviposition and feeding by H. halys were preferred by the wasp compared to clean plants, suggesting a possible activation of an indirect defense mechanism. Furthermore, T. japonicus females were attracted by cues from gravid females and mature males of H. halys but not from eggs. By contrast, naïve parasitoid females never responded to cues associated with P. maculiventris, although this non-target host is suitable for complete parasitoid development. Such lack of responses might reduce the probability of T. japonicus locating and parasitizing P. maculiventris under field conditions. Our experimental approach properly simulates the parasitoid host-location process and could be combined with the required host specificity tests for risk assessment in biological control programs.

Mating Status of an Herbivorous Stink Bug Female Affects the Emission of Oviposition-Induced Plant Volatiles Exploited by an Egg Parasitoid.

Insect parasitoids are under selection pressure to optimize their host location strategy in order to maximize fitness. In parasitoid species that develop on host eggs, one of these strategies consists in the exploitation of oviposition-induced plant volatiles (OIPVs), specific blends of volatile organic compounds released by plants in response to egg deposition by herbivorous insects. Plants can recognize insect oviposition via elicitors that trigger OIPVs, but very few elicitors have been characterized so far. In particular, the source and the nature of the elicitor responsible of egg parasitoid recruitment in the case of plants induced with oviposition by stink bugs are still unknown. In this paper, we conducted behavioral and molecular investigations to localize the source of the elicitor that attracts egg parasitoids and elucidate the role of host mating in elicitation of plant responses. We used as organism study model a tritrophic system consisting of the egg parasitoid Trissolcus basalis, the stink bug host Nezara viridula and the plant Vicia faba. We found that egg parasitoid attraction to plant volatiles is triggered by extracts coming from the dilated portion of the stink bug spermathecal complex. However, attraction only occurs if extracts are obtained from mated females but not from virgin ones. Egg parasitoid attraction was not observed when extracts coming from the accessory glands (mesadene and ectadene) of male hosts were applied, either alone or in combination to plants. SDS-PAGE electrophoresis correlated with olfactometer observations as the protein profile of the dilated portion of the spermathecal complex was affected by the stink bug mating status suggesting post-copulatory physiological changes in this reproductive structure. This study contributed to better understanding the host location process by egg parasitoids and laid the basis for the chemical characterization of the elicitor responsible for OIPV emission.

An invasive insect herbivore disrupts plant volatile-mediated tritrophic signalling

Plants respond to insect attack by emission of volatile organic compounds, which recruit natural enemies of the attacking herbivore, constituting an indirect plant defence strategy. In this context, the egg parasitoid Trissolcus basalis is attracted by oviposition-induced plant volatiles emitted by Vicia faba plants as a consequence of feeding and oviposition by the pentatomid host Nezara viridula. However, this local tritrophic web could be affected by the recent invasion by the alien pentatomid bug Halyomorpha halys, an herbivore that shares the same environments as native pentatomid pests. Therefore, we investigated in laboratory conditions the possible impact of H. halys on the plant volatile-mediated signalling in the local tritrophic web V. faba–N. viridula–T. basalis. We found that T. basalis wasps were not attracted by volatiles induced in the plants by feeding and oviposition activities of H. halys, indicating specificity in the wasps’ response. However, the parasitoid attraction towards plant volatiles emitted as a consequence of feeding and oviposition by the associated host was disrupted when host, N. viridula, and non-associated host, H. halys, were concurrently present on the same plant, indicating that invasion by the alien herbivore interferes with established semiochemical webs. These outcomes are discussed in a context of multiple herbivory by evaluating the possible influences of alien insects on local parasitoid foraging behaviour.

Volatile-mediated foraging behaviour of three parasitoid species under conditions of dual insect herbivore attack

Infochemicals play an important role in structuring intra- and interspecific interactions. Many parasitoid wasp species rely on herbivory or oviposition-induced plant volatiles (HIPVs/OIPVs) to locate their herbivorous hosts, and must cope with variation in the volatile blends due to factors such as plant/host species, herbivore density or attack by several herbivores. However, little is known about how dual herbivory or changes in herbivore density affect multiple parasitoid species, each attacking a different herbivore, in the same system. In a natural system, we investigated the effect of dual attack on the ability of three parasitoid species to differentiate between volatiles induced by hosts and those induced by a combination of hosts and nonhosts. Black mustard, Brassica nigra, plants were infested with eggs or caterpillars of Pieris brassicae, alone or in combination with different densities of Brevicoryne brassicae aphids. We determined the ability of three different parasitoid species that parasitize either P. brassicae eggs (Trichogramma brassicae), caterpillars (Cotesia glomerata) or B. brassicae aphids (Diaeretiella rapae) to discriminate between the induced volatiles, and analysed the plant volatile blends. Dual infestation did not affect the parasitoid species equally and aphid infestation altered, in a density-dependent manner, the volatile-mediated foraging of all three parasitoid species. Chemical analyses of the volatile blends revealed nonlinear emission patterns in relation to aphid density in both plants attacked by aphids alone and in plants attacked by a combination of aphids and caterpillars. Simple correlations between behaviour and volatile emissions in pairwise comparisons suggest the importance of certain volatiles explaining attraction, whereas dose-response type analyses reveal that these simple correlation analyses provide an incomplete picture.

Role of Large Cabbage White butterfly male-derived compounds in elicitation of direct and indirect egg-killing defenses in the black mustard.

To successfully exert defenses against herbivores and pathogens plants need to recognize reliable cues produced by their attackers. Up to now, few elicitors associated with herbivorous insects have been identified. We have previously shown that accessory reproductive gland secretions associated with eggs of Cabbage White butterflies (Pieris spp.) induce chemical changes in Brussels sprouts plants recruiting egg-killing parasitoids. Only secretions of mated female butterflies contain minute amounts of male-derived anti-aphrodisiac compounds that elicit this indirect plant defense. Here, we used the black mustard (Brassica nigra) to investigate how eggs of the Large Cabbage White butterfly (Pieris brassicae) induce, either an egg-killing direct [i.e., hypersensitive response (HR)-like necrosis] or indirect defense (i.e., oviposition-induced plant volatiles attracting Trichogramma egg parasitoids). Plants induced by P. brassicae egg-associated secretions expressed both traits and previous mating enhanced elicitation. Treatment with the anti-aphrodisiac compound of P. brassicae, benzyl cyanide (BC), induced stronger HR when compared to controls. Expression of the salicylic (SA) pathway- and HR-marker PATHOGENESIS-RELATED GENE1 was induced only in plants showing an HR-like necrosis. Trichogramma wasps were attracted to volatiles induced by secretion of mated P. brassicae females but application of BC did not elicit the parasitoid-attracting volatiles. We conclude that egg-associated secretions of Pieris butterflies contain specific elicitors of the different plant defense traits against eggs in Brassica plants. While in Brussels sprouts plants anti-aphrodisiac compounds in Pieris egg-associated secretions were clearly shown to elicit indirect defense, the wild relative B. nigra, recognizes different herbivore cues that mediate the defensive responses. These results add another level of specificity to the mechanisms by which plants recognize their attackers.

Egg parasitoid attraction toward induced plant volatiles is disrupted by a non-host herbivore attacking above or belowground plant organs.

Plants respond to insect oviposition by emission of oviposition-induced plant volatiles (OIPVs) which can recruit egg parasitoids of the attacking herbivore. To date, studies demonstrating egg parasitoid attraction to OIPVs have been carried out in tritrophic systems consisting of one species each of plant, herbivore host, and the associated egg parasitoid. Less attention has been given to plants experiencing multiple attacks by host and non-host herbivores that potentially could interfere with the recruitment of egg parasitoids as a result of modifications to the OIPV blend. Egg parasitoid attraction could also be influenced by the temporal dynamics of multiple infestations, when the same non-host herbivore damages different organs of the same plant species. In this scenario we investigated the responses of egg parasitoids to feeding and oviposition damage using a model system consisting of Vicia faba, the above-ground insect herbivore Nezara viridula, the above- and below-ground insect herbivore Sitona lineatus, and Trissolcus basalis, a natural enemy of N. viridula. We demonstrated that the non-host S. lineatus disrupts wasp attraction toward plant volatiles induced by the host N. viridula. Interestingly, V. faba damage inflicted by either adults (i.e., leaf-feeding) or larvae (i.e., root-feeding) of S. lineatus, had a similar disruptive effect on T. basalis host location, suggesting that a common interference mechanism might be involved. Neither naïve wasps or wasps with previous oviposition experience were attracted to plant volatiles induced by N. viridula when V. faba plants were concurrently infested with S. lineatus adults or larvae. Analysis of the volatile blends among healthy plants and above-ground treatments show significant differences in terms of whole volatile emissions. Our results demonstrate that induced plant responses caused by a non-host herbivore can disrupt the attraction of an egg parasitoid to a plant that is also infested with its hosts.

Olfactory Attraction of the Larval Parasitoid,Hyposoter horticola, to Plants Infested with Eggs of the Host Butterfly,Melitaea cinxia

Parasitoids locate inconspicuous hosts in a heterogeneous habitat using plant volatiles, some of which are induced by the hosts. Hyposoter horticola Gravenhost (Hymenoptera: Ichneumonidae) is a parasitoid of the Glanville fritillary butterfly Melitaea cinxia L. (Lepidoptera: Nymphalidae). Melitaea cinxia lays eggs in clusters on leaves of Plantago lanceolata L. (Lamiales: Plantaginaceae) and Veronica spicata L. (Lamiales: Plantaginaceae). The parasitoid oviposits into host larvae that have not yet hatched from the egg. Thus, though H. horticola is a parasitoid of Melitaea cinxia larvae, it must find host eggs on plants that have not been fed on by the larvae. Using a Y-tube olfactometer, the response of H. horticola to odors of Melitaea cinxia and extracts of the attacked plant species were tested. Three week-old eggs (near hatching) were attractive to young H. horticola, but one week-old eggs were attractive only to old or experienced H. horticola. Melitaea cinxia larvae were not attractive. A water extract of P. lanceolata was attractive, but ethanol or hexane extracts were not. None of the extracts of V. spicata were attractive. Leaves of V. spicata were attractive only if harboring eggs, but P. lanceolata leaves with eggs were not. Free flying H. horticola in a large outdoor enclosure were presented with host and plant cues. As in the olfactometer, V. spicata was attractive only when eggs were on it, and P. lanceolata was somewhat attractive with or without eggs. This study shows for the first time that a parasitoid of larvae uses egg volatiles or oviposition-induced plant volatiles, to find host larvae, and that Melitaea cinxia eggs or traces of oviposition induce the production of these volatiles by the plant. Based on the results, and given the natural distribution of the plants and M. cinxia eggs, parasitism of Melitaea cinxia eggs on P. lanceolata would be expected to be low. Instead, under natural conditions, a fraction of the eggs in virtually all egg clusters are parasitized on both plant species. The mismatch between the experimental results and the natural pattern of host-parasitoid interactions is discussed in terms of the expected coupling foraging cues with foraging success.

Unusual mechanisms involved in learning of oviposition-induced host plant odours in an egg parasitoid?

Parasitoids of herbivorous larvae are well known to be able to associatively learn a response to feeding-induced plant volatiles. However, little is known of the abilities of egg parasitoids to learn a response to herbivore oviposition-induced plant volatiles (OVIV). The eulophid wasp Chrysonotomyia ruforum is known to be attracted by OVIV from Scots pine released 3 days after host egg deposition. This study aimed to elucidate (1) the impact of the nutritional state of the parasitoid on its response to OVIV and (2) the effect of learning and response to the temporal change of OVIV. Supply of food as a reward during associative learning trials inevitably influences the nutritional state and thus leads to an experimental constraint. Nevertheless, the results indicate that the parasitoids have to associate OVIV with host presence and that they need to be supplied with food to learn and to respond to OVIV. Furthermore, the parasitoids' positive response to OVIV from pine twigs laden with eggs for 3 days was independent of the induction time of pine twigs experienced. However, a positive response was given only to OVIV from pine twigs with 3-day-old eggs, while OVIV from twigs with other induction times (1, 2, 4 days) were not attractive. Thus, our findings indicate an unusual learning process in this specialized egg parasitoid. We suggest that the parasitoid is using a learning strategy by which common information is filtered out of similar odour blends (here: 1- to 4-day-long induced pine twigs), but a behavioural response to this learned information becomes evident only in a specific odour context (here: 3-day-long induced twigs).

Early Herbivore Alert: Insect Eggs Induce Plant Defense

Plants are able to "notice" insect egg deposition and to respond by activating direct and indirect defenses. An overview of these defenses and the underlying mechanisms is given from a tritrophic perspective. First, the interface between plant and eggs is addressed with respect to the mode of attachment of eggs on the plant surface. It is elucidated which plant cells might respond to components from insect eggs or the egg deposition. The scarce knowledge on the elicitors associated with the eggs or the egg-laying female is outlined. Since endosymbiotic microorganisms are often present on the eggs, and microorganisms are also abundant on the leaf surface, the role of these hidden players for eliciting oviposition-induced plant responses is considered. Furthermore, the question of which physiological and molecular processes are induced within the plant in response to egg deposition is addressed. Second, studies on the response of the herbivorous insect to oviposition-induced plant defenses are outlined. Third, the importance of oviposition-induced plant volatiles and contact cues for host and prey location of parasitoids and predators is discussed in the context of other informative chemicals used by carnivores when searching for food. Finally, physiological and ecological costs of oviposition-induced plant responses are addressed.