Eric Conti

Insect oviposition induces volatile emission in herbaceous plants that attracts egg parasitoids

SUMMARY The egg parasitoid Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae) responded to synomones emitted by leguminous plants induced by feeding and oviposition activity of the bug Nezara viridula (L.) (Heteroptera: Pentatomidae). This was shown by laboratory bioassays using a Y-tube olfactometer. Broad bean leaves (Vicia faba L.) damaged by feeding activity of N. viridula and on which host egg mass had been laid produced synomones that attracted T. basalis. By contrast, undamaged leaves or feeding-damaged leaves without eggs did not attract wasp females. French bean plants (Phaseolus vulgaris L.) also emitted attractive synomones when they were damaged by host feeding and carrying egg masses. Thus, release of feeding- and oviposition-induced synomones does not seem to be plant-specific. Synomone production was shown to be a systemically induced plant physiological response to feeding damage and oviposition. Also, parts of the plant that were left undamaged and did not carry host eggs emitted attractive synomones when other parts of the plant were damaged by feeding and carrying eggs. Furthermore, wasps were not attracted by N. viridula egg masses offered alone or combined with damaged broad bean leaves. Thus, the attractiveness of feeding-damaged leaves carrying eggs is due to induction by feeding and oviposition rather than due to a combined effect of attractive volatiles released from eggs and damaged leaves. The production of synomones was influenced by the age of the host egg mass, because feeding-damaged leaves bearing egg masses attracted the parasitoid until the eggs were ∼72-96 h old but not once the larvae had hatched from the eggs (∼120 h old). These results show that annual plants are able to produce synomones as a consequence of feeding and egg mass oviposition by a sucking insect.

Characterization of the Complex Locus of Bean Encoding Polygalacturonase-Inhibiting Proteins Reveals Subfunctionalization for Defense against Fungi and Insects

Polygalacturonase-inhibiting proteins (PGIPs) are extracellular plant inhibitors of fungal endopolygalacturonases (PGs) that belong to the superfamily of Leu-rich repeat proteins. We have characterized the full complement of pgip genes in the bean (Phaseolus vulgaris) genotype BAT93. This comprises four clustered members that span a 50-kb region and, based on their similarity, form two pairs (Pvpgip1/Pvpgip2 and Pvpgip3/Pvpgip4). Characterization of the encoded products revealed both partial redundancy and subfunctionalization against fungal-derived PGs. Notably, the pair PvPGIP3/PvPGIP4 also inhibited PGs of two mirid bugs (Lygus rugulipennis and Adelphocoris lineolatus). Characterization of Pvpgip genes of Pinto bean showed variations limited to single synonymous substitutions or small deletions. A three-amino acid deletion encompassing a residue previously identified as crucial for recognition of PG of Fusarium moniliforme was responsible for the inability of BAT93 PvPGIP2 to inhibit this enzyme. Consistent with the large variations observed in the promoter sequences, reverse transcription-PCR expression analysis revealed that the different family members differentially respond to elicitors, wounding, and salicylic acid. We conclude that both biochemical and regulatory redundancy and subfunctionalization of pgip genes are important for the adaptation of plants to pathogenic fungi and phytophagous insects.

Chemical Cues from Murgantia histrionica Eliciting Host Location and Recognition in the Egg Parasitoid Trissolcus brochymenae

Host location and recognition by the egg parasitoid Trissolcus brochymenae were analyzed in terms of response to kairomones from several stages of its host, Murgantia histrionica. In a Y-tube olfactometer, parasitoid females responded by increasing residence time and/or reducing linear speed to chemical cues from gravid females, virgin females and males, fifth and third instars, and eggs. In an open arena, T. brochymenae females also responded to patches contaminated by chemicals from the host in the same stages, sexes, and/or physiological conditions as those tested in the olfactometer. The parasitoid displayed arrestment behavior, increased residence time, changed walking pattern, and intense substrate examination. When host egg clusters or glass dummies with a chemical egg extract were placed on the host-contaminated open arena, these elicited an orientation response in the parasitoid. In addition, the chemical egg extract without dummies elicited the same response, whereas dummies without extract did not influence parasitoid behavior. In a closed arena, the parasitoid females recognized and attempted to probe glass beads treated with chemical extracts of host eggs. There were no significant differences compared with their response to the host eggs, and they did not respond to untreated beads. Host recognition was elicited by chemicals from the follicular secretion used by the host to glue the eggs on the substrate. These results are discussed in relation to the level of the host selection sequence influenced by these cues.

Short-range allelochemicals from a plant–herbivore association: a singular case of oviposition-induced synomone for an egg parasitoid

SUMMARY Oviposition-induced plant synomones are semiochemical cues used by egg parasitoids during host selection, and are therefore considered important elements of plant defence. In this paper we show that, in the tritrophic system Brassica oleracea–Murgantia histrionica–Trissolcus brochymenae, the latter responded in a closed arena and in a static olfactometer to induced chemicals that are perceived from a very short range and after parasitoid contact with the leaf surface opposite the treated surface. An additive or synergistic effect due to (1) egg deposition, (2) feeding punctures and (3) chemical footprints of M. histrionica was observed. When all three phases were present, the parasitoid reacted to the induced synomone locally on the treated leaf area, at a close distance to the treated area, and on the leaf above the treated one, showing that there is also a systemic effect. When plants with host footprints combined with feeding punctures or with oviposition were tested, responses were obtained both locally and at a close distance, whereas in the remaining assays only local responses were observed. Induction time was less than 24 h, whereas signal duration was apparently related to the suitability of the host eggs, as parasitoids did not respond to plants carrying old or hatched eggs. These oviposition-induced short-range plant synomones might have an important role in the host location process after parasitoid landing on the plant, in different combinations with the host kairomones involved in the system studied here.

Molecular Cloning and in Situ Expression Patterns of Two New Pheromone-Binding Proteins from the Corn Stemborer Sesamia nonagrioides

We describe the identification and characterization of two new cDNAs encoding pheromone-binding proteins (PBPs) from the male antennae of Sesamia nonagrioides, a species where no PBPs have been identified to date. Because PBPs are thought to participate in the first step of odor detection in a specific manner, we focused our investigation on this olfactory protein family using reverse transcription–polymerase chain reaction strategies. The deduced amino acid sequences of SnonPBP1 and SnonPBP2 revealed mature proteins of 142 and 143 amino acids, respectively, with six cysteine residues in conserved positions relative to other known PBPs. The alignment of the two mature S. nonagrioides PBPs with other noctuid PBPs showed high sequence identity (70–80%) with other full-length sequences from GenBank. Sequence identity between SnonPBP1 and SnonPBP2 was only 46%, suggesting that the two proteins belong to different classes of PBPs already described from the Noctuidae. Furthermore, analyses of expression patterns of SnonPBP1 and SnonPBP2 were performed by in situ hybridization on antennae of both sexes, and these studies revealed the expression of the two PBPs at the bases of olfactory sensilla (basiconica or trichodea) from both sexes. The possible binding properties of these two new PBPs are discussed according to their homologies with other known PBPs and S. nonagrioides pheromone components.

Vicia faba-Lygus rugulipennis Interactions : Induced Plant Volatiles and Sex Pheromone Enhancement

The profiles of volatile chemicals emitted by Vicia faba plants damaged by Lygus rugulipennis feeding, and by feeding plus oviposition, were shown to be quantitatively different from those released by undamaged plants. Samples of volatile chemicals collected from healthy plants, plants damaged by males as a consequence of feeding, plants damaged by females as a consequence of feeding and oviposition, plants damaged by feeding with mated males still present, and plants damaged by feeding and oviposition with gravid females still present, showed significant differences in the emission of hexyl acetate, (Z)-β-ocimene, (E)-β-ocimene, (E)-β-caryophyllene, and methyl salicylate. In particular, treatments with mated females present on plants had a significant increase in emission levels of the above compounds, possibly due to eggs laid within plant tissues or active feeding, compared with undamaged plants and plants damaged by males feeding, with or without insects still present. Furthermore, the pheromonal blend released by mated L. rugulipennis females, mainly comprising hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal, was enhanced when females were active on broad bean plants, whereas such an increase was not observed in males. Both sexes gave electroantennogram responses to green leaf volatiles from undamaged plants and to methyl salicylate and (E)-β-caryophyllene emitted by Lygus-damaged plants, suggesting that these compounds may be involved in colonization of host plants by L. rugulipennis. In addition, mated males and females were responsive to hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal released by mated females on V. faba, indicating that these substances could have a dual function as a possible aggregation pheromone in female–female communication, and as a sex pheromone in female–male communication.

Influence of feeding and oviposition by phytophagous pentatomids on photosynthesis of herbaceous plants.

Feeding by herbivorous insects may change photosynthetic activity of host plants. We studied how feeding and oviposition by herbivorous stink bugs, Murgantia histrionica and Nezara viridula (Heteroptera: Pentatomidae), affect photosynthetic parameters of Brassica oleracea (savoy cabbage) and Phaseolus vulgaris (French bean). First, we measured photosynthetic gas exchange, chlorophyll fluorescence imaging, and emission of induced volatile organic compounds (VOC) immediately after feeding and during a post-feeding period. Photosynthesis decreased rapidly and substantially in B. oleracea and P. vulgaris infested by feeding bugs. Stomatal conductance did not decrease proportionally with photosynthesis; instead, inhibition of photosynthesis likely was due to a reduced diffusion of CO2 in the mesophyll. We also measured the impact of oviposition per se and oviposition associated with feeding on photosynthetic parameters. A surprisingly large inhibition of photosynthesis was detected in cabbage leaves in response to oviposition by M. histrionica, even when oviposition was not associated with feeding activity. High resolution chlorophyll fluorescence imaging revealed that the damage to photochemistry caused by feeding and oviposition was restricted to the attacked areas. By contrast, the photochemical yield increased temporarily in the unaffected areas of the attacked leaves, indicating the onset of a compensatory response. Measurement of volatile organic compounds (VOC) revealed that feeding-damaged plants did not emit detectable amounts of VOC, indicating cellular damage (methanol and green leaf volatiles). However, feeding by M. histrionica induced emission of mono- and sesquiterpenes in savoy cabbage leaves. The different time-course of the induction of these two classes of terpenes may reflect the induction of two different biosynthetic pathways and indicate different roles of these terpenoids in tritrophic interactions.

Chemical Ecology of Egg Parasitoids Associated with True Bugs

Parasitoids representing some 15 families of Hymenoptera develop in insect eggs; three of these families, Platygastridae (= Scelionidae), Mymaridae, and Encyrtidae, are associated with Heteroptera. Several species of heteropteran egg parasitoids are or may be important for biological pest control. Successful parasitism of insect herbivores by insect parasitoids arises through several phases of host searching, which lead female wasps to the vicinity of, or in contact with, their hosts. During the host location process, females encounter and explore a variety of stimuli, among which chemical cues (i.e., semiochemicals or infochemicals) play a pivotal role. Female parasitoids are under selection pressure to efficiently invest their limited time on the location and exploitation of host-derived stimuli. In general, the levels of reliability and detectability of a particular stimulus are inversely correlated. Female parasitic wasps adopt differing strategies to solve this dilemma. In this paper we focus on the various host selection strategies employed by heteropteran egg parasitoids and possible means whereby the chemically mediated behavior of these wasps may be exploited to enhance biological pest control.

Role of the plant–conspecific complex in host location and intra‐specific communication of Lygus rugulipennis

Abstract Wind tunnel and vertical open Y‐shaped olfactometer studies are used to test whether volatile cues from the host plant (Vicia faba), from conspecific bugs, and from a plant–conspecifics combination, would elicit behavioural responses in mated males and females of Lygus rugulipennis. In the olfactometer, females move towards volatiles from healthy plants but they do not respond to volatiles released by oviposition‐ and/or feeding‐damaged plants without conspecifics, nor to conspecifics alone. Both in the wind tunnel and olfactometer, females respond to volatiles emitted by the plant–insect complex. By contrast, in the wind tunnel, both sexes move significantly towards damaged host plants, even if the presence of conspecifics on these plants enhances only the female response. However, the presence of eggs from conspecifics on host plants reduces the responses of both sexes in the wind tunnel. Finally, males, as well as females, are less responsive to conspecifics alone compared with damaged plants, especially when conspecifics are present on the host plants (host plant–Lygus complex). The results suggest that volatiles emitted by plants and conspecifics influence L. rugulipennis behaviour, giving information to both sexes on the presence of suitable host plants that have been colonized by other conspecifics acting as pioneers, or providing information on the presence of an already exploited host plant (presence of eggs), thus preventing competition. Males also can use this information to increase the probability of encountering mature females.

Host Searching by Egg Parasitoids: Exploitation of Host Chemical Cues

Insect parasitoids are considered “keystone species” in many ecosystems in terms of biodiversity, ecological impact and economic importance (Vinson 1985, LaSalle and Gauld 1993, Hawkins et al. 1999). In the last decades, several reviews have been published on the relationships among plants, hosts and parasitoids, which reflect a strong interest in these insects both as models for behavioral ecologists and as important organisms for classical and applied biological control programs (Hawkins et al. 1999, Vet 1999, Bale et al. 2008). The majority of these studies have considered the larval parasitoid s, besides the extensive use of egg parasitoids in biological control (Hawkins et al. 1999). Insect eggs can be parasitized by about 15 families of Hymenoptera parasitoids, among which several may have potential for biological control application, such as Aphelinidae, Encyrtidae, Eulophidae, Eupelmidae, Mymaridae, Platygastridae, Pteromalidae, Scelionidae, Tetracampidae and Trichogrammatidae (Bin 1994). Three families, Mymaridae, Scelionidae and Trichogrammatidae are exclusively composed of egg parasitoids, whereas the other families are represented by species developing in different host stages, and they also include egg-larval parasitoids, egg-prepupal parasitoids and egg predators (Bin 1994, Vinson 1994). In this chapter we will focus only on egg parasitoids. Successful parasitism of herbivores by insect parasitoids arises through several phases during host searching , which lead wasp females into the close vicinity/contact of their hosts (Vinson 1998). During the host location process, females encounter and explore a great variety of stimuli, among which the chemical cues , named semiochemicals or infochemicals , play a relevant role (Godfray 1994, Vet and Dicke 1992, Vinson 1998). Female parasitoids are under selection pressure to efficiently invest their limited time on the location and exploitation of host derived stimuli, so that the appropriateness and usability of semiochemicals could be influenced by their reliability in indicating host presence and by the degree to which stimuli can be detected, as explained by the reliability-detectability theory (Vet and Dicke 1992). In developing this theory, it was argued that the level of reliability and detectability of a particular stimulus is inversely correlated, e.g. cues from the hosts may be highly reliable, but are less detectable compared to volatiles from plants, which have a much larger biomass. To get through the reliability-detectability dilemma, wasp females can adopt three different strategies based on the exploitation of either: (1) cues originated from stages different from the one attacked (infochemical detour); (2) cues originated from the interaction of the plant and the herbivore (host-induced synomones); or (3) reliable but poorly detectable cues which were linked, through associative learning, with more detectable but unreliable cues (Vet and Dicke 1992).