Novel pheromone-mediated reproductive behaviour in the stag beetle, Lucanus cervus.

The iconic European stag beetle (Lucanus cervus) (Coleoptera: Lucanidae) is one of the largest terrestrial beetles in Europe. Due to decreasing population numbers, thought to be a consequence of habitat loss, this beetle has become a near-threatened species across much of Europe, and a reliable monitoring system is required to measure its future population trends. As part of a programme aimed at conserving UK populations, we have investigated the chemical ecology of the beetle, with a view to developing an efficient semiochemical-based monitoring system. Such a scheme will be beneficial not only in the UK but across the European range of the species, where the beetle is of conservation concern. Here, we report on a surprising discovery of a male-produced pheromone, which provokes initial sexual receptivity in females, and which has not been previously identified in the animal kingdom. Furthermore, we assign sex pheromone function to a previously described female-specific compound.

Bumblebee electric charge stimulates floral volatile emissions in Petunia integrifolia but not in Antirrhinum majus.

The timing of volatile organic compound (VOC) emission by flowering plants often coincides with pollinator foraging activity. Volatile emission is often considered to be paced by environmental variables, such as light intensity, and/or by circadian rhythmicity. The question arises as to what extent pollinators themselves provide information about their presence, in keeping with their long co-evolution with flowering plants. Bumblebees are electrically charged and provide electrical stimulation when visiting plants, as measured via the depolarisation of electric potential in the stem of flowers. Here we test the hypothesis that the electric charge of foraging bumblebees increases the floral volatile emissions of bee pollinated plants. We investigate the change in VOC emissions of two bee-pollinated plants (Petunia integrifolia and Antirrhinum majus) exposed to the electric charge typical of foraging bumblebees. P. integrifolia slightly increases its emissions of a behaviorally and physiologically active compound in response to visits by foraging bumblebees, presenting on average 121 pC of electric charge. We show that for P. integrifolia, strong electrical stimulation (600-700 pC) promotes increased volatile emissions, but this is not found when using weaker electrical charges more representative of flying pollinators (100 pC). Floral volatile emissions of A. majus were not affected by either strong (600-700 pC) or weak electric charges (100 pC). This study opens a new area of research whereby the electrical charge of flying insects may provide information to plants on the presence and phenology of their pollinators. As a form of electroreception, this sensory process would bear adaptive value, enabling plants to better ensure that their attractive chemical messages are released when a potential recipient is present.

Chemical Identity and Functional Characterization of Semiochemicals That Promote the Interactions between Rice Plant and Rice Major Pest Nilaparvata lugens.

The interaction between food crops and insect pests is mediated by semiochemicals emitted from host plants. These semiochemicals are natural behavioral modifiers and act on the insect olfactory system to locate hosts and preys. In this study, eight rice neuroactive semiochemicals were identified from rice varieties by GC-EAG and GC-MS. Their ability to modify rice pest behaviors was further studied as individual chemicals and physiologically relevant blend. The total amount of each semiochemical and the expression of their biosynthesis genes were significantly higher in pest susceptible variety than in pest-resistant variety and upregulated by the infestation of the pest Nilaparvata lugens (BPH). The semiochemicals emitted by uninfested plants (UIRVs) were more attractive to BPHs. Interestingly, the attractiveness of UIRVs was significantly reduced by the addition of the blend that mimics the natural composition of these semiochemicals emitted by infested plants (IRVs). Our study suggests a mechanism for the spread of pest infestation from infested plants to uninfested plants nearby. UIRVs initially serve as attractive signals to rice insect pests. The pest infestation changes the rice semiochemical profile to be less attractive or even repellent, which pushes further colonization to uninfested plants nearby. The identified semiochemicals can be used for crop protection based on a push-pull strategy.

Three Aphid-Transmitted Viruses Encourage Vector Migration From Infected Common Bean (Phaseolus vulgaris) Plants Through a Combination of Volatile and Surface Cues.

Bean common mosaic virus (BCMV), bean common mosaic necrosis virus (BCMNV), and cucumber mosaic virus (CMV) are important pathogens of common bean (Phaseolus vulgaris), a crop vital for food security in sub-Saharan Africa. These viruses are vectored by aphids non-persistently, with virions bound loosely to stylet receptors. These viruses also manipulate aphid-mediated transmission by altering host properties. Virus-induced effects on host-aphid interactions were investigated using choice test (migration) assays, olfactometry, and analysis of insect-perceivable volatile organic compounds (VOCs) using gas chromatography (GC)-coupled mass spectrometry, and GC-coupled electroantennography. When allowed to choose freely between infected and uninfected plants, aphids of the legume specialist species Aphis fabae, and of the generalist species Myzus persicae, were repelled by plants infected with BCMV, BCMNV, or CMV. However, in olfactometer experiments with A. fabae, only the VOCs emitted by BCMNV-infected plants repelled aphids. Although BCMV, BCMNV, and CMV each induced distinctive changes in emission of aphid-perceivable volatiles, all three suppressed emission of an attractant sesquiterpene, α-copaene, suggesting these three different viruses promote migration of virus-bearing aphids in a similar fashion.

Responses of the putative trachoma vector, Musca sorbens, to volatile semiochemicals from human faeces.

The putative vector of trachoma, Musca sorbens, prefers to lay its eggs on human faeces on the ground. This study sought to determine whether M. sorbens females were attracted to volatile odours from human faeces in preference to odours from the faeces of other animals, and to determine whether specific volatile semiochemicals mediate selection of the faeces. Traps baited with the faeces of humans and local domestic animals were used to catch flies at two trachoma-endemic locations in The Gambia and one in Ethiopia. At all locations, traps baited with faeces caught more female M. sorbens than control traps baited with soil, and human faeces was the most successful bait compared with soil (mean rate ratios 44.40, 61.40, 10.50 [P<0.001]; 8.17 for child faeces [P = 0.004]). Odours from human faeces were sampled by air entrainment, then extracts of the volatiles were tested by coupled gas chromatography-electroantennography with laboratory-reared female M. sorbens. Twelve compounds were electrophysiologically active and tentatively identified by coupled mass spectrometry-gas chromatography, these included cresol, indole, 2-methylpropanoic acid, butanoic acid, pentanoic acid and hexanoic acid. It is possible that some of these volatiles govern the strong attraction of M. sorbens flies to human faeces. If so, a synthetic blend of these chemicals, at the correct ratios, may prove to be a highly attractive lure. This could be used in odour-baited traps for monitoring or control of this species in trachoma-endemic regions.

Identification of Semiochemicals from Cowpea, Vigna unguiculata, for Low-input Management of the Legume Pod Borer, Maruca vitrata.

Cowpea, Vigna unguiculata L. Walp. (Fabaceae), is one of the most important food legumes grown on the African continent, as it provides an affordable source of dietary protein. Yields of cowpea are significantly reduced through damage by legume pod-borer, Maruca vitrata (Lepidoptera: Crambidae), caterpillars to flowers, tender leaves and pods. Semiochemical-based strategies are considered as environmentally benign and affordable for pest management, particularly on smallholder farms. In this study, we investigated the importance of cowpea flower volatiles as host location cues for egg-laying M. vitrata, and herbivore-induced plant volatiles (HIPVs) as M. vitrata repellents and natural enemy (Apanteles taragamae and Phanerotoma syleptae parasitoid) attractants. In oviposition choice assays, M. vitrata laid more eggs on flowering cowpea plants than non-flowering plants. Coupled gas chromatography-electrophysiology (GC-EAG) analysis using the antennae of female M. vitrata and an extract of flower volatiles collected by dynamic headspace collection revealed the presence of five EAG-active components that were identified by coupled GC-mass spectrometry (GC-MS) analysis as benzaldehyde, benzyl alcohol, acetophenone, a vinylbenzaldehyde isomer and (E)-cinnamaldehyde. A synthetic blend of the identified compounds, prepared using 3-vinylbenzaldehyde, induced M. vitrata to lay as many eggs on non-flowering cowpea as on flowering plants. The moths also preferred laying eggs on intact plants compared to M. vitrata-infested plants. As the emission of EAG-active floral compounds was determined to be lower in the headspace of infested cowpea flowers, the role of HIPVs emitted by M. vitrata-damaged leaves was also investigated. Of the compounds induced by larval damage, (E)-DMNT, indole, n-hexyl acetate, 1-octen-3-ol and linalool were shown by GC-EAG to possess electrophysiological activity. A synthetic blend of the EAG-active compounds, using racemic 1-octen-3-ol and linalool, significantly reduced egg numbers on flowering cowpea. Larval and egg parasitoids, i.e. A. taragamae and Ph. syleptae, respectively, of M. vitrata both preferred the Y-tube olfactometer arm treated with synthetic (E)-DMNT, whereas preference for racemic linalool and (E)-nerolidol was dose-dependent in A. taragamae. Our results provide the platform for the development of future semiochemical-based pest management strategies against M. vitrata on smallholder farms in West Africa.

Molasses Grass Induces Direct and Indirect Defense Responses in Neighbouring Maize Plants.

Plants have evolved intricate defence strategies against herbivore attack which can include activation of defence in response to stress-related volatile organic compounds (VOCs) emitted by neighbouring plants. VOCs released by intact molasses grass (Melinis minutiflora), have been shown to repel stemborer, Chilo partellus (Swinhoe), from maize and enhance parasitism by Cotesia sesamiae (Cameron). In this study, we tested whether the molasses grass VOCs have a role in plant-plant communication by exposing different maize cultivars to molasses grass for a 3-week induction period and then observing insect responses to the exposed plants. In bioassays, C. partellus preferred non-exposed maize landrace plants for egg deposition to those exposed to molasses grass. Conversely, C. sesamiae parasitoid wasps preferred volatiles from molasses grass exposed maize landraces compared to volatiles from unexposed control plants. Interestingly, the molasses grass induced defence responses were not observed on hybrid maize varieties tested, suggesting that the effect was not simply due to absorption and re-emission of VOCs. Chemical and electrophysiological analyses revealed strong induction of bioactive compounds such as (R)-linalool, (E)-4,8-dimethyl-1,3,7-nonatriene and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene from maize landraces exposed to molasses grass volatiles. Our results suggest that constitutively emitted molasses grass VOCs can induce direct and indirect defence responses in neighbouring maize landraces. Plants activating defences by VOC exposure alone could realize enhanced levels of resistance and fitness compared to those that launch defence responses upon herbivore attack. Opportunities for exploiting plant-plant signalling to develop ecologically sustainable crop protection strategies against devastating insect pests such as stemborer C. partellus are discussed.

Different Plant Viruses Induce Changes in Feeding Behavior of Specialist and Generalist Aphids on Common Bean That Are Likely to Enhance Virus Transmission.

Bean common mosaic virus (BCMV), bean common mosaic necrosis virus (BCMNV), and cucumber mosaic virus (CMV) cause serious epidemics in common bean (Phaseolus vulgaris), a vital food security crop in many low-to-medium income countries, particularly in Sub-Saharan Africa. Aphids transmit these viruses "non-persistently," i.e., virions attach loosely to the insects' stylets. Viruses may manipulate aphid-host interactions to enhance transmission. We used direct observation and electrical penetration graph measurements to see if the three viruses induced similar or distinct changes in feeding behaviors of two aphid species, Aphis fabae and Myzus persicae. Both aphids vector BCMV, BCMNV, and CMV but A. fabae is a legume specialist (the dominant species in bean fields) while M. persicae is a generalist that feeds on and transmits viruses to diverse plant hosts. Aphids of both species commenced probing epidermal cells (behavior optimal for virus acquisition and inoculation) sooner on virus-infected plants than on mock-inoculated plants. Infection with CMV was especially disruptive of phloem feeding by the bean specialist aphid A. fabae. A. fabae also experienced mechanical stylet difficulty when feeding on virus-infected plants, and this was also exacerbated for M. persicae. Overall, feeding on virus-infected host plants by specialist and generalist aphids was affected in different ways but all three viruses induced similar effects on each aphid type. Specifically, non-specialist (M. persicae) aphids encountered increased stylet difficulties on plants infected with BCMV, BCMNV, or CMV, whereas specialist aphids (A. fabae) showed decreased phloem ingestion on infected plants. Probing and stylet pathway activity (which facilitate virus transmission) were not decreased by any of the viruses for either of the aphid species, except in the case of A. fabae on CMV-infected bean, where these activities were increased. Overall, these virus-induced changes in host-aphid interactions are likely to enhance non-persistent virus transmission, and data from this work will be useful in epidemiological modeling of non-persistent vectoring of viruses by aphids.

Attractiveness of host banana leaf materials to the banana weevil, Cosmopolites sordidus in Ghana for development of field management strategies.

BACKGROUND: The banana weevil, Cosmopolites sordidus, has been frequently cited as the most challenging constraint to banana and plantain production, particularly in small-scale (smallholder) farming. For the development of a new, low-cost weevil management technology based on attractive host plant material, we previously identified (2R,5S)-theaspirane as the active component of attractive senesced banana leaves. In this new study, we used behavioural (olfactometer) bioassays with adult weevils to compare the attractiveness of four different developmental stages of banana leaves, i.e. unfolding (pale green), matured green (deep green), matured yellowing and senesced, to determine which leaf developmental stage would be most appropriate for use in weevil management. We also investigated the attractiveness of senesced leaf extracts prepared using different solvents to determine which solvent would be most appropriate for local production of leaf extracts. Coupled gas chromatography-electroantennography (GC-EAG) was then used with adult weevils to confirm the presence of (2R,5S)-theaspirane in attractive leaf extracts. RESULTS: Of the leaf materials tested, only the odour of senesced leaf material was significantly attractive to adult weevils (P < 0.005). Furthermore, an extract of senesced material prepared using palm wine alcohol was significantly attractive (P < 0.05). Using coupled GC-EAG with weevil antennae, (2R,5S)-theaspirane was identified as a minor component with strong EAG activity within the palm wine alcohol extract. CONCLUSION: The results suggest that palm wine alcohol extracts of senesced banana leaf material could be used to lure adult C. sordidus to traps in the field, as part of an ethnobotanical-based approach for C. sordidus management on smallholder farms. © 2018 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Environmentally vulnerable noble chafers exhibit unusual pheromone-mediated behaviour.

Conserving populations of environmentally vulnerable insect species requires a greater understanding of the factors that determine their abundance and distribution, which requires detailed knowledge of their population and community ecology. Chemical ecological tools such as pheromones can be used for non-destructive monitoring of scarab beetle populations, enabling European countries to detect and, in some cases, map the range of some of these species, proving a valuable technique for monitoring elusive saproxylic beetles. In this paper, we investigated the behavioural and chemical ecology of the noble chafer, Gnorimus nobilis L., a model insect species of conservation concern across a Europe-wide distribution, and a red-listed UK Biodiversity Action Plan priority species. We identified a potential pheromone of adult beetles using electrophysiological recordings, behavioural measurements and field trials in the UK. Gnorimus nobilis is highly unusual in that although both sexes produce, at high metabolic cost, the natural product 2-propyl (E)-3-hexenoate, it only attracts males. This pattern of chemical signalling makes the classification of the compound, based on current semiochemical terminology, somewhat problematic, but in our view, it should be termed an aggregation pheromone as a consequence of the production pattern. Since both sexes emit it, but apparently only males respond positively to it, 2-propyl (E)-3-hexenoate may reflect an intermediate evolutionary stage towards developing into a sex-specific signal. From an applied perspective, our study provides a model for the non-invasive surveillance of cryptic vulnerable insect species, without the need for habitat searching or disturbance, and continuous human monitoring.

Host shift induces changes in mate choice of the seed predator Acanthoscelides obtectus via altered chemical signalling.

The mechanisms of host shift in phytophagous insects are poorly understood. Among the many proposed processes involved, sexual selection via semiochemicals has recently been suggested. This hypothesizes that sexual communication using pheromones is modified as a result of development on a new host, and such plant-induced phenotypic divergence in mate recognition cues can lead to reproductive isolation between host lines. We tested this hypothesis on Acanthoscelides obtectus, an oligophagous bruchid of Phaseolus vulgaris beans worldwide, which also develops in acceptable non-hosts, such as chickpea (Cicer arietinum L.). Male sex pheromone blends of the bean, chickpea and chickpea/bean host lines during artificially induced host shifts showed different composition. Bean-reared females did not distinguish between blends, whereas chickpea and chickpea/bean females preferred the chickpea male pheromone. However, electrophysiological (EAG) responses to male odour of antennae of the three female host lines were similar, all preferring bean-reared males. Egg-laying choice tests revealed a uniform preference for bean seeds across female host lines, even after multiple generations, whereas larvae did not distinguish between bean and chickpea seeds. We conclude that the development of divergent chemical signalling systems during host shifts does not facilitate the evolution of host races in A. obtectus, because oviposition preferences remain unaffected.

(2R,5S)-Theaspirane Identified as the Kairomone for the Banana Weevil, Cosmopolites sordidus, from Attractive Senesced Leaves of the Host Banana, Musa spp.

The principal active component produced by highly attractive senesced host banana leaves, Musa spp., for the banana weevil, Cosmopolites sordidus, is shown by coupled gas chromatography-electroantennography (GC-EAG), coupled GC-mass spectrometry (GC-MS), chemical synthesis and coupled enantioselective (chiral) GC-EAG to be (2R,5S)-theaspirane. In laboratory behaviour tests, the synthetic compound is as attractive as natural host leaf material and presents a new opportunity for pest control.

Isolation and identification of floral attractants from a nectar plant for the dried bean beetle, Acanthoscelides obtectus (Coleoptera: Chrysomelidae, Bruchinae).

BACKGROUND: The response of virgin females of the legume pest Acanthoscelides obtectus (Coleoptera: Bruchidae) to headspace extracts of volatiles collected from flowers of a nectar plant, Daucus carota, was investigated using behaviour (four-arm olfactometry) and coupled gas chromatography-electroantennography (GC-EAG). RESULTS: Odours from inflorescences were significantly more attractive to virgin female beetles than clean air. Similarly, a sample of volatile organic compounds (VOCs) collected by air entrainment (dynamic headspace collection) was more attractive to beetles than a solvent control. In coupled GC-EAG experiments with beetle antennae and the VOC extract, six components showed EAG activity. Using coupled GC-mass spectrometry (GC-MS) and GC peak enhancement with authentic standards, the components were identified as α-pinene (S:R 16:1), sabinene, myrcene, limonene (S:R 1:3), terpinolene and (S)-bornyl acetate. Females preferred the synthetic blend of D. carota EAG-active volatiles to the solvent control in bioassays. When compared directly, odours of D. carota inflorescences elicited stronger positive behaviour than the synthetic blend. CONCLUSION: This is the first report of behaviourally active volatiles linked to pollen location for A. obtectus, and development of the six-component blend is being pursued, which could underpin the design of semiochemical-based field management approaches against this major pest of stored products. © 2018 Society of Chemical Industry.

A maize landrace that emits defense volatiles in response to herbivore eggs possesses a strongly inducible terpene synthase gene.

Maize (Zea mays) emits volatile terpenes in response to insect feeding and egg deposition to defend itself against harmful pests. However, maize cultivars differ strongly in their ability to produce the defense signal. To further understand the agroecological role and underlying genetic mechanisms for variation in terpene emission among maize cultivars, we studied the production of an important signaling component (E)-caryophyllene in a South American maize landrace Braz1006 possessing stemborer Chilo partellus egg inducible defense trait, in comparison with the European maize line Delprim and North American inbred line B73. The (E)-caryophyllene production level and transcript abundance of TPS23, terpene synthase responsible for (E)-caryophyllene formation, were compared between Braz1006, Delprim, and B73 after mimicked herbivory. Braz1006-TPS23 was heterologously expressed in E. coli, and amino acid sequences were determined. Furthermore, electrophysiological and behavioral responses of a key parasitic wasp Cotesia sesamiae to C. partellus egg-induced Braz1006 volatiles were determined using coupled gas chromatography electroantennography and olfactometer bioassay studies. After elicitor treatment, Braz1006 released eightfold higher (E)-caryophyllene than Delprim, whereas no (E)-caryophyllene was detected in B73. The superior (E)-caryophyllene production by Braz1006 was positively correlated with high transcript levels of TPS23 in the landrace compared to Delprim. TPS23 alleles from Braz1006 showed dissimilarities at different sequence positions with Delprim and B73 and encodes an active enzyme. Cotesia sesamiae was attracted to egg-induced volatiles from Braz1006 and synthetic (E)-caryophyllene. The variation in (E)-caryophyllene emission between Braz1006 and Delprim is positively correlated with induced levels of TPS23 transcripts. The enhanced TPS23 activity and corresponding (E)-caryophyllene production by the maize landrace could be attributed to the differences in amino acid sequence with the other maize lines. This study suggested that the same analogous genes could have contrasting expression patterns in different maize genetic backgrounds. The current findings provide valuable insight not only into genetic mechanisms underlying variation in defense signal production but also the prospect of introgressing the novel defense traits into elite maize varieties for effective and ecologically sound protection of crops against damaging insect pests.

cis-Jasmone Elicits Aphid-Induced Stress Signalling in Potatoes.

Elicitation of plant defense signaling that results in altered emission of volatile organic compounds (VOCs) offers opportunities for protecting plants against arthropod pests. In this study, we treated potato, Solanum tuberosum L., with the plant defense elicitor cis-jasmone (CJ), which induces the emission of defense VOCs and thus affects the behavior of herbivores. Using chemical analysis, electrophysiological and behavioral assays with the potato-feeding aphid Macrosiphum euphorbiae, we showed that CJ treatment substantially increased the emission of defense VOCs from potatoes compared to no treatment. Coupled GC-electroantennogram (GC-EAG) recordings from the antennae of M. euphorbiae showed robust responses to 14 compounds present in induced VOCs, suggesting their behavioral role in potato/aphid interactions. Plants treated with CJ and then challenged with M. euphorbiae were most repellent to alate M. euphorbiae. Principal component analysis (PCA) of VOC collections suggested that (E)-2-hexenal, (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT), (E)-ß-farnesene, (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), methyl salicylate (MeSA), CJ, and methyl benzoate (MeBA) were the main VOCs contributing to aphid behavioral responses, and that production of TMTT, (E)-ß-farnesene, CJ, and DMNT correlated most strongly with aphid repellency. Our findings confirm that CJ can enhance potato defense against aphids by inducing production of VOCs involved in aphid-induced signalling.

Electrophysiological and behavioral responses of female African rice gall midge, Orseolia oryzivora Harris and Gagné, to host plant volatiles.

African rice gall midge, Orseolia oryzivora Harris and Gagné, is a major pest of rice in Africa. Depsite its economic importance, its chemical ecology is not well understood. Here, we assessed behavioral and electrophysiological responses of O. oryzivora to host plant volatiles. In olfactometer bioassays, mated female O. oryzivora were attracted to volatiles emitted from intact rice plants but were repelled by volatiles collected from plants infested by conspecifics. In a choice test, there was a preference for volatiles from uninfested plants over those from infested plants. Coupled gas chromatography-electroantennography analyses of panicle volatiles isolated four electrophysiologically active components: (S)-linalool, 4,8-dimethyl-1,3,7-nonatriene, (E)-caryophyllene, and (R/S)-(E)-nerolidol. A synthetic blend of volatiles at the same concentration and ratio as that from an intact plant was attractive to mated females, whereas a blend based on the ratio of volatiles from an infested plant was repellent. This suggests that O. oryzivora uses olfaction for host plant recognition. The identification of blends of volatiles emitted by plants that can both attract and repel O. oryzivora may aid the development of sustainable control measures.

Responses of the two-spotted oak buprestid, Agrilus biguttatus (Coleoptera: Buprestidae), to host tree volatiles.

BACKGROUND: Agrilus bigutattus (Fabricius) is a forest pest of increasing importance in the United Kingdom. The larvae damage weakened native oaks and are thought to contribute to premature tree death. Suspected links with acute oak decline (AOD) are not yet confirmed, but AOD-predisposed trees appear to become more susceptible to A. biguttatus attack. Thus, management may be necessary for control of this insect. To explore the possibility of monitoring beetle populations by baited traps, the host tree volatiles regulating A. biguttatus-oak interactions were studied. RESULTS: Biologically active volatile organic compounds in dynamic headspace extracts of oak foliage and bark were identified initially by coupled gas chromatography-electroantennography (GC-EAG) and GC-mass spectrometry (GC-MS), and the structures were confirmed by GC coinjection with authentic compounds. Of two synthetic blends of these compounds comprising the active leaf volatiles, the simpler one containing three components evoked strongly positive behavioural responses in four-arm olfactometer tests with virgin females and males, although fresh leaf material was more efficient than the blend. The other blend, comprising a five-component mixture made up of bark volatiles, proved to be as behaviourally active for gravid females as bark tissue. CONCLUSIONS: These initial results on A. biguttatus chemical ecology reveal aspects of the role of attractive tree volatiles in the host-finding of beetles and underpin the development of semiochemically based surveillance strategies for this forest insect.

The first crop plant genetically engineered to release an insect pheromone for defence.

Insect pheromones offer potential for managing pests of crop plants. Volatility and instability are problems for deployment in agriculture but could be solved by expressing genes for the biosynthesis of pheromones in the crop plants. This has now been achieved by genetically engineering a hexaploid variety of wheat to release (E)-ß-farnesene (Eßf), the alarm pheromone for many pest aphids, using a synthetic gene based on a sequence from peppermint with a plastid targeting amino acid sequence, with or without a gene for biosynthesis of the precursor farnesyl diphosphate. Pure Eßf was produced in stably transformed wheat lines with no other detectable phenotype but requiring targeting of the gene produced to the plastid. In laboratory behavioural assays, three species of cereal aphids were repelled and foraging was increased for a parasitic natural enemy. Although these studies show considerable potential for aphid control, field trials employing the single and double constructs showed no reduction in aphids or increase in parasitism. Insect numbers were low and climatic conditions erratic suggesting the need for further trials or a closer imitation, in the plant, of alarm pheromone release.

Responses of parasitoids to volatiles induced by Chilo partellus oviposition on teosinte, a wild ancestor of maize.

Maize, a genetically diverse crop, is the domesticated descendent of its wild ancestor, teosinte. Recently, we have shown that certain maize landraces possess a valuable indirect defense trait not present in commercial hybrids. Plants of these landraces release herbivore-induced plant volatiles (HIPVs) that attract both egg [Trichogramma bournieri Pintureau & Babault (Hymenoptera: Trichogrammatidae)] and larval [Cotesia sesamiae Cameron (Hymenoptera: Braconidae)] parasitoids in response to stemborer egg deposition. In this study, we tested whether this trait also exists in the germplasm of wild Zea species. Headspace samples were collected from plants exposed to egg deposition by Chilo partellus Swinhoe (Lepidoptera: Crambidae) moths and unexposed control plants. Four-arm olfactometer bioassays with parasitic wasps, T. bournieri and C. sesamiae, indicated that both egg and larval parasitoids preferred HIPVs from plants with eggs in four of the five teosinte species sampled. Headspace samples from oviposited plants released higher amounts of EAG-active compounds such as (E)-4,8-dimethyl-1,3,7-nonatriene. In oviposition choice bioassays, plants without eggs were significantly preferred for subsequent oviposition by moths compared to plants with prior oviposition. These results suggest that this induced indirect defence trait is not limited to landraces but occurs in wild Zea species and appears to be an ancestral trait. Hence, these species possess a valuable trait that could be introgressed into domesticated maize lines to provide indirect defense mechanisms against stemborers.

Novel olfactory ligands via terpene synthases.

A synthetic biology approach to the rational design of analogues of olfactory ligands by providing unnatural substrates for the enzyme synthesising (S)-germacrene D, an olfactory ligand acting as a plant derived insect repellent, to produce novel ligands is described as a viable alternative to largely unsuccessful ligand docking studies. (S)-14,15-Dimethylgermacrene D shows an unexpected reversal in behavioural activity.