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.

Solvent Extraction of PDMS Tubing as a New Method for the Capture of Volatile Organic Compounds from Headspace.

Polydimethylsiloxane (PDMS) tubing is increasingly being used to collect volatile organic compounds (VOCs) from static biological headspace. However, analysis of VOCs collected using PDMS tubing often deploys thermal desorption, where samples are considered as 'one-offs' and cannot be used in multiple experiments. In this study, we developed a static headspace VOC collection method using PDMS tubing which is solvent-based, meaning that VOC extracts can be used multiple times and can be linked to biological activity. Using a synthetic blend containing a range of known semiochemicals (allyl isothiocyanate, (Z)-3-hexen-1-ol, 1-octen-3-one, nonanal, (E)-anethol, (S)-bornyl acetate, (E)-caryophyllene and pentadecane) with differing chemical and physicochemical properties, VOCs were collected in static headspace by exposure to PDMS tubing with differing doses, sampling times and lengths. In a second experiment, VOCs from oranges were collected using PDMS sampling of static headspace versus dynamic headspace collection. VOCs were eluted with diethyl ether and analysed using gas chromatography - flame ionization detector (GC-FID) and coupled GC - mass spectrometry. GC-FID analysis of collected samples showed that longer PDMS tubes captured significantly greater quantities of compounds than shorter tubes, and that sampling duration significantly altered the recovery of all tested compounds. Moreover, greater quantities of compounds were recovered from closed compared to open systems. Finally, analysis of orange headspace VOCs showed no qualitative differences in VOCs recovered compared to dynamic headspace collections, although quantities sampled using PDMS tubing were lower. In summary, extraction of PDMS tubing with diethyl ether solvent captures VOCs from the headspace of synthetic blends and biological samples, and the resulting extracts can be used for multiple experiments linking VOC content to biological activity.

Effects of root inoculation of entomopathogenic fungi on olfactory-mediated behavior and life-history traits of the parasitoid Aphidius ervi (Haliday) (Hymenoptera: Braconidae).

BACKGROUND: Although most biological control programs use multiple biological agents to manage pest species, to date only a few programs have combined the use of agents from different guilds. Using sweet pepper (Capsicum annuum L.), the entomopathogenic fungus Akanthomyces muscarius ARSEF 5128, the tobacco peach aphid Myzus persicae var. nicotianae and the aphid parasitoid Aphidius ervi as the experimental model, we explored whether root inoculation with an entomopathogenic fungus is compatible with parasitoid wasps for enhanced biocontrol of aphids. RESULTS: In dual-choice behavior experiments, A. ervi was significantly attracted to the odor of M. persicae-infested C. annuum plants that had been inoculated with A. muscarius, compared to noninoculated infested plants. There was no significant difference in attraction to the odor of uninfested plants. Myzus persicae-infested plants inoculated with A. muscarius emitted significantly higher amounts of indole, (E)-nerolidol, (3E,7E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene and one unidentified terpene compared to noninoculated infested plants. Coupled gas chromatography-electroantennography, using the antennae of A. ervi, confirmed the physiological activity of these elevated compounds. Inoculation of plants with A. muscarius did not affect parasitism rate nor parasitoid longevity, but significantly increased the speed of mummy formation in parasitized aphids on fungus-inoculated plants. CONCLUSION: Our data suggest that root inoculation of C. annuum with A. muscarius ARSEF 5128 alters the olfactory-mediated behavior of parasitoids, but has little effect on parasitism efficiency or life-history parameters. However, increased attraction of parasitoids towards M. persicae-infested plants when inoculated by entomopathogenic fungi can accelerate host localization and hence improve biocontrol efficacy. © 2023 Society of Chemical Industry.

Investigating volatile semiochemical production from Bos taurus and Bos indicus as a novel phenotype for breeding host resistance to ixodid ticks.

Ticks and tick-borne diseases cause significant loss in livestock production with about 80% world's cattle at risk. The cost of chemical control is high and there is an ever-increasing tick resistance to chemical acaricides. Genetic selection as alternative long-term control strategy is constrained by laborious phenotyping using tick counts or scores. This study explored the use of host volatile semiochemicals that may be attractants or repellents to ticks as a phenotype for new tick resistance, with potential to be used as a proxy in selection programmes. Approximately 100 young cattle composed of Bos indicus and Bos taurus were artificially infested with 2,500 African blue tick, Rhipicephalus decoloratus larvae, with daily female tick (4.5 mm) counts taken from day 20 post-infestation. Volatile organic compounds were sampled from cattle before and after tick infestation by dynamic headspace collection, analysed by high-resolution gas chromatography (GC) and subjected to multivariate statistical analysis. Using 6-day repeated measure analysis, three pre-infestation GC peaks (BI938 - unknown, BI966 - 6-methyl-5-hepten-2-one and BI995 - hexyl acetate) and one post-infestation GC peak (AI933 - benzaldehyde / (E)-2-heptenal) were associated with tick resistance (P < 0.01 and P < 0.05 respectively). The high correlation coefficients (r = 0.66) between repeated records with all volatile compounds support the potential predictive value for volatile compounds in selective breeding programmes for tick resistance in cattle.

Impact of endophytic colonization by entomopathogenic fungi on the behavior and life history of the tobacco peach aphid Myzus persicae var. nicotianae.

Entomopathogenic fungi can adopt an endophytic lifestyle and provide protection against insect herbivores and plant pathogens. So far, most studies have focused on Beauveria bassiana to increase plant resistance against abiotic and biotic stresses, while only little is known for other entomopathogenic fungi. In this study, we investigated whether root inoculation of sweet pepper (Capsicum annuum L.) by the entomopathogenic fungi Akanthomyces muscarius ARSEF 5128 and B. bassiana ARSEF 3097 can improve resistance against the tobacco peach aphid Myzus persicae var. nicotianae. First, dual-choice experiments were performed to test the hypothesis that the fungi deter aphids via modifying plant volatile profiles. Next, we tested the hypothesis that endophytic colonization negatively affects aphid life history traits, such as fecundity, development and mortality rate. Aphids were significantly attracted to the odor of plants inoculated with A. muscarius over non-inoculated plants. Plants inoculated with A. muscarius emitted significantly higher amounts of ß-pinene than non-inoculated plants, and significantly higher amounts of indole than B. bassiana-inoculated and non-inoculated plants. Inoculation with the fungal strains also caused significantly higher emission of terpinolene. Further, both aphid longevity and fecundity were significantly reduced by 18% and 10%, respectively, when feeding on plants inoculated with A. muscarius, although intrinsic rate of population increase did not differ between inoculated and non-inoculated plants. Sweet pepper plants inoculated with B. bassiana ARSEF 3097 did not elicit a significant behavioral response nor affected the investigated life history traits. We conclude that endophytic colonization by entomopathogenic fungi has the potential to alter olfactory behavior and performance of M. persicae var. nicotianae, but effects are small and depend on the fungal strain used.

Companion and Smart Plants: Scientific Background to Promote Conservation Biological Control.

To attain sustainable agricultural crop protection, tools such as host plant resistance, enhanced ecosystem services (i.e. conserving natural enemies) and the deployment of companion plants should be promoted in pest management programmes. These agro system manipulations could be based on chemical ecology studies considering the interactions with natural enemies and pests, regarding specifically plant defence signalling. Further, new crop protection strategies might rise from widening the knowledge regarding how herbivore-induced plant volatiles can govern a multifaceted defence response including natural enemy recruitment, pest repellence or induced defence in neighbouring plants. It is crucial to use a multitrophic approach to understand better the interactions involving companion plants, herbivores and natural enemies in the field, increasing the knowledge to build more efficient and sustainable pest management strategies. In this review, we explore the perspectives of companion plants and their semiochemicals to promote conservation biological control according to the 'smart plants' concept. Further, we discuss the advantages and disadvantages of using companion plants and explore the application of companion plants in different agroecosystems using several case studies.

Identifying aphid resistance in the ancestral wheat Triticum monococcum under field conditions.

Wheat is an economically, socially, and nutritionally important crop, however, aphid infestation can often reduce wheat yield through feeding and virus transmission. Through field phenotyping, we investigated aphid resistance in ancestral wheat Triticum monococcum (L.). Aphid (Rhopalosiphum padi (L.), Sitobion avenae (F.) and Metopolophium dirhodum (Wlk.)) populations and natural enemy presence (parasitised mummified aphids, ladybird adults and larvae and lacewing eggs and larvae) on two naturally susceptible wheat varieties, Triticum aestivum (L.) var. Solstice and T. monococcum MDR037, and three potentially resistant genotypes T. monococcum MDR657, MDR045 and MDR049 were monitored across three years of field trials. Triticum monococcum MDR045 and MDR049 had smaller aphid populations, whereas MDR657 showed no resistance. Overall, natural enemy presence was positively correlated with aphid populations; however, MDR049 had similar natural enemy presence to MDR037 which is susceptible to aphid infestation. It is hypothesised that alongside reducing aphid population growth, MDR049 also confers indirect resistance by attracting natural enemies. The observed resistance to aphids in MDR045 and MDR049 has strong potential for introgression into commercial wheat varieties, which could have an important role in Integrated Pest Management strategies to reduce aphid populations and virus transmission.

Sex Pheromone of the Alfalfa Plant Bug, Adelphocoris lineolatus: Pheromone Composition and Antagonistic Effect of 1-Hexanol (Hemiptera: Miridae).

The sex pheromone composition of alfalfa plant bugs, Adelphocoris lineolatus (Goeze), from Central Europe was investigated to test the hypothesis that insect species across a wide geographical area can vary in pheromone composition. Potential interactions between the pheromone and a known attractant, (E)-cinnamaldehyde, were also assessed. Coupled gas chromatography-electroantennography (GC-EAG) using male antennae and volatile extracts collected from females, previously shown to attract males in field experiments, revealed the presence of three physiologically active compounds. These were identified by coupled GC/mass spectrometry (GC/MS) and peak enhancement as hexyl butyrate, (E)-2-hexenyl butyrate and (E)-4-oxo-2-hexenal. A ternary blend of these compounds in a 5.4:9.0:1.0 ratio attracted male A. lineolatus in field trials in Hungary. Omission of either (E)-2-hexenyl-butyrate or (E)-4-oxo-2-hexenal from the ternary blend or substitution of (E)-4-oxo-2-hexenal by (E)-2-hexenal resulted in loss of activity. These results indicate that this Central European population is similar in pheromone composition to that previously reported for an East Asian population. Interestingly, another EAG-active compound, 1-hexanol, was also present in female extract. When 1-hexanol was tested in combination with the ternary pheromone blend, male catches were reduced. This compound showed a dose-response effect with small doses showing a strong behavioral effect, suggesting that 1-hexanol may act as a sex pheromone antagonist in A. lineolatus. Furthermore, when (E)-cinnamaldehyde was field tested in combination with the sex pheromone, there was no increase in male catch, but the combination attracted both males and females. Prospects for practical application are discussed.

Iridoid Sex Pheromone Biosynthesis in Aphids Mimics Iridoid-Producing Plants.

Biosynthesis of (1R,4aS,7S,7aR)-nepetalactol (1) and (4aS,7S,7aR)-nepetalactone (2) in plants involves iridoid synthase (ISY), an atypical reductive cyclase that catalyses the reduction of 8-oxogeranial into the reactive enol of (S)-8-oxocitronellal, and cyclization of this enol intermediate, either non-enzymatically or by a nepetalactol-related short chain dehydrogenase enzyme (NEPS) that yields the nepetalactols. In this study, we investigated the biosynthesis in vivo of 1 and 2 in the pea aphid, Acyrthosiphon pisum, using a library of isotopically-labelled monoterpenoids as molecular probes. Topical application of deuterium-labelled probes synthesized from geraniol and nerol resulted in production of 2 H4 -lactol 1 and 2 H4 -lactone 2. However, deuterium incorporation was not evident using labelled probes synthesized from (S)-citronellol. These results suggest that iridoid biosynthesis in animals, specifically aphids, may follow a broadly similar route to that characterised for plants.

Identification and application of bacterial volatiles to attract a generalist aphid parasitoid: from laboratory to greenhouse assays.

BACKGROUND: Recent studies have shown that microorganisms emit volatile compounds that affect insect behaviour. However, it remains largely unclear whether microbes can be exploited as a source of attractants to improve biological control of insect pests. In this study, we used a combination of coupled gas chromatography-electroantennography (GC-EAG) and Y-tube olfactometer bioassays to identify attractive compounds in the volatile extracts of three bacterial strains that are associated with the habitat of the generalist aphid parasitoid Aphidius colemani, and to create mixtures of synthetic compounds to find attractive blends for A. colemani. Subsequently, the most attractive blend was evaluated in two-choice cage experiments under greenhouse conditions. RESULTS: GC-EAG analysis revealed 20 compounds that were linked to behaviourally attractive bacterial strains. A mixture of two EAG-active compounds, styrene and benzaldehyde applied at a respective dose of 1 µg and 10 ng, was more attractive than the single compounds or the culture medium of the bacteria in Y-tube olfactometer bioassays. Application of this synthetic mixture under greenhouse conditions resulted in significant attraction of the parasitoids, and outperformed application of the bacterial culture medium. CONCLUSION: Compounds isolated from bacterial blends were capable of attracting parasitoids both in laboratory and greenhouse assays, indicating that microbial cultures are an effective source of insect attractants. This opens new opportunities to attract and retain natural enemies of pest species and to enhance biological pest control.

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.

Inefficient weapon-the role of plant secondary metabolites in cotton defence against the boll weevil.

MAIN CONCLUSION: Cotton genotypes displayed similar volatile organic compound (VOC) profiles, but major differences in terpenoid aldehyde (TA) content. The differences in VOC production were minor among genotypes, but these differences are crucial for boll weevil attraction. Weevils did not display any preference in feeding behaviour towards cotton genotypes, suggesting physiological adaptation to cope with cotton chemical defence mechanisms. Plant cultivar selection for resistance to herbivore pests is an effective, environmentally safe and inexpensive method to implement in integrated pest management programmes. In this study, we evaluated seven cotton genotypes with respect to the production of volatile organic compounds (VOCs) and non-volatile compounds [terpenoid aldehydes (TAs)], and the attraction and feeding preference of adult boll weevils. Chemical analyses of VOCs from BRS-293, BRS-Rubi, CNPA TB-15, CNPA TB-85, CNPA TB-90, Delta Opal, and Empire Glandless showed that there were few qualitative and quantitative differences across the range of genotypes. In contrast, major differences in TA content were observed, with CNPA TB-15 and CNPA TB-85 producing higher levels of TAs compared to the other genotypes. Our results showed that boll weevil attraction to cotton genotypes varied, suggesting that the ratios and quantities of emitted cotton VOCs are important for host location. However, boll weevil feeding behaviour was neither positively nor negatively influenced by the terpenoid content (non-volatile compounds) of cotton genotypes. The results in this study suggest that boll weevils have adapted physiologically to cope with cotton chemical defence mechanisms.

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.

Species-specific alterations in Anopheles mosquito olfactory responses caused by Plasmodium infection.

Mosquitoes infected with malaria parasites have demonstrated altered behaviour that may increase the probability of parasite transmission. Here, we examine the responses of the olfactory system in Plasmodium falciparum infected Anopheles gambiae, Plasmodium berghei infected Anopheles stephensi, and P. berghei infected An. gambiae. Infected and uninfected mosquitoes showed differential responses to compounds in human odour using electroantennography coupled with gas chromatography (GC-EAG), with 16 peaks triggering responses only in malaria-infected mosquitoes (at oocyst, sporozoite or both stages). A selection of key compounds were examined with EAG, and responses showed differences in the detection thresholds of infected and uninfected mosquitoes to compounds including lactic acid, tetradecanoic acid and benzothiazole, suggesting that the changes in sensitivity may be the reason for differential attraction and biting at the oocyst and sporozoite stages. Importantly, the different cross-species comparisons showed varying sensitivities to compounds, with P. falciparum infected An. gambiae differing from P. berghei infected An. stephensi, and P. berghei infected An. gambiae more similar to the P. berghei infected An. stephensi. These differences in sensitivity may reflect long-standing evolutionary relationships between specific Plasmodium and Anopheles species combinations. This highlights the importance of examining different species interactions in depth to fully understand the impact of malaria infection on mosquito olfactory behaviour.

Identification of a non-host semiochemical from tick-resistant donkeys (Equus asinus) against Amblyomma sculptum ticks.

Amblyomma sculptum is a tick affecting animal and human health across Argentina, Bolivia, Paraguay and Brazil. Donkeys, Equus asinus, are known to be resistant to A. sculptum, suggesting that they can produce non-host tick semiochemicals (allomones), as already demonstrated for some other vertebrate host/pest interactions, whereas horses, Equus caballus, are considered as susceptible hosts. In this study, we tested the hypothesis that donkeys produce natural repellents against A. sculptum, by collecting sebum from donkeys and horses, collecting the odour from sebum extracts, and identifying donkey-specific volatile compounds by gas chromatography (GC) and coupled GC-mass spectrometry (GC-MS). From the complex collected blends, five main compounds were identified in both species. Hexanal, heptanal and (E)-2-decenal were found predominantly in donkey extracts, whilst ethyl octanoate and ethyl decanoate were found predominantly in horse extracts. One compound, (E)-2-octenal, was detected exclusively in donkey extracts. In Y-tube olfactometer bioassays 36 different A. sculptum nymphs were tested for each extract, compound and concentration. The dry sebum extracts and the compounds identified in both species induced neither attraction nor repellency. Only (E)-2-octenal, the donkey-specific compound, displayed repellency, with more nymphs preferring the arm containing the solvent control when the compound was presented in the test arm across four concentrations tested (p < 0.05, Chi-square test). A combination of a tick attractant (ammonia) and (E)-2-octenal at 0.25 M also resulted in preference for the control arm (p < 0.05, Chi-square test). The use of semiochemicals (allomones) identified from less-preferred hosts in tick management has been successful for repelling brown dog ticks, Rhipicephalus sanguineus sensu lato from dog hosts. These results indicate that (E)-2-octenal could be used similarly to interfere in tick host location and be developed for use in reducing A. sculptum numbers on animal and human hosts.

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.

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.

Maize Chlorotic Mottle Virus Induces Changes in Host Plant Volatiles that Attract Vector Thrips Species.

Maize lethal necrosis is one of the most devastating diseases of maize causing yield losses reaching up to 90% in sub-Saharan Africa. The disease is caused by a combination of maize chlorotic mottle virus (MCMV) and any one of cereal viruses in the Potyviridae group such as sugarcane mosaic virus. MCMV has been reported to be transmitted mainly by maize thrips (Frankliniella williamsi) and onion thrips (Thrips tabaci). To better understand the role of thrips vectors in the epidemiology of the disease, we investigated behavioral responses of F. williamsi and T. tabaci, to volatiles collected from maize seedlings infected with MCMV in a four-arm olfactometer bioassay. Volatile profiles from MCMV-infected and healthy maize plants were compared by gas chromatography (GC) and GC coupled mass spectrometry analyses. In the bioassays, both sexes of F. williamsi and male T. tabaci were significantly attracted to volatiles from maize plants infected with MCMV compared to healthy plants and solvent controls. Moreover, volatile analysis revealed strong induction of (E)-4,8-dimethyl-1,3,7-nonatriene, methyl salicylate and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene in MCMV-infected maize seedlings. Our findings demonstrate MCMV induces changes in volatile profiles of host plants to elicit attraction of thrips vectors. The increased vector contact rates with MCMV-infected host plants could enhance virus transmission if thrips feed on the infected plants and acquire the pathogen prior to dispersal. Uncovering the mechanisms mediating interactions between vectors, host plants and pathogens provides useful insights for understanding the vector ecology and disease epidemiology, which in turn may contribute in designing integrated vector management strategies.