Non-host Volatiles Research Articles

Direct and mediated impacts of mixed forests on Norway spruce infestation by European bark beetle Ips typographus

Climate change-induced windfalls and droughts exacerbate bark beetle outbreaks, severely impacting forest ecosystems. Despite extensive research on bark beetle infestation patterns, the question regarding the optimal spatial grid (SG) size for analyzing this phenomenon remains unresolved. The protective potential of natural forest complexity against bark beetles has been underestimated. We used remotely sensed data to fit Generalized Linear Models and structural equation models to explore the direct and environment-mediated effects of mixed forest cover (MFC) on Norway spruce forest infestation in central Europe. We assessed the effectiveness of various spatial grids (from 100 m to 1 km) for studying infestation by Ips typographus. We found a strong non-linear decline in infestation with increasing MFC across different spatial scales. The relationship between infestation and temperature was positive, while elevation had a negative effect on infestation, with higher infestation rates observed below 900 m. Direct effects of environmental predictors on infestation were significant at SGs of 100 m, 300 m, 400 m, and 500 m, but insignificant at 200 m and 1000 m SGs. Slope positively influenced infestation at 300 m and 400 m SGs. MFC exhibited significant indirect effects on infestation mediated by elevation, temperature, potential evapotranspiration, slope, and heat load index. Landscape variables played a significant role in the models at high-resolution spatial grids, whereas climate variables were influential in models at lower spatial grids of infestation data. We argue that mixed forest facilitates cooling, preserves water, enriches symbiotic fungal community, alleviating tree drought stress and mitigating infestation risk. Broad-leave trees’ non-host volatiles presumably disrupt olfactory signals, impeding beetles’ ability to locate host trees effectively. Our results underscore the potential of increasing mixed forest cover as a self-sustainable silvicultural measure for forest protection against I. typographus and for mitigating the effects of climate change.

Deorphanizing an odorant receptor tuned to palm tree volatile esters in the Asian palm weevil sheds light on the mechanisms of palm tree selection

The Asian palm weevil, Rhynchophorus ferrugineus, is a tremendously important agricultural pest primarily adapted to palm trees and causes severe destruction, threatening sustainable palm cultivation worldwide. The host plant selection of this weevil is mainly attributed to the functional specialization of odorant receptors (ORs) that detect palm-derived volatiles. Yet, ligands are known for only two ORs of R. ferrugineus, and we still lack information on the mechanisms of palm tree detection. This study identified a highly expressed antennal R. ferrugineus OR, RferOR2, thanks to newly generated transcriptomic data. The phylogenetic analysis revealed that RferOR2 belongs to the major coleopteran OR group 2A and is closely related to a sister clade containing an R. ferrugineus OR (RferOR41) tuned to the non-host plant volatile and antagonist, α-pinene. Functional characterization of RferOR2 via heterologous expression in Drosophila olfactory neurons revealed that this receptor is tuned to several ecologically relevant palm-emitted odors, most notably ethyl and methyl ester compounds, but not to any of the pheromone compounds tested, including the R. ferrugineus aggregation pheromone. We did not evidence any differential expression of RferOR2 in the antennae of both sexes, suggesting males and females detect these compounds equally. Next, we used the newly identified RferOR2 ligands to demonstrate that including synthetic palm ester volatiles as single compounds and in combinations in pheromone-based mass trapping has a synergistic attractiveness effect to R. ferrugineus aggregation pheromone, resulting in significantly increased weevil catches. Our study identified a key OR from a palm weevil species tuned to several ecologically relevant palm volatiles and represents a significant step forward in understanding the chemosensory mechanisms of host detection in palm weevils. Our study also defines RferOR2 as an essential model for exploring the molecular basis of host detection in other palm weevil species. Finally, our work showed that insect OR deorphanization could aid in identifying novel behaviorally active volatiles that can interfere with weevil host-searching behavior in sustainable pest management applications.

An Antennae-Enriched Odorant-Binding Protein EonuOBP43 Mediate the Behavioral Response of the Tea Green Leafhopper, Empoasca onukii Matsuda to the Host and Nonhost Volatiles.

Olfaction is crucial for Empoasca onukii Matsuda to recognize odors from the host and nonhost plants, and it has been proposed that odorant binding proteins are directly required for odorant discrimination and represent potential targets of interest for pest control. Here, we cloned EonuOBP43 and expressed the recombinant EonuOBP43 protein. Furthermore, competitive fluorescence binding assays with 19 ligands indicated that terpenoids and alkanes showed a relatively higher than for other classes of chemicals. Additionally, ligand docking and site-directed mutagenesis results revealed that seven hydrophobic residues, including Val-86, Met-89, Phe-90, Ile-104, Ile-105, Leu-130, and Val-134, played a key role in the binding of EonuOBP43 to plant volatiles. In olfactometer tests, E. onukii were significantly attracted to α-farnesene and repelled to β-caryophyllene, and dsOBP43 treated adult lost response to α-farnesene and β-caryophyllene. In summary, our results demonstrated that EonuOBP43 may function as a carrier in the process of sensing plant compounds of E. onukii.

A meta-analytic investigation of the potential for plant volatiles and sex pheromones to enhance detection and management of Lepidopteran pests.

Effective early detection, monitoring and management methods are critical for reducing the impacts of insect pests in agriculture and forestry. Combining host plant volatiles with sex pheromones could enhance trapping methodologies, whilst the use of non-host volatiles could improve the effectiveness of pest management through repellency effects. In this meta-analysis approach, we analysed 51 studies that used electroantennograms (EAG), wind tunnels and/or field traps to evaluate the antennal and behavioural responses of Lepidoptera to sex pheromones combined with attractant or repellent plant volatiles. Proposed attractant plant volatiles had a positive association with female Lepidoptera responses to sex pheromone, but effects on males were highly variable, with unexpected repellency reported in some studies. Proposed repellent plant volatiles were significantly or near-significantly negatively associated with male attraction to sex pheromones but were scarcely studied. Sub-group analysis identified that male responses to sex pheromone were reduced when the dose of attractant plant volatile relative to sex pheromone was increased. Green-leaf volatiles were associated with the strongest positive effects for males in field traps. Multiple-compound attractant plant volatile blends were less effective than single compounds in field studies. Our analysis demonstrates, (i) the potential value of combining host plant volatiles with sex pheromones to capture females rather than only males, (ii) the importance of identifying appropriate host plant volatiles and optimal relative doses, and (iii) the potential for non-host plant volatile use in pest management strategies.

How Effective Are Push-Pull Semiochemicals as Deterrents for Bark Beetles? A Global Meta-Analysis of Thirty Years of Research.

Bark beetles (Coleoptera: Curculionidae: Scolytinae) are among the most damaging tree pests globally. Rising temperatures, drought, fire, storms, cyclones, and poor forest management cause stress and loss of vigour in trees, and these conditions favour bark beetle outbreaks. While research has been conducted on push-pull strategies to deter bark beetles, using attractive and deterrent semiochemicals, the potential of this strategy to reduce bark beetle populations, particularly in the genera Dendroctonus and Ips, remains uncertain. Here, we conducted a global meta-analysis of 52 research articles to quantify the effects of semiochemical treatments on managing different species of Dendroctonus and Ips for forest protection. Based on this analysis, we found that push-pull semiochemicals can significantly reduce Dendroctonus and Ips populations measured by a reduction in the attraction to lure/trap catches, tree mortality, and attacks on trees. The overall efficacy of the push-pull semiochemical treatment shows a 66% reduction for Ips compared to control and a 54% reduction compared to control for Dendroctonus, while, at the species level, there was a 69% reduction for Dendroctonus ponderosae (Hopkins) and a 94% reduction in Ips perturbatus (Eichhoff), and a 93% reduction in Ips latidens (LeConte). Interestingly, among different treatment sources, the efficacy of conspecific semiochemicals in combination with heterospecific semiochemicals and non-host volatiles showed a 92% reduction in Dendroctonus spp., and conspecific semiochemicals in combination with non-host volatiles showed a 77% significant reduction in Ips spp., while the efficacy of heterospecific semiochemicals in reducing Ips population was about 69%, and 20% in Dendroctonus. Among different ecological regions, the use of a push-pull strategy showed a 70% reduction in Dendroctonus in central-west North America, and Ips showed a 75% reduction in southwest North America. Our results demonstrate that semiochemical-based push-pull techniques have the potential to reduce Dendroctonus and Ips bark beetle populations. Furthermore, based on our analysis, the efficacy of such eco-friendly interventions could be further improved and provide a good tool for forest managers to control these pests, at least under some circumstances.

Potential of Scots Pine for a Push Strategy against the European Spruce Bark Beetle Ips typographus

Ips typographus, the European spruce bark beetle, is a destructive forest insect that attacks spruce trees. It poses a significant threat to the forestry industry, causing widespread tree mortality and economic losses, and its management through chemical ecology is a promising approach. Attraction relies on aggregation pheromones while repellency is achieved via antiaggregation pheromones combined with mixtures of nonhost volatiles (NHVs), but an effective push and pull strategy has not been established yet. In this study, we observed the beetle colonization dynamic and density comparing pure Norway spruce stacks and Norway spruce stacks with 20% Scots pine logs. We collected the spruce bark volatiles before the beetle colonization and at the end of the trial, and tested the behavioral effects on the beetles of both spruce and pine essential oils in olfactometer trials. In the stacks with Scots pine, we observed a delayed primary colonization and a 65% reduction of the secondary colonization, confirming the colonization disruption mediated by NHVs. Volatile analysis showed a significant change in the bark emission profiles, and the reaching of the antiaggregation phase was marked by the release of verbenone. Behavioral experiments confirmed the attractivity of spruce essential oil and the strong repellency of the pine essential oil. The study supports the use of complex NHVs blends for the beetle behavioral manipulation in the field. Further studies are needed to assess if the same results can be achieved by deploying essential oils in forest stands.

Characterization of olfactory sensory neurons in the striped ambrosia beetle Trypodendron lineatum.

Introduction: The striped ambrosia beetle Trypodendron lineatum (Coleoptera, Curculionidae, Scolytinae) is a major forest pest in the Holarctic region. It uses an aggregation pheromone and host and non-host volatiles to locate suitable host trees, primarily stressed or dying conifer trees. The beetles bore into the xylem and inoculate spores of their obligate fungal mutualist Phialophoropsis ferruginea inside their excavated egg galleries, with the fungus serving as the main food source for the developing larvae. Olfactory sensory neuron (OSN) responses to pheromones and host volatiles are poorly understood in T. lineatum and other ambrosia beetles, and nothing is known about potential responses to fungal volatiles. Methods: We screened responses of OSNs present in 170 antennal olfactory sensilla using single sensillum recordings (SSR) and 57 odor stimuli, including pheromones, host and non-host compounds, as well as volatiles produced by P. ferruginea and fungal symbionts of other scolytine beetles. Results and Discussion: Thirteen OSN classes were characterized based on their characteristic response profiles. An OSN class responding to the aggregation pheromone lineatin was clearly the most abundant on the antennae. In addition, four OSN classes responded specifically to volatile compounds originating from the obligate fungal mutualist and three responded to non-host plant volatiles. Our data also show that T. lineatum has OSN classes tuned to pheromones of other bark beetles. Several OSN classes showed similar response profiles to those previously described in the sympatric bark beetle Ips typographus, which may reflect their shared ancestry.

Exploring plant volatile-mediated interactions between native and introduced plants and insects

In invasion scenarios, native and introduced species co-occur creating new interactions and modifying existing ones. Many plant–plant and plant–insect interactions are mediated by volatile organic compounds (VOCs), however, these have seldom been studied in an invasion context. To fill this knowledge gap, we explored some interactions mediated by VOCs between native and introduced plants and insects in a New Zealand system. We investigated whether a native plant, Leptospermum scoparium (mānuka), changes its volatile profile when grown adjacent to two European introduced plants, Calluna vulgaris (heather) and Cytisus scoparius (Scotch broom), in a semi-field trial using potted plants without above- or below-ground physical contact. We also investigated the influence of plant cues on the host-searching behaviour of two beetles, the native Pyronota festiva (mānuka beetle), and the introduced biocontrol agent Lochmaea suturalis (heather beetle), by offering them their host-plant and non-host volatiles versus clean air, and their combination in a Y-tube olfactometer. As a follow-up, we performed preference/feeding tests in Petri dishes with fresh plant material. Results of the semi-field experiment show a significant reduction in green leaf volatiles, sesquiterpenes and total volatile emissions by mānuka plants neighbouring heather. In the Y-tube assays, the native beetle P. festiva performed poorly in discriminating between host and non-host plants based on plant volatile cues only. However, it performed relatively well in the Petri dish tests, where other cues (i.e., visual, gustatory or tactile) were present. In contrast, the introduced beetle L. suturalis showed high host-specificity in both Y-tube and Petri dish assays. This study illustrates the importance of VOCs in mediating interactions between introduced and native species, suggesting that invasive plants can disrupt native plants’ communication and affect the host-searching behaviour of native insects. It also reinforces the relevance of regular host testing on introduced weed biocontrol agents to avoid unwanted host shifts or host-range expansion.

Identification of Repellents from Four Non-Host Asteraceae Plants for the Root Knot Nematode, Meloidogyne incognita

Olfactory cues guide plant parasitic nematodes (PPNs) to their host plants. We tested the hypothesis that non-host plant root volatiles repel PPNs. To achieve this, we compared the olfactory responses of infective juveniles (J2s) of the PPN Meloidogyne incognita to four non-host Asteraceae plants, namely, black-jack (Bidens pilosa), pyrethrum (Chrysanthemum cinerariifolium), marigold (Tagetes minuta), and sweet wormwood (Artemisia annua), traditionally used in sub-Saharan Africa for the management of PPNs. Chemical analysis by coupled gas chromatography-mass spectrometry (GC/MS) combined with random forest analysis, followed by behavioral assays, identified the repellents in the root volatiles of B. pilosa, T. minuta, and A. annua as (E)-β-farnesene and 1,8-cineole, whereas camphor was attractive. In contrast, random forest analysis predicted repellents for C. cinerariifolium and A. annua as β-patchoulene and isopropyl hexadecanoate. Our results suggested that terpenoids generally account for the repellency of non-host Asteraceae plants used in PPN management.

Protecting forest edges using trap logs – Limited effects of associated push-pull strategies targeting Ips typographus

Bark beetles can cause epidemic outbreaks and kill millions of cubic meters of economical and ecologically important forests around the world. It is well known what attracts and what repels different species of bark beetle, and these chemical cues can be used to protect trees and catch the beetles without using pesticides. Applying this knowledge, we investigated the use of push–pull strategies with trap logs along susceptible edges of a Swedish boreal spruce forest. The repellents (push) used were non-host volatiles (NHV) attached to tree trunks at the forest edge, and the attractants (pull) was a commercial aggregation pheromone attached to trap logs. The aim was to test whether the Ips typographus catch could be significantly increased by combining a push–pull system with traditional trap logs, thereby providing additional protection. The experiment was performed over two years and included the main flight period of I. typographus. The study sites were clear-cuts that had been harvested the preceding winter, and sun-exposed forest edges of mature spruce were targeted for protection. A full factorial setup was used comprising two treatments (repellent and attractant) and a control. Seven replicates of the trap logs were used, three during the first year and four during the second. The number of established I. typographus maternal galleries per square meter of log surface was used as the response variable. The trap logs captured large numbers of I. typographus, at an average density of 353 and 169 maternal galleries per m2 during year 1 and year 2, respectively, over all treatments. Based on the catch data, with a sufficient number of trap logs, the risk of tree mortality at forest edges may be reduced and we recommend its general use. However, we did not see any significant effect of either the repellent or the attractant on the density of maternal galleries. Hence, we cannot recommend the addition of chemical cues to improve the efficiency of trap logs. Although trap logs are efficient in capturing bark beetles and hence may protect forest edges, it does not imply that they can provide protection on a larger scale. In line with other studies, we hence recommend that forest management to target nature-based solutions that strengthen the resilience of forest stands, by using mixed forest stands and resistant plant species, and nurture habitats for natural predators of I. typographus.

Non-palm Plant Volatile α-Pinene Is Detected by Antenna-Biased Expressed Odorant Receptor 6 in the Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae).

The majority of insects rely on a highly complex and precise olfactory system to detect various volatile organic compounds released by host and non-host plants in environments. The odorant receptors (ORs) are considered to play an important role in odor recognition and the molecular basis of ORs, particularly in coleopterans they are relatively poorly understood. The red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae), is one of the most destructive pests of the global palm industry. Although feeding and egg oviposition behaviors of RPW can be repelled by some non-palm plant volatiles, such as α-pinene, geraniol, or 1-octen-3-ol, there is limited understanding of how RPW recognizes the non-host plant volatiles. In this study, three candidate RferOrs were identified from the Rfer-specific clade, and the tissue expression analysis used was mainly expressed in the antennae of both sexes. Functional characterization of RferOr6, RferOr40, and RferOr87 was analyzed by using the Xenopus oocyte expression system, and the results indicated that RferOr6/RferOrco was narrowly tuned to α-pinene. The behavioral experiment showed that α-pinene at the concentrations of 10 and 100 μg/μl can cause a significantly repelled behavioral response of RPW. In conclusion, this study reveals that RferOr6 is an antenna-biased expressed OR used by RPW to detect the volatile compound α-pinene in non-palm plants, and our results provide a foundation for further in vivo functional studies of Or6 in RPW, including in vivo knockout/knockdown and feeding/ovipositing behavioral studies of RPW and further pest control.

Changes in host basal area explain associational resistance of mixed forests to primary pests

Tree species diversity generally has positive effects on forest primary productivity and resistance to natural perturbations, but diversity-function relationships can vary with site conditions. Recently, studies in forest diversity experiments have shown that tree diversity and local climate, in particular drought intensity, interactively affect insect herbivory. On the other hand, many studies focused on the response of forests to drought in terms of tree growth but without analysing the concomitant effects on susceptibility to pests. It is of particular interest to understand the combined effects of drought and tree diversity on the growth of the host tree, since host resource concentration is a determining factor of a pest’s host choice.We used a tree diversity experiment where tree species diversity and drought conditions were both manipulated to evaluate their interactive effects on the susceptibility of maritime pine (Pinus pinaster Aït.) forests to two primary pests (i.e. infesting healthy trees): the pine stem borer (PSB) Dioryctria sylvestrella, and the pine processionary moth (PPM; a leaf chewer), Thaumetopoea pityocampa. Using structural equation models, we investigated the direct and indirect effects (i.e. mediated by host resources) of the presence of birch and drought on the total number of attacks of PPM and PSB, in the same plots and in the same year.We showed that pine-birch plots were more resistant to both PPM and PSB attacks than pine monocultures. Furthermore, we found that this associational resistance pattern was due to direct effects of birch trees on attacks, possibly related to disrupting non-host volatiles (NHVs), but also to indirect, resource-mediated effects whereby the presence of birch trees reduced the amount of host pine resources available to the pests. Drought conditions modulated birch mediated effects on resistance of maritime pine forests only for PSB attacks. Overall, our work improves our understanding of tree diversity-herbivory relationships and helps explain how climate might modulate such relationships.

Disruption of coniferophagous bark beetle (Coleoptera: Curculionidae: Scolytinae) mass attack using angiosperm nonhost volatiles: from concept to operational use

AbstractAlthough the use of nonhost plants intercropped among host crops has been a standard agricultural practice for reducing insect herbivory for millennia, the use of nonhost signals to deter forest pests is much more recent, having been developed over the past several decades. Early exploratory studies with synthetic nonhost volatile semiochemicals led to targeted electrophysiological and trapping experiments on a variety of bark and ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) across three continents. This work disclosed a suite of antennally and behaviourally active nonhost volatiles, which are detected in common across a range of coniferophagous bark beetles. It also established the fact that dispersing bark and ambrosia beetles detect nonhost signals while in flight and avoid nonhost trees without necessarily landing on them. Later work showed that groups of synthetic nonhost volatiles, sometimes combined with insect-derived antiaggregants, are effective in protecting individual trees and forest stands. Further work in this system may lead to the development of a variety of new and useful tactics for use in various integrated pest management strategies.

Efficacy of verbenone and a blend of verbenone and nonhost volatiles for protecting lodgepole pine from mountain pine beetle (Coleoptera: Curculionidae)

Abstract Significant research has been devoted to the development of semiochemical repellents for mountain pine beetle Dendroctonus ponderosae Hopkins. Several formulations of the beetle's antiaggregation pheromone verbenone (4,6,6‐trimethylbicyclo[3.1.1]hept‐3‐en‐2‐one) are registered to protect pine trees and stands. We compared the efficacy of verbenone and Verbenone Plus for protecting individual lodgepole pine Pinus contorta Dougl. ex Loud. and small stands of P. contorta from mortality attributed to D. ponderosae. Verbenone Plus is a blend of semiochemicals [(−)‐verbenone, acetophenone, and (E)‐2‐hexen‐1‐ol + (Z)‐2‐hexen‐1‐ol] originally developed to inhibit aggregation of western pine beetle D. brevicomis LeConte. It is the only effective semiochemical repellent that protects ponderosa pine P. ponderosa Dougl. ex Laws. from mortality attributed to D. brevicomis, but has yet to be registered or commercialized. Both verbenone and Verbenone Plus were effective for protecting individual P. contorta and small stands of P. contorta from mortality attributed to D. ponderosae. Verbenone alone was as effective as Verbenone Plus. An obstacle to the broader use of semiochemical repellents for the management of bark beetles is the relatively high cost and specificity of the semiochemicals involved. A semiochemical repellent with efficacy across multiple bark beetle species and hosts, such as Verbenone Plus, would be desirable, assuming the additional components are not cost‐prohibitive or limit registration.

Chemical signal interactions of the bark beetle with fungal symbionts, and host/non-host trees

The symbiosis between the bark beetle (Ips subelongatus) and its fungal symbiont (Endoconidiophora fujiensis) poses a serious threat to larch forests. However, the signaling pathways between these symbiotic partners and their host/non-host trees are not fully understood. Inoculation of the host larch (Larix principis-rupprechtii) with two strains of E. fujiensis induced a rapid and long-term release of monoterpenes. Although the fungi had a level of tolerance to these compounds, many monoterpenes inhibited fungal growth in culture. Moreover, monoterpenes with stronger inhibitory effects on fungal growth exhibited weaker synergistic effects on the attraction of I. subelongatus to aggregation pheromone. Surprisingly, individual isomers of aggregation pheromone components promoted fungal symbiont growth in a culture medium. Non-host volatiles (NHVs) were tested and shown to completely inhibit the growth of fungal symbionts in culture but had no effects on beetle responses to aggregation pheromone, with the exception of (Z)-3-hexen-1-ol. These results reveal convergence and mutualism patterns in the evolution of I. subelongatus and E. fujiensis with respect to host tree volatiles but not in response to NHVs. Ultimately, we put forward a hypothesis that host plants are ecological and evolutionary determinants of bark beetle-fungus symbioses in terms of their complex signaling interactions.

Interactive Effects of Tree Mixing and Drought on a Primary Forest Pest

Climate change and biodiversity erosion are two major threats to the vitality of the world's forests. However, it is difficult to predict the extent to which tree diversity and environmental conditions interact to modify forest health, and in particular resistance and/or tolerance to insect pests. We used a tree diversity experiment with an irrigation treatment to investigate the effect of mixing and diluting host pines (Pinus pinaster) amongst broadleaved trees on pine stem borer (Dioryctria sylvestrella) infestations under contrasting drought conditions. We further tested whether the attack patterns of this primary pest resulted from direct effects of tree diversity and drought, or was indirectly mediated by their combined effects on pine vigor. The total number of stem borer attacks per plot significantly decreased with tree species richness, i.e. increased with the density of pines, being maximum in pure plots. There were more attacks in irrigated plots. Mixing pines with birches resulted in lower plot infestations but only in irrigated plots. The probability of individual tree being attacked was higher in irrigated plots and decreased with increasing pine density. More vigorous trees, i.e. with higher radial growth, were more likely to be attacked. We suggest that the pine stem borer is attracted by the volatile organic compounds released by the resin exuding from bark cracks. Bark fissuring increases with radial growth, which is enhanced by irrigation and reduced intraspecific competition in low-density pine plots. The presence of birch limits plot infestation, probably because it disrupts host-finding behavior through repellence by non-host volatiles. This phenomenon is mainly observed when pines are vigorous, i.e. when they are irrigated, and particularly attractive to the stem borer. Our results confirm that more severe droughts decrease the attacks by primary pests feeding on tree trunks by reducing host tree vigor and that focal tree species can obtain protection against specialized insect pests when mixed with non-host species. The functional characteristics of herbivorous insects are therefore to be taken into account in predicting the interactive effects of climate change and loss of diversity on forest health.

Antennal olfactory sensory neurones responsive to host and nonhost plant volatiles in gorse pod moth Cydia succedana

AbstractWe investigate the response profiles of the antennal olfactory sensory neurones (OSNs) in male and female gorse pod moth Cydia succedana to host and nonhost volatiles, using the single sensillum recording technique. Eight different classes of olfactory sensilla are identified in female C. succedana and five different classes of olfactory sensilla in males. Nineteen different classes of OSNs are identified from the sensilla in females, and nine different classes of OSNs in the male sensilla. All classes of sensilla, except class F7 and class M1 sensilla, co‐compartmentalize two or three OSNs in each sensillum, and the OSNs present in the same sensillum are specialized for different volatiles. Most plant‐volatile OSNs exhibit phasic‐tonic type of temporal responses, whereas the pheromone OSNs in male C. succedana show rather phasic responses. The majority of OSNs identified in C. succedana display highly specialized responses to a narrow range of volatiles, whereas only a small proportion of OSNs show broad response spectra. Two most abundant classes of OSNs exhibit highly specialized responses to β‐myrcene and (E)‐β‐ocimene, two major volatiles released by gorse (Ulex europaeus), the main host of C. succedana. By contrast, several other classes of OSNs exhibit highly specialized responses to geraniol, (Z)‐3‐hexen‐1‐ol, (±)‐α‐terpineol, citral and benzyl acetate, which are produced by various nonhost plants. Taking the results of the present study together, we suggest that C. succedana use the combinational input from a set of highly specialized OSNs for host plant volatiles and another set of highly specialized OSNs for nonhost volatiles to discriminate between hosts and nonhosts.

The Ratio between Field Attractive and Background Volatiles Encodes Host-Plant Recognition in a Specialist Moth.

Volatiles emitted by plants convey an array of information through different trophic levels. Animals such as host-seeking herbivores encounter plumes with filaments from both host and non-host plants. While studies showed a behavioral effect of non-host plants on herbivore host location, less information is available on how a searching insect herbivore perceives and flies upwind to a host-plant odor plume within a background of non-host volatiles. We hypothesized here that herbivorous insects in search of a host-plant can discriminate plumes of host and non-host plants and that the taxonomic relatedness of the non-host have an effect on finding the host. We also predicted that the ratio between certain plant volatiles is cognized as host-plant recognition cue by a receiver herbivorous insect. To verify these hypotheses we measured the wind tunnel response of the moth Argyresthia conjugella to the host plant rowan, to non-host plants taxonomically related (Rosaceae, apple and pear) or unrelated to the host (Pinaceae, spruce) and to binary combination of host and non-host plants. Volatiles were collected from all plant combinations and delivered to the test insect via an ultrasonic sprayer as an artificial plume. While the response to the rowan as a plant was not affected by the addition of any of the non-host plants, the attraction to the corresponding sprayed headspace decreased when pear or apple but not spruce were added to rowan. A similar result was measured toward the odor exiting a jar where freshly cut plant material of apple or pear or spruce was intermixed with rowan. Dose-response gas-chromatography coupled to electroantennography revealed the presence of seven field attractive and seven background non-attractive antennally active compounds. Although the abundance of field attractive and of some background volatiles decreased in all dual combinations in comparison with rowan alone, an increased amount of the background compounds (3E)-4,8-Dimethyl-1,3,7-nonatriene ((E)-DMNT) and (Z)-3-hexenyl acetate was found in the rowan-apple and rowan-pear but not in the rowan-spruce headspace. A higher ratio between the abundance of each field attractive component and that of (E)-DMNT and (Z)-3-hexenyl acetate was measured for rowan and rowan-spruce in contrast to rowan-pear and rowan-apple headspaces. Our result suggests that the ratio between field attractive and background antennaly active volatiles encodes host-plant recognition in our study system.

Sensory specificity and speciation: a potential neuronal pathway for host fruit odour discrimination in Rhagoletis pomonella.

Behavioural changes in habitat or mate choice can trigger population divergence, leading to speciation. However, little is known about the neurological bases for such changes. Rhagoletis pomonella (Diptera: Tephritidae) is a model for ecological speciation via host plant shifts. Within the past 180 years, Rhagoletis flies infesting hawthorn (Crataegus spp.) shifted to attack domesticated apple (Malus pumila). The two populations differ in their olfactory preferences for apple versus hawthorn fruit. Here, we looked for patterns of sensory organization that may have contributed to this shift by characterizing the morphology, specificity and distribution of olfactory sensory neurons (OSNs) on the antennae of Rhagoletis responding to host fruit and non-host volatiles. Of 28 OSN classes identified, two colocalized OSN pairs were found that specifically responded to the major behavioural attractant and antagonist volatiles for each fly population. A reversal in the response of these OSNs to fruit volatiles, either through a switch in receptor expression between these paired neurons or changes in neuronal projections in the brain, could therefore account for the behavioural difference between apple and hawthorn flies. The finding supports the hypothesis that relatively minor changes in olfactory sensory pathways may contribute to rapid host shifting and divergence in Rhagoletis.

Olfactory Cues, Visual Cues, and Semiochemical Diversity Interact During Host Location by Invasive Forest Beetles

Plant-feeding insects use visual and olfactory cues (shape, color, plant volatiles) for host location, but the relative importance of different cues and interactions with non-host-plant volatiles in ecosystems of varying plant biodiversity is unclear for most species. We studied invasive bark beetles and wood borers associated with pine trees to characterize interactions among color, host and non-host volatiles, by employing traps that mimic tree trunks. Cross-vane flight intercept traps (black, green, red, white, yellow, clear) and black funnel traps were used with and without attractants (α-pinene + ethanol), repellents (non-host green leaf volatiles, 'GLV'), and attractant/repellent combinations in four pine forests in New Zealand. We trapped 274,594 Hylurgus ligniperda, 7842 Hylastes ater, and 16,301 Arhopalus ferus. Trap color, attractant, and color × attractant effects were highly significant. Overall, black and red traps had the highest catches, irrespective of the presence of attractants. Alpha-pinene plus ethanol increased trap catch of H. ligniperda 200-fold but only 6-fold for H. ater and 2-fold for A. ferus. Green leaf volatiles had a substantial repellent effect on trap catch of H. ligniperda but less on H. ater and A. ferus. Attack by H. ligniperda was halved when logs were treated with GLV, and a similar effect was observed when logs were placed among broadleaved understory shrubs emitting GLV. Overall, H. ligniperda was most strongly affected by the olfactory cues used, whereas H. ater and A. ferus were more strongly affected by visual cues. Collectively, the results support the semiochemical diversity hypothesis, indicating that non-host plant volatiles from diverse plant communities or artificial dispensers can contribute to resistance against herbivores by partly disrupting host location.