Host Plant Location Research Articles

Identification of attractants for adult Spodoptera litura based on the interaction between odorant-binding protein 34 and host volatiles

Odorant-binding proteins (OBPs) play key roles in host plant location by insects, and can accordingly serve as important targets for the development of attractants. In this study, we detected the high expression of SlitOBP34 in male antennae of Spodoptera litura. Subsequently, the fluorescence competitive binding experiments displayed that the SlitOBP34 protein has binding affinity for different ligands. Then, protein-ligand interaction analyses found the presence of six amino acid residues may serve as key recognition sites. Further electroantennographic and biobehavioral assessments revealed that the electrophysiological responses of male antennae were evoked in response to stimulation with the six identified host volatiles, and that these volatiles attracted male moths to varying extents. Notably, low concentrations of benzaldehyde, 1-hexanol, and cis-3-hexenyl acetate were found to have significant attractant effects on male moths, thereby identifying these three host volatiles as potential candidates for the development of male attractants. These findings advance our current understanding of the olfactory-encoded mechanisms of host plants selection in S. litura and have enabled us to develop novel adult attractants for controlling the pest in the future.

Identification of odorant-binding proteins and functional analysis of antenna-specific BhorOBP28 in Batocera horsfieldi (Hope).

The important wood-boring pest Batocera horsfieldi has evolved a sensitive olfactory system to locate host plants. Odorant-binding proteins (OBPs) are thought to play key roles in olfactory recognition. Therefore, exploring the physiological function of OBPs could facilitate a better understanding of insect chemical communications. In this research, 36 BhorOBPs genes were identified via transcriptome sequencing of adults' antennae from B. horsfieldi, and most BhorOBPs were predominantly expressed in chemosensory body parts. Through fluorescence competitive binding and fluorescence quenching assays, the antenna-specific BhorOBP28 was investigated and displayed strong binding affinities forming stable complexes with five volatiles, including (+)-α-Pinene, (+)-Limonene, β-Pinene, (-)-Limonene, and (+)-Longifolene, which could also elicit conformation changes when they were interacting with BhorOBP28. Batocera horsfieldi females exhibited a preference for (-)-Limonene, and a repellent response to (+)-Longifolene. Feeding dsOBP19 produced by a bacteria-expressed system with a newly constructed vector could lead to the knockdown of BhorOBP28, and could further impair B. horsfieldi attraction to (-)-Limonene and repellent activity of (+)-Longifolene. The analysis of site-directed mutagenesis revealed that Leu7, Leu72, and Phe121 play a vital role in selectively binding properties of BhorOBP28. By modeling the molecular mechanism of olfactory recognition, these results demonstrate that BhorOBP28 is involved in the chemoreception of B. horsfieldi. The bacterial-expressed dsRNA delivery system gains new insights into potential population management strategies. Through the olfactory process concluded that discovering novel behavioral regulation and environmentally friendly control options for B. horsfieldi in the future. © 2024 Society of Chemical Industry.

Antennal Sensitivity of Spotted Lanternflies, Lycorma delicatula: Differential Electrophysiological Responses of Males and Females to Compounds Derived from Host Plants and Conspecifics.

In herbivorous insects, antennae play a crucial role in chemical communication and orientation when locating host plants and mates. To evaluate antennal sensitivity in response to odor stimuli, electroantennography (EAG) has been a practical technique. In the current study of the invasive spotted lanternfly (SLF), Lycorma delicatula (Hemiptera: Fulgoridae), we evaluated and compared their antennal sensitivity to a series of volatile chemicals collected from their bodies, honeydew, and host plants. To do this, we exposed the antennae of SLF fourth-instar and adult males and females to individual chemicals at a fixed dose of 50 ng. Further, a series of dose-response tests were carried out within a range of 0.5 to 100 ng. Although the amplitude of antennal responses varied among stages and sexes, adult males generated the strongest antennal responses in both experiments. In dose-response experiments, increased doses of chemicals up to 50 ng revealed the saturation points except in adult females which required a higher dose (100 ng) to reveal the saturation point. Although EAG does not provide any information on behavioral responses, our results are consistent with the olfactory bioassays in previous publications in which adult males, not females, were attracted to natural volatiles of their conspecifics. EAG indicated a higher sensitivity of adult male antennae to odor stimuli, particularly conspecific volatiles, than female antennae and highlighted sexual differences in the perception of chemical cues in SLF.

Visual and chemical cues in the host plant selection of the melon ladybird Chnootriba elaterii (Coleoptera: Coccinellidae)

Phytophagous insects employ various sensory cues in the host plant location such as visual, olfactory, gustatory, and tactile stimuli, which are perceived by sensory systems and integrated in higher brain centres to trigger the correct behavioural responses. In the present study, the host location process of the oligophagous species Chnootriba elaterii (melon ladybird) is investigated in controlled conditions using both a Y-Tube olfactometer and an open Y-Track olfactometer. Olfactory and visual cues from the host plant act synergistically to lead the ladybirds towards the host plant. Females of C. elaterii are not able to discriminate between host and non-host plants using either olfactory or visual cues alone. Visual cues, particularly those associated with the colour green, are of higher relative importance compared to olfactory cues in the host location process. Green dummy plants made of cardboard represent strong supernormal stimuli for C. elaterii females. The results of the present study can shed light on the host location process of Coccinellidae and can help to develop visual or chemical traps which can be useful in monitoring and controlling this important crop pest of the Mediterranean Basin.

Plant volatiles mediated the orientation preference of slugs to different plant species.

Slugs mechanically damage plant leaves, resulting in significant economic losses. However, there are limited cost-efficient strategies available in slug management. By studying how slugs utilize plant volatiles to locate host plants, we can gain insights into the design of attractants and repellents. Bioassay results suggest slugs (Agriolimax agrestis) prefer to orientate to lettuce (Lactuca sativa), cabbage (Brassica oleracea L.), and young tobacco seedlings, compared with old tobacco seedlings. We analyzed the volatomics of lettuce, cabbage, young and old tobacco seedlings. 2-(2-butoxyethoxy)-ethanol acetate (2EA) had high abundance while nonanal, decanal, and β-cylocitral had relatively low content in volatiles. Old tobacco seedlings released significantly more hexanal but fewer 1,4-dihydro-4-oxopyridazine (DO). In olfactory tests, hexanal, nonanal, decanal, and β-cylocitral showed strong repellency to slugs, while DO at a dose of 500 ng/μL and 2EA at a dose of 1% were attractive to slugs. The two alkanes, hexadecane and heptadecane, had no effect on slug orientating to host plants. DO and 2EA can thus alleviate the repellency of hexanal, nonanal, decanal and β-cylocitral. The high emission of hexanal in old tobacco seedlings helps repel slugs, while 2EA and DO attract slugs to lettuce and cabbage. These findings suggest that these chemicals can be utilized in the design of repellents and attractants, and contribute to constructing a push-pull system for slug control. © 2023 Society of Chemical Industry.

Antennae-enriched expression of candidate odorant degrading enzyme genes in the turnip aphid, Lipaphis erysimi.

Aphids heavily rely on their olfactory system for foraging behavior. Odorant-degrading enzymes (ODEs) are essential in preserving the olfactory acuity of aphids by removing redundant odorants in the antennae. Certain enzymes within this group stand out as being enriched and/or biased expressed in the antennae, such as carboxylesterases (CXEs), cytochrome P450 (CYPs), glutathione S-transferases (GSTs), and UDP-glycosyltransferases (UGTs). Here, we performed a comparative transcriptome analysis of antennae and body tissue to isolate the antennal ODE genes of turnip aphid Lipaphis erysimi. A dataset of one CXE, seven CYPs, two GSTs, and five UGTs enriched in the antennae was identified and subjected to sequence analysis. Furthermore, qRT-PCR analyses showed that 13 ODE genes (LeCXE6, LeCYP4c1, LeCYP6a2, LeCYP6a13, LeCYP6a14.2, LeCYP6k1, LeCYP18a1, LeGST1, LeUGT1-7, LeUGT2B7, LeUGT2B13, LeUGT2C1.1, and LeUGT2C1.2) were specifically or significantly elevated in antennal tissues. Among these antennae-enriched ODEs, LeCYP4c1, LeCYP6a2, LeCYP6a13, LeCYP6a14.2, LeCYP18a1, LeUGT2B7, and LeUGT2B13 were found to exhibit significantly higher expression levels in alate aphids compared to apterous and nymph aphids, suggesting their putative role in detecting new host plant location. The results presented in this study highlight the identification and expression of ODE genes in L. erysimi, paving the path to investigate their functional role in odorant degradation during the olfactory processes.

A Review of the Host Plant Location and Recognition Mechanisms of Asian Longhorn Beetle.

The Asian longhorn beetle (ALB), Anoplophora glabripennis Motschulsky, is a polyphagous xylophage with dozens of reported host tree species. However, the mechanisms by which individuals locate and recognize host plants are still unknown. We summarize the current knowledge of the host plant list, host kairomones, odorant-binding proteins (OBPs) and microbial symbionts of this beetle and their practical applications, and finally discuss the host localization and recognition mechanisms. A total of 209 species (or cultivars) were reported as ALB host plants, including 101 species of higher sensitivity; host kairomones were preferentially bound to ALB recombinant OBPs, including cis-3-hexen-1-ol, δ-3-carene, nonanal, linalool, and β-caryophyllene. In addition, microbial symbionts may help ALB degrade their host. Complementarity of tree species with different levels of resistance may reduce damage, but trapping effectiveness for adults was limited using a combination of host kairomones and sex pheromones in the field. Therefore, we discuss host location behavior from a new perspective and show that multiple cues are used by ALB to locate and recognize host plants. Further research into host resistance mechanisms and visual signal recognition, and the interaction of sex pheromone synthesis, symbiont microbiota, and host plants may help reveal the host recognition mechanisms of ALBs.

Sizing up spotted lanternfly nymphs for instar determination and growth allometry.

A major ongoing research effort seeks to understand the behavior, ecology and control of the spotted lanternfly (SLF) (Lycorma delicatula), a highly invasive pest in the U.S. and South Korea. These insects undergo four nymphal stages (instars) before reaching adulthood, and appear to shift host plant preferences, feeding, dispersal and survival patterns, anti-predator behaviors, and response to traps and chemical controls with each stage. However, categorizing SLF life stage is challenging for the first three instars, which have the same coloration and shape. Here we present a dataset of body mass and length for SLF nymphs throughout two growing seasons and compare our results with previously-published ranges of instar body lengths. An analysis using two clustering methods revealed that 1st-3rd instar body mass and length fell into distinct clusters consistently between years, supporting using these metrics to stage nymphs during a single growing season. The length ranges for 2nd-4th instars agreed between years in our study, but differed from those reported by earlier studies for diverse locations, indicating that it is important to obtain these metrics relevant to a study's region for most accurate staging. We also used these data to explore the scaling of SLF instar bodies during growth. SLF nymph body mass scaled with body length varied between isometry (constant shape) and growing somewhat faster than predicted by isometry in the two years studied. Using previously published data, we also found that SLF nymph adhesive footpad area varies in direct proportion to weight, suggesting that footpad adhesion is independent of nymphal stage, while their tarsal claws display positive allometry and hence disproportionately increasing grasp (mechanical adhesion). By contrast, mouthpart dimensions are weakly correlated with body length, consistent with predictions that these features should reflect preferred host plant characteristics rather than body size. We recommend future studies use the body mass vs length growth curve as a fitness benchmark to study how SLF instar development depends on factors such as hatch date, host plant, temperature, and geographic location, to further understanding of life history patterns that help prevent further spread of this invasive insect.

Complex host/symbiont integration of a multi-partner symbiotic system in the eusocial aphid Ceratovacuna japonica.

Some hemipteran insects rely on multiple endosymbionts for essential nutrients. However, the evolution of multi-partner symbiotic systems is not well-established. Here, we report a co-obligate symbiosis in the eusocial aphid, Ceratovacuna japonica. 16S rRNA amplicon sequencing unveiled co-infection with a novel Arsenophonus sp. symbiont and Buchnera aphidicola, a common obligate endosymbiont in aphids. Both symbionts were housed within distinct bacteriocytes and were maternally transmitted. The Buchnera and Arsenophonus symbionts had streamlined genomes of 432,286bp and 853,149bp, respectively, and exhibited metabolic complementarity in riboflavin and peptidoglycan synthesis pathways. These anatomical and genomic properties were similar to those of independently evolved multi-partner symbiotic systems, such as Buchnera-Serratia in Lachninae and Periphyllus aphids, representing remarkable parallelism. Furthermore, symbiont populations and bacteriome morphology differed between reproductive and soldier castes. Our study provides the first example of co-obligate symbiosis in Hormaphidinae and gives insight into the evolutionary genetics of this complex system.

Olfaction in phytophagous ladybird beetles: antennal sensilla and sensitivity to volatiles from host plants in Chnootriba elaterii

Olfaction and chemical ecology in phytophagous ladybird beetles have been largely ignored so far. The present paper describes for the first time basiconic, grooved, and trichoid porous olfactory sensilla on the antennae of the phytophagous ladybird Chnootriba elaterii (Epilacnini) and demonstrates the sensitivity of the antennae to Volatile Organic Compounds (VOCs) from host plants, belonging to common and less common Cucurbitaceae crops and wild species (Citrullus lanatus, Cucumis melo, Cucumis sativus, Cucurbita pepo, Cucurbita moschata, Lagenaria siceraria, Luffa cylindrica, and Ecballium elaterium). Epilachnini ladybirds are phytophagous, with adult and larvae feeding on various Cucurbitaceae, Fabaceae, and Solanaceae. Notwithstanding some species are serious pests in crops, their chemical ecology has been largely disregarded so far. The present paper discusses the ultrastructure of antennal sensilla (scanning electron microscopy-SEM) and the electroantennographic recordings (EAG) from the olfactory sensilla responding to the proposed chemicals, also considering the literature on chemical ecology and olfaction of aphidophagous ladybugs. In particular, antennal sensilla are similar to those described in predatory species but in Epilachnini they are much more numerous on the apical flagellum, probably because involved in host plant location and selection. This research can be considered a first step in developing semiochemical-based and sustainable control strategies for melon ladybird in crops.

Ultrastructure Characteristics and Sexual Dimorphism of Antennal Sensilla in Tirathaba rufivena (Lepidoptera: Pyralidae).

Simple SummaryTo elucidate the external morphology and internal structures of antennal sensilla that could play a role in locating host plants and mates, we describe and further characterize the ultrastructure, abundance and distribution of nine sensilla types in both sexes using scanning electron microscopy and transmission electron microscopy. Sexual dimorphism mainly occurs in variation in the length of sensilla basiconica, sensilla chaetica, sensilla coeloconica 1 and Böhm sensilla, and the abundance of sensilla basiconica, sensilla auricillica 1 and sensilla auricillica 2. The possible structural function of each kind of sensillum is discussed. These results provide direct morphological evidence that the antennae of Tirathaba rufivena possess structures that can play a role in locating mates and host plants.Tirathaba rufivena Walker, a major insect pest of Areca catechu L., has severely threatened areca nut cultivation in Hainan, China. To improve our understanding of the communication mechanism in host plant seeking and mate-finding for T. rufivena, we described and further characterized the external morphology and internal sensilla structures using scanning electron microscopy and transmission electron microscopy in this study. The antennal morphology was similar between males and females, and there was no significant difference in length between the two sexes. In total, nine sensilla types were identified: sensilla trichodea (Str), sensilla chaetica (Sch), sensilla basiconica (Sba), sensilla auricillica (Sau), sensilla coeloconica (Sco), sensilla styloconica (Sst), Böhm sensilla (Bs), uniporous peg sensilla (Ups) and sensilla squamiformia (Ssq). Sexual dimorphism mainly occurs in variation in the length of Sba, Sch, Sco1 and Bs, and the abundance of Sba, Sau1 and Sau2. The Sba had larger size and numbers on female antennae than that on males, suggesting that these sensilla might have important roles in locating host plants. Both Sau1 and Sau2 were significantly more abundant in females and were probably associated with the detection of mates and host plant for oviposition. These data were important for ongoing studies on host plant seeking and mate-finding behavior in T. rufivena and provided a theoretical foundation to further studies of semiochemical control for this pest.

Identification and functional analysis of a chemosensory protein from Bactrocera minax (Diptera: Tephritidae).

Olfaction has an indispensable role in insect behavior, enabling location of suitable host plants and oviposition sites, finding mates and evasion of natural enemies. Chemosensory proteins (CSPs) function to screen external odorants and transport them to olfactory receptor neurons, thereby increasing the sensitivity of the olfactory system. At present, CSP genes have been identified in many insect species, but there are relatively few studies on the function of CSP, especially in Tephritidae. In this study, we sequenced and analyzed 12 transcriptomes of Bactrocera minax and identified five CSP genes. The results of polymerase chain reactions with reverse transcription showed that BminCSP3 was highly expressed only in antennae. Results from competitive binding experiments showed that BminCSP3 has good binding ability to citral compared with 23 other volatile organic compounds. The docking model with citral showed hydrogen bond formation with residues (ARG97); however, no hydrogen bonds were formed in the docking of five other ligands (furfuryl alcohol, linalool, cis-3-hexenyl acetate, (R)-(+)-limonene and (+)-carvone). Electroantennogram (EAG) analyses revealed that citral was active in B. minax at the antennal level, and the EAG response value of female adults was significantly higher than that of male adults. Furthermore, the results of behavioral bioassays showed that females were significantly attracted to citral. Our results suggest that BminCSP3 plays an important role in the recognition of citral by B. minax adults. © 2022 Society of Chemical Industry.

Different Binding Affinities of Three General Odorant-Binding Proteins in Grapholita funebrana (Treitscheke) (Lepidoptera: Tortricidae) to Sex Pheromones, Host Plant Volatiles, and Insecticides.

Insect general odorant-binding proteins (GOBPs) play irreplaceable roles in filtering, binding, and transporting host odorants to olfactory receptors. Grapholita funebrana (Treitscheke) (Lepidoptera: Tortricidae), an economically important pest of fruit crops, uses fruit volatiles as cues to locate host plants. However, the functions of GOBPs in G. funebrana are still unknown. Three GOBP genes, namely, GfunGOBP1, GfunGOBP2, and GfunGOBP3, were cloned, and their expression profiles in different tissues were detected by the method of real-time quantitative PCR (RT-qPCR). The binding properties of recombinant GfunGOBPs (rGfunGOBPs) to various ligands were investigated via fluorescence binding assays. The three GfunGOBPs were mainly expressed in the antennae of both male and female moths. All these three rGfunGOBPs could bind to sex pheromones, while having varying affinities toward these pheromones. The three rGfunGOBPs also displayed a wide range of ligand-binding spectrums with tested host odorants. The rGfunGOBP1, rGfunGOBP2, and rGfunGOBP3 bound to 34, 33, and 30 out of the 41 tested odorants, respectively. Three rGfunGOBPs had overlapping binding activities to β-myrcene, (-)-α-phellandrene, and ethyl isovalerate with the Ki less than 3.0 μM. The rGfunGOBP1 and rGfunGOBP3 could selectively bind to several insecticides, whereas rGfunGOBP2 could not. Three rGfunGOBPs had the dual functions of selectively binding to sex pheromones and host odorants. Moreover, the rGfunGOBP1 and rGfunGOBP3 can also serve as 'signal proteins' and bind to different insecticides. This study contributed to elucidating the potential molecular mechanism of the olfaction for G. funebrana, and thereby promotes the development of effective botanical attractants or pheromone synergists to control G. funebrana.

Transcriptome Analysis of Antennal Chemosensory Genes in Curculio Dieckmanni Faust. (Coleoptera: Curculionidae).

The olfactory system plays a key role in regulating insect behaviors, such as locating host plants, spawning sites, and mating partners and avoiding predators. Chemosensory genes are required for olfactory recognition in insects. Curculio dieckmanni Faust. (Coleoptera: Curculionidae) damages hazelnuts and causes severe economic losses. There are no effective control measures, but understanding the olfaction mechanisms of this insect could lead to a new approach for population management. However, the genes that perform chemosensory functions in C. dieckmanni are still unclear. Using high-throughput sequencing, we assembled the antennal transcriptome of C. dieckmanni and annotated the major chemosensory gene families. Of the chemosensory gene families, we found 23 odorant-binding proteins, 15 chemosensory proteins, 2 sensory neuron membrane proteins, 15 odorant receptors, 23 ionotropic receptors, and nine gustatory receptors. Using Blast sequence alignment and phylogenetic analysis, the sequences of these proteins were identified. Male- and female-specific chemosensory genes involved in odorant detection and recognition were validated by qRT-PCR. Among the chemosensory genes, we found significant differences in the expression of CdieOBP8, CdieOBP9, CdieOBP19, CdieOBP20, CdieOBP21, CdieCSP15, CdieOR13, and CdieOR15 between adult male and female C. dieckmanni. A total of 87 expressed chemosensory proteins were found in C. dieckmanni. Investigating these proteins will help reveal the molecular mechanism of odorant recognition in C. dieckmanni and may aid the development of novel control strategies for this species.

Identification and functional characterization of ApisOr23 in pea aphid Acyrthosiphon pisum

Pea aphid, Acyrthosiphon pisum, is a serious pest of many different leguminous plants, and it mainly relies on its odorant receptors (Ors) to discriminate among host species. However, less is known about the role that Ors play in the host plant location. In this study, we identified a novel conserved odorant receptor clade by phylogenetic analysis, and conducted the functional analysis of ApisOr23 in A. pisum. The results showed that the homologous Ors from A. pisum, Aphis glycines and Aphis gossypii share 94.28% identity in amino acid sequences. Moreover, conserved motifs were analyzed using the annotated homologous Or23 from eight aphid species, providing further proof of the high conservation level of the Or23 clade. According to the tissue expression pattern analysis, ApisOr23 was mainly expressed in the antennae. Further functional study using a heterologous Xenopus expression system revealed that ApisOr23 was tuned to five plant volatiles, namely trans-2-hexen-1-al, cis-2-hexen-1-ol, 1-heptanol, 4´-ethylacetophenone, and hexyl acetate. Among them, trans-2-hexen-1-al, which is one of the main volatile organic compounds released from legume plants, activated the highest response of ApisOr23. Our findings suggest that the conserved Or23 clade in most aphid species might play an important role in host plant detection.

More phylogenetically diverse polycultures inconsistently suppress insect herbivore populations.

Because the diet of many herbivorous insects is restricted to closely related taxa with similar chemistry, intercropping with diverse plant communities may reduce both pest populations and reliance on chemical pesticides in agroecosystems. We tested whether the effectiveness of intercropping against herbivorous insects depends on the phylogenetic relatedness of neighboring crops, using butternut squash (Cucurbita moschata) as a focal crop species in a series of different intercropping combinations. We found that increased phylogenetic divergence of neighboring plants could reduce abundance of herbivorous insects, but the effect was only detectable mid-season. In addition, we tested two hypothesized mechanisms for a negative association between phylogenetic distance of neighboring plants and reduced herbivore populations: one, we tested using Y-tube olfactometer and choice cage trials whether diverse volatile cues impede host-plant location by the dominant pest of butternut squash in our experiment, striped cucumber beetle Acalymma vittatum. Two, we recorded predator and parasitoid abundance relative to crop phylodiversity to test whether diverse crops support larger natural-enemy populations that can better control pest species. Our results, however, did not support either hypothesis. Striped cucumber beetles preferentially oriented toward non-host-plant volatiles, and predator populations more often decreased with phylodiversity than increased. Thus, the mechanisms driving associations in the field between phylogenetic divergence and herbivore populations remain unclear.

The Adelphocoris lineolatus OBP4: Support for evolutionary and functional divergence of a mirid pheromone-binding protein from that found in lepidopteran moths.

Pheromone-binding proteins (PBPs) have been extensively investigated in lepidopteran moths, but their evolution and function in hemipteran species remain unclear. Our previous study demonstrated that an odorant-binding protein, OBP4, of the mirid bug Adelphocoris lineolatus functions as a candidate hemipteran PBP but clustered with lepidopteran antennae-binding proteins (ABPs) rather than in the PBP/general odorant-binding protein (GOBP) clade. In this study, we hypothesized that origin and function of PBPs in hemipteran bugs may differ from those of lepidopteran moths. To test this hypothesis, we first constructed a phylogenetic tree using insect OBPs from sister hemipteran and holometabolous lineages, and the results indicated that neither OBP4 nor other types of candidate PBPs of mirid bugs clustered with the lepidopteran PBP/GOBP clade. Then, a fluorescence competitive binding assay was employed to determine binding affinities of recombinant OBP4 protein to host plant volatiles, with functional groups different from A. lineolatus sex pheromone components. The results revealed that OBP4 highly bound the female adult attractant 3-hexanone and 15 other mirid bug biologically active plant volatiles. Finally, we examined cellular expression profiles of OBP4 in putative antennal sensilla that are related to female A. lineolatus host plant location. The fluorescence in situ hybridization and immunocytochemical labeling assay showed that the OBP4 gene was highly expressed in the multiporous olfactory sensilla medium-long sensilla basiconica rather than in the short sensilla basiconica or uniporous sensilla chaetica. These results, together with those of our previous studies, indicate that OBP4 not only functions in recognition of bug-produced sex pheromones in males, but is probably involved in detection of host plant volatiles in both A. lineolatus sexes. Our findings support the hypothesis that the origin and function of PBPs in hemipteran bugs differ from those of well-known PBPs in lepidopteran moths, which provides a novel perspective on evolutionary mechanisms of sex pheromone communication across insect orders.

Plant-associated CO2 mediates long-distance host location and foraging behaviour of a root herbivore.

Insect herbivores use different cues to locate host plants. The importance of CO2 in this context is not well understood. We manipulated CO2 perception in western corn rootworm (WCR) larvae through RNAi and studied how CO2 perception impacts their interaction with their host plant. The expression of a carbon dioxide receptor, DvvGr2, is specifically required for dose-dependent larval responses to CO2. Silencing CO2 perception or scrubbing plant-associated CO2 has no effect on the ability of WCR larvae to locate host plants at short distances (<9 cm), but impairs host location at greater distances. WCR larvae preferentially orient and prefer plants that grow in well-fertilized soils compared to plants that grow in nutrient-poor soils, a behaviour that has direct consequences for larval growth and depends on the ability of the larvae to perceive root-emitted CO2. This study unravels how CO2 can mediate plant-herbivore interactions by serving as a distance-dependent host location cue.

Herbivore-induced plant volatiles mediate behavioral interactions between a leaf-chewing and a phloem-feeding herbivore

Plants respond adaptively to herbivore stress in order to maintain fitness. Upon herbivore attack, plants emit blends of volatile organic compounds (VOCs) that differ from those that are constitutively emitted. These defense responses are typically specific to the identity of the attacking herbivore and often linked to the herbivore's feeding guild (e.g. chewing, phloem-feeding). Herbivores use plant volatiles to locate suitable host plants and changes in volatile emissions can affect host-plant location. Therefore, herbivores from separate feeding guilds can interact indirectly through the modulation of plant responses. In this study we tested how damage by an herbivore from one feeding guild affected the host-plant choice of an herbivore from a separate feeding guild, and vice versa. A chewing herbivore, the Colorado potato beetle (Leptinotarsa decemlineata), and a phloem feeding herbivore, the green peach aphid (Myzus persicae), were assayed in olfactometers to assess behavioral responses to odors emitted by potato plants (Solanum tuberosum) that were damaged by herbivores from the other feeding guild. Leptinotarsa decemlineata oriented more frequently towards undamaged plants compared to M. persicae damaged plants. Surprisingly, M. persicae preferred plants that were damaged by L. decemlineata, although previous studies had shown that they perform worse on these plants. Distinct differences were detected in the volatile profiles of herbivore-damaged and undamaged plants. Leptinotarsa decemlineata induced stronger volatile emissions compared to undamaged control plants, while M. persicae tended to suppress volatile emissions. These herbivores demonstrate contrasting induction of plant volatiles and behavioral responses. Exploring the nature of co-occurring herbivores and how they perceive potential hosts can play a significant role in understanding the ecological functions and community dynamics of plant plasticity and interactions with a variety of herbivores.

Fine Morphology of Antennal and Ovipositor Sensory Structures of the Gall Chestnut Wasp, Dryocosmus kuriphilus.

Simple SummaryHymenoptera encompass a large group of insects with different habits, ranging from phytophagy to parasitic/predatory lifestyles. This is also true in the superfamily Cynipoidea, where phytophagy becomes highly specialized towards the exploitation of specific plant tissues (i.e., buds), leading to the induction of galls. In this paper, we investigated the organization of antennal and ovipositor sensory structures in the chestnut gall wasp, Dryocosmus kuriphilus. This insect became a major threat to chestnut production in Italy in the last 15 years. We investigated only females (this is a parthenogenetic species with thelytoky), and on the antennae we found several sensilla with the clear functional specialization to different groups of stimuli, with almost no overlapping among each sensilla. Similarly, specialization was also found on the ovipositor where groups of gustatory and mechanoreceptive sensilla were observed. This information represents an advancement in the knowledge of this pest, which may be useful to understand the biological role of plant derived chemical cues or to implement new control methods.Dryocosmus kuriphilus is a gall-inducing insect, which can cause significant damage on plants of the genus Castanea Mill., 1754. Antennae and ovipositor are the main sensory organs involved in the location of suitable oviposition sites. Antennal sensilla are involved in the host plant location, while ovipositor sensilla assess the suitability of the ovipositional bud. On both organs, diverse sensillar organs are present. Here, the distribution and ultrastructural organization of the sensilla were investigated by scanning and transmission electron microscopy. The antennae of D. kuriphilus are filiform and composed of 14 antennomeres, with the distal flagellomere bearing the highest number of sensilla. On the antennae, 6 sensilla types were found; sensilla chaetica, campaniformia, coeloconica-I, coeloconica-II, trichoidea and placoidea. The sensilla placoidea and trichoidea were the most abundant types. On the external walls of the ovipositor, gustatory and mechanoreceptive sensilla were observed. Internally, the egg channel hosted two additional sensory structures. The putative functional role of each sensilla in the context of insect’s ecology is discussed as well as the ovipositional mechanism used by this insect.