Electroantennogram Research Articles

Expression Profiling and Functional Analysis of Candidate Odorant Receptors in Galeruca daurica.

Simple SummaryOdorant receptors (ORs) play an important role in the olfactory system in insects. However, there is no functional research on the odorant receptors of Galeruca daurica. In this study, 21 OR genes were identified from the transcriptome database of G. daurica adults. Most GdauORs were mainly expressed in antennae, and the expression levels of GdauORs in adults were affected by age. When GdauOR4, GdauOR15, and GdauORco were silenced by RNAi, the electrophysiological responses to host plant volatiles were significantly decreased.Galeruca daurica (Joannis) is an oligophagous pest in the grasslands of Inner Mongolia, China, which feed mainly on Allium spp. Odorant receptors (ORs) play an important role in the olfactory system in insects, and function together with olfactory co-receptor (ORco). In this study, 21 OR genes were identified from the transcriptome database of G. daurica adults, and named GdauOR1-20 and GdauORco. The expression profiles were examined by RT-qPCR and RNA interference (RNAi) and electroantennogram (EAG) experiments were conducted to further identify the olfactory functions of GdauOR4, GdauOR11, GdauOR15, and GdauORco. It was found that 15 GdauORs (OR1, OR3-6, OR8, OR11-13, OR15, OR17-20, and ORco) were mainly expressed in antennae, and the expression levels of GdauORs in adults were affected by age. When GdauOR4, GdauOR15, and GdauORco were silenced by RNAi, the electrophysiological responses to host plant volatiles were significantly decreased in G. daurica. This study lays a necessary foundation for clarifying the mechanism on finding host plants in G. daurica.

Electrophysiological and behavioural responses of Stegobium paniceum to volatile compounds from Chinese medicinal plant materials.

BACKGROUND Stegobium paniceum (Coleoptera, Anobiidae) is an important pest of stored products causing severe damage to dried Chinese medicinal plant materials (CMPMs). Plant volatiles play an important role in host‐searching of insects. The olfactory responses of S. paniceum to the most abundant volatile components of some drugstore attractant CMPMs such as Panax notoginseng, Angelica sinensis, Gastrodia elata and Peucedanum praeruptorum, namely falcarinol, 3‐n‐butylphthalide, p‐cresol and β‐pinene, respectively, were studied by electroantennography (EAG) and behavioural bioassays in six‐ and four‐arm olfactometers.RESULTSEAG recordings showed that male and female antennae are able to perceive the test compounds in a wide range of concentrations and in a dose‐dependent manner. Moreover, for each dose of different compounds tested, no significant differences were found between the mean male and female EAG responses. In six‐arm olfactometer bioassays, S. paniceum exhibited positive responses to falcarinol, 3‐n‐butylphthalide, p‐cresol and β‐pinene at doses of 1, 10, 100, 500 and 1000 μg. The most attractive dose was 500 μg for falcarinol, 100 μg for 3‐n‐butylphthalide, 500 μg for p‐cresol and 1000 μg for β‐pinene. Olfactory preferences of S. paniceum, based on comparison of these four compounds at their optimally attractive concentrations in a four‐arm olfactometer, were 3‐n‐butylphthalide > p‐cresol > falcarinol > β‐pinene.CONCLUSIONThe results indicated that the four volatiles of CMPMs are perceived by the peripheral olfactory system of S. paniceum adults and are able to individually elicit a positive chemotaxis in S. paniceum adults confirming the role of chemical cues in host‐plant detection and selection of this pest. Further field studies are needed to evaluate the potential of the attractive compounds identified in this study, particularly 3‐n‐butylphthalide, to be applied as a novel monitoring and control tool against this storage‐beetle pest. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Abnormal Antennal Olfactory Sensilla Phenotypes Involved in Olfactory Deficit in Bactrocera correcta (Diptera: Tephritidae).

Simple SummaryTephritidae fruit flies sense odorants mainly through antennal olfactory sensilla with nanopores. Therefore, theoretically, the development of nanopore-targeted pest control technologies is an important direction in the future. Here, we report naturally occurring abnormal antennal trichoid and basiconic olfactory sensilla phenotypes consisting of abnormal bulges and reduced nanopore numbers in a long-term laboratory rearing colony of the guava fruit fly Bactrocera correcta, and further find that the reduction of nanopore numbers in these sensilla led to an olfactory deficit. Our findings provide a basis for developing nanopore-targeted pest control technologies in the future.The guava fruit fly, Bactrocera correcta, is one of the most destructive pests in the genus Bactrocera and detects environmental odorants mainly through antennal olfactory sensilla phenotypes with nanopores. However, it is unclear whether there are naturally occurring abnormal antennal olfactory sensilla phenotypes that affect olfaction. Here, we found that there were abnormal bulges besides nanopores on the surface of trichoid and basiconic olfactory sensilla in the antennal flagellum of long-term laboratory rearing colony (LTC), and that nanopore number in these olfactory sensilla was also remarkably reduced. Notably, the electroantennogram (EAG) responses of LTC insects to methyl eugenol or β-caryophyllene were inhibited, and their behavioral responses elicited by the same odorants were also impaired. These results revealed naturally occurring abnormal antennal olfactory sensilla phenotypes which were involved in olfactory deficit in B. correcta, providing a platform to further study nanopore-targeted pest control technologies in the future.

Odor-Pollution From Fungicides Disrupts Learning and Recognition of a Common Floral Scent in Bumblebees (Bombus impatiens)

Background and AimsBumblebees provide vital pollination services to both natural and agricultural ecosystems. Consequently their declines in species-diversity and population size over the last five decades is alarming. Direct contributors to these declines include pesticides, habitat loss, and disease. However, given that colony fitness is linked to foraging success, successful conservation requires mitigation of any anthropogenic practices that negatively impact foraging. Previous work has shown that agrochemical odor-pollution, including that of fungicides, can modulate bumblebee foraging behavior. This study investigates how odor pollution from three common fungicides (Safer® Brand Garden Fungicide II, Scotts® Lawn Fungus Control, and Reliant® Systemic Fungicide) affects Bombus impatiens’ floral-odor learning and recognition using an associative learning paradigm.MethodsThe effects of fungicide-odor pollution were tested in three ways: (1) background pollution during floral-odor learning; (2) background pollution during floral-odor recognition; and (3) point (localized) pollution during floral-odor recognition. Electroantennogram (EAG) recordings from B. impatiens confirmed the salience of all odor-stimuli and examined impacts of background fungicide-odor on antennal responses to floral-odor. To better understand how fungicide-odor structure related to behavioral data, scents were sampled (Solid Phase Microextraction) and analyzed using gas chromatography–mass spectrometry. Odors were then characterized using the Compounds Without Borders (CWB) vectorization method.ConclusionAll fungicides tested disrupted floral-odor learning and recognition for at least one concentration tested, and Scotts® was universally disruptive at all tested concentrations. All fungicides induced EAG responses, indicating they provide perceivable odor stimuli. Interestingly, two of three tested fungicides (Scotts® and Reliant®) inhibit antennal responses to Monarda fistulosa odor. Odor characterization supports previous findings that sulfurous scents could be disruptive to odor-driven foraging behaviors. Inability for foraging bumblebees to associate to rewarding floral odors in the presence of fungicidal odor pollution could have negative large-scale implications for colony health and reproductive fitness.

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.

Evaluation of Selected Plant Volatiles as Attractants for the Stick Tea Thrip Dendrothrips minowai in the Laboratory and Tea Plantation.

Simple SummaryThe stick tea thrip Dendrothrips minowai is a key pest in tea plantations in China. In recent years, plant-derived semiochemicals have attracted considerable attention as promising attractants for the management of thrips, due to their safety and low cost. In this study, compounds that have been reported to attract other thrips or emitted from tea plants were evaluated for their electroantennogram (EAG), behavioral tests and field trapping efficacy for D. minowai. The EAG relative response value of D. minowai evoked by p-anisaldehyde, 3-methyl butanal, (E)-β-ocimene, farnesene, nonanal, eugenol, (+)-α-pinene, limonene, (−)-α-pinene, and γ-terpinene was significantly higher than the other compounds. Meanwhile, p-anisaldehyde, eugenol, farnesene, methyl benzoate, 3-methyl butanal, (E)-β-ocimene, (−)-α-pinene, (+)-α-pinene, and γ-terpinene led to attraction or repellency responses of female D. minowai. In addition, trap capture numbers of female D. minowai on sticky traps baited with p-anisaldehyde, eugenol, farnesene, and 3-methyl butanal were significantly higher than the control in tea plantations. Overall, our results highlight the potential application of plant volatiles in the development of effective, eco-friendly lure formulations for use in the monitoring and management of thrips.The stick tea thrip (Dendrothrips minowai Priesner) is the main pest thrip in tea (Camellia sinensis) plantations in China, and seriously affects the quality and yield of tea. Plant-derived semiochemicals provide an alternative to pheromones as lures and these compounds possess powerful attractiveness. In this study, we selected 20 non-pheromone semiochemicals, including compounds that have been reported to attract other thrips and some volatiles emitted from tea plants as the potential attractant components for D. minowai. In electroantennogram (EAG) assays, 10 synthetic compounds (p-anisaldehyde, 3-methyl butanal, (E)-β-ocimene, farnesene, nonanal, eugenol, (+)-α-pinene, limonene, (−)-α-pinene, and γ-terpinene) elicited significant antennal responses in female D. minowai. In addition, a two-choice H-tube olfactometer bioassay showed that D. minowai displayed significant positive responses to eight compound dilutions (p-anisaldehyde, eugenol, farnesene, methyl benzoate, 3-methyl butanal, (E)-β-ocimene, (−)-α-pinene, and (+)-α-pinene) when compared with the solvent control at both 1 and 2 h. Moreover, γ-terpinene exhibited a significantly deterrent effect on D. minowai. Finally, trap catches of four compounds (p-anisaldehyde, eugenol, farnesene, and 3-methyl butanal, respectively) significantly increase in tea plantations. Among these, the maximum number of D. minowai collected by blue sticky traps baited with p-anisaldehyde was 7.7 times higher than the control. In conclusion, p-anisaldehyde, eugenol, farnesene, and 3-methyl butanal could significantly attract D. minowai in the laboratory and under field conditions, suggesting considerable potential as commercial attractants to control D. minowai populations.

Electroantennographic activity of 21aliphatic compounds that bind well to a locust odorant-binding protein.

Odorants that bind well to odorant-binding proteins (OBPs) often trigger olfactory responses and have important biological significance. The locust Locusta migratoria (Meyen) (Orthoptera: Acrididae) is a serious agricultural pest. Twenty-one saturated aliphatic compounds with carbon-oxygen bonds and straight chains of 10-17carbon atoms bind well to anL. migratoria OBP. In this study, olfactory activities of these aliphatic compounds on L. migratoria adult males were tested by electroantennography (EAG) and comparatively analyzed. Four alcohols (undecanol, dodecanol, tridecanol, and tetradecanol), two ketones (2-dodecanone and 2-tridecanone), and two esters (ethyl octanoate and ethyl nonanoate) triggered strong EAG responses, and there was no significant difference between them. The results suggest that the eight compounds are more likely to have important biological significance than the other compounds. Moreover, we found that there is not necessarily a positive correlation between the olfactory activity of odorants and their binding ability with OBP. The study contributes to understanding the odorants with biological significance for L. migratoria and the molecular mechanism of the locust's olfaction.

The Correlation between Volatile Compounds Emitted from Sitophilus granarius (L.) and Its Electrophysiological and Behavioral Responses.

The granary weevil Sitophilus granarius (L.) is one of the most serious primary insect pests of stored products. When S. granarius present in grains, various volatile organic compounds are released as chemical signals which can be used to detect the insects. In this study, volatile chemical compounds released from S. granarius were analyzed using the headspace solid phase micro-extraction (HS-SPME) coupled with gas chromatography (GC)-mass spectrometry (MS) techniques. Two key compounds, 3-hydroxy-2-butanone and 1-pentadecene, were identified from mixed gender of S. granarius adults at high density. Moreover, both male and female adults showed dose-dependent electroantennography (EAG) responses to 3-hydroxy-2-butanone. In behavioral assays, S. granarius was attracted to 3-hydroxy-2-butanone at 0.001 µg/10 µL but repelled at 10 µg/10 µL or higher. S. granarius was consistently repelled by 1-pentadecene at concentrations at 0.001 and 1000 µg/10 µL. 3-hydroxy-2-butanone and 1-pentadecene have considerable potential to offer in the development of new approaches for the monitoring and management of this destructive stored grain insect pest.

Impact of Chronic Exposure to Two Neonicotinoids on Honey Bee Antennal Responses to Flower Volatiles and Pheromonal Compounds.

Volatile compounds provide important olfactory cues for honey bees (Apis mellifera L.), which are essential for their ecology, behavior, and social communication. In the external environment bees locate food sources by the use of floral scents, while inside the hive, pheromones such as the queen mandibular pheromone (QMP) and alarm pheromones serve important functions in regulating colony life and inducing aggressive responses against intruders and parasites. Widely reported alterations of various behaviors in- and outside the hive following exposure to pesticides could therefore be associated with a disturbance of odor sensitivity. In the present study, we tested the effects of neonicotinoid pesticides at field concentrations on the ability of honey bees to perceive volatiles at the very periphery of the olfactory system. Bee colonies were subjected to treatments during the summer with either Imidacloprid or Thiacloprid at sublethal concentrations. Antennal responses to apple (Malus domestica L.) flower volatiles were studied by GC-coupled electro-antennographic detection (GC-EAD), and a range of volatiles, a substitute of the QMP, and the alarm pheromone 2-heptanone were tested by electroantennography (EAG). Short-term and long-term effects of the neonicotinoid treatments were investigated on bees collected in the autumn and again in the following spring. Treatment with Thiacloprid induced changes in antennal responses to specific flower VOCs, with differing short- and long-term effects. In the short term, increased antennal responses were observed for benzyl-alcohol and 1-hexanol, which are common flower volatiles but also constituents of the honey bee sting gland secretions. The treatment with Thiacloprid also affected antennal responses to the QMP and the mandibular alarm pheromone 2-heptanone. In the short term, a faster signal degeneration of the response signal to the positive control citral was recorded in the antennae of bees exposed to Thiacloprid or Imidacloprid. Finally, we observed season-related differences in the antennal responses to multiple VOCs. Altogether, our results suggest that volatile-specific alterations of antennal responses may contribute to explaining several behavioral changes previously observed in neonicotinoid-exposed bees. Treatment effects were generally more prominent in the short term, suggesting that adverse effects of neonicotinoid exposure may not persist across generations.

Encounter With a Selfish Virus Sabotages Its Vector to Orient Toward Requisite Host Plant: A Case Study With Chili Leaf Curl Virus-Whitefly

Interactions of a virus with its vector and host plant have challenged entomologists, pathologists and biologists alike. Phytophagous insects depend on specific host volatile cues to locate suitable host plants for feeding and oviposition. Several studies have revealed that plant viruses modify their insect vector’s orientation toward specific host plants to facilitate their spread and survival. The ecological and molecular basis of this vector behavior modification remains largely unknown and was therefore explored in this study. Interestingly, host volatile preference for non-viruliferous female whiteflies [Bemisia tabaci (Genn.)] was found to be preferentially oriented toward infected chili plant [with chili leaf curl (ChLCV)] volatiles, while viruliferous whiteflies preferred healthy chili plant (Capsicum annum L.) volatiles in olfactometer. The electrophysiological studies involving electroantennogram (EAG) assays exhibited similar trend in EAG response amplitudes. Gas Chromatography linked electroantennodetection (GC EAD) revealed specific plant volatile cues responsible for altered host orientation behavior of the vector. Transcriptome profiling of the viruliferous and non-viruliferous whiteflies and Realtime qPCR validation showed differential expression of certain odorant binding proteins (OBPs) in viruliferous whiteflies. Our results suggest that there is a plant virus mediated altered chemoecological behavior in the vector with respect to orientation toward its host plant. Based on the findings we speculate that the virus mediates such change in the vector for a continued transmission success to the host.

Molecular characterization and its binding properties of general odorant binding protein 2 in Dioryctria abietella

AbstractDioryctria abietella is a major pest of conifer cones that can initiate specific behaviours (locating hosts, finding mates and selecting oviposition sites) through sensitive olfactory systems. General odorant‐binding proteins are involved in the first step in odorant perception in insects, but their specific functional roles in D. abietella chemoreception have not been fully elucidated. In this study, quantitative real‐time polymerase chain reaction (PCR) examinations showed that DabiGOBP2 is predominantly expressed in the antennae of female adults. Western blot experiments further confirmed that these proteins show biased expression in the tissues. Binding property analysis demonstrated that DabiGOBP2 exhibited ligand specificity to terpenes, especially myrcene (Ki = 8.19 ± 0.05). Electroantennogram (EAG) and Y‐tube assays showed that three plant volatiles elicited high EAG responses, but only dodecanal showed a strong attraction to females. However, myrcene and isobutyl benzoate were significantly repellent to males at high doses in the behaviour test. In conclusion, exploring the functions of DabiGOBP2 can help in better understanding the mechanism of the olfactory system of D. abietella and developing novel strategies to control this pest.

Acute ozone exposure impairs detection of floral odor, learning, and memory of honey bees, through olfactory generalization

Air pollution stemming from human activities affects the environment in which plant and animal species live and interact. Similar to primary air pollutants which are emitted, secondary air pollutants, such as tropospheric ozone (O3) formed from nitrogen oxides, are also harmful to human health and plant physiology. Yet, few reports studied the effects of O3 on pollinators' physiology, despite that this pollutant, with its high oxidative potential, likely affects pollinators behaviors, especially the perception of signals they rely on to navigate their environment. Volatile Organic Compounds (VOCs) released by plants are used as signals by different animals. For pollination services, VOCs attract different insects to the flowers and strengthen these interactions. Here, we used the honey bee Apis mellifera as a model to characterize the effects of acute exposure to different realistic mixing ratios of O3 (80-, 120-, and 200-ppb) on two crucial aspects: first, how exposed honey bees detect VOCs; and second, how O3 affects these pollinators' learning and memory processes. With electroantennogram (EAG) recordings, we showed that increasing O3 mixing ratios had a biphasic effect: an initial 25% decrease of the antennal activity when bees were tested directly after exposure (O3 direct effect), followed by a 25% increase in activity and response when bees were allowed a two-hour rest after exposure (O3 delayed effect). In parallel, during olfactory conditioning, increasing O3 mixing ratios in both exposure protocols scarcely affected olfactory learning, followed by a decrease in recall of learned odors and an increase of response to new odors, leading to a higher generalization rate (i.e., discrimination impairment). These results suggest a link between O3-related oxidative stress and olfactory coding disturbance in the honey bee brain. If ozone affects the pollinators' olfaction, foraging behaviors may be modified, in addition with a possible long-term harmful effect on pollination services.

CRISPR-mediated mutagenesis of the odorant receptor co-receptor (Orco) gene disrupts olfaction-mediated behaviors in Bactrocera dorsalis.

Olfaction plays an essential role in insect behavior such as host location, foraging, mating, and oviposition. The odorant receptor co-receptor (Orco) is an obligatory odorant receptor and indispensable in odor perception. Here, we characterized the Orco gene from the oriental fruit fly, Bactrocera dorsalis (Hendel), a notorious agriculture pest. The olfactory deficiency mutants were generated by editing the BdorOrco gene using the CRISPR/Cas9 system. Electroantennograms (EAG) and olfactory preference assays confirmed that BdorOrco-/- mutant flies had reduced perception of methyl eugenol, β-caryophyllene, and ethyl acetate. Oviposition bioassays showed that the eggs laid by BdorOrco-/- females mediated by benzothiazole and 1-octen-3-ol were significantly decreased. In addition, BdorOrco-/- mutant flies took a significantly longer time to locate the food source compared with wild type (WT) flies. Altogether, our data indicated that Orco is essential for multiple physiological processes in B. dorsalis, and it expands our understanding of the function of insect Orco.

Expression profiles and functional prediction of ionotropic receptors in Asian corn borer, Ostrinia furnacalis (Lepidoptera: Crambidae)

Genes involved in chemosensation are essential for odorant-mediated insect behaviors. Odorant receptors (ORs) bind and respond to pheromones and plant volatiles, regulating insect behaviors such as mating and host-plant selection, while ionotropic receptors (IRs), which are present at lower levels in insects than ORs, influence ion channels, especially in agricultural pests. Asian corn borer, Ostrinia furnacalis, is the main pest of maize that causes huge economic losses in Asia. Twenty-one OfurIRs have been identified, but none has been characterized. In this study, tissue-specific expression profiling, phylogenetic analysis, and electroantennography (EAG) analysis were applied to characterize the evolution, expression, and the potential function of OfurIRs. It was found that 20 OfurIRs were highly expressed in the antennae, except for OfurIR75p3, whereas 10 and nine OfurIRs were highly expressed in the proboscis and genitalia, respectively, indicating that these OfurIRs were functionally associated with feeding and oviposition. EAG results showed that seven acids elicited responses in the antennae of O. furnacalis and that 2-oxopentanoic acid displayed a significant female-biased response. Combined with the phylogenetic analysis, 10 OfurIRs in clade 4 were roughly predicted to be candidate receptors for 2-oxopentanoic acid and other tested acids. These results provide basic information about OfurIRs and may help advance the knolwedge on the olfactory system of O. furnacalis.

Biosynthesis of artemisinic acid in engineered Saccharomyces cerevisiae and its attractiveness to the mirid bug Apolygus lucorum

Artemisia annua is an important preferred host of the mirid bug Apolygus lucorum in autumn. Volatiles emitted from A. annua attract A. lucorum. Volatile artemisinic acid of A. annua is a precursor of artemisinin that has been widely investigated in the Chinese herbal medicine field. However, little is known at this point about the biological roles of artemisinic acid in regulating the behavioral trends of A. lucorum. In this study, we collected volatiles from A. annua at the seedling stage by using headspace solid phase microextraction (HS-SPME). Gas chromatography-mass spectrometry (GC-MS) analysis showed that approximately 11.03±6.00 and 238.25±121.67 ng h−1 artemisinic acid were detected in volatile samples and milled samples, respectively. Subsequently, a key gene for artemisinic acid synthesis, the cytochrome P450 gene cyp71av1, was expressed in engineered Saccharomyces cerevisiae to catalyze the production of artemisinic acid. After the addition of exogenous artemisinic alcohol or artemisinic aldehyde, artemisinic acid was identified as the product of the expressed gene. In electroantennogram (EAG) recordings, 3-day-old adult A. lucorum showed significant electrophysiological responses to artemisinic alcohol, artemisinic aldehyde and artemisinic acid. Furthermore, 3-day-old female bugs were significantly attracted by artemisinic acid and artemisinic alcohol at a concentration of 10 mmol L−1, whereas 3-day-old male bugs were attracted significantly by 10 mmol L−1 artemisinic acid and artemisinic aldehyde. We propose that artemisinic acid and its precursors could be used as potential attractant components for the design of novel integrated pest management strategies to control A. lucorum.

The Inhibitory Effect of Powdery Mildew-Induced Volatiles from Rose on Host Selection Behavior of Beet Armyworm Moths (Lepidoptera: Noctuidae)

Abstract Plant-mediated indirect interactions (PMIIs) between phytopathogenic fungi and herbivorous insects on shared host plants occur in nature. Knowledge of PMIIs is critical in plant molecular breeding and integrated pest management. We studied the response and chemical mechanism of beet armyworm, Spodoptera exigua (Hübner), adults to rose plants, Rosa chinensis Jacquin, infected with rose powdery mildew, Podosphaera pannosa (Wallr.: Fr.) de Bary. Using gas chromatography-electroantennographic detection (GC-EAD) coupled with electroantennogram (EAG), we found that beet armyworm antennae responded to 8, 11, and 3 volatile organic compounds (VOCs) from noninfected roses, mildew-infected roses, and mildew alone, respectively. The EAG analyses showed 11 chemicals (e.g., limonene [1], 2-ethyl-1-hexanol [2], linalool [3], nonanal [4], (E)-β-caryophyllene [5], 1-dodecanol [7], nhexadecane [9], 1-hexadecanol [11], methyl palmitate [12], 1-octadecanol [14], and n-butyl hexadecanoate [15]) elicited electrophysiological responses of beet armyworm antennae with significant dose-response relationships (P < 0.05). The EAG responses to the three chemicals (3, 11, and 15) were greater than that to the reference chemical [i.e., (E)-2-hexenal] at 0.5, 5.0, and 50.0 mg/ml. Olfactory and ovipositional behavior assays indicated that three chemicals (2, 3, and 5) significantly attracted beet armyworm females and four chemicals (7, 11, 14, and 15) strongly repelled females. Chemicals 2, 3, and 5 from healthy roses appear to be responsible for the attraction of beet armyworm moths to healthy roses, whereas chemicals 7, 11, 14, and 15 from mildew-infected roses play key roles in inhibiting attraction of moths. VOCs from mildew alone did not attract or repel beet armyworm moths.

Optimization and field evaluation of sex-pheromone of potato tuber moth, Phthorimaea operculella Zeller (Lepidoptera: Gelechiidae).

The potato tuber moth, Phthorimaea operculella is a key pest of potato and tobacco in China. We compared the sex pheromone titers released by P. operculella female, and the electroantennogram (EAG) responses of male antennae to pheromone compounds under laboratory conditions. Then, the optimal sex pheromone ratio was screened in the field. The P. operculella sex pheromone feeding on potato or tobacco was extracted by solvent-extraction method. Main sex pheromone compounds including E4, Z7-13: AC (PTM1) and E4, Z7, Z10-13: AC (PTM2) were found in gland. The titer and relative ratio of P. operculella sex pheromone compounds secreted by female adult moths differ between host populations, while the relative EAG responses trend of P. operculella males to the pheromone components were the same when directly stimulated. In field trials, PTM1:PTM2=8:1-4:1 and PTM1:PTM2=1:4-1:7 was the most attractive ratio to P. operculella males in tobacco and potato fields, respectively. During 2019-2020, after 8 weeks of mass trapping for each year, the sex pheromone could reduce the population of P. operculella in the potato and tobacco fields. In this study, fixed property and quantity analyzing methods were adopted to compare sex pheromones from feeding on potato versus tobacco. EAG tests then were carried out on P. operculella males. Finally, we optimized the ratio of two sex pheromone compounds in potato and examined how this influenced field trapping. By further improving the parameters of sex pheromone application in the field, we demonstrate that deployment of sex pheromones can provide effective control of P. operculella. © 2021 Society of Chemical Industry.

Identification of Sex Pheromone Components of Korean Dioryctria abietella (Lepidoptera: Pyralidae) Population and Synergism of Pheromone and Pine Cone Volatile Blends.

(9Z,11E)-tetradecadienyl acetate (9Z,11E-14:OAc) and (3Z,6Z,9Z,12Z,15Z)-pentacosapentaene (C25 pentaene), known as the sex pheromone of the European Dioryctria abietella population, were identified in the Korean D. abietella population. The ratio of 9Z,11E-14:OAc to C25 pentaene extracted from female pheromone glands was 1:23. The electroantennogram (EAG) response of male antennae to 9Z,11E-14:OAc was stronger than to C25 pentaene, whereas an opposite EAG response was observed in female antennae. Major volatile compounds in mature Korean pine cone (Pinus koraiensis), such as limonene, myrcene, and (-)-α-pinene, elicited dose-dependent EAG responses of male and female antennae. In field attraction testing, traps baited with 100:1,000, 100:2,000, and 100:3,000 ug of 9Z,11E-14:OAc to C25 pentaene were the most attractive to male D. abietella. More males were attracted to traps baited with 9Z,11E-14:OAc+C25 pentaene+limonene than traps baited with 9Z,11E-14:OAc+C25 pentaene. This showed that limonene acted as a synergist to sex pheromone. Delta traps, wing traps, and diamond traps were superior to bucket traps for capturing D. abietella males. This study showed that pheromone traps baited with 100:1,000 ~ 100:3,000 ug of 9Z,11E-14:OAc to C25 pentaene and limonene will be useful for the exact monitoring of D. abietella flight phenology in Pinus koraiensis Sieb. & Zucc. (Pinales: Pinaceae) forests.

The short neuropeptide F receptor regulates olfaction-mediated foraging behavior in the oriental fruit fly Bactrocera dorsalis (Hendel)

The short neuropeptide F (sNPF) signaling system, consisting of sNPF and its receptor (sNPFR), influences many physiological processes in insects, including feeding, growth and olfactory memory. We previously showed that sNPF regulates olfactory sensitivity in the oriental fruit fly Bactrocera dorsalis (Hendel) during starvation. However, the functional analysis of sNPFR is constrained by the failure of RNA interference in this species. Here, we generated a null sNPFR mutant using the CRISPR/Cas9 system to investigate the physiological roles of this receptor in more detail. G0 adults were produced at a frequency of 60.8%, and sNPFR−/− mutants were obtained after several generations of backcrossing followed by self-crossing among heterozygous flies. We found that the mutants were significantly less successful at foraging for certain foods and showed increased foraging latency. Electroantennogram (EAG) assays indicated that the mutants had significantly lower electrophysiological responses to three tested odorants. Furthermore, qPCR data revealed the inhibition of several olfactory receptor genes, including Orco. Immunohistochemistry showed that BdsNPFR was localized in cells under the sensillum on the antennae. Based on their shape and size, the BdsNPFR+ cells differ from odorant receptor neurons (ORNs), which were labeled using a Drosophila melanogaster Orco antibody. Our data suggest that sNPFR regulates olfaction-mediated foraging behavior by mediating interactions between BdsNPFR+ cells and selected ORNs.

Olfactory Response of the Spotted Asparagus Beetle, Crioceris duodecimpunctata (L.) to Host Plant Volatiles.

The spotted asparagus beetle, Crioceris duodecimpunctata (L.) is an invasive host-specific pest of asparagus cultivations. To contribute to the understanding of the role of plant volatiles in host-finding by this species, behavioural and electrophysiological tests were carried out. Y-tube olfactometer bioassays, testing intact or mechanically-damaged cladophylls vs. clean air, revealed sexually-dimorphic responses with males being the only sex attracted to both plant materials. Electroantennographic (EAG) assays showed that antennae of both sexes can perceive a wide range of asparagus volatiles. Male and female EAG profiles were almost similar and (Z)-3-hexen-1-ol was by far the most EAG-active compound. (E)-2-hexenal, (±)-linalool, and 3-heptanone elicited the strongest EAG amplitude within the corresponding chemical groups. Eight of the most EAG-active compounds elicited dose-dependent responses indicating the sensitivity of male and female olfactory systems to changes in stimulus concentration. In a Y-tube olfactometer bioassay, (Z)-3-hexen-1-ol at the doses of 1, 10, and 50μg did not elicit female attraction whereas a significant attraction at the 10μg dose and a repellent effect at the 50μg dose was induced in males. Sexual dimorphism of male behavioural response to host plant volatiles is discussed. This study provides a basis for future investigations that could contribute to the development of semiochemical-based monitoring and management strategies for this pest.