Electroantennogram Responses Research Articles

Isolation and identification of the main components of garlic, Allium sativum, volatiles and their determination of repellent or toxic activities on green peach aphid, Myzus persicae

AbstractTo determine the control mechanism of garlic on Myzus persicae, volatiles from garlic plants were collected, and the components were determined by GC–MS. Then, the volatile extracts and standard components were screened by electroantennogram (EAG) apparatus and Y‐tube olfactometer, and the toxic effects were determined by the immersion method. The results showed the main components from garlic plant volatile were 2‐methoxyphenol, 2,4‐di‐tert‐butylphenol, diallyl disulfide, butyl phthalate esters, (1‐hydroxycyclohexyl) phenyl ketone, and 2,6‐di‐tert‐butylp‐cresol. The volatile extracts and the standard components had significant repellent effects (P ≤ 0.05) on M. persicae when the volume ratio was 1:100. The relative values of EAG from volatile extracts and diallyl disulfide were significantly higher than those of the other substances (P ≤ 0.05) when the volume ratio was 1:100. The toxicity of garlic volatile extracts to M. persicae significantly (P ≤ 0.05) increased with increasing concentration. After 72 h (treated), the adjusted mortality of the extract to M. persicae was 36.99% at 90 ml/L. The standard components had the significant toxic effect on M. persicae at 30 mg/mL, and the rates of mortality were over 78.95%. Thus, the results suggested that the volatile extracts and the four standard components from garlic plants had significant repellent effects and electroantennogram responses for M. persicae, and the toxic effects were stronger than the repellent effects for M. persicae.

Environmental exposure to cadmium induces olfactory neurotoxicity in fire ants and the molecular basis

Cadmium (Cd) exhibits widely olfactory toxicity to animals. We previously reported that Cd exposure induces the transcriptional dysregulation of olfactory marker proteins (OMPs) of the red imported fire ant Solenopsis invicta. However, it is still unclear how environmental Cd exposure-induced deregulation of OMPs affects the olfactory signal transduction and olfaction-driven social behavior of S. invicta. Here, we showed that S. invicta displayed dull sensory perception on bait in Cd-contaminated areas and dietary Cd ingestion by S. invicta reduced the bait search efficiency. We hypothesize that deregulation of OMPs by Cd exposure blocks the olfactory signal transduction in fire ants. Our results indicated the odor binding protein 14 (SiOBP14) was consistently inhibited in antennal sensilla of fire ants across Cd exposure at 0.5, 5 and 50 mg/kg. Function analysis in vitro and in vivo demonstrated that SiOBP14 is essential in perception of S. invicta to bait odorants. Cd-exposed fire ants showed weak odorant receptor neurons (ORNs) chemosensory signaling and electroantennogram (EAG) response. Moreover, Cd exposure repeals the preference of S. invicta to the active bait odorants, including 2-methyltetrahydrofuran-3-one, 2-methyl-3-furanthiol and 4,5-dimethylthiazole, and even triggers a behavioral transition from preference to repellence. These results indicate that Cd exposure inhibits the specific OMP expression and disrupts olfactory signal transduction, thereby inducing dull sensory perception of S. invicta to bait odorants. The findings provide new implications for monitoring and control of agricultural insect pests in heavy metal polluted areas.

Binding characterization of odorant-binding protein BhorOBP29 in Batocera horsfieldi (Hope) with host-plant volatiles

Odorant binding proteins (OBPs) are involved in odorant discrimination and act as the first filter in the peripheral olfactory system. Previous studies have shown that BhorOBP29 is potentially involved in olfactory perception in an important wood-boring pest Batocera horsfieldi (Hope) (Coleoptera: Cerambycidae), however, its function remains unclear. Here, we investigated the ligand-binding profiles of recombinant BhorOBP29 with 22 compounds from its host plant using fluorescence competitive binding assays and fluorescence quenching assays. The results showed that BhorOBP29 could bind to five ligands relying mainly on hydrophobic interactions. Molecular docking analysis indicated that residues Ile48, Leu51, Met52, Trp57, Asn105, and Val119 were extensively involved in the interactions between BhorOBP29 and the five ligands. Furthermore, the site-directed mutagenesis analysis revealed that Leu51 and Met52 residues were indispensable for BhorOBP29-ligands binding. Finally, electroantennogram (EAG) assays confirmed that hexanal, (−)-limonene, and 2-methylbutyraldehyde elicited a concentration-dependent EAG response with a maximum at the concentration of 1/10 v/v. These findings suggest that BhorOBP29 may play a significant role in the perception of host plant volatiles by B. horsfieldi. This study may help to discover novel behavioral regulation and environmentally friendly strategies for controlling B. horsfieldi in the future.

RpedOBP1 plays key roles in aggregation pheromones reception of the Riptortus pedestris

Riptortus pedestris (Hemiptera: Alydidae) is a notable soybean pest, with diapause and non-diapause individuals showing different sensitivities to aggregation pheromones. This study aimed to investigate how R. pedestris detects aggregation pheromones through electroantennogram (EAG) and behavioral experiments, transcriptome sequencing and qRT-PCR, as well as competitive fluorescence-binding assay. Results indicated that diapausing females and males of R. pedestris exhibited a heightened EAG response and were more attracted to the aggregation pheromone components compared to their non-diapause counterparts. Transcriptome sequencing and qRT-PCR analyses revealed significantly higher expression of RpedOBP1 in the antennae of diapause females and males compared to non-diapausing R. pedestris. The competitive fluorescence-binding assay demonstrated that RpedOBP1 displayed the strongest binding affinity to E2HE2H, suggesting its crucial role in recognizing the aggregation pheromone. These findings have the potential to inform the development of integrated pest management strategies utilizing behavioral approaches for bean bug control.

Behavioral and electrophysiological responses of the male medfly, Ceratitis capitata, to thymol and carvacrol ethers.

The Mediterranean fruit fly, Ceratitis capitata, is one of the most economically important insect pests attacking fruits and vegetables in tropical and subtropical areas of the world. Semiochemical-based pest management programs are being used to provide environmentally friendly control methods for medflies. The goals of the current study were to discover potential new, attractive, kairomones by designing, synthesizing, and testing simplified ethers of thymol and carvacrol along with their ether derivatives in short-range attraction assays and electroantennogram (EAG) assays with male C. capitata. To the best of our knowledge, this study represents the first investigation of thymol and carvacrol, and their respective ethers for attractancy to C. capitata, a major agricultural pest worldwide. In short-range attraction bioassays, parent compounds, thymol and carvacrol, along with their propyl, butyl, benzyl, and octyl ethers captured the most male C. capitata. The attraction patterns changed over time and captures were only significant if they were greater than the positive controltea tree oil (TTO) at 90 min. In EAG assays, thymol benzyl, octyl ethers, and carvacrol benzyl ether evoked significantly greater antennal responses than their parent compounds. The EAG responses did not correlate with short-range male attraction. The aliphatic side chains of thymol and carvacrol had a small effect on the activity. Future studies will investigate the long-range attraction of the ethers that elicited large EAG responses. This report provides new information for discovering potential kairomones through synthesis and structure-activity studies for sterile male medflies. Thymol, carvacrol, and several of their ether derivatives displayed improved longevity of attraction compared with TTO (a strong medfly attractant), with significantly higher captures than TTO observed at 90 min in laboratory bioassays. Further chemical synthesis of thymol and carvacrol ethers within this series may lead to the development of ethers that are more attractive or persistent than their parent compounds, thymol and carvacrol. © 2024 Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Repellency, toxicity, and physiological actions of low molecular weight basic amines in mosquitoes.

This study investigated the behavioral responses and toxicity of three basic amines: 1-methylpiperazine, 1-methylpyrrolidine, and triethylamine (TEA), compounds suggested previously to be anosmic in vapor exposures to caged mosquitoes. These compounds showed repellency of Aedes aegypti mosquitoes, followed by flightlessness, knockdown, and paralysis, all increasing with exposure time and dosage. Electrophysiological experiments showed a blocking effect on nerve discharge of the Drosophila melanogaster larval central nervous system (CNS) with little evidence of hyperexcitation. Blockage of voltage-gated (Kv2) potassium channel currents under patch clamp occurred at similar concentrations. Involvement of K+ channels in the action of basic amines was supported by behavior and CNS recordings of a Shaker Kv1 mutant exposed to TEA, where instead of blockage, a hyperexcitation of nerve firing was observed. Experiments on cockroach leg mechanoreceptors demonstrated neuronal excitation and on mosquito antennae strong electroantennogram (EAG) signals with an augmentation of blank air responses after a single puff of basic amine. The neurophysiological effects of basic amines are consistent with K+ channel block, whereas the antennal EAG response was not obviously associated with anosmia. The low-dose effects of basic amines appear to be repellency and bradykinesia. Overall, the findings provide key insights into the mechanisms underlying the biological activity of basic amines. © 2024 Society of Chemical Industry.

Functional characterization of two odorant‐binding proteins to sex pheromones in Spodoptera frugiperda

AbstractThe primary function of odorant‐binding proteins (OBPs) is thought to be binding and transporting odorants molecules. The research on the molecular mechanisms of SfruOBPs binding with sex pheromones is beneficial to the prevention and control of Spodoptera frugiperda (S. frugiperda). Herein, two screened SfruOBPs (Sfru10933 and Sfru21187) were knocked down by RNA interference, and then the changes in electroantennogram (EAG) and behavioral responses of male mutants to their main sex pheromone components were recorded. Upon interfering with Sfru10933, the EAG response and behavioural preferences of males to sex pheromones were significantly reduced or eliminated. The interference with Sfru21187 resulted in a weakening of the behavioural preference, while the EAG response remained unchanged. However, the co‐interference of Sfru10933/Sfru21187 led to a significant reduction in the EAG activity of male to sex pheromones. These results reveal that Sfru10933 and Sfru21187 play a crucial role in sex pheromone recognition, and there may be a synergistic effect between them. Furthermore, these results will help clarify the perception of sex pheromones by S. frugiperda, which may provide new targets for the development of mating inhibitors in the future.

Electrophysiological and behavioral responses of Tamarixia radiata (Hymenoptera: Eulophidae) to volatiles of nymphal Diaphorina citri (Hemiptera: Liviidae)

Abstract Huanglongbing (HLB), a devastating citrus disease caused by Candidatus Liberibacter asiaticus, is efficiently vectored by the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae). Tamarixia radiata (Waterston) plays a crucial role as an ectoparasitoid, preying on D. citri nymphs. By collecting and identifying headspace volatiles from fifth instar nymphs of D. citri using a gas chromatograph–mass spectrometer (GC–MS), we obtained a collection of 9 volatile compounds. These compounds were subsequently chosen to investigate the electrophysiological and behavioral responses of female T. radiata. At a concentration of 10 μg/μl, 9 compounds were compared with cis-3-hexen-1-ol (control), resulting in trans-2-nonenal inducing the highest relative electroantennogram (EAG) value, followed by hexanal, heptanal, n-heptadecane, tetradecanal, n-tetradecane, n-pentadecane, 1-tetradecanol, and 1-dodecanol. The top 5 EAG responses of female T. radiata to these compounds were further investigated through EAG dose–response experiments. The results showed positive dose–responses as concentrations increased from 0.01 to 10 μg/μl. In Y-tube olfactometer bioassays, female T. radiata exhibited a preference for specific compounds. They were significantly attracted to tetradecanal at a concentration of 10 µg/µl and trans-2-nonenal at 0.01 µg/µl, while no significant attraction was observed toward hexanal, heptanal, or n-heptadecane. Our report is the first to demonstrate that volatiles produced by D. citri nymphs attract T. radiata, which suggests that this parasitoid may utilize nymph volatiles to locate its host.

Electrophysiological and behavioural responses of fall armyworm Spodoptera frugiperda: A potential phytovolatile technology for its management

The fall armyworm (FAW), scientifically known as Spodoptera frugiperda (J.E. Smith) (Lepidoptera:Noctuidae), is an invasive and polyphagous pest associated with maize (Zea mays L.). An experiment was conducted during 2020–21 at South Eastern University of Sri Lanka, Oluvil, Sri Lanka and the Coconut Research Institute of Sri Lanka Lunuwila, Sri Lanka to investigate the olfactory perception of 22 host plant volatiles by electroantennogram (EAG) and behavioural assays for males, virgin and mated females of FAW. The findings indicated that trans-2-Hexen-1-ol, Benzaldehyde, Nonanoic acid, cis-3-Hexen-1-ol, and 4-Phenyl-2-butanone elicited elevated EAG responses in FAW antennae. Additionally, β-Citronellol and ethyl butyrate elicited amplified responses specifically in mated females and males, respectively. Dose-response study revealed that the observed responses remained statistically non-significant (P>0.05), regardless gender or physiological condition of females across the 1%, 5%, 10% and 15% dose intervals. In behavioural study, observed that 1% Benzaldehyde attracted a substantial number of moths to the treatment side for both virgin and mated females, whereas 1% β-Citronellol displayed repellent properties specifically for mated females of FAW. Therefore, these findings suggest that volatile plant compounds may serve as a theoretical basis for devising novel control strategies for FAW. This includes the potential development of attractants, repellents, and mass capture killers effective for both males and females of FAW, in addition to the currently used female sex pheromone.

The different responses of Batocera horsfieldi adults to Juglans regia bark and leaf volatile compounds

AbstractBatocera horsfieldi (Hope) (Coleoptera: Cerambycidae) is an important pole borer pest. It causes serious harm to various hosts, particularly Juglans regia L. (Juglandaceae). In this study, headspace solid‐phase microextraction (HS‐SPME), gas chromatography–mass spectrometry (GC–MS), and electroantennogram (EAG) responses were combined to examine the mechanism by which B. horsfieldi adults locate their host, J. regia. The results showed that J. regia contained 65 volatile compounds in all, with 36 and 42 volatile substances in bark and leaves, respectively. Moreover, terpenes accounted for the largest relative content among the volatile compounds of bark and leaves, 43.4 and 78.9%, respectively. Ten of 19 selected volatile components elicited significant EAG responses in adult B. horsfieldi, which indicated that the pest may prefer certain volatile compounds when selecting a host. Sex and volatile concentration affected the responses of B. horsfieldi. Male adults had the strongest EAG responses to 1‐octen‐3‐ol, whereas female adults had the strongest EAG responses to linalool. Host selection in B. horsfieldi was somewhat positively associated with the concentrations of the effective volatile compounds in the host. Furthermore, B. horsfieldi showed varying preferences for isomers. This work provided data support for the development of insect attractants, and laid a theoretical foundation for the ecological protection of local forestry.

Development of terpenoid repellents against Aedes albopictus: a combined study of biological activity evaluation and computational modelling

ABSTRACT To explore novel terpenoid repellents, 22 candidate terpenoid derivatives were synthesized and tested for their electroantennogram (EAG) responses and repellent activities against Aedes albopictus. The results from the EAG experiments revealed that 5-(2-hydroxypropan-2-yl)-2-methylcyclohex-2-en-1-yl formate (compound 1) induced distinct EAG responses in female Aedes albopictus. At concentrations of 0.1, 1, 10, 100, and 1000 mg/L, the EAG response values for compound 1 were 179.59, 183.99, 190.38, 193.80, and 196.66 mV, demonstrating comparable or superior effectiveness to DEET. Repellent activity analysis indicated significant repellent activity for compound 1, closest to the positive control DEET. The in silico assessment of the ADMET profile of compound 1 indicates that it successfully passed the ADMET evaluation. Molecular docking studies exhibited favourable binding of compound 1 to the active site of the odorant binding protein (OBP) of Aedes albopictus, involving hydrophobic forces and hydrogen bond interactions with residues in the OBP pocket. The QSAR model highlighted the influential role of hydrogen-bonding receptors, positively charged surface area of weighted atoms, polarity parameters of molecules, and maximum nuclear-nuclear repulsion force of carbon-carbon bonds on the relative EAG response values of the tested compounds. This study holds substantial significance for the advancement of new terpenoid repellents.

1-nonene plays an important role in the response of maize-aphid-ladybird tritrophic interactions to nitrogen.

Plant volatile organic compounds (VOCs) are the key distress signals involved in tritrophic interactions, by which plants recruit predators to protect themselves from herbivores. However, the effect of nitrogen fertilization on VOCs that mediate tritrophic interactions remains largely unidentified. In this study, a maize (Zea mays)-aphid (Rhopalosiphum padi)-ladybird (Harmonia axyridis) tritrophic interaction model was constructed under high-nitrogen (HN) and low-nitrogen (LN) regimens. H. axyridis had a stronger tendency to be attracted by aphid-infested maize under HN conditions. Then, volatiles were collected and identified from maize leaves on which aphids had fed. All of the HN-induced volatiles (HNIVs) elicited an electroantennogram (EAG) response from H. axyridis. Of these HNIVs, 1-nonene was attractive to H. axyridis under simulated natural volatilization. Furthermore, our regression showed that the release of 1-nonene was positively correlated with H. axyridis visitation rates. Supplying 1-nonene to maize on which aphids had fed under LN enhanced attractiveness to H. axyridis. These results supported the conclusion that 1-nonene was the active compound that mediated the response to nitrogen in the tritrophic interaction. In addition, the 1-nonene synthesis pathway was hypothesized, and we found that the release of 1-nonene might be related to the presence of salicylic acid (SA) and abscisic acid (ABA). This research contributes to the development of novel environmentally friendly strategies to optimize nitrogen fertilizer application and to improve pest control in maize crops.

The greening-causing agent alters the behavioral and electrophysiological responses of the Asian citrus psyllid to a putative sex pheromone

The Asian Citrus Psyllid (ACP), Diaphorina citri, is a vector of the pathological bacterium Candidatus Liberibacter asiaticus (CLas), which causes the most devastating disease to the citrus industry worldwide, known as greening or huanglongbing (HLB). Earlier field tests with an acetic acid-based lure in greening-free, ‘Valencia’ citrus orange groves in California showed promising results. The same type of lures tested in São Paulo, Brazil, showed unsettling results. During the unsuccessful trials, we noticed a relatively large proportion of females in the field, ultimately leading us to test field-collected males and females for Wolbachia and CLas. The results showed high rates of Wolbachia and CLas infection in field populations. We then compared the olfactory responses of laboratory-raised, CLas-free, and CLas-infected males to acetic acid. As previously reported, CLas-uninfected males responded to acetic acid at 1 µg. Surprisingly, CLas-infected males required 50 × higher doses of the putative sex pheromone, thus explaining the failure to capture CLas-infected males in the field. CLas infection was also manifested in electrophysiological responses. Electroantennogram responses from CLas-infected ACP males were significantly higher than those obtained with uninfected males. To the best of our knowledge, this is the first report of a pathogen infection affecting a vector’s response to a sex attractant.

Electrophysiological and Behavioral Responses of Plodia interpunctella (Hübner) Females to Aldehyde Volatiles from Dried Fruits.

Plodia interpunctella (Lepidoptera: Pyraloidea) is a notorious pest of stored grain globally. The dried fruits (Ziziphus jujuba, Malus pumila, and Fragaria ananassa) can strongly attract P. interpunctella. However, specific volatile compounds responsible for such effects have not been identified. Volatiles were analyzed by using headspace solid-phase microextraction (HS-SPME) and chromatography-mass spectrometry (GCMS) techniques. Five aldehyde compounds were selected for electroantennogram (EAG), single sensillum recording (SSR), and behavioral response assays. The three chemicals that elicited the strongest EAG responses to mated females at 100 μg/μL include hexanal (1.13 mV), heptanal (0.92 mV), and octanal (0.73 mV). In SSR experiments, the basiconic sensilla of the antennae responded to these aldehyde compounds. The results of behavioral responses showed that all aldehydes exhibited dose-dependent responses, with hexanal having the highest attractant rate of 74.56%. These compounds have the potential to be used for monitoring P. interpunctella and its integrated management program.

Transcription factor CncC regulates the expression of antennal CYP6MU1 gene responsible for trans-2-hexen-1-al and nonanal recognition in Locusta migratoria

Cytochrome P450 monooxygenases (P450s) are a superfamily of multifunctional heme-containing proteins and could function as odorant-degrading enzymes (ODEs) in insect olfactory systems. In our previous study, we identified a P450 gene from the antennal transcriptome of Locusta migratoria, LmCYP6MU1, which could be induced by a variety of volatiles. However, the regulatory mechanisms of this gene in response to volatiles remain unknown. In current study, we investigated the tissues and development stages expression patterns of LmCYP6MU1 and determined its olfactory function in the recognition of the main host plant volatiles which induced LmCYP6MU1 expression. The results showed that LmCYP6MU1 was antenna-rich and highly expressed throughout the antennal developmental stages of locusts. LmCYP6MU1 played important roles in the recognition of trans-2-hexen-1-al and nonanal. Insect CncC regulates the expression of P450 genes. We tested whether LmCncC regulates LmCYP6MU1 expression. It was found that LmCncC knockdown in the antennae resulted in the downregulation of LmCYP6MU1 and repressed the volatiles-mediated induction of LmCYP6MU1. LmCncC knockdown reduced the electroantennogram (EAG) and behavioral responses of locusts to volatiles. These results suggested that LmCncC could regulate the basal and volatiles-mediated inducible expression of LmCYP6MU1 responsible for the recognition of trans-2-hexen-1-al and nonanal. These findings provide an original basis for understanding the regulation mechanisms of LmCncC on LmCYP6MU1 expression and help us better understand the LmCncC-mediated olfactory plasticity.

An odorant receptor tuned to an attractive plant volatile vanillin in Spodoptera litura

The insect olfaction plays crucial roles in many important behaviors, in which ORs are key determinants for signal transduction and the olfactory specificity. Spodoptera litura is a typical polyphagous pest, possessing a large repertoire of ORs tuning to broad range of plant odorants. However, the specific functions of those ORs remain mostly unknown. In this study, we functionally characterized one S. litura OR (OR51) that was highly expressed in the adult antennae. First, by using Xenopus oocyte expression and two-electrode voltage clamp recording system (XOE-TEVC), OR51 was found to be strongly and specifically responsive to vanillin (a volatile of S. litura host plants) among 77 tested odorants. Second, electroantennogram (EAG) and Y-tube behavioral experiment showed that vanillin elicited significant EAG response and attraction behavior especially of female adults. This female attraction was further confirmed by the oviposition experiment, in which the soybean plants treated with vanillin were significantly preferred by females for egg-laying. Third, 3D structural modelling and molecular docking were conducted to explore the interaction between OR51 and vanillin, which showed a high affinity (−4.46 kcal/mol) and three residues (Gln163, Phe164 and Ala305) forming hydrogen bonds with vanillin, supporting the specific binding of OR51 to vanillin. In addition, OR51 and its homologs from other seven noctuid species shared high amino acid identities (78–97%) and the same three hydrogen bond forming residues, suggesting a conserved function of the OR in these insects. Taken together, our study provides some new insights into the olfactory mechanisms of host plant finding and suggests potential applications of vanillin in S. litura control.

The Aversive Response of the Locust Locusta migratoria to 3-Octanone, an Odorant from Fungal Pathogens, Is Mediated by a Chemosensory Protein

(1) Locusts are important agricultural pests. Identifying harmful substances and avoiding them is important for locusts’ survival; their abilities to do so remain to be clarified. (2) We examined the electrophysiological (electroantennogram (EAG) and single sensillum recording (SSR)) and behavioral responses (preference behavior in a T-maze) of locusts to 18 different compounds; (3) Of these 18 compounds, 9 elicited strong EAG responses, and 3 elicited SSR responses of neurons expressing locust odorant receptor 3 (LmigOR3). The 11 chemicals that elicited stronger EAG or SSR responses were selected for evaluation of the behavioral responses of locusts. Only 2-heptanone induced significant attraction responses in locusts at the tested concentration. RNA interference (RNAi) of LmigOR3 and SSR experiments revealed that LmigOR3 could detect 2-heptanone and 3-octanone. However, in behavioral experiments, RNAi of LmigOR3 did not alter 2-heptanone-induced attraction but increased attraction by 3-octanone. (4) Our results suggest that the broadly tuned receptor expressed in a heterologous expression system exhibits a narrow electrophysiological response spectrum, and the aversive response of locusts to 3-octanone, an odorant from fungal pathogens, natural enemies, and non-host plants, is mediated by LmigOR3. These findings enhance our understanding of the complex olfactory recognition mechanism in insects.

Foraging specificity of Tetralonia (Thygatina) macroceps (Hymenoptera: Apidae: Anthophorinae) on Argyreia cuneata (Convolvulaceae)

Floral specificity is a behavior that evolved due to mutualistic interactions between the plant-pollinator community. Flowers advertise themselves using visual or chemical cues to attract pollinators and gain reproductive success through pollination. Pollinators forage for rewards such as nectar or pollen produced by the flowers. We found that an anthophorid bee, Tetralonia macroceps, foraged specifically on Argyreia cuneata flowers. No visitation was observed on the flowers of A. nervosa though both belong to Convolvulaceae. T. macroceps was the most abundant floral visitor (5.21 bees/flower/5 min) on A. cuneata and did not visit A. nervosa. Mass flowering and narrow tubular flower structure with easy access to pollen in A. cuneata were the traits that accounted for the foraging specificity of T. macroceps. The present study investigates the preference of T. macroceps for the flowers and floral extracts of A. cuneata and A. nervosa. The bee visited 10.16 flowers/5 min of A. cuneata. T. macroceps were highly attracted to the flowers of A. cuneata. No bees were attracted to A. nervosa. The floral abundance of A. cuneata was relatively higher compared to A. nervosa. Pollen analysis of foraging bees of T. macroceps revealed the selective preference towards the pollen of A. cuneata. The highest number of bees preferred the extract of A. cuneata (7.75) compared to A. nervosa (0.50) in the Y-olfactory maze. Floral extract of A. cuneata caused the highest neuronal electroantennogram (EAG) response (1.48 mV) than A. nervosa (0.36 mV). Our preliminary studies indicated the presence of specific volatile organic compounds (VOCs) nonacosane (13.26%), hexatriacontane (12.06%), and beta farnesene (6.19%) observed in A. cuneata were absent in congener A. nervosa.

Behavioral and Electrophysiological Study on Eight Japanese Papilio Species with Five Hostplant Volatiles and Linalool

An electroantennogram (EAG) technique compared the antennal olfactory responses by both sexes of eight Japanese Papilio species with known host plants in laboratory experiments. Papilio species were collected from Honshû and Kyûshû (Japanese islands). The behavioral responses to volatile leaf substances from Citrus deliciosa, Zanthoxylum ailanthoides, Phellodendron amurense, Orixa japonica, and Foeniculum vulgare were examined in laboratory experiments. Individual EAG reactions were recorded. The results were very similar to the empirical field observations. The electrophysiological results of both sexes showed that the volatile substances released from non-preferred plants mainly elicited more significant EAG responses than the volatile substances from preferred host plants. Moreover, we performed behavioral experiments using eight female butterflies and their responses to five host plant species. An association between host plant selection behavior and taxonomical classification exists within the Papilio genus. The EAG responses were small when exposed to the plants with high scores in the behavioral experiments. Host plant preference patterns seem to be related to the volatile substances within the host plants. The butterflies responded to Linalool in both the behavioral and electrophysiological experiments.

Attraction of Male Pine Sawflies, Diprion jingyuanensis, to Synthetic Pheromone Candidates: Synergism between Two Stereoisomers

The pine sawfly Diprion jingyuanensis Xiao and Zhang (Hymenoptera: Diprionidae) is a serious pest of Pinus tabulaeformis Carr. in the Shanxi, Gansu, and Inner Mongolia provinces in P. R. China. The sex pheromone of D. jingyuanensis was shown to be the propionate ester of 3,7-dimethyl-2-tridecanol. Virgin females contained an approximate 1:3 blend of the pheromone precursors erythro-(2S,3S,7R/S and 2R,3R,7R/S)-3,7-dimethyl-2-tridecanol and threo-(2S,3R,7R/S and 2R,3S,7R/S)-3,7-dimethyl-2-tridecanol, but the exact stereoisomers were not determined. Males responded the strongest to the propionate ester of the two threo-isomers, (2S,3R,7R) and (2S,3R,7S), in electroantennogram (EAG) recordings, followed by a significant EAG response to the (2S,3R,7R) propionate of diprionol (pheromone component of D. similis), whereas the remaining two isomers (2S,3S,7S and 2S,3S,7R) of the propionate ester of 3,7-dimethyl-2-tridecanol and the acetate of the (2S,3R,7R) isomer (one of the two pheromone components of D. pini) did not elicit any significant increase in antennal response. In the field, the strongly EAG-active (2S,3R,7R)-isomer alone was only weakly (but significantly) attractive to D. jingyuanensis males at 100 µg, while the equally EAG- active (2S,3R,7S)-isomer alone at the same loading was 8–14 times more attractive than was the (2S,3R,7R)-isomer alone. Traps baited with the same amounts of the two threo-isomers ((2S,3R,7R) and (2S,3R,7S), 100 µg + 100 µg) caught significantly more males than did traps baited with other isomers, either of the two isomers alone or other proportions of the two isomers. Thus, the (2S,3R,7S)-isomer is considered as a strong and essential sex-attractant component for D. jingyuanensis males, whereas the (2S,3R,7R)-isomer is a weak but synergistic sex-attractant. This is one of the few examples of a pine sawfly responding significantly stronger to a binary blend of stereoisomers in a synergistic fashion than to a single stereoisomer alone.