Male-produced Aggregation Pheromone Research Articles

Red palm weevil olfactory proteins annotated from the rostrum provide insights into the essential role in chemosensation and chemoreception

Red palm weevil (RPW), Rhynchophorus ferrugineus (Coleoptera: Curculionidae), is rapidly infesting palm trees (Arecaceae) in several countries, threatening coconut, date, and oil cultivations. The male-produced aggregation pheromone in palm weevils has been reported to be secreted through the mouth to the rostrum, a snout-like projection key for pheromone emission and dispersion. The olfactory mechanisms that underlie peripheral odorant detection in RPW have been addressed at the antennal level. However, the rostrum remains unexplored. Through RNA-seq, 27 odorant-binding proteins (OBPs), 6 chemosensory proteins (CSPs), 4 sensory neuron membrane proteins (SNMPs), 21 gustatory receptors (GRs), 25 odorant receptors (ORs) (including one odorant receptor coreceptor, Orco) and 10 ionotropic receptors (IRs), were identified. We reported 27 novel rostrum-specific olfactory proteins (4 IRs, 11 GRs, 2 CSPs, 3 OBPs, and 7 ORs) in R. ferrugineus (Rfer). The OBPs (RferSOBPs) [Rfer with “S” indicating “snout” (rostrum)] were the most abundant transcripts compared with the rest of the olfactory proteins. We identified several rostrum OBPs, which predominately emerged through gene duplication, and were found expressed in both rostrum and antennae. Noticeably, we found R. ferrugineus pheromone-binding protein (RferOBP1768) paralog in the rostrum (RferOBP14) and mapped it in the same scaffold at a different position on the RPW genome as a recent duplicate. We found that an OR (RferSOR1) was the most abundant for both field-collected and lab-reared RPWs, in the rostrum and antennae. Likewise, up-regulated olfactory-related proteins were established in field conditions compared with those from laboratory-reared. We found a rostrum-specific, highly expressing RferSIR1 in IR93a-clade related to hygrosensation. The role of these olfactory proteins as targets for identifying more specific and powerful semiochemicals is discussed in the context of pest management.

Vittatalactone is the Male-Produced Aggregation Pheromone of the Western Striped Cucumber Beetle, Acalymma trivittatum.

We found that vittatalactone, specifically (3R,4R)-3-methyl-4-[(1S,3S,5S)-1,3,5,7-tetramethyloctyl]oxetan-2-one, is the male-produced aggregation pheromone of the western striped cucumber beetle, Acalymma trivittatum (Mannerheim), as was previously shown for the striped cucumber beetle, Acalymma vittatum (F.) (Coleoptera: Chrysomelidae). A synthetic mixture containing 9% of the authentic natural pheromone, is attractive to both sexes of both species in the field, as demonstrated by trapping using baited and unbaited sticky panels in California and earlier in Maryland. Females of both species do not produce detectable vittatalactone. This finding expands the usefulness of the synthetic vittatalactone mixture for pest management throughout the range of both A. vittatum and A. trivittatum. Development of vittatalactone time-release formulations combined with cucurbitacin feeding stimulants offer the potential for selective and environmentally-friendly cucurbit pest management tactics.

Exploration of Candidate Genes Involved in the Biosynthesis, Regulation and Recognition of the Male-Produced Aggregation Pheromone of Halyomorpha halys

The aggregation pheromone of the brown marmorated stink bug, Halyomorpha halys (Stål), is produced by adult males, and plays an important role in the behavioral regulation of H. halys. However, information on the molecular mechanisms underlying this pheromone's biosynthesis is limited. In this study, HhTPS1, a key candidate synthase gene in the aggregation pheromone biosynthesis pathway of H. halys, was identified. Then, through weighted gene co-expression network analysis, the candidate P450 enzyme genes in the biosynthetic downstream of this pheromone and the related candidate transcription factor in this pathway were also identified. In addition, two olfactory-related genes, HhCSP5 and HhOr85b, involved in the recognition of the aggregation pheromone of H. halys, were detected. We further identified the key amino acid sites of HhTPS1 and HhCSP5 that interact with substrates by using molecular docking analysis. This study provides basic information for further investigations into the biosynthesis pathways and recognition mechanisms of aggregation pheromones in H. halys. It also provides key candidate genes for bioengineering bioactive aggregation pheromones necessary for the development of technologies for the monitoring and control of H. halys.

Spotted cucumber beetle (Diabrotica undecimpunctata howardi) is attracted to vittatalactone, the pheromone of striped cucumber beetle (Acalymma vittatum)

Spotted cucumber beetle (Diabrotica undecimpunctata howardi; also known as southern corn rootworm) is a pest feeding on many crops, including cucurbits, corn (maize), peanuts, beans, and other crops and wild plants in North America. On cucurbit crops, where it co-occurs with striped cucumber beetle (Acalymma vittatum), both are key pests that vector Erwinia tracheiphila, a destructive pathogen that causes bacterial wilt. Here we deployed synthetic vittatalactone, the male-produced aggregation pheromone of A. vittatum, as lures in the field, to measure attraction of the target and non-target insects. Both sexes of D. u. howardi were strongly attracted to the vittatalactone lures (5–7.5-fold compared to blank trap) throughout the growing season in Maryland and Virginia, USA. Moreover, D. u. howardi was more attracted to live A. vittatum males feeding on squash plants than to plants with females or without any beetles, confirming the genuine organismal cross-attraction between these two key pests. The kairomonal attraction to vittatalactone suggests that it functions as a “keystone semiochemical,” mediating the colonization of cucurbits by multiple herbivore species. Such cross attraction could allow for use of vittatalactone in management of multiple pest species, with potential reduction of pesticide use in cucurbit crops.

Ophiostomatoid fungi synergize attraction of the Eurasian spruce bark beetle, Ips typographus to its aggregation pheromone in field traps.

Eurasian spruce bark beetle, Ips typographus is a destructive pest of the Norway spruce (Picea abies). Recent outbreaks in Europe have been attributed to global warming and other anthropogenic impacts. Bark beetles are guided by multiple complex olfactory cues throughout their life cycle. Male-produced aggregation pheromones, comprising 2-methyl-3-buten-2-ol and cis-verbenol, have been identified as the most powerful attractants for dispersing conspecifics. In addition to host trees, bark beetles interact with multiple organisms, including symbiotic ophiostomatoid fungi, which may promote beetle colonization success and offspring development. Previously, in a short-distance laboratory assay, we demonstrated that I. typographus adults are attracted to the volatile organic compounds (VOCs) produced by three symbiotic fungi: Grosmannia penicillata, Endoconidiophora polonica, and Leptographium europhioides. Furthermore, the abundant fusel alcohols and their acetates were found to be the most attractive odorants in the fungal VOC profile. In this study, using a long-distance field-trapping experiment, we analyzed the role of fungal VOCs as attractants for dispersing I. typographus. Two types of fungal lures were tested in combination with pheromones in traps: (1) live cultures of fungi grown on potato dextrose agar (PDA) and (2) dispensers containing synthetic fusel alcohols and their acetates in equal proportions. Subsequently, the composition of VOCs emitted from live fungal lures were analyzed. We found that the symbiotic fungi synergistically increased the attraction of beetles to pheromones in field traps and the attractiveness of live fungal lures depended on the fungal load. While one Petri dish with E. polonica, when combined with pheromones synergistically increased trapping efficiency, three Petri dishes with L. europhioides were required to achieve the same. The synthetic mix of fungal fusel alcohols and acetates improved the catch efficiency of pheromones only at a low tested dose. VOC analysis of fungal cultures revealed that all the three fungi produced fusel alcohols and acetates but in variable composition and amounts. Collectively, the results of this study show that, in addition to pheromones, bark beetles might also use volatile cues from their symbiotic fungi to improve tree colonization and reproductive success in their breeding and feeding sites.

Vibrational calling signals improve the efficacy of pheromone traps to capture the brown marmorated stink bug

Halyomorpha halys (Stål, 1855), the brown marmorated stink bug (BMSB), is an invasive species that has become a key agricultural pest in its invaded range. Commercial traps available for BMSB monitoring rely on male produced aggregation pheromones as lure, with two possible shortcomings: trap spillover and low detection precision. In this study, we assessed if vibrational signals can increase the attractiveness of pheromone traps by testing the optimized vibration-based lure (Female Song 2, FS2) associated with a specifically designed trap (i.e., the vibrotrap). We evaluated the efficacy of this bimodal trap (i.e., pheromones + vibrations) on females, males and nymphs in controlled conditions (greenhouse) and in the field, in two sites at the margin of two commercial vineyards. In the field, bimodal vibrotraps were compared to three unimodal (i.e., only pheromone) trap types. Both experiments showed that the vibrotrap is highly attractive for BMSB, and the optimized FS2 signal significantly improved its effectiveness. Even though FS2 was selected to target males, the number of trapped females increased as well. Overall, the presented findings show a feasible improvement to future commercial BMSB traps through the synergic use of semiophysicals and semiochemicals. Further research is needed to evaluate the effectiveness of vibrotraps for both early detection and mass trapping.

Identification of Components of the Aggregation Pheromone of the Guam Strain of Coconut Rhinoceros Beetle, Oryctes rhinoceros, and Determination of Stereochemistry

The coconut rhinoceros beetle, Oryctes rhinoceros (Linnaeus 1758) (Coleoptera: Scarabaeidae: Dynastinae) (CRB), is endemic to tropical Asia where it damages both coconut and oil palm. A new invasion by CRB occurred on Guam in 2007 and eradication attempts failed using commonly applied Oryctes rhinoceros nudivirus (OrNV) isolates. This and subsequent invasive outbreaks were found to have been caused by a previously unrecognized haplotype, CRB-G, which appeared to be tolerant to OrNV. The male-produced aggregation pheromone of the endemic, susceptible strain of O. rhinoceros (CRB-S) was previously identified as ethyl 4-methyloctanoate. Following reports from growers that commercial lures containing this compound were not attractive to CRB-G, the aim of this work was to identify the pheromone of CRB-G. Initial collections of volatiles from virgin male and female CRB-G adults from the Solomon Islands failed to show any male- or female-specific compounds as candidate pheromone components. Only after five months were significant quantities of ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by males but not by females. No other male-specific compounds could be detected, in particular methyl 4-methyloctanoate, 4-methyl-1-octanol, or 4-methyl-1-octyl acetate, compounds identified in volatiles from some other species of Oryctes. Ethyl 4-methyloctanoate elicited a strong electroantennogram response from both male and female CRB-G, but these other compounds, including 4-methyloctanoic acid, did not. The enantiomers of ethyl 4-methyloctanoate and 4-methyloctanoic acid were conveniently prepared by enzymatic resolution of the commercially-available acid, and the enantiomers of the acid, but not the ester, could be separated by gas chromatography on an enantioselective cyclodextrin phase. Using this approach, both ethyl 4-methyloctanoate and 4-methyloctanoic acid produced by male CRB-G were shown to be exclusively the (R)-enantiomers whereas previous reports had suggested male O. rhinoceros produced the (S)-enantiomers. However, re-examination of the ester and acid produced by male CRB-S from Papua New Guinea showed that these were also the (R)-enantiomers. In field trapping experiments carried out in the Solomon Islands, both racemic and ethyl (R)-4-methyloctanoate were highly attractive to both male and female CRB-G beetles. The (S)-enantiomer and the corresponding acids were only weakly attractive. The addition of racemic 4-methyloctanoic acid to ethyl 4-methyloctanoate did significantly increase attractiveness, but the addition of (R)- or (S)-4-methyloctanoic acid to the corresponding ethyl esters did not. Possible reasons for the difference in assignment of configuration of the components of the CRB pheromone are discussed along with the practical implications of these results.

Social communication activates the circadian gene Tctimeless in Tribolium castaneum

Chemical communication via pheromones is an integral component in insect behavior, particularly for mate searching and reproduction. Aggregation pheromones, that attract conspecifics of both sexes, are particularly common and have been identified for hundreds of species. These pheromones are among the most ecologically selective pest suppression agents. In this study, we identified an activating effect of the aggregation pheromone of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenibroidae) on a highly conserved circadian clock gene (Tctimeless). Tribolium castaneum is one of the most damaging cosmopolitan pest of flour and other stored food products. Its male produced aggregation pheromone, 4,8-dimethyldecanal (DMD), attracts both conspecific males and females and is used for pest management via monitoring and mating disruption. The Tctimeless gene is an essential component for daily expression patterns of the circadian clock and plays vital roles in eclosion, egg production, and embryonic development. In this study, we demonstrate that constant exposure to the species-specific aggregation pheromone led to Tctimeless up-regulation and a different pattern of rhythmic locomotive behavior. We propose that changing the well-adapted "alarm clock", using DMD is liable to reduce fitness and can be highly useful for pest management.

Exploiting Thrips Aggregation Pheromones to Develop a Lure-and-Kill Strategy for the Management of the Bean Flower Thrips

The potential of semiochemicals to lure insect pests to a trap where they can be killed with biopesticides has been demonstrated as an eco-friendly pest management alternative. In this study, we tested two recently characterized male-produced aggregation pheromones of the bean flower thrips Megalurothrips sjostedti (Trybom), namely (R)-lavandulyl 3-methylbutanoate (major) and (R)-lavandulol (minor), for their field efficacy. Moreover, compatibility of these pheromones and two other thrips attractants, Lurem-TR and neryl (S)-2-methylbutanoate, with the entomopathogenic fungus (EPF) Metarhizium anisopliae ICIPE 69 has been determined. Our study revealed that the M. sjostedti aggregation pheromones have dose-dependent antifungal effects on the EPF viability, but showed no fungistatic effect at a field-realistic dose for attraction of thrips. (R)-lavandulyl 3-methylbutanoate had similar antifungal effects as neryl (S)-2-methylbutanoate 8 days after exposure; whereas, Lurem-TR had a stronger antifungal effect than other thrips attractants. In the semi-field experiments, all autoinoculation devices maintained at least 86% viability of M. anisopliae conidia after 12 days of exposure. Field trials demonstrated for the first time that (R)-lavandulyl 3-methylbutanoate increases trap catches. Our findings pave a way for designing a lure-and-kill thrips management strategy to control bean flower thrips using autoinoculation devices or spot spray application.

Olfactory attraction to aggregation pheromone is mediated by disti-flagellum of antennal segments in Riptortus pedestris

Riptortus pedestris (Hemiptera: Alydidae) is a serious pest of soybean and sweet persimmon and uses male produced aggregation pheromone, (E)-2-hexenyl (Z)-3-hexenoate, (E)-2-hexenyl (E)-2-hexenoate, and tetradecyl isobutyrate to facilitate food location and recognition by conspecifics. Using electroantennogram (EAG) and greenhouse bioassay, we determined which antennal segment is involved in the detection of their aggregation pheromone. In the first EAG test using individual antennal segments, significant EAG responses to 1:1:1 mixture of the aggregation pheromone were observed only from the disti-flagellum segments of both male and female antennae at both pheromone doses tested (1 µg and 100 µg). In the following EAG tests using gradually removed antennal segment(s), EAG response was still maintained when the distal half of a disti-flagellum was surgically removed, while EAG response was lost when whole segment of disti-flagellum or other whole segments were gradually removed from intact antenna of both sexes. In greenhouse experiment, removing one or both segment(s) of disti-flagellum from male or female antennae resulted in significant reduction in their attraction to the aggregation pheromone. Together, these findings support that the disti-flagellum of R. pedestris houses olfactory neurons associated with attraction to their aggregation pheromone.

Trapping Failure Leads to Discovery of Potent Semiochemical Repellent for the Walnut Twig Beetle.

The walnut twig beetle, Pityophthorus juglandis Blackman, and its associated fungal pathogen that causes thousand cankers disease, currently threaten the viability of walnut trees across much of North America. During a 2011 assessment of seasonal flight patterns of P. juglandis with yellow sticky traps baited with the male-produced aggregation pheromone component, 3-methyl-2-buten-1-ol, dramatically reduced catches were recorded when Tree Tanglefoot adhesive was used to coat the traps. In summer 2011, two trap adhesives were tested for potential repellency against P. juglandis in a field trapping bioassay. SuperQ extracts of volatiles from the most repellent adhesive were analyzed by gas chromatography-mass spectrometry, and limonene and α-pinene were identified as predominant components. In field-based, trapping experiments both enantiomers of limonene at a release rate of ~700 mg/d conferred 91-99% reduction in trap catches of P. juglandis to pheromone-baited traps. (+)- and (‒)-α-Pinene reduced trap catch by 40 and 53%, respectively, at the highest release rate tested. While a combination of R-(+)-limonene and (+)-α-pinene resulted in a 97% reduction in the number of P. juglandis caught, the combination did not consistently result in greater flight trap catch reduction than individual limonene enantiomers. The repellent effect of limonene may be valuable in the development of a semiochemical-based tool for management of P. juglandis and thousand cankers disease.

The male-produced aggregation pheromone of the bean flower thrips Megalurothrips usitatus in China: identification and attraction of conspecifics in the laboratory and field.

Thrips, Megalurothrips usitatus, usually display aggregation behavior, which is probably mediated by a male-produced aggregation pheromone. Aggregation pheromones are species-specific, and can be used to develop commercial lures for monitoring and mass-trapping of pests. The active components of the aggregation pheromone for four thrips species have been identified. However, the components of M. usitatus-produced aggregation pheromone are still not clear. Y-tube olfactometer assays showed that both male and virgin female M. usitatus were significantly attracted to male but not female volatiles. This was additionally supported by electroantennogram (EAG) assays. Coupled gas chromatography-electroantennogram detection (GC-EAD) showed that one component of male-specific odors elicited a significant electrophysiological response. This compound was characterized as (2E,6E)-farnesyl acetate, which is structurally different from the active components of the aggregation pheromones of other reported thrips species. Electroantennal responses of M. usitatus increased with increasing doses of synthetic (2E,6E)-farnesyl acetate. Additionally, this compound significantly attracted adults in laboratory behavioral bioassays. Under field conditions, sticky traps with synthetic (2E,6E)-farnesyl acetate caught 1.5-7-fold more M. usitatus than controls, and this effect of the compound at a dose of 60 μg lasted at least 6 days. (2E,6E)-Farnesyl acetate was identified as the male-produced aggregation pheromone of M. usitatus. It could attract this thrips species under laboratory and field conditions, suggesting considerable potential as a commercial application to control M. usitatus populations. © 2020 Society of Chemical Industry.

The Effects of Weather on the Flight of an Invasive Bark Beetle, Pityophthorus juglandis.

The walnut twig beetle, Pityophthorus juglandis Blackman (Coleoptera: Scolytidae), vectors the fungus Geosmithia morbida, which has been implicated in thousand cankers disease of walnut. Little is known about the flight behavior of the insect across seasons, or about the variability in its flight patterns with weekly fluctuations in weather. We sampled flying adults weekly over a 142-week period (from 29 August, 2011 to 2 June, 2014) with 12-unit black plastic multiple funnel traps baited with a male-produced aggregation pheromone in California, USA. Up to 5000 beetles were captured per trap per week, although catches in most weeks were less than 100 insects. Trap catches were regressed against terms for precipitation, solar radiation, vapor pressure, air temperature, relative humidity, wind speed, and trap catches in preceding weeks. The number of beetles captured in each of the preceding two weeks explained most variation in a current week’s catch. This strong temporal autocorrelation was present in regression models developed for males, females, and both sexes pooled. These models were improved by including two environmental variables. Captures of P. juglandis increased with mean weekly air temperature and decreased with increasing mean minimum relative humidity. The percentage of variation in male, female, or total trap catch explained by the temporal variables and the two environmental variables in these multiple regression models ranged from 72% to 76%. While the flight of this invasive insect will likely be affected by site-specific factors as it spreads to new areas, the strong temporal correlation present in this system may provide a useful starting point for developing flight models for newly invaded areas.

Attractiveness of Pheromone Components With and Without the Synergist, Methyl (2E,4E,6Z)-2,4,6-Decatrienoate, to Brown Marmorated Stink Bug (Hemiptera: Pentatomidae).

The brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) is attracted to its male-produced aggregation pheromone, a ~3.5:1 mixture of (3S,6S,7R,10S)- and (3R,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol (SSRS and RSRS respectively), and also to the pheromone of its Asian sympatric species Plautia stali Scott (Hemiptera: Pentatomidae), methyl (2E,4E,6Z)-2,4,6-decatrienoate (MDT). A stereoisomeric mixture of (7R) 10,11-epoxy-1-bisabolen-3-ols (= mixed murgantiols) is used together in commercialized products with MDT because the latter is a synergist for H. halys attraction to mixed murgantiols. However, the optimal ratio for MDT combination with mixed murgantiols, and the sensitivity of bug captures to variation in ratio of the two pheromone components, have not been tested to date. Using black pyramid traps at two sites (in Maryland and West Virginia, United States), different ratios of mixed murgantiols to MDT were tested over two entire seasons. Also, captures using various ratios of the two active pheromone stereoisomers were undertaken in month-long trials with and without MDT. Results showed that H. halys adult and nymphal captures were relatively insensitive to the ratio of synthetic pheromone (mixed murgantiols) to MDT, as long as each was present in the trap. Captures of adults and nymphs were responsive to the lure loading of the SSRS isomer, but relatively insensitive to levels of the minor component, RSRS. The relative insensitivity of H. halys to these attractant ratios gives flexibility to development of more cost-efficient synthesis and trapping as well as other semiochemical-based management tactics.

Acaricidal activity of the aggregation pheromone of Japanese pine sawyer against two-spotted spider mite

Abstract Japanese pine sawyer (Monochamus alternatus) (Coleoptera: Cerambycidae), a vector of pine wilt disease, produces male-produced aggregation pheromone called monochamol, 2-(1-undecyloxy)-1-ethanol (C11OEtOH). After the identification of unique nematicidal activity of the synthetic monochamol and its homologues (ROEtOH: R = C7-C11) against Bursaphelenchus xylophilus nematode, their acaricidal activities were evaluated against two spotted spider mite (TSSM, Tetranychus urticae Koch) (Acari: Tetranychidae). In a leaf disc bioassay method, 100 mgL−1 of monochamol and 2-(1-decyloxy)-1-ethanol (C10OEtOH) exhibited 100% adult mortality and zero fecundity. Along with C11OEtOH and C10OEtOH, 2-(1-octyloxy)-1-ethanol (C8OEtOH) and 2-(1-nonyloxy)-1-ethanol (C9OEtOH) were also 100% acaricidal with complete oviposition inhibition at increased concentration of 1000 mgL−1. When the TSSM eggs on leaf discs were immersed in 1000 mgL−1 solutions, none of the components were ovicidal. Egg hatching was completely inhibited by three components (C9OEtOH, C10OEtOH and C11OEtOH) at 6000 mgL−1 and by four components (C8OEtOH, C9OEtOH, C10OEtOH and C11OEtOH) at 8000 mgL−1. However, none of the compounds showed acaricidal effects in 0.5 h and 24 h residual toxicity tests at 1000 mgL−1. This study highlights the potential acaricidal activity of monochamol and its homologues. Notably, the monochamol and its C10-homologue could be listed as efficient acaricidal agents.

Male-Produced Aggregation Pheromone of the Sloe Bug, Dolycoris baccarum L. (Hemiptera: Heteroptera: Pentatomidae).

The sloe bug, Dolycoris baccarum L. (Hemiptera: Heteroptera: Pentatomidae), is a Palearctic species and an important polyphagous pest. Previous studies have reported that male D. baccarum produce an aggregation pheromone, to which conspecific adults of both sexes respond. We used solid-phase microextraction (SPME) to collect volatiles released by sexually mature virgin males and females, and identified potential components of the male-produced aggregation pheromone. Analysis of the SPME samples by gas chromatography-mass spectrometry (GC-MS) revealed three male-specific components, α-bisabolol, trans-α-bergamotene, and β-bisabolene, at a ratio of 100:10:3. A field trapping experiment showed that (±)-α-bisabolol alone attracted D. baccarum adults, and it was synergized by trans-α-bergamotene, but not (S)-β-bisabolene. However, neither trans-α-bergamotene nor (S)-β-bisabolene was attractive individually. A binary blend of (±)-α-bisabolol and trans-α-bergamotene in a 10:1 ratio was less attractive than caged live males, suggesting that further study is necessary to determine the absolute configuration or steroisomeric composition of the natural pheromone components. This is the first report of α-bisabolol and trans-α-bergamotene as semiochemicals among Pentatomidae, and the first to identify α-bisabolol from insects.

Orientation of rusty grain beetles, Cryptolestes ferrugineus (Coleoptera: Laemophloeidae), to semiochemicals in field and laboratory experiments

Cryptolestes ferrugineus is a common beetle pest of stored grain for which techniques to monitor dispersing beetles are limited. Early research found several male-produced aggregation pheromones, but there has been little related work since that time. This paper reports experiments on orientation of C. ferrugineus in response to synthetic pheromones and other semiochemicals via flight and walking. Field trapping studies showed that flying beetles were caught on the western sides of grain bins in Kansas compared to the other three cardinal directions. Work with synthetic formulations of the two male-produced aggregation pheromones found that flying beetles were attracted to traps with the pheromone and wheat compared to traps with wheat only. Walking bioassays in the laboratory determined that either of the two aggregation pheromones, known as Cucujolide I and Cucujolide II, were attractive whether deployed singly or in combination. Laboratory bioassays showed that volatiles from grains, other grain-based materials and two commercial food attractants used in traps were attractive to C. ferrugineus. Further laboratory assays demonstrated that C. ferrugineus would orient to synthetic pheromones of three other beetle species and one moth species, all common pests of stored products. These new data on semiochemicals for C. ferrugineus suggest future work that could be done toward developing new tools for detecting and monitoring this serious pest.

Characterization of Male-Produced Aggregation Pheromone of the Bean Flower Thrips Megalurothrips sjostedti (Thysanoptera: Thripidae)

Aggregation of the bean flower thrips, Megalurothrips sjostedti (Trybom) (Thysanoptera: Thripidae), has been observed on cowpea, Vigna unguiculata (L.) Walp. To understand the mechanism underpinning this behavior, we studied the responses of M. sjostedti to headspace volatiles from conspecifics in a four-arm olfactometer. Both male and female M. sjostedti were attracted to male, but not to female odor. Gas chromatography/mass spectrometry (GC/MS) analyses revealed the presence of two distinct compounds in male M. sjostedti headspace, namely (R)-lavandulyl 3-methylbutanoate (major compound) and (R)-lavandulol (minor compound); by contrast, both compounds were only present in trace amounts in female headspace collections. A behavioral assay using synthetic compounds showed that male M. sjostedti was attracted to both (R)-lavandulyl 3-methylbutanoate and (R)-lavandulol, while females responded only to (R)-lavandulyl 3-methylbutanoate. This is the first report of a male-produced aggregation pheromone in the genus Megalurothrips. The bean flower thrips is the primary pest of cowpea, which is widely grown in sub-Saharan Africa. The attraction of male and female M. sjostedti to these compounds offers an opportunity to develop ecologically sustainable management methods for M. sjostedti in Africa.

Isophorone derivatives as a new structural motif of aggregation pheromones in Curculionidae

The beetle Homalinotus depressus (Coleoptera: Curculionidae) is a major pest of coconuts in the Northern region of Brazil, for which environmentally friendly methods of control are desired. Behavioral responses of H. depressus to airborne volatile extracts from conspecifics suggested the presence of a male-produced aggregation pheromone. GC analyses of these extracts showed the presence of four male-specific compounds. Analytical data in combination with the synthesis of standards led to the identification of the male-released semiochemicals as epoxyisophorone (1), isophorone (2), homalinol (3), and 2-hydroxyisophorone (4), of which (3) was the major constituent. The configuration of homalinol was determined to be cis on the basis of retention times of synthetic cis and trans synthetic standards. Enantiomers of cis-homalinol were obtained in high enantiomeric excess by using biocatalysis. Their separation on a GC enantioselective column (β-Dex325®), allowed us to unambiguously determine that the absolute configuration of natural homalinol was (1R,2R,6S). Field bioassays demonstrated that both the synthetic major compound per se and mixtures of all four male-specific compounds were attractive to H. depressus.

Olfactory responses of banana pseudostem weevil Odoiporus longicollis Olivier to pheromone and host plant volatiles

Banana pseudostem weevil Odoiporus longicollis Olivier (Coleoptera: Curculionidae) is an important pest of banana. In this study, its male produced aggregation pheromone was identified as 2-methy-4-heptanol (2M4H) which attracts both male and female. In order to achieve maximum attraction for field trapping, an aggregation pheromone (2M4H), host plant extract (HPE) and combination of 2M4H + HPE were evaluated under laboratory conditions with wind tunnel bioassay. Maximum weevil attraction was observed in the descending order of: 2M4H + HPE > 2M4H > HPE, suggesting a better synergistic effect of host plant extract on pheromone.