Olfactometer Bioassays Research Articles

Olfactory responses of Argentine stem weevil to herbivory and endophyte-colonisation in perennial ryegrass

Argentine stem weevil adults (ASW, Listronotus bonariensis) feed on the leaves of agricultural grasses and their larvae mine the pseudostem, causing extensive damage that can result in plant death. Plants emit volatiles that serve as signals to host-searching insects and these odours can be altered by both herbivory and fungal endophyte-infection. This study investigated whether ASW adults utilise olfaction to identify their host plants, perennial ryegrass (Lolium perenne), and if conspecific herbivory or the presence of Epichloë festucae var. lolii fungal endophytes (strain wild-type or AR1) influenced such responses. Results from olfactometer bioassays established that ASW adults were able to utilise their olfactory response to orient towards volatiles released by perennial ryegrass and further, the weevils displayed a preference for plants previously damaged by conspecific weevils. However, there was no evidence that weevils had the ability to distinguish between endophyte-infected and endophyte-free plants using olfaction alone. Using a push–pull extraction technique, thirteen volatile compounds were identified in the blend released by perennial ryegrass. Endophyte and herbivory were found to alter these volatile compounds and quantities emitted by this forage grass. This study suggests that despite observing differences in the plant volatile blend, ASW do not perceive endophyte (wild-type and AR1) using olfaction alone and must rely on other cues, e.g. contact chemoreception or post-ingestional malaise, to detect the presence of a bioactive endophyte in an otherwise acceptable host plant.

Herbivore-induced plant volatiles do not affect settling decisions by synanthropic spiders

An underlying assumption of optimal foraging models is that animals are behaviorally, morphologically, and physiologically adapted to maximize their net energy intake. Here we explored whether this concept applies to web-building spiders in a multi-trophic context. If a spider were to build her web next to herbivore-fed-on plants that signal the herbivores’ enemies for help by emitting herbivore-induced plant volatiles (HIPVs), that spider may maximize web captures in the short term. However, she would also risk predation by generalist predators that “listen” to signaling plants to find both herbivore and spider prey, likely resulting in lower overall reproductive fitness for the spider. We tested the hypothesis that HIPVs trigger avoidance responses by web-building spiders. We selected seven common HIPVs and one HIPV elicitor, and in two-choice olfactometer bioassays tested their effect on four synanthropic spider species (false black widow, Steatoda grossa; common cellar spider, Pholcus phalangioides; hobo spider, Eratigena agrestis; western black widow, Latrodectus hesperus). The 8-component HIPV/HIPV elicitor blend had a weak deterrent effect on S. grossa, but the effect did not extend to P. phalangioides, E. agrestis, and L. hesperus. Our findings imply that there was insufficient selection pressure for these spiders to recognize HIPVs in a multi-trophic context, where spiders themselves could become prey if generalist predators or spider-hunting parasitoid wasps were to respond to signaling plants.

Chemically-mediated colonization of black cherry by the peach bark beetle, Phloeotribus liminaris.

The peach bark beetle (Phloeotribus liminaris Harris, PBB) affects the health, quality, and value of black cherry (Prunus serotina Ehrh.) within the Central Hardwoods Forest Region of North America. When colonized by adult beetles, black cherry trees produce a defensive exudate, or 'gum', staining the wood and decreasing its value up to 90%. Current management tactics are inadequate to avoid extensive damage to most veneer-sized black cherry in the region. We test the hypothesis that PBB colonization behavior is chemically-mediated and determine the extent to which PBB is attracted to compounds associated with wounded or PBB-infested cherry wood. Through olfactometer and field bioassays, we determined that adult PBB were attracted to cherry branches infested with female beetles. We then used dynamic headspace sampling to collect volatiles associated with wounded and infested bolts of black cherry. The volatile benzaldehyde dominated these collections and was more abundant in aerations of female-infested bolts than other odor sources. In subsequent field bioassays, we evaluated the bioactivity of benzaldehyde, as well as α-longipinene, in combination with several chemical carriers. Traps baited with benzaldehyde captured more PBB than all other treatments, irrespective of other lure components. Moreover, PBB were not attracted to traps baited solely with ethanol, a common attractant for bark beetles that colonize hardwood trees. This is the first report of benzaldehyde as an attractant for a species of bark beetle and could aid in developing semiochemical-based management tactics for this important pest.

Dufour\u2019s gland analysis reveals caste and physiology specific signals in Bombus impatiens

Reproductive division of labor in insect societies is regulated through multiple concurrent mechanisms, primarily chemical and behavioral. Here, we examined if the Dufour’s gland secretion in the primitively eusocial bumble bee Bombus impatiens signals information about caste, social condition, and reproductive status. We chemically analyzed Dufour’s gland contents across castes, age groups, social and reproductive conditions, and examined worker behavioral and antennal responses to gland extracts. We found that workers and queens each possess caste-specific compounds in their Dufour’s glands. Queens and gynes differed from workers based on the presence of diterpene compounds which were absent in workers, whereas four esters were exclusive to workers. These esters, as well as the total amounts of hydrocarbons in the gland, provided a separation between castes and also between fertile and sterile workers. Olfactometer bioassays demonstrated attraction of workers to Dufour’s gland extracts that did not represent a reproductive conflict, while electroantennogram recordings showed higher overall antennal sensitivity in queenless workers. Our results demonstrate that compounds in the Dufour’s gland act as caste- and physiology-specific signals and are used by workers to discriminate between workers of different social and reproductive status.

Plant volatiles guide the new pest Dichelops furcatus to feed on corn seedlings.

Recently, in temperate and neotropical regions of South America the generalist stink bug Dichelops furcatus (Hemiptera: Pentatomidae) became a new pest of corn (Zea mays) seedlings. Implementation of no-tillage cultivation system left organic matter covering the soil, which shelters adults of stink bugs during winter. In spring, corn is sowed under soybean stubble and D. furcatus adults start to feed on seedlings. To determine corn-derived volatile organic compounds (VOCs) that attract this stink bug species, we evaluated stink bug preferences from two corn hybrids with contrast germplasm backgrounds, a temperate and a tropical hybrid. Stink bugs preferred to feed on temperate seedlings rather than on the tropical ones. GC-MS and PCA analysis of VOCs suggested that hybrids emitted contrasting blends. Linalool represented 68% of total VOCs emitted from temperate corn, while in the tropical hybrid this compound represented 48%. Olfactometer experiments demonstrated that linalool was attractive to stink bugs. However, 2 h of D. furcatus attack induced emission of 14 additional VOCs in temperate seedlings, and olfactometer bioassay and blend of VOCs emission suggested that perceived volatiles by stink bugs induced feeding avoidance. The increment of VOCs emission was associated with the induction of JA, JA-Ile, ABA, and IAA, and decreasing of SA concentrations. This is the first time showing a complete profile of defensive phytohormones induced by stink bugs feeding on corn, and further demonstrating that a blend of corn seedling-associated VOCs, mainly composed by linalool, modulates D. furcatus adults' behavior and feeding preferences. © 2021 Society of Chemical Industry.

The Role of Trialeurodes vaporariorum-Infested Tomato Plant Volatiles in the Attraction of Encarsia formosa (Hymenoptera: Aphelinidae).

Natural enemies locate their herbivorous host and prey through kairomones emitted by host plants and herbivores. These kairomones could be exploited to attract and retain natural enemies in crop fields for insect pest control. The parasitoid Encarsia formosa preferentially parasitises its whitefly host, Trialeurodes vaporariorum, a major pest of tomato Solanum lycopersicum, thus offering an effective way to improve whitefly control. However, little is known about the chemical interactions that occur in E. formosa-T. vaporariorum-S. lycopersicum tritrophic system. Using behavioural assays and chemical analyses, we investigated the kairomones mediating attraction of the parasitoid to host-infested tomato plants. In Y-tube olfactometer bioassays, unlike volatiles of healthy tomato plants, those of T. vaporariorum-infested tomato plants attracted E. formosa, and this response varied with host infestation density. Coupled gas chromatography/mass spectrometric analyses revealed that host infestation densities induced varying qualitative and quantitative differences in volatile compositions between healthy and T. vaporariorum adult-infested tomato plants. Bioassays using synthetic chemicals revealed the attractiveness of 3-carene, β-ocimene, β-myrcene and α-phellandrene to the parasitoid, and the blend of the four compounds elicited the greatest attraction. Our results suggest that these terpenes could be used as an attractant lure to recruit the parasitoid E. formosa for the control of whiteflies in tomato crop fields.

Odorants of Capsicum spp. Dried Fruits as Candidate Attractants for Lasioderma serricorne F. (Coleoptera: Anobiidae).

Simple SummaryThe cigarette beetle, Lasioderma serricorne F. (Coleoptera: Anobiidae), is an important pest of stored products. It can be monitored using pheromone traps with or without a food source as a synergistic attractant. The study objective was to assess the response of L. serricorne to volatile organic compounds (VOCs) from different chili fruit powders to identify semiochemicals involved in this attraction which could be used as synthetic co-attractants in pheromone traps. Olfactometer results indicated that Capsicum annuum and C. frutescens VOCs elicit attraction toward L. serricorne adults, while C. chinense VOCs elicit no attraction. Chemical analysis and behavioral assays indicated a primary role for polar compounds in the attraction toward these sources. α-Ionone and β-ionone, compounds abundant in the attractive species and which elicited positive results in the olfactometer bioassays, may be promising candidates as attractant and/or pheromone synergist in monitoring traps for L. serricorne.The cigarette beetle, Lasioderma serricorne F. (Coleoptera: Anobiidae) is an important food storage pest affecting the tobacco industry and is increasingly impacting museums and herbaria. Monitoring methods make use of pheromone traps which can be implemented using chili fruit powder. The objective of this study was to assess the response of L. serricorne to the volatile organic compounds (VOCs) from different chili powders in order to identify the main semiochemicals involved in this attraction. Volatiles emitted by Capsicum annuum, C. frutescens, and C. chinense dried fruit powders were tested in an olfactometer and collected and analyzed using SPME and GC-MS. Results indicated that C. annuum and C. frutescens VOCs elicit attraction toward L. serricorne adults in olfactometer, while C. chinense VOCs elicit no attraction. Chemicals analysis showed a higher presence of polar compounds in the VOCs of C. annuum and C. frutescens compared to C. chinense, with α-ionone and β-ionone being more abundant in the attractive species. Further olfactometer bioassays indicated that both α-ionone and β-ionone elicit attraction, suggesting that these compounds are candidates as synergistic attractants in pheromone monitoring traps for L. serricorne.

Interrelation between Surface Wax Alkanes from Red Kidney Bean (Phaseolus vulgaris L.) Seeds and Adzuki Bean Weevil [Callosobruchus chinensis (F.)] (Coleoptera: Bruchidae)

Background: Callosobruchus chinensis (Fabricius) (Coleoptera: Bruchidae) is one of the major insect pests of Phaseolus vulgaris L. grains, commonly known as rajma seeds, in Europe and Asia including India. Infestations of these insects destroy majority of legume seeds including rajma which causes a great economic loss. Hence, a proper sustainable pest management measures are necessary for storage of rajma seeds. For this, the study aims to identify and quantify the n-alkane profile from the surface waxes of rajma seeds and their role as olfactory cue in C. chinensis. Individual synthetic alkane followed by the synthetic blends mimicking rajma seed surface wax n-alkanes as olfactory cue was also evaluated.Methods: Collected rajma seeds were solvent extracted to isolate surface waxes. The extract then fractioned by thin-layer chromatography and followed by gas chromatography-mass spectrometry analyses to purify, quantify and identify n-alkanes.Result: Rajma seeds’ surface waxes analysis revealed 18 n-alkanes between n-C15 and n-C33. The predominant alkanes were n-octacosane and n-hexadecane. n-Octadecane was the least abundant alkane in seeds. Total alkane content was 3502.67±12.82µg from 100 g (number 200 ± 5.13) seeds. Adult female C. chinensis elicited attraction towards the surface wax alkanes at concentrations of 0.5, 1, 2, 4 and 6 seed(s) equivalent of rajma seed(s) in the Y-tube olfactometer bioassay, but the highest attraction was observed at 6 seeds equivalent. Hence, a synthetic alkane blend resembling of 6 seeds equivalent, present in seeds’ surface wax alkane or a combination of nine (which elicited positive response) synthetic alkane blend resembling 6 seeds equivalent could be used as lures in developing baited trap in insect pest management programme.

Rosmarinus officinialis L. (Lamiales: Lamiaceae), a Promising Repellent Plant for Thrips Management.

A number of thrips species are among the most significant agricultural pests globally. Use of repellent intercrop plants is one of the key components in plant-based 'push-pull' strategies to manage pest populations. In this study, the behavioral responses of three thrips species, Frankliniella occidentalis (Pergande), Frankliniella intonsa (Trybom), and Thrips palmi Karny (Thysanoptera: Thripidae) to Rosmarinus officinalis were investigated in Y-tube olfactometer bioassays and cage experiments. In addition, the major volatile compounds from rosemary were identified and the effect of the individual compounds on thrips behavior was evaluated. Females and males of the three thrips species were significantly repelled by the volatiles from cut rosemary leaves. The presence of rosemary plants significantly reduced settlement of females of the three thrips species and eggs laid by F. occidentalis females on target host plants. In total, 47 compounds were identified in the volatiles collected from the cut leaves of rosemary plants. The responses of the three thrips species to 10 major volatile compounds showed significant differences. However, α-pinene, the most abundant volatile, was repellent to F. occidentalis and F. intonsa. Eucalyptol, the second most abundant volatile, showed significant repellent activity to all the three thrips species. Our findings showed that rosemary is a promising repellent plant against the three thrips pests we tested, which could be a good candidate for 'push' plants in plant-based 'push-pull' strategies. The identified volatile compounds that accounted for the repellent activity could be developed as repellents for sustainable thrips management.

Oviposition-Site Selection of Phlebotomus papatasi (Diptera: Psychodidae) Sand Flies: Attraction to Bacterial Isolates From an Attractive Rearing Medium

Phlebotomine sand flies are worldwide vectors of Leishmania parasites as well as other bacterial and viral pathogens. Due to the variable impact of traditional vector control practices, a more ecologically based approach is needed. The goal of this study was to isolate bacteria from the most attractive substrate to gravid Phlebotomus papatasi Scopoli sand flies and determine the role of bacterial volatiles in the oviposition attractancy of P. papatasi using behavioral assays. We hypothesized that gravid sand flies are attracted to bacterially derived semiochemical cues associated with breeding sites. Bacteria were isolated from a larvae-conditioned rearing medium, previously shown to be highly attractive to sand flies. The isolated bacteria were identified by amplifying and sequencing 16S rDNA gene fragments, and 12 distinct bacterial species were selected for two-choice olfactometer bioassays. The mix of 12 bacterial isolates elicited strong attraction at the lower concentration of 107 cells per ml and significant repellence at a high concentration of 109 cells per ml. Three individual isolates (SSI-2, SSI-9, and SSI-11) were particularly attractive at low doses. In general, we observed dose-related effects, with some bacterial isolates stimulating negative and some positive dose-response curves in sand fly attraction. Our study confirms the important role of saprophytic bacteria, gut bacteria, or both, in guiding the oviposition-site selection behavior of sand flies. Identifying the specific attractive semiochemical cues that they produce could lead to development of an attractive lure for surveillance and control of sand flies.

Impact of the exotic fall armyworm on larval parasitoids associated with the lepidopteran maize stemborers in Kenya

Exotic invasive insect herbivores have the potential to interfere with existing herbivore-natural enemy interactions in new environments. Fall armyworm (FAW), Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae), is a new invasive pest in maize fields in Africa. Understanding the acceptability and suitability of FAW to existing maize stemborer-parasitoid interactions is the first step in elucidating the impact that this exotic insect pest can have on the existing natural enemies used in biological control of maize stemborers in Kenya. The most commonly used larval parasitoids for biological control programs against maize stemborer communities in East Africa are Cotesia flavipes Cameron (Hymenoptera: Braconidae) and two populations of the native Cotesia sesamiae (Cs-Inland and Cs-Coast) Cameron (Hymenoptera: Braconidae). All these parasitoid species attacked FAW larvae but none yielded offspring, although they induced high non-reproductive host mortality when compared to natural mortality. Furthermore, the parasitoids that inserted their ovipositor into FAW larvae exhibited no significant preference between FAW larvae and their respective stemborer hosts under dual-choice bioassays. In olfactometer bioassays, the parasitoids were more attracted to plants infested by FAW than uninfested plants and even showed a marked preference for the odours of plants infested by FAW over those of plants infested by their natural host counterparts. This study illustrates that exotic pests, such as FAW, can impact existing stemborer-parasitoid interactions associated with maize, even if they cannot be used as hosts by parasitoids associated with these stemborers. Although additional studies are needed, FAW might therefore have a negative impact on stemborer biological control existing before its invasion.

Identification and application of bacterial volatiles to attract a generalist aphid parasitoid: from laboratory to greenhouse assays.

Recent studies have shown that microorganisms emit volatile compounds that affect insect behaviour. However, it remains largely unclear whether microbes can be exploited as a source of attractants to improve biological control of insect pests. In this study, we used a combination of coupled gas chromatography-electroantennography (GC-EAG) and Y-tube olfactometer bioassays to identify attractive compounds in the volatile extracts of three bacterial strains that are associated with the habitat of the generalist aphid parasitoid Aphidius colemani, and to create mixtures of synthetic compounds to find attractive blends for A. colemani. Subsequently, the most attractive blend was evaluated in two-choice cage experiments under greenhouse conditions. GC-EAG analysis revealed 20 compounds that were linked to behaviourally attractive bacterial strains. A mixture of two EAG-active compounds, styrene and benzaldehyde applied at a respective dose of 1 μg and 10 ng, was more attractive than the single compounds or the culture medium of the bacteria in Y-tube olfactometer bioassays. Application of this synthetic mixture under greenhouse conditions resulted in significant attraction of the parasitoids, and outperformed application of the bacterial culture medium. Compounds isolated from bacterial blends were capable of attracting parasitoids both in laboratory and greenhouse assays, indicating that microbial cultures are an effective source of insect attractants. This opens new opportunities to attract and retain natural enemies of pest species and to enhance biological pest control.

Behavioral response of the greenhouse whitefly (Trialeurodes vaporariorum) to plant volatiles of Ocimum basilicum and Tagetes minuta

The use of chemical pesticides as a main pest control strategy has been highly criticised due to environmental pollution and negative effects on natural enemies of pests. In modern farming, it is essential to implement integrated pest management approaches that seek to control insect pests without causing environmental damage, e.g. the use of companion plants. Basil and Mexican marigold are often used as companion plants to attract greenhouse whiteflies, hence reducing damage to solanaceous crops, but the mechanism and role of volatile cues in crop protection strategies are unknown. This study found that both flowering basil and marigold were preferred to tomato by the greenhouse whitefly (Trialeurodes vaporariorum) in Y-tube olfactometer bioassays. PCA revealed that some volatiles were more correlated to one stage than to another. The dominant volatile constituents of Mexican marigold are limonene, dihydrotagetone, (Z)-β-ocimene, α-pinene, (Z)-3-hexenyl acetate, and those from basil are linalool, 1,8-cineole, eugenol and β-elemene. Among these dominant compounds, 1,8-cineole and (Z)-3-hexenyl acetate elicited strong attraction in greenhouse whitefly at 0.01%, whereas (Z)-β-ocimene and linalool elicited strong repellence at 0.1% and 1% dosages. This suggested that the basil flowering stage attraction is due to 1,8-cineole. These volatiles demonstrated potential as lures or bio-repellents and could be used in a “push–pull” semiochemical approach for greenhouse whitefly management.

Host-Induced Plant Volatiles Mediate Ability of the Parasitoid Microplitis croceipes to Discriminate Between Unparasitized and Parasitized Heliothis virescens Larvae and Avoid Superparasitism.

In solitary endoparasitoids, oviposition in a host previously parasitized by a conspecific (superparasitism) leads to intraspecific competition, resulting in the elimination of all but one parasitoid offspring. Therefore, avoidance of parasitized hosts presents a strong selective advantage for such parasitoid species. Parasitoids use herbivore-induced plant volatiles (HIPVs) to find their hosts. In this study, we evaluated the ability of Microplitis croceipes (Hymenoptera: Braconidae) to discriminate between unparasitized and parasitized Heliothis virescens (Lepidoptera: Noctuidae) larvae using cotton plant odors as cues. A combination of behavioral and analytical techniques were used to test two hypotheses: (i) parasitoids will show preference for plant odors induced by unparasitized hosts over odors induced by parasitized hosts, and (ii) the parasitism status of herbivores affects HIPV emission in plants. Heliothis virescens larvae were parasitized for varying durations (0, 2 and 6-days after parasitism (DAP)). In four-choice olfactometer bioassays, female M. croceipes showed greater attraction to plant odors induced by unparasitized hosts compared to plant odors induced by parasitized hosts (2 and 6-DAP). Comparative gas chromatography-mass spectrometry analyses of cotton volatiles indicated reduced emission of 10 out of 21 identified compounds from plants infested by parasitized hosts compared with plants infested by unparasitized hosts. The results suggest that changes in plant volatile emission due to the parasitism status of infesting herbivores affect recruitment of parasitoids. Avoidance of superparasitism using plant odors optimizes host foraging in M. croceipes, and this strategy may be widespread in solitary parasitoid species.

Identification of Lathyrus sativus plant volatiles causing behavioral preference of Aphis craccivora.

The viviparous aphid Aphis craccivora Koch (Hemiptera: Aphididae) is a serious threat to the crop yield of Lathyrus sativus L. (Fabaceae), commonly known as grass pea. The synthetic insecticides applied to control this insect pest are not safe for the environment. Hence, it is necessary to find volatile organic compounds (VOCs) from two cultivars [BIO L 212 Ratan (BIO) and Nirmal B-1 (NIR)] of L. sativus plants causing behavioral preference of A. craccivora. The VOCs from undamaged (UD), insect-damaged (ID) [plants on which 50 or 100 adults of A. craccivora were fed for 4 h (ID 50 or ID 100)], and mechanically damaged (MD) plants were identified and quantified by gas chromatography-mass spectrometry and gas chromatography-flame ionization detection analyses, respectively. Total VOCs were higher in ID plants compared to UD plants of each cultivar. However, total VOCs were higher in NIR cultivar compared to BIO cultivar for both UD and ID plants. Benzyl alcohol was predominant in volatile extracts of all treatments. In Y-tube olfactometer bioassays, females showed preference towards volatile extracts of UD, ID, and MD plants of each cultivar compared to the control solvent (CH2 Cl2 ). Insects preferred certain synthetic blends comparable to volatile extracts of UD, ID, and MD plants of each L. sativus cultivar against the control solvent. Females preferred a synthetic blend of benzyl alcohol, 1,3-diethylbenzene, thymol, and 1-hexadecene at ratios of 142.49: 62.03:1.18:1 dissolved in 25 μL of CH2 Cl2 in olfactometer bioassays, which could be used in developing lures to control this insect pest.

Behavioral Responses of Thrips hawaiiensis (Thysanoptera: Thripidae) to Volatile Compounds Identified from Gardenia jasminoides Ellis (Gentianales: Rubiaceae)

Thrips hawaiiensis is a common thrips pest of various plant flowers with host preference. Plant volatiles provide important information for host-searching in insects. We examined the behavioral responses of T. hawaiiensis adults to the floral volatiles of Gardenia jasminoides Ellis, Gerbera jamesonii Bolus, Paeonia lactiflora Pallas, and Rosa chinensis Jacq. in a Y-tube olfactometer. T. hawaiiensis adults showed significantly different preferences to these four-flower plants, with the ranking of G. jasminoides > G. jamesonii > P. lactiflora ≥ R. chinensis. Further, 29 components were identified in the volatile profiles of G. jasminoides, and (Z)-3-hexenyl tiglate (14.38 %), linalool (27.45 %), and (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (24.67 %) were the most abundant. Six-arm olfactometer bioassays showed that T. hawaiiensis had significant positive responses to (Z)-3-hexenyl tiglate, linalool, and (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene tested at various concentrations, with the most attractive ones being 10−3 μL/μL, 10−2 μL/μL and 100 μg/μL for each compound, respectively. In pairing of these three compounds at their optimal concentrations, T. hawaiiensis showed the preference ranking of (Z)-3-hexenyl tiglate > linalool > (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene. Large numbers of T. hawaiiensis have been observed on G. jasminoides flowers in the field, which might be caused by the high attraction of this pest to G. jasminoides floral volatiles shown in the present study. Our findings shed light on the olfactory cues routing host plant searching behavior in T. hawaiiensis, providing important information on how T. hawaiiensis targets particular host plants. The high attractiveness of the main compounds (e.g., linalool, (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene, particular (Z)-3-hexenyl tiglate) identified from volatiles of G. jasminoides flowers may be exploited further to develop novel monitoring and control tools (e.g., lure and kill strategies) against this flower-inhabiting thrips pest.

Bacterial phylogeny predicts volatile organic compound composition and olfactory response of an aphid parasitoid

There is increasing evidence that microorganisms emit a wide range of volatile compounds (mVOCs, microbial volatile organic compounds) that act as insect semiochemicals, and therefore play an important role in insect behaviour. Although it is generally believed that phylogenetically closely related microbes tend to have similar phenotypic characteristics and therefore may elicit similar responses in insects, currently little is known about whether the evolutionary history and phylogenetic relationships among microorganisms have an impact on insect–microbe interactions. In this study, we tested the hypothesis that phylogenetic relationships among 40 Bacillus strains isolated from diverse environmental sources predicted mVOC composition and the olfactory response of the generalist aphid parasitoid Aphidius colemani. Results revealed that phylogenetically closely related Bacillus strains emitted similar blends of mVOCs and elicited a comparable olfactory response of A. colemani in Y‐tube olfactometer bioassays, varying between attraction and repellence. Analysis of the chemical composition of the mVOC blends showed that all Bacillus strains produced a highly similar set of volatiles, but often in different concentrations and ratios. Benzaldehyde was produced in relatively high concentrations by strains that repel A. colemani, while attractive mVOC blends contained relatively higher amounts of acetoin, 2,3‐butanediol, 2,3‐butanedione, eucalyptol and isoamylamine. Overall, these results indicate that bacterial phylogeny had a strong impact on mVOC compositions and as a result on the olfactory responses of insects.

The importance of leaf surface wax as short‐range attractant and oviposition stimulant in a generalist Lepidoptera

AbstractGreen gram, Vigna radiata (L.) Wilczek, is an important pulse crop of Asia. Severe attack by the larvae of Spilosoma obliqua Walker (Lepidoptera: Arctiidae) causes defoliation of green gram and reduces seed yield. Females lay eggs on the leaf surface, and therefore, surface wax plays an important role as short‐range attractant and oviposition stimulant. So, we have attempted to find out whether leaf surface wax compounds (alkanes and free fatty acids) from three green gram cultivars (PDM 54, PUSA BAISAKHI and SAMRAT) could act as short‐range attractant and oviposition stimulant in females. The TLC, GC‐MS and GC‐FID analyses of n‐hexane extracts revealed 20 n‐alkanes from n‐C15 to n‐C36 and 13 free fatty acids from C12:0 to C21:0, whilst linoleic acid was unique in SAMRAT. Pentacosane was the predominant amongst n‐alkanes in the leaf surface waxes of three cultivars. Heneicosanoic acid and palmitoleic acid were the predominant free fatty acids in the leaf surface waxes of PDM 54, and PUSA BAISAKHI and SAMRAT, respectively. Females were attracted towards one leaf equivalent surface wax of three green gram cultivars against solvent controls (n‐hexane) in Y‐tube olfactometer bioassays. A synthetic blend of pentacosane, heptacosane, nonacosane, hexatriacontane, palmitoleic acid, linolenic acid and stearic acid, a synthetic blend of pentacosane, hexatriacontane and stearic acid, and a synthetic blend of hexatriacontane, linolenic acid and stearic acid resembling in amounts present in one leaf equivalent surface wax of PDM 54, PUSA BAISAKHI and SAMRAT, respectively, served as short‐range attractant and oviposition stimulant in females. Females showed equal preference for egg laying towards the above three synthetic blends when these blends were tested against each other, and hence, these blends could be employed in development of baited traps in pest management strategies.

Camphor and Borneol as the Male-Produced Sex Pheromone of the Shield Bug, Orsilochides leucoptera (Hemiptera: Scutelleridae).

The ability of stink bugs to release high amounts of strong-smelling and irritating defensive compounds is related to their metathoracic gland (MTG), which is an exocrine gland with defensive, sexual, alarm and aggregation signal functions. Orsilochides leucoptera (Scutelleridae) is a widespread species in the Neotropical region that feeds on plants of the families Malvaceae, Poaceae and Euphorbiaceae. A series of compounds (ketones, alcohols and esters) have been identified in the MTGs among the three species of Scutelleridae whose MTG secretions have been investigated thus far; however, no sex pheromone compounds have been described for any scutellerid species. The aim of this work was to study sex pheromone communication within this family of stink bugs, and identify the compounds present in the MTG of O. leucoptera. Analysis by gas chromatography/mass spectrometry (GC/MS) revealed two male specific compounds identified as (R)-camphor (1) and (R)-borneol (2), which were attractive to females in Y-tube olfactometer bioassays. Also, GC/MS analysis of secretions from MTG of males and females of O. leucoptera, identified eight additional compounds: 2-(E)-hexenal (3), (E)-4-oxo-2-hexenal (4), (E)-hex-2-enyl acetate (5), (R)-linalool (6), (R)-α-terpineol (7), dodecane (8), 1-tridecene (9) and n-tridecane (10). From these, (R)-α-terpineol was detected only in the gland of males, and is probably a biosynthetic intermediate of the pheromone components. Most of the MTG compounds identified in O. leucoptera have been identified in other heteropteran species. Camphor is often a toxic and repellent compound for insects. However, we report it, for the first time, as a sex pheromone component of an insect.

Host preference in parasitic phorid flies: response of Pseudacteon curvatus and P. obtusus to venom alkaloids of native and imported Solenopsis fire ants

Pseudacteon curvatus and P. obtusus are two species of parasitic phorid flies (Diptera: Phoridae), introduced in the southern USA for biological control of Solenopsis fire ants (Hymenoptera: Formicidae). The preference of both parasitoid species (P. curvatus and P. obtusus) for native Solenopsis fire ants (S. geminata complex) versus imported fire ants, S. saevissima complex (S. invicta and S. richteri) was investigated in four-choice olfactometer assays. We hypothesized that differences in the venom alkaloid profiles of fire ant species mediate host preference in phorid flies. In olfactometer bioassays, female P. obtusus showed preference for the venom alkaloid (cis + trans alkaloid mixture) of the imported S. invicta fire ant over the venom alkaloid of the native geminata fire ant. Similarly, P. obtusus showed preference for the venom alkaloid of the S. richteri fire ant over that of the native S. geminata ants. Pseudacteon curvatus also showed preference for the venom alkaloid of the S. invicta and S. richteri fire ants over that of the native geminata ants in separate bioassays. The results suggest that host preference in P. curvatus and P. obtusus is mediated, at least in part, by differences in the venom alkaloid profile of various fire ant species. This finding may explain why Pseudacteon species associated with either the S. geminata or S. saevissima complexes have not switched to the other group despite the overlap of native and imported fire ants in some areas of their distributions.