Courtship behavior and identification of a sex pheromone in Ibalia leucospoides (Hymenoptera: Ibaliidae), a larval parasitoid of Sirex noctilio (Hymenoptera: Siricidae)

Background Ibalia leucospoides (Hymenoptera: Ibaliidae) is a larval parasitoid that has been widely introduced as a biological control agent for the invasive woodwasp,Sirex noctilio (Hymenoptera: Siricidae) in the Southern Hemisphere. In this study, the courtship behavior and identificaion of sex pheromones are described for I. leucospoides under laboratory conditions. Methods For courtship behavior, both sexes were observed in a wire mesh observation cylinder (75 cm length ×10 cm diameter) for 15 minutes. The female body washes were analyzed using Gas Chromatography- Electroantennographic Detection (GC-EAD). Then the EAD-active compounds were tentatively identified using GC-Mass Spectrometry (GC-MS) and examined in olfactometer assays. Results The courtship behavior included rhythmic lateral movements, mounting, head-nodding cycles in males, and wing-fanning in females. GC-EAD analysis of female body washes with male antennae revealed seven compounds which elicited antennal responses, four of which are straight-chain alkanes (C23, C25, C26, and C27). The identities of these alkanes were confirmed by matching the retention times, mass spectra, and male antennal activity to those of commercially obtained chemicals. In olfactometer assays, a blend of the four straight-chain alkanes was attractive to I. leucospoides males, and there was no response to blends that lacked any of these four compounds. Female body wash was no more attractive than the four-component blend. The ratios of EAD-active components differ between hydrocarbon profiles from males and females. Conclusion This study is the first investigation of cuticular hydrocarbons in the family Ibaliidae. It provides evidence that the ubiquitous alkanes (C23, C25, C26, and C27) in sex-specific ratios attract I. leucospoides males.

Neural and behavioural responses of the pollen-specialist bee Andrena vaga to Salix odours

An effective means of finding food is crucial for organisms. Whereas specialized animals select a small number of potentially available food sources, generalists use a broader range. Specialist (oligolectic) bees forage on a small range of flowering plants for pollen and use primarily olfactory and visual cues to locate their host flowers. So far, however, little is known about the specific cues oligoleges use to discriminate between hosts and non-hosts and how floral scent compounds of hosts and non-hosts are processed in the bees’ olfactory system. In this study, we recorded physiological responses of the antennae (electroantennographic detection coupled to gas chromatography GC-EAD) and in the brain (optical imaging, GC-imaging), and studied host-finding behaviour of oligolectic Andrena vaga bees, a specialist on Salix plants. In total, we detected 37 physiologically active compounds in host and non-host scents. 4-Oxoisophorone, a common constituent in the scent of many Salix species, evoked strong responses in the antennal lobe glomeruli of A. vaga, but not the generalist honeybee Apis mellifera. The specific glomerular responses to 4-oxoisophorone in natural Salix scents reveals a high degree of specialization in A. vaga for this typical Salix odorant component. In behavioural experiments, we found olfactory cues to be the key attractants for A. vaga to Salix hosts, which are also used to discriminate between hosts and non-hosts, and demonstrated a behavioural activity for 4-oxoisophorone. A high sensitivity to floral scents enables the specialized bees to effectively find flowers and it appears that A. vaga bees are highly tuned to 4-oxoisophorone at a very low concentration.

Generalized olfactory detection of floral volatiles in the highly specialized Greya-Lithophragma nursery pollination system

Volatiles are of key importance for host-plant recognition in insects. In the pollination system of Lithophragma flowers and Greya moths, moths are highly specialized on Lithophragma, in which they oviposit and thereby pollinate the flowers. Floral volatiles in Lithophragma are highly variable between species and populations, and moths prefer to oviposit into Lithophragma flowers from populations of the local host species. Here we used gas chromatography coupled with electroantennographic detection (GC-EAD) to test whether Greya moths detect specific key volatiles or respond broadly to many volatiles of Lithophragma flowers. We also addressed whether olfactory detection in Greya moths varies across populations, consistent with a co-evolutionary scenario. We analyzed flower volatile samples from three different species and five populations of Lithophragma occurring across a 1400 km range in the Western USA, and their sympatric female Greya politella moths. We showed that Greya politella detect a broad range of Lithophragma volatiles, with a total of 23 compounds being EAD active. We chemically identified 15 of these, including the chiral 6, 10, 14-trimethylpentadecan-2-one (hexahydrofarnesyl acetone), which was not previously detected in Lithophragma. All investigated Lithophragma species produced the (6R, 10R)-enantiomer of this compound. We showed that Greya moths detected not only volatiles of their local Lithophragma plants, but also those from allopatric populations/species that they not encounter in local populations. In conclusion, the generalized detection of volatiles and a lack of co-divergence between volatiles and olfactory detection may be of selective advantage for moths in tracking hosts with rapidly evolving, chemically diverse floral volatiles.

Electrophysiological and Behavioral Responses of Dasychira baibarana (Lepidoptera: Lymantriidae) to Tea Plant Volatiles

Tea black tussock moth, Dasychira baibarana (Matsumura) (Lepidoptera: Lymantriidae), is a devastating pest species of the tea plant in China. Here, we evaluated the responses of D. baibarana to tea plant volatiles using gas chromatography coupled electroantennographic detection (GC–EAD), eleclectroantennography (EAG), and a Y-tube olfactometer. In total, 11 of 18 analyzed compounds elicited GC–EAD responses from test insects. GC–EAD bio-active compounds were further investigated using EAG and behavioral responses. In the EAG analysis, male moths had significantly greater responses to four compounds [(Z)-3-hexenyl butyrate, (Z)-3-hexen-1-ol, ocimene and benzyl alcohol] than female moths. For females, maximum EAG amplitudes, were recorded in response to linalool, (Z)-3-hexenyl hexanoate and (Z)-jasmone. In EAG and behavioral bio-assays, the responses of both sexes were dose independent. In behavioral bio-assays male moths responding significantly to (Z)-3-hexen-1-ol, ocimene, (Z)-3-hexenyl butyrate, linalool, benzyl alcohol, and (Z)-jasmone at various concentrations. For females, significant behavioral responses were observed to (Z)-3-hexenyl hexanoate, followed by (Z)-jasmone, linalool, ocimene, and benzyl alcohol. However, neither sex was sensitive to 4 of the 11 tested compounds, phenyethyl alcohol, phenylacetonitrile, (E)-nerolidol, and indole. The present results showed that tea plant volatiles influenced the behavior of D. baibarana moths, which will greatly contribute in developing eco-friendly control strategies for D. baibarana, through the application of a blend of compounds that showed significant EAG and behavioral responses or a blend combined with female-produced sex pheromones.

Antennal and Behavioral Responses of Drosophila suzukii to Volatiles from a Non-Crop Host, Osyris wightiana

Drosophila suzukii (Diptera: Drosophilidae) infests a variety of commercial fruits, including cherries and other soft-skinned fruits. After the cropping season of most cultivated crop hosts, it heavily infests the fruit of a wild host-plant, Osyris wightiana in southwest China. Here, we employ gas chromatography-electroantennographic detection (GC-EAD) together with behavioral bioassays and a trapping experiment to identify volatile semiochemicals emitted by O. wightiana that are involved in D. suzukii attraction. GC-EAD recordings of D. suzukii antenna showed responses to 13 compounds, including α-pinene, 3-methylbutyl acetate, 2-hexanol, E-β-ocimene, Z-3-hexenol, β-caryophyllene, α-humulene, and six unidentified compounds. The flies were attracted by seven individual EAD-active compounds at low doses (0.01 and 0.1 μg), but were repelled at high doses (10 and 100 μg). In a similar manner, a blend of seven EAD-active compounds at low doses (0.1 and 1 μg) was attractive to female flies, but became repulsive at high doses (10 μg). The low dose of the blend was as attractive as the fruit volatiles, although both were less attractive than the fruits. The blend attracted both female and male D. suzukii and other Drosophila flies. The percentage of D. suzukii out of all flies captured by the blend was significantly greater than that captured by the control. These results indicate that the EAD-active volatile compounds emitted by fruits of O. wightiana play an important role in D. suzukii attraction, and have the potential to be used for management of D. suzukii.

Electrophysiological and Behavioral Responses of Holotrichia parallela to Volatiles from Peanut

Holotrichia parallela (Coleoptera: Scarabaeidae: Melolonthinae) is a notorious pest of many crops, especially peanuts. In this study, volatiles from peanut plants were analyzed using both gas chromatographic-electroantennographic detection (GC-EAD) and gas chromatography/mass spectrometry (GC/MS) techniques, and tested for adult attraction with field trapping bioassays in Hebei Province, China. GC-EAD analyses indicated that H. parallela antennae strongly responded to twelve GC peaks, including eight identified compounds, (Z)-β-ocimene, hexanal, 6-methyl-5-hepten-2-one, nonanal, dihydromyrcenol, linalool, β-caryophyllene, methyl salicylate, and four unidentified compounds. When tested individually in field conditions from 24 to 31 July, 2020, β-caryophyllene and hexanal significantly attracted both sexes of H. parallela, whereas all other compounds were unattractive. A blend of β-caryophyllene and hexanal at a ratio of 2:1, close to the natural ratio of these two compounds from the intact peanut plant, was most attractive to the beetles. The remaining identified compounds, (Z)-β-ocimene, 6-methyl-5-hepten-2-one, nonanal, dihydromyrcenol, linalool, and methyl salicylate had no synergistic effects on H. parallela attraction when tested in combination with the blend of β-caryophyllene and hexanal. These results demonstrated that β-caryophyllene and hexanal in the volatiles from peanut plants have strong attraction to H. parallela. These two compounds have the potential to be used for monitoring H. parallela and its management programs.

Semiochemical-based Reproductive Isolation Among Sympatric Species of Trypodendron (Coleoptera: Curculionidae: Scolytinae)

Trypodendron retusum (LeConte) (Coleoptera: Curculionidae: Scolytinae) females excised from newly attacked trembling aspen, Populus tremuloides Michaux (Salicaceae), were shown for the first time to produce the aggregation pheromone (+)-lineatin. Coupled gas chromatographic-electroantennographic detection analysis (GC-EAD) disclosed that the antennae of T. retusum, as well as the antennae of three sympatric species, native T. lineatum (Olivier) and T. rufitarsus (Kirby) and exotic T. domesticum (L.), respond to synthetic (+)-lineatin, but not the (−) enantiomer. In contrast, the antennae of T. betulae Swaine responded to SR- and RR-linalool oxide pyranoid and did not detect lineatin. GC-EAD analysis of volatiles from host and nonhost tree species revealed that conifer-produced α-pinene and angiosperm-produced conophthorin and salicylaldehyde were perceived by the antennae of all three native lineatin-perceiving species, suggesting behavioral activity. Field trapping experiments showed that salicylaldehyde synergized the response of coastal, but not interior, T. retusum to lineatin and inhibited the response of T. lineatum and T. rufitarsus. In the absence of salicylaldehyde, α-pinene appeared to inhibit the response of interior T. retusum to lineatin, while for T. lineatum and T. rufitarsus it had an apparent positive additive or synergistic effect. No behavioral response occurred to conophthorin. The results provide evidence for semiochemical-based reproductive isolation between T. retusum and T. betulae, and between these two angiosperm-infesting species and the two conifer-infesting species. They do not explain how isolation could be maintained between T. lineatum and T. rufitarsus.

The Attraction of the Dung Beetle Anoplotrupes stercorosus (Coleoptera: Geotrupidae) to Volatiles from Vertebrate Cadavers

During decomposition, vertebrate carrion emits volatile organic compounds to which insects and other scavengers are attracted. We have previously found that the dung beetle, Anoplotrupes stercorosus, is the most common dung beetle found on vertebrate cadavers. Our aim in this study was to identify volatile key compounds emitted from carrion and used by A. stercorosus to locate this nutritive resource. By collecting cadaveric volatiles and performing electroantennographic detection, we tested which compounds A. stercorosus perceived in the post-bloating decomposition stage. Receptors in the antennae of A. stercorosus responded to 24 volatiles in odor bouquets from post-bloating decay. Subsequently, we produced a synthetic cadaver odor bouquet consisting of six compounds (benzaldehyde, DMTS, 3-octanone, 6-methyl-5-hepten-2-ol, nonanal, dodecane) perceived by the beetles and used various blends to attract A. stercorosus in German forests. In field assays, these beetles were attracted to a blend of DMTS, 3-octanone, and benzaldehyde. Generalist feeding behavior might lead to the super-dominant occurrence of A. stercorosus in temperate European forests and have a potentially large impact on the exploitation and rapid turnover of temporally limited resources such as vertebrate cadavers.

Behavioral Responses of Scolytus schevyrewi (Coleoptera: Curculionidae: Scolytinae) to Volatiles From Apricot Tree (Rosales: Rosaceae)

Scolytus schevyrewi Semenov (Coleoptera: Curculionidae: Scolytinae) is native to China and Central Asia. Damage by the adults and larvae weakens local apricot trees (Armeniaca spp.), often causing death to many host trees. In previous studies, freshly cut apricot logs were found to be highly attractive to S. schevyrewi adults. To explore the possibility of trapping and monitoring this bark beetle, we evaluated the effect of the apricot tree volatiles on S. schevyrewi behavior. Volatiles from the apricot logs were collected by headspace sampling and subjected to coupled gas chromatography-electroantennographic detection (GC-EAD) analysis. Behavioral responses to EAD-active compounds were assessed using two-choice Y-tube olfactometers. The results showed that the antennae of females responded to 21 volatile compounds from apricot logs. Seventeen compounds were confirmed with authentic compounds. The Y-olfactometer bioassays showed that at a stimulation dose of 100 µg, four compounds [(1S)-(−)-α-pinene, (±)-limonene, (1S)-(+)-3-carene, and 1-hexanol], and some binary mixtures of the four compounds [(1S)-(−)-α-pinene plus (±)-limonene; (1S)-(−)-α-pinene plus (1S)-(+)-3-carene; (1S)-(−)-α-pinene plus camphene; (1S)-(−)-α-pinene plus (±)-limonene, (1S)-(+)-3-carene, and 1-hexanol] were significantly attractive to both sexes (except (±)-limonene and (1S)-(+)-3-carene for males), suggesting that these compounds may play a role in host tree selection by S. schevyrewi and should be evaluated as lures for population monitoring. In contrast, octanal, nonanal, decanal, linalool and N,N-diethylformamide appeared to repel S. schevyrewi adults in Y-tube at the concentration tested.