Aphids, predators and parasitoids.

A number of studies have demonstrated the role of herbivore-induced release of plant volatiles in mediating foraging behaviour of aphid parasitoids, particularly with the parasitoid Aphidius ervi, its aphid host Acyrthosiphon pisum and the aphid food plant Vicia faba. These studies have shown that feeding by the aphid alters the composition of volatiles released by the plant and that these compounds act as synomones for the foraging parasitoid. Of particular interest is the species-specificity of the herbivore-induced synomones associated with different aphids feeding on V. faba. Aphids employ various pheromones that mediate behaviour, particularly mating and alarm responses. These pheromones play important roles in reproduction and defence against predation and parasitism. Many species of aphids reproduce sexually on their primary hosts during the autumn and the sexual females produce a pheromone that attracts males. The sex pheromones for a number of aphid species have been identified and laboratory and field studies have shown that synthetic material can act as a kairomone in attracting predators and parasitoids. The aphid alarm pheromone is released from the cornicles of aphids when they are attacked by predators or parasitoids. The activity of the main alarm pheromone component, (E)-beta-farnesene, is inhibited by the related sesquiterpene hydrocarbon beta-caryophyllene, which is reported to attract the lacewing Chrysoperla carnea. In addition, electrophysiological studies have shown that the seven-spot ladybird, Coccinella septempunctata, possesses specific olfactory receptors for (E)-beta-farnesene and beta-caryophyllene. Laboratory studies show these compounds to have behavioural activity with C. septempunctata, suggesting that they may be involved in prey location.

Treating hop plants with (Z)-jasmone increases colonization by Phorodon humuli (Hemiptera: Aphididae) spring migrants.

Hop plants were sprayed with (Z)-jasmone, at a rate of 50 g ha(-1), during the spring migration of the damson-hop aphid Phorodon humuli (Schrank) in 2002 and 2003. Numbers of P. humuli spring migrants colonizing hop plants, Humulus lupulus L., 2-6 and 7-11 days after applying this treatment were assessed in both years. During the first five-day period, significantly more spring migrants were found on hop plants treated with (Z)-jasmone, in comparison with control plants, in 2002. By contrast, no significant difference was evident in the second five-day period. Although the migration in 2003 was much lighter than in 2002, greater numbers of migrants were again removed from treated plants. Indeed, more spring migrants were removed from plants sprayed with (Z)-jasmone in this year during both five-day periods (11 and 44%, respectively) compared with the 23% greater numbers removed in the first five-day period in 2002. Therefore, unlike some other species of aphid, where numbers were consistently lower on plots sprayed with (Z)-jasmone, spraying the secondary host of P. humuli with this compound appears to increase colonization by spring migrants.

Diel periodicity in the production of green leaf volatiles by wild and cultivated host plants of stemborer moths, Chilo partellus and Busseola fusca.

The volatile chemicals produced by four poaceous plant species, blue thatching grass, Hyparrhenia tamba, Napier grass, Pennisetum purpureum, sorghum, Sorghum bicolor, and maize, Zea mays, which are host plants for the lepidopterous stemborers, Chilo partellus and Busseola fusca, were collected by air entrainment and analyzed by gas chromatography. The total quantities of volatiles collected hourly, over a 9-hr period, from P. purpureum and H. tamba showed an approximately hundredfold increase in the first hour of the scotophase. Thereafter, the amount decreased rapidly to levels present during photophase. Although onset of scotophase also triggered an increase in quantities of volatiles collected from two cultivars of S. bicolor and two out of three cultivars of Z. mays, these increases were less dramatic than in the two wild grasses, being only up to 10 times as much as in the last hour of photophase. Analysis showed that up to 95% of the increase in volatiles at the onset of the scotophase was due to just four compounds, the green leaf volatiles hexanal, (E)-2-hexenal, (Z)-3-hexen-1-ol, and (Z)-3-hexen-1-yl acetate, with the latter dominating the volatile profile. Volatiles from P. purpureum were also collected at 10-min intervals for 70 min spanning the transition from light to dark. The vast increase in production of the green leaf volatiles in this species occurs in the first 10 min of the scotophase followed by a rapid decline within the next 20 min. The relevance of these results to the control of stemborers in a "push-pull" strategy is discussed.

Electrophysiological responses of the lepidopterous stemborers Chilo partellus and Busseola fusca to volatiles from wild and cultivated host plants.

The stemborers Chilo partellus and Busseola fusca are major pests of subsistence cereal farming in Africa. Volatiles released by two cultivated hosts, sorghum and maize (Sorghum bicolor and Zea mays), and two wild grass hosts, Pennisetum purpureum and Hyparrhenia tamba, were collected by air entrainment. Electrophysiologically active components in these samples were detected by coupled gas chromatography-electroantennography (GC-EAG), and the active peaks identified by gas chromatography-mass spectrometry. A total of 41 compounds were identified from the four plant species, all of which, as well as two unidentified compounds, elicited an electrophysiological response from one or both of the stemborers. The compounds included a number of green leaf volatiles and other aliphatic aldehydes, ketones, and esters, mono- and sesquiterpenoids, and some aromatic compounds. EAG studies with authentic samples, conducted at two discriminating doses for all compounds, and dose-response curves for 14 of the most highly EAG-active compounds, showed significant differences in relative responses between species. The compounds that elicited large responses in both species of moths included linalool, acetophenone, and 4-allylanisole, while a number of compounds such as the aliphatic aldehydes octanal, nonanal, and decanal elicited a large response in B. fusca, but a significantly smaller response in C. partellus. Furthermore, the wild hosts produced higher levels of physiologically active compounds compared with either of the cultivated hosts. These differences are discussed in relation to the differential attraction/oviposition of the two stemborers observed in the field and, particularly for eastern African small-scale farming systems, in the context of using a push-pull strategy for their control.

Male-produced aggregation pheromone in pea and bean weevil,Sitona lineatus (L.).

The attraction ofS. lineatus to live baits comprisingS. lineatus feeding onVicia faba (L.) was studied in a field experiment in the early spring. There was clear evidence that maleS. lineatus produced an aggregation pheromone which attracted approximately equal numbers of both sexes from overwintering sites. No evidence was obtained for the production, in the spring, of any semiochemical by female weevils.

Responses of the olfactory receptors ofScolytus scolytus (F.) (Coleoptera: Scolytidae) to the stereoisomers of 4-methyl-3-heptanol.

Electroantennogram (EAG) and single-cell recording techniques have been used to demonstrate the presence of separate receptors for (-)-threo- and (-)-erythro-4-methyl-3-heptanol on the antenna ofS. scolytus. The majority of single-cell recordings showed spikes of two different amplitudes. The cell giving spikes of larger amplitude responded to the (-)-threo stereoisomer while the cell with the small-amplitude spikes responded to (-)-erythro-4-methyl-3-heptanol. It is suggested that in most recordings the two cells are associated with a single sensillum basiconicum.

Absolute configuration of mosquito oviposition attractant pheromone, 6-acetoxy-5-hexadecanolide.

6-Acetoxy-5-hexadecanolide (Ia) in the oviposition attractant pheromone released from egg apical droplets of the mosquitoCulex pipiens fatigans Wied. is shown to be the (-)-(5R,6S)- enantiomer. Identification was by chromatography of the 6-trifluoroacetoxy derivatives of the natural pheromone and of the synthetic (-)-(5R,6S)- (Ib) and (+)-(5S,6R)- (IIb) enantiomers on a capillary column having a chiral stationary phase comprising a derivative of (1S,3S)-chrysanthemic acid. The synthetic (-)-(5R,6S)- enantiomer (Ia) attracted oviposition of four fold more mosquito egg rafts than the control (P < 0.01) whereas for the (5S,6R)- enantiomer (IIa) there was no statistically significant oviposition attraction.

Methyl salicylate and (-)-(1R,5S)-myrtenal are plant-derived repellents for black bean aphid,Aphis fabae Scop. (Homoptera: Aphididae).

Methyl salicylate and (-)-(1R,5S)-myrtenal stimulate specific olfactory cells in the primary rhinaria on the sixth and fifth antennal segments, respectively, of the black bean aphid.Aphis fabae. In behavioral studies employing a linear track olfactometer, both compounds were repellent toA. fabae and also inhibited attraction to volatiles from its host, broad bean (Vicia faba). Methyl salicylate is associated with secondary metabolite-based defense in plants, and the monoterpenoid (-)-(1R,5S)-myrtenal is metabolically related to (-)-(1S,5S)-α-pinene, an abundant component of defensive resins produced by gymnosperms. It is argued that these two compounds are employed byA. fabae as indicators of nutritionally unsuitable or nonhost plants.

Sex pheromone of aphid parasitoidPraon volucre (Hymenoptera, Braconidae).

Mating in the aphid parasitoidPraon volucre is mediated by a female-produced sex pheromone, which laboratory behavioral studies suggest is released from the abdomen. Gas chromatography coupled with electrophysiological and behavioral assays on males showed one area of activity in an air entrainment sample of virgin females. However, levels of pheromone production are very low and this has so far precluded its identification.

Convenient synthesis of mosquito oviposition pheromone and a highly fluorinated analog retaining biological activity.

A simple three-step synthesis is described for 6-acetoxy-5-hexadecanolide, the oviposition pheromone of the mosquitoCulex quinquefasciatus Say and others in that genus. An aldol condensation between 1-trimethylsilyloxycyclopent-1-ene and undecanal, followed by Baeyer-Villiger ring expansion and acetylation, gave the required compound as a 1∶1 mixture of diastereoisomers in high overall yield (>80%). This synthetic approach is readily adapted for synthesis of analogs. The heptadecafluoro compound, in which then-octyl group is replaced by perfluorooctyl, retained high biological activity.

Asymmetrical generalisation between pheromonal and floral odours in appetitive olfactory conditioning of the honey bee (Apis mellifera L.).

The capacity to generalise between similar but not identical olfactory stimuli is crucial for honey bees, allowing them to find rewarding food sources with varying volatile emissions. We studied bees' generalisation behaviour with odours having different biological values: typical floral odours or alarm compounds. Bees' behavioural and peripheral electrophysiological responses were investigated using a combined proboscis extension response conditioning-electroantennogram assay. Bees were conditioned to pure linalool (floral) or to pure isoamyl acetate (alarm) and were tested with different concentrations of both compounds. Electrophysiological responses were not influenced by conditioning, suggesting that the learning of individual compounds does not rely on modulations of peripheral sensitivity. Behaviourally, generalisation responses of bees conditioned to the alarm compound were much higher than those of bees conditioned to the floral odour. We further demonstrated such asymmetrical generalisation between alarm and floral odours by using differential conditioning procedures. Conditioning to alarm compounds (isoamyl acetate or 2-heptanone) consistently induced more generalisation than conditioning to floral compounds (linalool or phenylacetaldehyde). Interestingly, generalisation between the two alarm compounds, which are otherwise chemically different, was extremely high. These results are discussed in relation to the neural representation of compounds with different biological significance for bees.

Antennal perception of oilseed rape,Brassica napus (Brassicaceae), volatiles by the cabbage seed weevilCeutorhynchus assimilis (Coleoptera, Curculionidae).

The response of theCeutorhynchus assimilis antenna to volatiles in air entrainment-derived extracts of oilseed rape,Brassica napus, was studied using coupled gas chromatography (GC)-electroantennography (EAG) and coupled GC-single cell recording (SCR). By means of these techniques and coupled gas chromatography-mass spectrometry (GC-MS), 25 active compounds were identified, including isoprenoids and compounds derived from fatty acids and amino acids. Some of the latter, the isothiocyanates and goitrin, and probably indole and benzyl cyanide, are catabolites of glucosinolates. The electrophysiological activity of the identified compounds was confirmed by EAG using a physiologically discriminating dose, and by SCR studies. The importance of the combined use of the EAG and SCR techniques was demonstrated, since specific olfactory cells were located for five compounds that did not elicit significant EAG responses. The majority of the olfactory cells from which single cell recordings were obtained showed very high specificity, and in numerous recordings there were consistent pairings of specific cell types.

Spring migration of damson-hop aphid,Phorodon humuli (Homoptera, Aphididae), and summer host plant-derived semiochemicals released on feeding.

Behavioral studies using an olfactometer demonstrated that spring migrants of the damson-hop aphid,Phorodon humuli, respond to semiochemicals released by spring migrants feeding on hop leaves. Samples of the total volatiles released on feeding were analyzed by coupled gas chromatographysingle cell recording techniques and showed the presence of several active components. Three compounds were identified, using coupled gas chromatography-mass spectrometry, as methyl salicylate, (E)-2-hexenal andß-caryophyllene. These three compounds elicited responses from separate olfactory receptors on the antenna. In the olfactometer, both (E)-2-hexenal andß-caryophyllene gave positive responses from spring migrants, and a mixture of the two compounds in the natural ratio was more attractive than (E)-2-hexenal alone. Addition of methyl salicylate eliminated the response to the active binary mixture.

Plant-derived synergists of alarm pheromone from turnip aphid,Lipaphis (Hyadaphis) erysimi (Homoptera, Aphididae).

The turnip aphid,Lipaphis (Hyadaphis) erysimi, responds weakly to (E)-ß-farnesene, the main component of the alarm pheromone, but the response is substantially increased by incorporating plant-derived isothiocyanates, identified in aphid volatiles by coupled gas chromatography-single-cell recording.

Volatiles from whitefly-infested plants elicit a host-locating response in the parasitoid, Encarsia formosa.

The blend of volatile compounds emitted by bean plants (Phaseolus vulgaris) infested with greenhouse whitefly (Trialeurodes vaporariorum) has been studied comparatively with undamaged plants and whiteflies themselves. Collection of the volatiles and analysis by gas chromatography revealed more than 20 compounds produced by plants infested with whitefly. Of these, 4 compounds, (Z)-3-hexen-1-ol, 4,8-dimethyl-1,3,7-nonatriene, 3-octanone, and one unidentified compound were emitted at higher levels than from the undamaged control plants. Synthetic (Z)-3-hexen-1-ol, 4,8-dimethyl-1,3,7-nonatriene, or 3-octanone all elicited a significant increase in oriented flight and landing on the source by the parasitoid, Encarsia formosa, in wind tunnel bioassays. Two-component mixtures of the compounds and the three-component mixture all elicited a similar or, in most cases, a better response by the parasitoid, the most effective being a mixture of (Z)-3-hexen-1-ol and 3-octanone. These results demonstrate that E. formosa uses volatiles from the plant-host complex as olfactory cues for host location.

Response of the ladybird parasitoid Dinocampus coccinellae to toxic alkaloids from the seven-spot ladybird, Coccinella septempunctata.

Electrophysiological and behavioral responses of the ladybird parasitoid Dinocampus coccinellae to volatiles from the seven-spot ladybird, Coccinella septempunctata, were investigated to identify semiochemicals involved in host location. Coupled gas chromatography-electroantennography (GC-EAG) with D. coccinellae located a small peak of prominent activity in an extract of volatiles from adult C. septempunctata. The active compound was identified by coupled GC-mass spectrometry and by comparison with an authentic sample as the free-base alkaloid precoccinelline, which forms part of the toxic defense of this ladybird. Behavioral studies in an olfactometer showed that D. coccinellae was significantly attracted to the volatile extract and also to the alkaloid. Myrrhine, a stereoisomer of precoccinelline found in low amounts in C. septempunctata and in other ladybird species, was shown to be electrophysiologically active and significantly attractive. Perception of ladybird alkaloids by D. coccinellae is a rare example of toxicants acting as aerially transmitted cues for interactions between the third and fourth trophic levels.

Field response of the dutch elm disease vectors,Scolytus multistriatus (Marsham) andS. scolytus (F.) (Coleoptera: Scolytidae) to 4-methyl-3-heptanol baits containing α-, ß-,δ-, or δ-multistriatin.

The field responses of English populations of the Dutch elm disease vectors,Scolytus multistriatus andS. scolytus to baits containing 4-methyl-3-heptanol, a host synergist [(-)-α-cubebene or (-)-limonene] and (±)-α-, (+)-ß-, (-)-ß-, (±)-γ-, or (±)-δ-multistriatin were examined. (±)-α-Multistriatin, released at 5-10 µg/day, enhanced the response ofS. multistriatus to baits containing 4-methyl-3-heptanol and either of the host synergists but had no effect on the capture ofS. scolytus. The release of larger amounts (57 or 365 µg/day) of (±)-α-multistriatin interrupted the response of both species to the 4-methyl-3-heptanol baits. It appears that α-multistriatin has multiple functions as a behavior-modifying substance for the two beetles. The (+)-ß-, (-)-ß-, (±)-γ-, and (±)-δ-multistriatins were inactive when released at 5-10 µg/day. The results of these field experiments suggest that one bait can be formulated to capture both species.

Winter host component reduces colonization by bird-cherry-oat aphid,Rhopalosiphum padi (L.) (homoptera, aphididae), and other aphids in cereal fields.

Methyl salicylate, a volatile component ofPrunus padus, the winter host ofRhopalosiphum padi, was found to reduce colonization of the summer host by this aphid. The compound was identified by gas chromatographic analysis coupled with recordings from cells in the primary rhinarium on the sixth antennal segment of the aphid. Methyl salicylate eliminated the attractancy of oat leaves to spring migrants in olfactometer tests. In Sweden, this compound significantly decreased colonization of field grown cereals byR. padi and in the U.K., populations ofSitobion avenae andMetopolophium dirhodum were significantly lower on treated plots.

Discrimination of oilseed rape volatiles by honey bee: Novel combined gas chromatographic-electrophysiological behavioral assay.

A novel technique for the simultaneous monitoring of electroan-tennogram (EAG) and conditioned proboscis extension (CPE) responses of honey bees to the effluent from a gas chromatograph (GC) was developed to locate biologically active components in blends of plant volatiles and to investigate odor recognition at the peripheral and behavioral levels. A six-component mixture, comprising compounds previously identified as oilseed rape floral volatiles, was used as the stimulus. Standard CPE and EAG recordings were done as a reference. EAG responses were elicited from unconditioned bees by all the components presented either in the coupled or the standard mode. Conditioned bees gave larger EAG responses than unconditioned bees, suggesting that antennal sensitivity is enhanced by conditioning. At the behavioral level, in both the standard and the coupled modes, only conditioned bees showed the proboscis extension response, with the majority of individuals responding to linalool, 2-phenylethanol, and benzyl alcohol.