Deposition and longevity of oviposition-deterring pheromone in the cabbage seed weevil

Abstract The cabbage seed weevil ( Ceutorhynchus assimilis Payk.) lays eggs singly into pods of oilseed rape ( Brassica napus L.) through punctures bored with the mouthparts, preferring pods not recently used for oviposition. A simple new choice test has been used to test individual components of egg-laying behaviour for their effect on oviposition site selection. It is confirmed that an oviposition-deterring pheromone (ODP) is deposited during abdomen brushing of the pod which follows egg-laying. Neither pin punctures, weevil feeding punctures, oviposition punctures nor eggs had any deterrent effect. Pods walked on by female weevils were not avoided by those laying eggs. Observations suggest that the ODP is sensed by contact chemoreceptors on the antennae. The deterrent effect lasted only 1–2 h. The implications of these findings on the adaptive significance of the pheromone and its possible use in pest control are discussed.     

Evolution of cannibalism and female's response to oviposition-deterring pheromone in aphidophagous predators

1.  Egg cannibalism by larvae is common in Coccinellidae and is known to be advantageous for the cannibals. Furthermore, larvae of aphidophagous ladybirds usually produce an oviposition-deterring pheromone (ODP), which inhibits oviposition by adult females. It has been proposed that the response to ODP has evolved because of the high costs of cannibalism. However, this has never been formally proved.
2.  In this paper, we study the theoretical evolution of this system. We first look at the conditions under which cannibalism and the response to ODP can evolve. Subsequently, we examine the occurrence of polymorphism both in the production of larval tracks and in the sensitivity of females to specific pheromones.
3.  The models predict that the amount of cannibalism should not depend on prey density and that evolution should lead to a continuous increase in cannibalism, and consequently larvae should always cannibalize eggs when possible. In response to the cost of cannibalism, ODP recognition can evolve, so that females avoid laying eggs in patches of prey already occupied by conspecific larvae. The result is an arms race between larvae and adult females, which favours a diversification of ODP pheromones. Our models show that: (i) females should be able to recognize mixtures of hydrocarbons rather than a single molecule; and (ii) females should be more sensitive to the tracks of their own offspring than those of non-related larvae.     

(1S,3S,7R)-3-methyl-alpha-himachalene from the male sandfly Lutzomyia longipalpis (Diptera: Psychodidae) induces neurophysiological responses and attracts both males and females

Lutzomyia longipalpis adult males form leks on or near hosts and release (1S,3S,7R)-3-methyl-alpha-himachalene from their tergal glands to lure females to the same site for mating and feeding. Here we have examined whether the male-produced attractant could also serve as a male aggregation stimulus. High resolution chiral capillary gas chromatography analysis of male tergal gland extracts, synthetic (1S,3S,7R)-3-methyl-alpha-himachalene, and a synthetic mixture of all isomers of 3-methyl-alpha-himachalene, was coupled to electrophysiological recordings from ascoid sensillum receptor cells in antennae of male and female sandflies. Receptor cells of both sexes responded only to the main component of the male tergal gland extract that eluted at the same retention time as (1S,3S,7R)-3-methyl-alpha-himachalene. Furthermore, of the eight 3-methyl-alpha-himachalene isomers in the synthetic mixture only the fraction containing (1S,3S,7R)-3-methyl-alpha-himachalene, co-eluting with an isomer of (1S*,3S*,7S*)-3-methyl-alpha-himachalene, elicited an electrophysiological response from male and female ascoid sensillum receptor cells. Both males and females flew upwind in a wind tunnel towards a filter paper disk treated with either 4-6 male equivalents of the tergal gland extract, pure (1S,3S,7R)-3-methyl-alpha-himachalene or the synthetic mixture of eight isomers. This indicates that (1S,3S,7R)-3-methyl-alpha-himachalene derived from L. longipalpis males may have a dual function in causing male aggregation as well as serving as a sex pheromone for females.     

Contact chemoreception related to host selection and oviposition behaviour in the monarch butterfly, Danaus plexippus

Abstract. Behavioural events during host selection by ovipositing monarch butterflies (Danaus plexippus (L.), Danainae, Nymphalidae) include tapping the leaf surface with fore-tarsi and touching this surface with mid-tarsi (‘drumming’) and antennae. Flavonoids identified from host plant extracts are known to stimulate oviposition. Scanning electron microscopy revealed the presence of contact-chemoreceptor sensilla on all appendages that contact the leaf surface. This electrophysiological study was conducted to identify the contact chemoreceptors that are sensitive to the known oviposition stimuli and are therefore probably involved in host recognition. Receptor cells of conspicuous sensilla grouped in clusters on fore-tarsi of females were sensitive to the behaviourally active butanol fraction of host plant (Asclepias curassavica) extract. However, these receptors generally had low sensitivity to three oviposition-stimulating flavonoids identified from this fraction, but they were also sensitive to the butanol fraction of a non-host (Brassica oleracea). Chemoreceptors in sensilla of the tarsomers 2–4 of the mid-legs also responded to the behaviourally active fraction of host plant extract and showed some sensitivity to two of the flavonoids that stimulate oviposition. Similar results were obtained from receptor cells in sensilla on the tip of the antennae. Most of these sensilla had cells responding to the butanol fraction of A. curassavica extract but only 25% of them were also sensitive to one of the behaviourally active flavonoids. These electrophysiological results, in combination with behavioural observations, suggest that host selection in monarch butterflies relies on a complex pattern of peripheral sensory information from several types of tarsal and antennal contact chemoreceptors.     

Behavioural effects of minor sex pheromone components in Brazilian apple leafroller Bonagota cranaodes (Lep., Tortricidae)

Abstract:  The behavioural response of Brazilian apple leafroller males, Bonagota cranaodes (Meyrick), to natural and synthetic sex pheromone was studied in a wind tunnel. Calling females elicited upwind flights followed by landing and wingfanning at the source in 72% of the males tested. Female gland extracts, with the main compound ( E , Z )3,5dodecadienyl acetate released at 100 pg/min, attracted 57% of the males to the source. Few males (1%) were attracted to the main compound alone, released at the same rate. Even a synthetic blend of all five gland compounds eliciting an antennal response, formulated according to their proportion in female gland extracts, was barely attractive (7%). Comparison of this synthetic blend and female gland extracts indicates a behavioural role of other gland compounds. Male attraction was significantly increased (34%) in response to a 100 : 5 : 5 : 5-blend of the main compound and three minor gland compounds, ( Z )-5-dodecenyl acetate, ( E , Z )-3,5-tetradecadienyl acetate, and ( Z )-9-hexadecenyl acetate.     

Effect of estrogen and its antagonist on the expression of arginine vasotocin (AVT) and its oxytocic-like receptor VT3 in the shell gland of Japanese quail, Coturnix coturnix japonica

Avian neurohypophysial hormone arginine vasotocin (AVT) is known to regulate shell gland contractility during oviposition. While studying the role of estrogen in the expression and regulation of AVT and its oxytocic-like receptor VT3, using in situ hybridization and immunohistochemistry, it was observed that the expression of AVT and its receptor was not detected in the shell gland of sexually immature Japanese quail. However, administration of estrogen to these birds not only stimulates the growth and activity (as assessed by increased mucosal fold length, total protein content and alkaline phosphatase level) of the shell gland but also upregulates the expression of AVT and VT3. Further, administration of estrogen antagonist tamoxifen to sexually mature bird shows opposite results. On the other hand, localization of ir-AVT, observed in the ovary of sexually mature bird, was not detected in the estrogen treated sexually immature quail. It is concluded that estrogen not only affects the growth and differentiation of avian oviduct, but also regulates the expression of shell gland AVT and its receptor VT3. Present findings suggest that the locally synthesized AVT acts in a paracrine way to upregulate VT3 receptor and thus facilitates the endocrine function of neurohypophysial AVT during oviposition.     

A dihydroxy-gamma-lactone as an oviposition stimulant for the swallowtail butterfly, Papilio bianor, from the rutaceous plant, Orixa japonica

The oviposition response of the Rutaceae-feeding swallowtail butterfly, Papilio bianor, was induced by a methanolic extract from leaves of its major host, Orixa japonica. Several components were responsible for this oviposition response. One of the stimulants was isolated and identified as (-)-2-C-methyl-D-erythrono-1,4-lactone. The compound was inactive alone, but elicited oviposition behavior when mixed with other fractions.     

Relationship between sex pheromone elicited behaviour and response of single olfactory receptor neurones in a wind tunnel

Responses from pheromone‐specific receptor neurones in male Agrotis segetum (Denis & Schiffermüller) (Lepidoptera: Noctuidae) were recorded in a laboratory wind tunnel. Stimuli were: (1) rubber septum dispensers loaded with single components or a four‐component pheromone blend, (2) excised glands from female A. segetum, (3) constrained A. segetum females with extruded glands. Dose–response curves for three neurone‐types with different specificity were established. The neurones were specifically tuned to respond to either one of the two pheromone components (Z)‐5‐decenyl acetate and (Z)‐7‐dodecenyl acetate, or to the behavioural antagonist (Z)‐5‐decenol. In parallel, a behavioural dose–response curve with males flying upwind to a four‐component pheromone blend was established. There was a clear correlation between behavioural arrestment of upwind flight and maximum spiking activity in Z5–10:OAc‐specific neurones. The pheromone release rates of individual females and synthetic dispensers were compared. A load of 50–200 ng of Z5–10:OAc on a rubber septum elicited approximately the same neural response as one female gland.     

Sex pheromone of the butterbur borer, Ostrinia zaguliaevi

The sex pheromone blend of the butterbur borer, Ostrinia zaguliaevi (Lepidoptera: Pyralidae) was analyzed by means of gas chromatography-electroantennographic detection (GC-EAD), GC-mass spectrometry and a series of wind-tunnel bioassays. Four EAD-active compounds were detected in the female sex pheromone gland extract, and these were identified as tetradecyl acetate (14:OAc), (Z)-9-tetradecenyl acetate (Z9-14:OAc), (E)-11-tetradecenyl acetate (E11-14:OAc) and (Z)-11-tetradecenyl acetate (Z11-14:OAc). The average amounts ± s.d. of the four compounds in a single sex pheromone gland were 7.9±3.7 ng, 10.1±3.2 ng, 1.1±0.5 ng and 11.6±5.1 ng, respectively. In a wind-tunnel bioassay, the ternary blend of Z9-, E11- and Z11-14:OAc at a ratio found in the sex pheromone gland (45:5:50) elicited the same behavioral responses from the males as did virgin females and pheromone gland extract. Removal of any single compound from the ternary blend significantly diminished the pheromonal activity, whereas addition of 14:OAc to the ternary blend had no effect on the males' behavioral responses. Therefore, it was concluded that the sex pheromone blend of O. zaguliaevi is composed of Z9-14:OAc, E11-14:OAc and Z11-14:OAc at a ratio of 45:5:50.     

Behavioral and electrophysiological studies on the sexually biased synergism between oligosaccharides and phospholipids in gustatory perception of nuptial secretion by the German cockroach

Females of Blattella germanica feed compulsively on a nuptial secretion from the male's eighth tergal gland (TG-8) during courtship. Using TG-8 extract and its essential ingredients, maltotriose (MT) and 1,2-dioleoylphosphatidylcholine (PC), we investigated the perception of the secretion by gustatory sensilla in both sexes. Female-biased chemosensitivity was found in the feeding responses to the TG-8 extract. The TG-8 extract induced specific impulses in four functionally different receptor cells: the sugar, salt and other two types of receptor cells in a single gustatory sensillum on the paraglossae in both sexes. The impulse frequencies of the sugar receptor cell and a receptor cell of unknown type were significantly higher in females than those in males. There were no sexual differences in the behavioral and electrophysiological responses to MT alone; no responses were elicited by PC. However, a mixture of MT and PC elicited the behavioral responses more strongly than MT in females. The impulse frequency of sugar receptor cells toward the mixture of MT and PC also increased in females but not in males. These results suggest that the synergistic effect of PC on ‘sweetness’ of MT in the female cockroach contributes to her compulsive nuptial feeding. The female-biased gustatory sensitivity seems to ensure successful coupling during the nuptial feeding.     

Behavioural plasticity in support of a benefit for aggregation pheromone use in Drosophila melanogaster

We explored behavioural plasticity in the use of aggregation pheromone in the fruit fly Drosophila melanogaster Meigen (Diptera: Drosophilidae). Based on previous field observations, we formulated two hypotheses on a benefit of using aggregation pheromone for aggregated oviposition. One hypothesis concerns a benefit to the females themselves, where reduced harassment by males can enhance oviposition rate; the other concerns a benefit to their offspring, where larvae can exploit arduous substrates more efficiently. We derive contrasting expectations on the strength of the behavioural response to pheromone for substrates that differ in nutritional quality to larvae. High quality substrates relax the strength of larval competition, which allows for stronger aggregative responses of the females, but conversely, it may yield aggregation less necessary when the benefit is related to resource exploitation by the larvae. In indoor and outdoor dual choice set-ups, we tested the behavioural responses of the adults to the aggregation pheromone with substrates of varying quality, and examined oviposition behaviour. The response of adults to the aggregation pheromone was strong and robust for low quality substrates, but significantly weaker for a high quality substrate. This supports the hypothesis on a benefit to the larvae. Females retained aggregation pheromones in the absence of oviposition substrates for at least 24 h. In the outdoor set-up, substrate with aggregation pheromone received more than three times as many eggs as control substrates, and this was directly related to the number of adults that visited each substrate. Per capita, oviposition rates were not different for differently sized aggregations, and consequently, no evidence was found in support of the hypothesis on reduced harassment. The combination of aggregation pheromone possession and a mutualistic relationship with micro-organisms for Drosophila and other insects is discussed.     

Evidence for an oviposition-deterring pheromone in Tephritis bardanae (Schrank) (Diptera: Tephritidae)

Summary Female Tephritis bardanae (Schrank) exhibit a characteristic ovipositor-dragging behaviour immediately after laying eggs into a flowerbud of Arctium minus (Hill) Bernh. To establish whether this behaviour is associated with an oviposition-deterring pheromone (ODP), female T. bardanae were presented with clean, unattacked flowerbuds and fly activity was monitored using video recording equipment. The distribution of oviposition and fly behaviour were analysed. Avoidance of attacked buds was revealed by the oviposition pattern becoming overdispersed. Spacing out of eggs continued until, on average, each bud contained one egg-batch, after which the distribution became more random or clumped. Analysis of fly behaviour showed that attacked and unattacked buds received similar numbers of visits, but fewer oviposition attempts were made on attacked buds. This behaviour was consistent with use of a contact pheromone deterring repeated oviposition and applied to the bud surface during ovipositor-dragging. Avoidance of attacked buds should increase the efficiency of resource use by T. bardanae populations in the field; however, distributions of egg-batches in flowerhead samples collected from Monks Wood NNR, Cambridgeshire, during 1983–1985, were highly clumped and did not provide supportive evidence. This suggests that the effectiveness of oviposition deterrence in the field is reduced, because the active life of ODP under natural conditions is short, and/or because wild females frequently experience high oviposition drive due to scarcity of suitable flowerbuds.     

The orientation inducer pheromone of the fire ant Solenopsis invicta

Abstract Foraging ants recruit nestmate workers to food sources by a variety of mechanisms. We report that one behavioural subcategory of the recruitment pheromone complex of Solenopsis invicta Buren involves orientation induction. The orientation inducer pheromone exerts its effects by changing the physiological state of the recipient rather than by releasing a measurable behaviour. Some ant species use a physical 'waggle' behaviour to motivate (change physiological state) nestmate workers to follow their chemical trail. The orientation inducer pheromone can be interpreted as a chemical analogue of the physical 'waggle' inducing effects. This behaviour is not elicited by the recruitment pheromone components responsible for orientation and/or attraction. Each of these behavioural categories is mediated by a different blend of chemicals from the Dufour's gland. Activity-concentration thresholds indicate that the attraction and inducer part of the recruitment pheromone require about 250 times more worker equivalents for a response than the orientation pheromone. Therefore, the recruitment sub-categories are differentially activated by the amount of Dufour's gland material released.     

Source of the host marking pheromone in the egg parasitoid Trissolcus basalis (Hymenoptera: Scelionidae)

After oviposition, Trissolcus basalis females always mark the host's surface, depositing host marking substances for herself and to warn other ovipositing females. The perception of these host marking substances, probably through the antennae, can induce the female to leave and seek healthy hosts. Parasitoid females exposed to conspecific parasitized egg masses left the host egg masses significantly more often than when exposed to non-parasitized egg masses. More egg mass leaving behavior also was observed when the egg masses were treated with Dufour's gland secretion but not when treated with secretion from the common oviducts. The common oviduct has a secretory epithelium that produces electron-dense vesicles, probably containing proteinaceous substances. The secretory cells of the accessory gland, Dufour's gland, contain electron-lucid vesicles, whose secretion appears to be a lipid similarly to that found in pheromone secreting glands. Ultrastructural and behavioral evidence suggests that Dufour's gland is the host marking pheromone source.     

Behavioural and electrophysiological responses of grape berry moth (Lep., Tortricidae) to selected plant extracts

Abstract:  Four non-host plant extracts from Bifora radians , Arctium lappa , Humulus lupulus and Xanthium strumarium , were tested against adult grape berry moths, Paralobesia viteana , for their potential as repellents or oviposition deterrents. Responses were compared with those elicited by the major component of the P. viteana sex pheromone. Moths of both sexes exhibited varying electroantennogram (EAG) responses from 1.7 to 2.4 mV to volatile odours from plant extracts, with the greatest response to extracts of H. lupulus (2.4 mV). A multi-arm olfactometer was used to assay the behavioural response of moths to the same treatments. Male moths responded significantly to H. lupulus extract, although their strongest response was to the sex pheromone (30.0%). Female moths showed no behavioural response to the sex pheromone, attracting only 6.7% of moths, but they were attracted to extracts of H. lupulus , X. strumarium and A. lappa , with 25.0%, 21.7% and 15.0% of the released insects responding to these treatments, respectively. Choice tests were conducted to compare oviposition by P. viteana on untreated grapes and on grapes treated with one of the extracts. Despite the attraction in the olfactometer assays, H. lupulus extract significantly reduced egg laying; only 12.5% of the total deposited eggs were laid on berries treated with this extract. The extract of B. radians provided complete protection against oviposition.     

Male accessory gland substance of Melanoplus sanguinipes: An oviposition stimulant under the control of the corpus allatum

The accessory glands of male Melanoplus sanguinipes contain an oviposition stimulant. Injection of gland extracts from mature males induces oviposition in 75 per cent of capable virgin females within 24 hr. Injection of gland extracts from allatectomized males produces no stimulatory effect. Gland extracts from mature males contain two antigens which cannot be detected in gland extracts from allatectomized males. However, both antigens can be detected in gland extracts from allatectomized males 3 days after treatment with juvenile hormone.Anion-exchange chromatography of mature gland extracts yielded two fractions which, when injected into virgin females, induced oviposition in 71 per cent of capable insects within 24 hr. These two fractions are immunologically identical to the two antigens which are absent from gland extracts of allatectomized males. We suggest that synthesis of the oviposition stimulant in Melanoplus is controlled by the corpus allatum.Injection of brain extract also induces oviposition in 100 per cent of capable virgin females within 24 hr. A possible rôle for the brain in the oviposition process is discussed.     

Presence of eupyrene spermatozoa in vestibulum accelerates oviposition in the silkworm moth,Bombyx mori

In the silkmoth, Bombyx mori, mating accelerates egg laying. Sterilized males did not induce females to oviposit, whereas artificial insemination was enough to accelerate oviposition. These findings showed that mechanical stimulation of the outer reproductive organs and the bursa copulatrix (BC) had no effect on oviposition. Factors included in seminal fluids also did not stimulate oviposition because neither implantation of female internal reproductive organs nor injection of the spermatophore extract into virgin females activated oviposition. Preventing sperm precedence by experimental removal of the BC or spermatheca inhibited activation of egg laying. More precise surgeries on the spermatheca showed that the main factor for eliciting oviposition was the presence of matured and fertile eupyrene spermatozoa in the vestibulum. Sperm were not effective unless they reached the vestibulum. Nerve transection and electrophysiological studies revealed that nerve 4 of the terminal abdominal ganglion was the pathway by which the mating signal was transmitted toward the higher central nervous system.     

Pre-exposure modulates attraction to sex pheromone in a moth

In behavioural experiments we investigated the influence of previous short exposure to sex pheromone on subsequent response of male Spodoptera littoralis moths to sex pheromone. We found that pre-exposed males showed increased sensitivity to female sex pheromone after a single exposure to a pheromone plume compared to that found in na?ve males. The increased responsiveness lasted for at least 27 h after the exposure, showing that it was not just a short-term sensitization of the males. Exposure to the odour source without upwind movement towards the source was enough to increase the responsiveness. Physical activation without exposure to odour did not affect responsiveness. The increase in responsiveness after exposure was higher when the males were pre-exposed to natural female pheromone gland extract than when they were exposed to a higher dose of the main component, even though both odour sources elicited similar upwind attraction in na?ve males. Thus, the quality of the pheromone mixture to which males were exposed influenced the subsequent response.     

Hormonal influence on the olfactory response to a female-attracting pheromone, sodefrin, in the newt, Cynops pyrrhogaster

Sodefrin is a female-attracting pheromone isolated from the abdominal glands of male newts, Cynops pyrrhogaster. Previously, the preference of conspecific female newts for sodefrin was shown to be completely abolished by plugging the bilateral nostrils, indicating that it acts on the olfactory organ. To determine the sensitivity of the olfactory receptor cells to sodefrin, electro-olfactograms (EOGs) in response to sodefrin solution were recorded from the ventral nasal epithelium of sexually developed female newts. Sodefrin elicited marked EOG responses in a dose-dependent manner on the epithelium of the lateral nasal sinus (LNS) region, a putative vomeronasal organ. In ovariectomized females, treatment with prolactin (PRL) and estrogen markedly enhanced the EOG response to sodefrin. The EOG response to the pheromone was also enhanced considerably by treatment with either PRL or estrogen alone. A slight but significant elevation was observed in castrated males receiving PRL plus estrogen or estrogen alone. It was concluded that the main site of action of sodefrin resides in the lateral sinus region and that sensitivity to sodefrin is under the control of PRL and estrogen. The presence of a sex difference in olfactory sensitivity to the hormones and/or pheromone was also suggested.     

Effects of photoperiod and temperature on the rate of larval development, food conversion efficiency, and imaginal diapause in Leptinotarsa decemlineata

Larvae of Leptinotarsa decemlineata developed faster and consumed less food under short-day (Sd, 12:12 h light:darkness) than under long-day (Ld, 18:6 h L:D) conditions. The average index of food conversion efficiency was 5.4 in the Ld (25 degrees C), and 7.2 and 11.9 (at 20 and 25 degrees C, respectively) in the Sd insects. Pupae were smaller under the Ld conditions due to a greater loss of biomass during the prepupal period that was nearly twice longer than in the Sd insects. Virgin Ld females laid eggs for 6 months and survived 13 months. The lack of oviposition, reduced food intake, and behavioural changes characterised diapause in the Sd adults. Application of 100 microg JH III to newly ecdysed adults was used to probe diapause intensity. At 25 degrees C, the treatment elicited oviposition most effectively in females that were just transferred from the Ld to the Sd conditions. A distinctly lower response occurred in insects that had been kept under Sd conditions since hatching; their transfer to Ld conditions at the time of treatment had little effect on JH sensitivity. JH application to Sd females reared at 20 degrees C caused enlargement of the germaria but no eggs were formed.