Jean Pouzat

Localization and fine structure of the female sex pheromone-producing glands in Bruchidius atrolineatus (PIC) (Coleoptera : Bruchidae)

Abstract The sex pheromone glands of female Bruchidius atrolineatus (Coleoptera: Bruchidae) have been localized by recording the electric response of the antenna of males subjected to a stream of air, containing the volatile sex pheromone (electroanntennography-EAG). Some 50 unicellular glands are distributed irregularly in the ventral and dorsal intersegmental membranes situated at the extremity of the pygidium, each gland containing a short ductule with an evacuation pore, 0.5–1 μm in diameter. The receiving canal is composed of a network of fine epicuticular filaments. The glands are type 3. The ultrastructure of these sex pheromone-producing glands is described in females whose production-emission activity had been previously verified with EAG. Deep basal invaginations and inflated intercellular spaces indicate the transport of substances from the hemolymph to the gland cells. The presence of numerous elongated mitochondria, diverse inclusions, vesicles containing crystalline bodies, and abundant apical microvilli, all reveal elevated cellular activity, which is never observed in young or diapausing females that do not produce sex pheromone. The ultrastructural differences in different types of females (sexually active or diapausing), combined with comparative EAG recordings obtained with intact females or those in which the suspected glandular zone was masked, made it possible to localize the glands.

Electroantennogram responses of Zabrotes subfasciatus to odours of the sexual partner

Odours of female Zabrotes subfasciatus Boh, elicit strong electroantennogram (EAG) responses in their sexual partners. Virgin female insects used in experiments were of different ages and given seeds, or not, of the host plant, Phaseolus vulgaris L. The odours of females which did not get host plant seeds, do not elicit EAG responses in virgin males, until they are at least 6 days old. On the other hand, the release of the sex pheromone as perceived by the male antennae, occurs earlier in the life of females when they are in contact with the host plants seeds. This suggests that the sexual attractiveness of the females of this specialist insect is probably closely correlated with vitellogenesis. The latter is indeed highly dependent on host plant stimulation.

Polypeptides of Acrolepiopsis assectella cocoon (Lepidoptera: Yponomeutoidea): an external host-acceptance kairomone for the parasitoid Diadromus pulchellus (Hymenoptera: Ichneumonidae).

Contact kairomones are essential for host-acceptance behaviour by female parasitoids. In the solitary endoparasitoid wasp, Diadromus pulchellus, this behaviour depends mainly on compound(s) in the cocoon of their host, Acrolepiopsis assectella pupae. Extracts of empty cocoons and polypeptides extracted from cocoons were tested in acceptance behaviour assays using cotton fibre lures bearing extracts. Extractions with solvents of increasing polarity indicated that the active compounds were polar, while SDS-PAGE showed that four glycopolypeptides contained enough information to trigger host-acceptance behaviour in female wasps. This kairomonal activity was found to be due to the protein moieties, and was independent of any glycosylation. These four glycopolypeptides might be two variants of two soluble sericin-like polypeptides differing in their degree of glycosylation.

Pheromone release by the two forms of Callosobruchus maculatus females: effects of age, temperature and host plant

Abstract. The influence of age, temperature and host plant on pheromone release was investigated in females of two forms of Callosobruchus maculatus (Coleoptera: Bruchidae), referred to as the flightless form and the flight form. The emission of pheromone seemed to be correlated with reproductive status: the flightless‐form females which were sexually mature at emergence began to release pheromone on the first day following emergence. The flight‐form females which exhibited no reproductive activity at emergence also showed a delayed emission of pheromone. The temperature conditions and the availability of host plant seeds had no apparent effect on pheromone release by the flightless form. In flight‐form females pheromone release was stimulated by the presence of host plant seeds. Temperature seemed to affect the age at which females initiated pheromone release for the first time after emergence, pheromonal emission being brought forward slightly by higher temperature. All these results are discussed relative to the specific environment of each form.

Location and ultrastructure of sex pheromone glands in female Callosobruchus maculatus (Fabricius) (Coleoptera : Bruchidae)

Abstract The ultrastructure of the female sex pheromone glands in Callosobruchus maculatus (Coleoptera : Bruchidae) were localized using a masking technique, combined with eiectro-antennography and by a comparison of the glandular cells of sexually active (flightless) females and non-sexually active (flight-form) females. Each unicellular gland is an invagination of the integumental membrane capped by a single secretory cell. These glands are situated on the fine intersegmental membrane, which joins the pygidium to the ovipositor. The secretory cells of the glands of active females are characterized by well-developed microvilli, with many elongated mitochondria among the latter. The high metabolic activity of these cells is revealed by the presence of heterogeneous secretion vesicles, some of which contain abundant crystallized material. Deep basal invaginations indicate the uptake of substances from the haemolymph. The receptor canal is a network of fine cuticular filaments which have the same structure regardless of the females sexual status. Cells from the glands of non-sexually active females are underdeveloped and show no invaginations of the basal membrane and very few microvilli. The localization of these glands was made possible by the use of SEM, TEM and EAG as well as by masking the suspected zones and by comparing females in different physiological states: flightless females, which were sexually active and producing pheromones; and flight-form females, non-sexually active and producing no sex pheromones. Only by adopting such a stringent method was it possible to confirm the function of the glands whose ultrastructure was studied.

Behavioural and olfactory variations in the leek moth, Acrolepiopsis assectella, after several generations of rearing under diverse conditions

Three strains of the leek moth, Acrolepiopsis assectella, were compared: two reared on leek leaves (Allium porrum) for which generations 10 to 12 (L) or generation 2 (G2) were used, the third one (D) reared on an artificial diet incorporating leek powder, for which generation ca 140 was examined. The walking behaviour (L and D) and the antennal responsiveness (L, G2 and D) were compared between individuals subjected to leek leaf odour and 4 volatiles identified in these leaves: 2 specific sulphur ones (dipropyl thiosulphinate and dipropyl disulphide) and 2 ‘general green leaf volatiles’ (cis‐3‐hexen‐1‐ol and trans‐2‐hexen‐1‐ol). The walking behavioural thresholds to leek odour, dipropyl thiosulphinate, and cis‐3‐hexen‐1‐ol were lower for L than for D females. The chemoanemotactic responses to dipropyl disulphide are almost identical, and trans‐2‐hexen‐1‐ol has nearly no activity. Accordingly, the olfactory sensitivity (assessed by EAG) to sulphur volatiles and cis‐3‐hexen‐1‐ol was higher for L and G2 females than for D ones. The threshold for thiosulphinate was at a concentration (v/v) of 4 ×10−4 M/l for D females, 4 × 10−5 M/l for L females and 4 × 10−8 M/l for G2 ones. It may be hypothesised that laboratory rearing conditions cause variations in the thresholds of antennal sensitivity and walking behaviour to specific and more efficient leek volatiles. This may result from the selective pressures experienced by the larvae reared in the laboratory.