Perching mate-locating strategy in Paysandisia archon (Lepidoptera: Castniidae): behavioral and morpho-physiological investigations.

We studied Paysandisia archon (Burmeister) (Lepidoptera: Castniidae: Castniinae) courtship behavior to provide a detailed qualitative and quantitative description of male and female behaviors. Moreover, to investigate the role of antennal olfaction and visual stimuli in mate-recognition, bioassays with antennectomized adults and dummies were performed. To assess the presence of a pheromone gland in the ovipositor, morphological (using light and scanning electron microscopic techniques), electrophysiological, and chemical investigations were carried out. We observed perching mate-locating behavior of P. archon males, with the female triggering the courtship sequence by approaching the perching male first. The stereotyped courtship sequence is made up of five main steps: female flight, pair flight, alighting close, copulation attempt, and clasping. Our findings suggest that visual cues are important in P. archon courtship behavior, and the role of chemical cues is also discussed. Moreover, we observed a higher antenna cleaning frequency in females than in males. Ovipositor extrusions during courtship appeared not to be related to calling behavior, and histological investigations showed no evidence of glandular tissues involved in sex pheromone production. The P. archon ovipositor consists of the eighth uromere, which forms the ovipositor base, and the 9th and 10th uromeres are fused together and connected to the base by an intersegmental membrane. Overall, 24 compounds were identified from extracts obtained from ovipositors: 9 compounds were detected only in extracts from the ovipositors of 24-h-old virgin females, and not from those of 1-h-old females. None of these compounds elicited any significant electrophysiological responses from male antennae.

Natural phytoplasma infection of four phloem-feeding Auchenorrhyncha across vineyard agroecosystems in central-eastern Italy.

The seasonal variations of grapevine yellow phytoplasma were investigated in four phloem-feeding planthopper and leafhopper species that are vectors of plant disease agents. In total, 1,148 wild specimens were collected from three vineyard agroecosystems in the Marche region (central-eastern Italy), from May to September 2008, and analyzed using polymerase chain reaction-restriction fragment-length polymorphism methods. Of 525 Euscelis lineolatus Brullé, 25.1% were positive for aster yellow phytoplasma (16SrI-C, 16SrI-B subgroups) and stolbur phytoplasma (16SrXII-A subgroup; Vergilbungskrankheit type I [VK-I]). Of 368 Hyalesthes obsoletus Signoret, 19.3% were positive for the 16SrXII-A subgroup (VK-I, VK-II; mainly according to their host plant). Of 146 Neoaliturus fenestratus (Herrich-Schäffer), 15.1% were positive for the 16SrI-C and 16SrI-B subgroups, and 7.3% of 109 Psammotettix alienus (Dahlbom) were positive for the 16SrI-B subgroup. The total inoculation efficiency in the feeding medium assays was 57.1% for P. alienus, 44.7% for E. lineolatus, 44.4% for N. fenestratus and 33.9% for H. obsoletus. All of the phytoplasma subgroups identified in the insect bodies were also detected in their feeding media. Detection of stolbur phytoplasma in E. lineolatus feeding media strengthens the hypothesis that it is a candidate vector of Bois noir disease causal agent. The phytoplasma subgroups detected in the Auchenorrhyncha species showed variations according to season and/or vineyard agroecosystem. This study highlights the different specificities of these phytoplasma-Auchenorrhyncha species relationships, and suggests a primary role of the entire vineyard agroecosystem in the epidemiology of grapevine yellow phytoplasma diseases.

Sex-dispersal differences of four phloem-feeding vectors and their relationship to wild-plant abundance in vineyard agroecosystems.

Sex-dispersal differences of four phloem-feeding vectors and their relationship to wild-plant abundance were analyzed over 2 yr in two vineyard agroecosystems. Adults of Euscelis lineolatus Brullè, Neoaliturus fenestratus (Herrich-Schäffer), Psammotettix alienus (Dahlbom), and Hyalesthes obsoletus Signoret were sampled by using yellow sticky traps at two heights (20 and 150 cm) above the ground in a regular geo-referenced grid. Wild vegetation was surveyed around each sampling point. The most abundant species collected was N. fenestratus, followed by P. alienus, H. obsoletus, and E. lineolatus. Low traps were more effective in capturing the females of N. fenestratus and P. alienus and the males of all of the species, except N. fenestratus at one site. Sex ratio was male-biased for N. fenestratus in both agroecosystems at both trap heights, and only in low traps for H. obsoletus and P. alienus at one site. Insect abundance, temporal population dynamics, and dispersal patterns showed considerable fluctuations according to vineyard agroecosystems and years. In the current study, all of the four phloem-feeding vectors were not found homogeneously in the vineyards, and they were aggregated in correspondence with their herbaceous wild-plant hosts. Insects tended to disperse from the borders toward the vineyard, or vice versa Colonizing pattern was strongly affected by border vegetation, fallow fields, and vineyard ground-cover vegetation. Different plant communities were identified inside the agroecosystems, and significant correlations among insect and plant-species abundances were recorded. Data highlight the importance of understanding the dispersal patterns of vector species for the application of correct integrated pest-management strategies.

Development and morphology of teratocytes in Encarsia berlesei and Encarsia citrina: first record for Chalcidoidea.

In several species of hymenopteran parasitoids of the superfamilies of Ichneumonoidea and Platygastroidea, the membrane enveloping the parasitoid embryo dissociates at hatching into a number of cells, called teratocytes, which autonomously develop in the host haemolymph. In this work we report for Encarsia berlesei and Encarsia citrina (Hymenoptera: Chalcidoidea), the dissociation of the extraembryonic membrane into cells whose morphological and embryological features correspond to those of teratocytes. In E. berlesei the membrane dissociated at hatching into 4-9 larger cells (100 microm diameter) and about 10 smaller cells (60 microm), which scarcely doubled their size during maturation. In E. citrina the membrane dissociated into five large cells (250 microm) which did not grow appreciably. Ultrastructural investigation of the dissociated cells in E. berlesei revealed that their surface was covered by microvilli, whose density and length increased from the egg stage to the 12 h following hatching. During the same period, rough endoplasmic reticulum evolved from a parallel profile to that of the cisternal type, while abundant vesicles represented the dominant cytological feature. The ploidy level of these cells ranged between 8c and 140c at hatching, but increased to 40c-350c at maturation. These findings provide the first clear evidences for the presence of teratocytes in the superfamily Chalcidoidea.

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.

Antennal multiporous sensilla: their gustatory features for host recognition in female parasitic wasps (insecta, Hymenoptera: Platygastroidea).

In parasitic wasps, various kinds of antennal plates have been interpreted as olfactory organs due to the presence of numerous pores. However, on the basis of ultrastructural investigations, some of these multiporous plates were revealed as being release sites of exocrine glands while others were postulated to have a gustatory function. Such sensilla, present only on female antennae, show unique features, found exclusively in Platygastroidea, with some differences between Scelionidae and Platygastridae. The cuticular apparatus consists of a short basiconic shaft with an apical multiporous area, the pores of which are covered by movable structures. The cellular components are made up of a remarkable number of sensory neurons, from 200 to 220, and an undefined number of sheath cells. These multiporous sensilla have tubular accessory glands that release their secretion through the socket sensillum only in scelionids. These morphological characters, combined with behavioral observations, strongly suggest a gustatory function, although electrophysiological studies are still needed to confirm this. A new terminology for antennal multiporous plates is discussed in relation to their different functions.

Host castration by Aphidius ervi venom proteins.

The braconid Aphidius ervi Haliday (Hymenoptera, Braconidae) is an endophagous parasitoid of the pea aphid, Acyrthosiphon pisum (Harris) (Homoptera, Aphididae). Parasitized host aphids show different degrees of castration, a response that is total when parasitoid oviposition takes place in first instar hosts. Deleterious effects on the host reproductive system are already evident by 24h following parasitization, before egg hatching. The effect of parasitoid venom on A. pisum ovaries has been studied by performing microinjections in non-parasitized host aphids and observing the cellular alterations of the apical germaria of ovarioles. Venom injection reproduced the same alterations observed in parasitized aphids, while injections of saline solution did not induce any detectable change. By 24h, the germarial cells of both venom-treated aphids and parasitized aphids showed the absence of the nucleolus and of electron-dense material around the nucleus, frequently referred to as "nuage material". By 48h more evident signs of degeneration were observed, suggesting the possible occurrence of apoptosis. The bioactive component of the venom was both heat- and protease-sensitive. The activity was found in purified fractions that were highly enriched in two proteins with an approximate molecular mass of 21kD and 36kD, respectively. These macromolecules are the most abundant components of A. ervi venom and, unlike many venom proteins of studied parasitic Hymenoptera, are not glycosylated and appear to be subunits of an oligomeric protein. The adaptive significance of host castration is discussed.

Ultrastructure and physiology of the CO2 sensitive sensillum ampullaceum in the leaf-cutting ant Atta sexdens.

The sensilla ampullacea on the apical antennomere of the leaf-cutting ant Atta sexdens were investigated regarding both their responses to CO2 and their ultrastructure. By staining the sensillum during recording, we confirmed that the sensilla ampullacea are responsible for CO2 perception. We showed that the sensory neurons of the sensilla ampullacea are continuously active without adaptation during stimulation with CO2 (test duration: 1 h). This feature should enable ants to assess the absolute CO2 concentration inside their nests. Sensilla ampullacea have been found grouped mainly on the dorso-lateral side of the distal antennal segment. Scanning and transmission electron microscopic investigations revealed that the external pore opens into a chamber which connects to the ampulla via a cuticular duct. We propose protection against evaporation as a possible function of the duct. The ampulla houses a peg which is almost as long as the ampulla and shows cuticular ridges on the external wall. The ridges are separated by furrows with cuticular pores. The peg is innervated by only one sensory neuron with a large soma. Its outer dendritic segment is enveloped by a dendritic sheath up to the middle of the peg. From the middle to the tip numerous dendritic branches (up to 100) completely fill the distal half of the peg. This is the first report of a receptor cell with highly branched dendrites and which probably is tuned to CO2 exclusively.

Identification of the gland secreting oviposition-deterring pheromone in the cabbage seed weevil, Ceutorhynchus assimilis, and the mechanism of pheromone deposition.

After laying an egg into a pod of Brassica napus, the female cabbage seed weevil, Ceutorhynchus assimilis, brushes the caudal setae of the eighth abdominal tergite (VIII UT) on the host pod as she walks along it, depositing oviposition-deterring pheromone (ODP). The VIII UT is periodically extended and withdrawn, thus repeatedly rubbing against the posterior fold of the seventh urotergite (VII UT) which bears the individual outlets of glandular epidermal cells. In post-diapause, sexually mature, gravid (i.e. oviferous) females (virgin or mated) the cells of this VII UT gland were hypertrophic, showing intense secretory activity. Extracts of VII UT from these individuals elicited strong electrophysiological responses from antennal club gustatory sensilla and deterred oviposition. In pre-diapause (sexually immature) females, the cells of the VII UT gland were neither hypertrophic nor active and an extract of their VII UT elicited no significant electrophysiological or behavioural response. Extract of female rectum was a less potent oviposition deterrent than VII UT extract and elicited an electrophysiological response similar to male rectum extract. An extract of ovarian calyces and ovaries elicited no behavioural response. We conclude that ODP is secreted by the epidermal cells of the VII UT posterior fold.