Marco Valerio Rossi Stacconi

The sensory structures of the antennal flagellum in Hyalesthes obsoletus (Hemiptera: Fulgoromorpha: Cixiidae): A functional reduction?

Despite their relevance as harmful pests on plants of economic importance, Hemiptera Fulgoromorpha have been poorly studied as regards their antennal sensory structures. In particular, the flagellum has been neglected and, therefore, to date there are no data on its structural organization and sensory equipment. In order to fill this gap, we carried out a study on the sensillum types and distribution on the flagellum of the planthopper Hyalesthes obsoletus Signoret, an efficient vector of the stolbur phytoplasma, the cause of various crop diseases. In this cixiid species the antenna is composed of three segments, the scape, an enlarged pedicel and a long flagellum. This latter is made of a single segment and presents a basal, bulb-like enlargement from which two processes arise, a short spur and a long arista. Combining scanning electron microscopy, transmission electron microscopy and focused ion beam investigations, we discovered the presence of a total number of 6 sensilla, belonging to 4 different types: a single scolopidium extending from the bulb to the arista, three sensilla styloconica within the cuticular spur and two different sensilla coeloconica inside the bulb. As far as structural data can suggest, these sensilla might be involved in the perception of mechanical stimuli (possibly air-borne vibrations), temperature and humidity variations and CO(2) concentration. The strong reduction in sensillum number in this species is discussed as possible functional specialization of the flagellum itself. The ultrastructure of the sensilla in the flagellum of a species of Fulgoromorpha is here presented for the first time.

Olfactory responses of Drosophila suzukii females to host plant volatiles

Drosophila suzukii Matsumura, an endemic pest in southeast Asia, has invaded Europe and the U.S.A. Unlike most of its closely related sibling species, the serrated ovipositor of D. suzukii permits ovipositing in undamaged fresh fruits. In the present study, volatiles are identified from host plants that are potentially involved in D. suzukii host recognition and oviposition behaviour. It is shown that mated females are attracted to volatiles emitted from intact fruits. The antennally‐active suite of compounds released from the fresh fruits is identified by gas chromatography coupled with electroantennographic detection, as well as gas chromatography‐mass spectrometry. In olfactometer bioassays, mated females are significantly attracted to an electroantennographically active volatile, isoamyl acetate, when tested at 10 µg of synthetic compound in a rubber septa, which has a release rate comparable to that of fresh fruits. In addition, a genomic survey shows that D. suzukii not only possesses the full repertoire of genes encoding odorant receptors activated by isoamyl acetate in D. melanogaster, but also that one of the genes, OR67a, is represented by five duplicated copies. These results indicate that D. suzukii uses olfactory cues to select oviposition sites. The identification of volatiles emitted by host fruits that attract D. suzukii may aid in the development of a selective and efficient synthetic lure for monitoring this pest. As a close relative of Drosophila melanogaster, D. suzukii provides a unique opportunity for understanding the physiological mechanisms involved in the shift of this species from use of rotten to ripe fruits for oviposition.

The process of pair formation mediated by substrate-borne vibrations in a small insect.

The ability to identify and locate conspecifics depends on reliable transfer of information between emitter and receiver. For a majority of plant-dwelling insects communicating with substrate-borne vibrations, localization of a potential partner may be a difficult task due to their small body size and complex transmission properties of plants. In the present study, we used the leafhopper Scaphoideus titanus as a model to investigate duetting and mate searching associated with pair formation. Studying these insects on a natural substrate, we showed that the spatio-temporal structure of a vibrational duet and the perceived intensity of partners signals influence the mating behaviour. Identification, localization and courtship stages were each characterized by a specific duet structure. In particular, the duet structure differed in synchronization between male and female pulses, which enables identification of the partner, while the switch between behavioural stages was associated with the male-perceived intensity of vibrational signals. This suggests that males obtain the information about their distance from the female and optimize their strategy accordingly. More broadly, our results show that even in insects smaller than 1cm, vibrational signals provide reliable information needed to find a mating partner.

Perception of Host Plant Volatiles in Hyalesthes obsoletus: Behavior, Morphology, and Electrophysiology

The Palearctic planthopper Hyalesthes obsoletus is the natural vector of the grapevine yellow disease Bois noir. Grapevine is an occasional host plant of this polyphagous planthopper. To deepen our knowledge of the role of plant volatile organic compounds for H. obsoletus host plant searching, we carried out behavioral, morphological, and electrophysiological studies. We tested the attraction of H. obsoletus to nettle, field bindweed, hedge bindweed, chaste tree, and grapevine by using a Y-shaped olfactometer. The results showed a significant attraction of male H. obsoletus to chaste tree, and of the females to nettle. Male H. obsoletus were repelled by odor from hedge bindweed. Ultrastructural studies of the antennae showed at least two types of olfactory sensilla at the antennal pedicel: plaque organs and trichoid sensilla. Volatile organic compounds from nettle and chaste tree were collected, and the extracts were analyzed by coupling gas-chromatography to both mass-spectrometry and electroantennography. The volatile organic compounds that elicited electrophysiological responses in male and female antennae were identified. These findings are discussed with respect to behavior of H. obsoletus males and females in the field.

Mapping and ultrastructure of antennal chemosensilla of the wheat bug Eurygaster maura

Antennae of the wheat stink bug Eurygaster maura L. (Hemiptera: Scutelleridae) were investigated to elucidate structure and distribution of antennal chemosensilla in females. Five type of sensilla were identified and characterized using a scanning electron microscope (SEM) and a transmission electron microscope (TEM). Type 1 sensilla are mechanical and contact chemoreceptors with a single apical pore. Types 2 and 3 sensilla are multiporous chemoreceptors both with typical features of olfactory sensilla. Type 4 are multiporous peg‐like sensilla, short and with a grooved surface. Type 5 are sensilla coeloconica with a smooth and aporous peg completely inserted in a sub‐cuticular chamber. All types are distributed on the two flagellar segments, but we considered only the apical flagellomere in which the largest number of sensilla are located. The most abundant sensilla are type 3, while the less numerous are type 5. All types, except type 2, decreased in number from the tip to the base of the segment. The lower density of sensilla was recorded on the dorsal‐internal part of the apical antennomere, while the higher density was recorded on the opposite side (external‐ventral).

Antennal sensory structures in Scaphoideus titanus Ball (Hemiptera: Cicadellidae).

Scaphoideus titanus Ball (Hemiptera: Cicadomorpha) is a leafhopper vector of a phytoplasma disease, the Flavescence dorée (FD), belonging to the vine yellows group. A scanning and transmission electron microscope study has been carried out to investigate the antennal sensory structures. The first two segments, the scape and the pedicel, are short, covered by cuticular scales and devoid of sensilla, with exception of some scattered hairs on the pedicel. The flagellum consists of a unique, elongated segment in which numerous subunits can be recognized, being separated by a sort of cuticular crown. The proximal five subunits bear most of the sensilla. We discovered the presence of single‐ and double‐walled coeloconic sensilla, campaniform sensilla, basiconic sensilla, and trichoid sensilla. A scolopidium is located within the proximal region of the flagellum. Ultrastructural investigations suggest that the antennal sensilla could be involved in the perception of air‐borne vibrations, temperature, and humidity variations. The most relevant feature is the extreme reduction of the olfactory sensilla, both in terms of number of sensory structures and sensory neurons per sensillum. The strong reduction in antennal olfactory sensilla to which this specie has undergone is discussed as possible consequence of the specificity toward the host plant. Microsc. Res. Tech. 2011.

Use of substrate-borne vibrational signals to attract the Brown Marmorated Stink Bug, Halyomorpha halys

Despite the increasing number of studies on the use of acoustic stimuli to control agricultural pests, this approach is still theoretical. Many insect pests, in particular hemipterans, use vibrational signals for mating communication, and therefore the application of a control strategy based on acoustic interference is a promising option. The Brown Marmorated Stink Bug, Halyomorpha halys, is causing severe economic damage to many crops in the USA and Italy. We tested a female vibrational signal, female signal 2 (FS2), to attract males in different settings, such as natural substrates, arenas and a cage representing an acoustic trap. We used video-tracking analysis and described the vibrational amplitude field around the individuals to study the male behavior. We found that FS2 can attract more than 50% of males to the source point and has a strong “loitering” effect on searching males that tend to remain in the stimulated area. We concluded that FS2 exhibits good attractiveness to H. halys males and that its potential use as a tool integrated into the currently existing pheromone traps should be tested in the field.

Comparative neuroanatomy of the antennal lobes of 2 homopteran species

We compared the morphology of the primary olfactory center, the antennal lobe (AL), in 2 homopteran insects, Hyalesthes obsoletus Signoret (Homoptera: Cixiidae) and Scaphoideus titanus Ball (Homoptera: Cicadomorpha). The comparison between the ALs of the 2 species is particularly interesting considering that, although both use volatile cues to locate their host plants, their feeding behavior differs considerably: specifically, H. obsoletus is a highly polyphagous species, whereas S. titanus is strictly monophagous (on grapevine). Our investigation of the AL structure using immunocytochemical staining and antennal backfills did not reveal any sexual dimorphism in either the size of the ALs or in the size of individual glomeruli for either species. Instead, the AL of H. obsoletus displayed numerous and well-delineated glomeruli (about 130 in both sexes) arranged in a multilayered structure, whereas the smaller AL of S. titanus contained fewer than 15 glomerular-like structures. This difference is likely to reflect the comparatively reduced olfactory abilities in S. titanus, probably as a consequence of the reduced number of volatiles coming from the single host plant. Instead, in H. obsoletus, the ability to distinguish among several host plants may require a more complex olfactory neuronal network.