Electrophysiological responses of Philaenus spumarius and Neophilaenus campestris females to plant volatiles

The spittlebugs Philaenus spumarius and Neophilaenus campestris (Hemiptera: Aphrophoridae) are xylem-feeder insects that have been identified as vectors of Xylella fastidiosa in Europe. In the present study, we aim to identify volatile organic compounds (VOCs) that may act as semiochemicals for these species. Using the dynamic headspace technique, we collected VOCs from Olea europaea L. and Polygala myrtifolia L., highly susceptible plant species to X. fastidiosa, Pinus halepensis Mill., a common plant where N. campestris is found during summer, and from host plant species that are used as cover crops or exist as natural vegetation in olive orchards, such as Cistus creticus L., Medicago sativa L., Cynodon dactylon (L.) Pers., Lolium arundinaceum (Schreb.) Darbysh., Apium graveolens L. and Petroselinum crispum (Mill.) Fuss. We tested the response of female antennae on those blends with Gas Chromatography-Electroantennographic Detection (GC-EAD). The chemical profile of C. creticus and P. halepensis was rich in terpenes, alcohols, aldehydes, and esters. In the O. europaea profile, the main compounds were terpenes. As for P. myrtifolia, the volatile profile consisted mostly of alkanes and their substitutes. In the volatile profile of M. sativa, C. dactylon and L. arundinaceum, common volatile compounds were detected. Petroselinum crispum and Apium graveolens chemical profiles were dominated by terpenes. Several compounds elicited a consistent response to the female antennae of both species. In total, 65 compounds elicited consistent EAD responses for P. spumarius and 16 compounds for N. campestris. α-pinene was found in all tested plants and elicited consistent EAD responses of P. spumarius in five plants. In addition, antennae of P. spumarius females responded to camphor, limonene, 4-methyl octane and sabinene. These compounds were found in the volatile profile of at least 5 out of 8 examined plant species. Behavioral bioassays using Y-tube olfactometry were performed on volatiles that elicited antennal responses during electrophysiological studies. Among the compounds tested in behavioral studies, namely (-)-α-pinene, (+)-α-pinene, sabinene, (-)-S-limonene and (1R)-(+)-camphor, only the last one elicited a significant attraction response by P. spumarius females. The results achieved shed light on the VOCs from selected host plant species of X. fastidiosa that are perceived by two important insect vectors and a non-host plant, P. crispum. The identification of semiochemicals for manipulating spittlebugs' behavior contribute to the development of efficient monitoring tools for X. fastidiosa vectors.