In this study, we present the first characterization of herbivore-induced plant volatiles (HIPVs) released from infested olive leaves. The gall midge Dasineura oleae is a specific pest of Olea europaea and endemic of the Mediterranean Basin, an area in which severe outbreaks currently occurred. Little is known about the damage caused by the pest and the relationship with its host. Since gall formation and larval feeding activity may lead to the release of specific plant volatile compounds, we investigated the volatile profiles emitted from infested plants compared with healthy plants under both laboratory and field conditions. Additionally, the volatiles emitted from mechanically damaged plants were considered. A blend of 12 volatiles was emitted from olive trees infested by D. oleae. Of these, β-copaene, β-ocimene, cosmene, unknown 1 and unknown 3 were found to be exclusively emitted in infested plants. The emission of germacrene-D, (E,E)-α-farnesene, and (Z,E)-α-farnesene, α-copaene, (E)-4,8-dimethylnona-1,3,7-triene, (E)-β-guaiene and heptadecane significantly increased in infested trees. Linalool, β-copaen-4-α-ol, β-bourbonene, β-cubebene, β-elemene, β-copaene and δ-amorphene were found only in the field trial and showed differences depending on the level of infestation and the plant stage. (Z)-3-Hexenol, (E)-4-oxohen-2-enal, and 2-(2-butoxyethoxy)-ethanol, were exclusively emitted from the leaves after mechanical damage. In a field trial in Italy, we also demonstrated spring synchronization between adults of D. oleae and O. europaea trees. Analyses of morphoanatomical malformations of gall leaves showed that tissue alterations occur at the spongy parenchyma causing an increase of the leaf blade thickness. We speculate that tissue alterations may lead to HIPV release, in turn potentially attracting D. oleae natural enemies.

We tested the enemy hypothesis for gall morphology on a model system comprising two Diplolepis rose gall wasp species and their associated parasitoids. The enemy hypothesis predicts both that gall traits will influence parasitoid attack rates within species, and that galls with contrasting morphologies will support different parasitoid communities. This hypothesis is supported by studies at both intraspecific and broader taxonomic levels (i.e. between genera), but patterns remain to be explored in closely related species. Our aims were to explore the relationships between aspects of gall morphology (number of larval chambers, overall gall size and thickness of the gall wall) in each of Diplolepis mayri and D. rosae, and to explore correlations between these traits and both the presence/absence (=incidence) and attack rates imposed by parasitoids. We found in both galls that chamber number is positively correlated with gall size. In galls of D. mayri, parasitoid incidence was negatively correlated with thickness of the wall of the larval chamber, but there was no significant correlation between parasitoid attack rates and overall gall size. In D. rosae galls, parasitoid incidence was positively correlated with chamber wall thickness, but parasitoid attack rates were negatively correlated with gall size, suggesting that selection may favour the induction of galls containing more larval chambers. These results confirm that gall extended phenotypes can significantly influence enemy attack rates, consistent with the ‘enemy hypothesis’. Further, differences in gall morphology between the two Diplolepis species may underlie differences in their associated parasitoid communities – further research is required to test this hypothesis.