Insect egg deposition induces defence responses inPinus sylvestris: characterisation of the elicitor

Egg deposition by the phytophagous sawfly Diprion pini L. (Hymenoptera, Diprionidae) is known to induce locally and systemically the emission of volatiles in Scots pine (Pinus sylvestris L.) that attract the egg parasitoid Chrysonotomyia ruforum Krausse (Hymenoptera, Eulophidae). The egg parasitoids kill the eggs and thus prevent damage to the plant from feeding sawfly larvae. The elicitor inducing the pine's response is known to be located in the oviduct secretion which the female sawfly applies to the eggs when inserting them into a slit in the pine needle using the sclerotized ovipositor valves. In this study we have characterized the elicitor. The elicitor was still active when isolated from the oviduct and applied directly to slits made in the pine needles. However, as soon as the oviduct secretion was dissolved in Aqua dest. and stored for 3 h at room temperature or kept frozen at -80 degrees C, its activity was lost. In contrast, oviduct secretion kept its eliciting activity, when dissolved in Ringer solution (pH 7.2) both after storage at room temperature and after freezing. The activity of the elicitor vanished after treatment of the oviduct secretion with proteinase K, which destroyed all proteins. This suggests that the elicitor in the oviduct secretion is a peptide or protein, or a component bound to these. SDS-PAGE revealed a similar, but not identical protein pattern from hemolymph and oviduct secretion. Hemolymph itself has no eliciting effect. The elicitor in the oviduct secretion is only active when transferred to slit pine needles, since its application on undamaged needles did not induce the emission of attractive volatiles.