Plant volatile organic compounds play a role in selection of host plants by herbivorous insects. Once the insect reaches the plant, contact cues determine host acceptance. Although the willow sawfly Nematus oligospilus (Hymenoptera: Tenthredinidae) can differentiate among willow genotypes, no knowledge is available on the cues used by this insect to seek and accept the host plant. In this study, we recorded behavioral orientation in a Y-tube olfactometer of willow sawfly females to volatiles of the highly preferred genotype Salix nigra and the non-preferred genotype S. viminalis. The volatiles released by undamaged willows of each genotype were analyzed by coupled gas chromatography-mass spectrometry. Contact cues were evaluated first by oviposition preference bioassays after selective leaf wax removal, and then by studying the micromorphology of abaxial and adaxial leaf surfaces and their chemical composition. Willow sawfly females oriented preferentially to S. nigra volatiles, which contained more than 3 times the amount of volatiles than that collected from S. viminalis. Analysis of volatiles showed significant differences in amounts of (Z) and (E)-β-ocimene, undecane, decanal, and β-caryophyllene. The adaxial leaf surface of S. nigra was less preferred after wax removal, suggesting a role of cuticular waxes for oviposition acceptance. No differences were found among the micromorphology of leaf surfaces between preferred and non-preferred genotypes. The chemical analysis of cuticular waxes showed that the abaxial leaf surface of S. viminalis, which is completely avoided for oviposition, possessed 97% of alkanes. The accepted leaf surfaces contained a more diverse wax profile including alcohols, acids, and esters. Thus, non-alkane wax compounds might be related to oviposition. In sum, our study suggests that several cues act in concert to provide oviposition cues for the sawfly N. oligospilus: females are attracted to volatiles from a distance, and once alighting on the plant, they seek specific chemical contact cues in order to lay eggs.

SUMMARYFlooding tolerance should be included as a trait in breeding programs in forested areas where extreme flooding events lasting several months can occur. In this context, it is difficult to carry out controlled, long-term flooding experiments with big trees due to the large number of specimens to screen. The hypothesis in this work was that a relatively short-term flooding experiment in a greenhouse would be useful to select clones capable of enduring long term flooding events in the field. The survival of 4 willow clones in a long term, 12 years field trial that was subjected twice to long term flooding events, was compared with the survival, growth and gas exchange of those same clones subjected to flooding in the greenhouse for three months. There were differences in flooding survival in the field: clone AN4 showed a 100 % survival, followed by clone 395 (43 %), clone 131-27 (18 %) and clone 13-44 (14 %). In the greenhouse, all plants survived and only flooded plants of clone 13-44 experienced a statistically significant growth and stomatal conductance reduction. The flood tolerance ranking in the field and in the greenhouse only partially overlapped; the worst clone in the field could be identified in the short-term greenhouse experiment, though the best one could not. It was not possible to identify clones with tolerance to long-term flooding episodes in the field. Nevertheless, the greenhouse results were useful to identify clones showing flooding sensitivity and tolerance to short term flooding episodes.Key words: Salix spp., survival, growth, Paraná River Delta. RESUMENEs necesario incluir la tolerancia a la inundación como un carácter en programas de mejoramiento para zonas forestadas donde eventos extremos de inundación que pueden durar meses son comunes. En este contexto, es difícil llevar a cabo experimentos controlados de larga duración con árboles de gran tamaño, debido al elevado número de ejemplares que es necesario evaluar. La hipótesis de este trabajo fue que un experimento relativamente corto de inundación en el invernáculo sería útil para seleccionar clones capaces de soportar eventos de inundación de varios meses en el campo. Se comparó la supervivencia de cuatro clones de sauces en un ensayo a campo que experimentó dos episodios largos de inundación a lo largo de 12 años, y la supervivencia, crecimiento e intercambio gaseoso de los mismos clones inundados por tres meses en invernáculo. Se encontraron diferencias en la supervivencia en el ensayo a campo, del clon AN4 sobrevivieron el 100 % de las plantas, seguido por el clon 395 (43%), clon 131-27 (18%) y clon 13-44 (14%). En el invernáculo, todas las plantas sobrevivieron y sólo las plantas inundadas de 13-44 experimentaron una reducción significativa del crecimiento y la conductancia estomática. El ranking de tolerancia en el campo y en invernáculo sólo coincidió parcialmente, el clon menos tolerante en el campo pudo ser identificado en el invernáculo, pero no el clon más tolerante. Si bien no fue posible identificar clones con tolera...

After an insect herbivore has reached its host plant, contact cues from the leaf surface often determine host acceptance. We studied contact cues during oviposition behavior of a willow pest, the sawfly Nematus oligospilus (Hymenoptera: Tenthredinidae), a specialist feeder on Salix (Salicaceae) trees, and how it determines oviposition preference in lab and field conditions. We described the sequence of behaviors that lead to egg laying on the most and least preferred willow species. Then we studied the morphology of chemosensory structures present on the female antenna, cerci and ovipositor. Since phenolic glycosides (PGs) are the main secondary metabolites present in Salicaceae species, we investigated their role in host acceptance. We quantified these compounds in different willow species and correlated PG content with oviposition preference under lab and natural field conditions. We demonstrated a major role for contact cues in triggering N. oligospilus egg laying on the leaf surface of preferred willow genotypes. Firstly cues are sensed by antennae, determining to leave or stay on the leaf. After that, sensing is performed by abdominal cerci, which finally triggers egg laying. The lack of PGs in non-preferred species and the significant correlation observed between PGs, natural damage and oviposition preference suggest a role for these compounds in host selection. Our study suggests that in specialist feeders, secondary compounds normally acting as defenses can actually act as a susceptibility factor by triggering specific insect behavior for oviposition. These defensive compounds could be selected against to increase resistance.

Your article is protected by copyright and all rights are held exclusively by Springer Science+Business Media B.V., part of Springer Nature. This e-offprint is for personal use only and shall not be self-archived in electronic repositories. If you wish to self-archive your article, please use the accepted manuscript version for posting on your own website. You may further deposit the accepted manuscript version in any repository, provided it is only made publicly available 12 months after official publication or later and provided acknowledgement is given to the original source of publication and a link is inserted to the published article on Springer's website. The link must be accompanied by the following text: "The final publication is available at link.springer.com".

No abstract