Plants under dual attack : consequences for plant chemistry and parasitoid behavior

Though immobile, plants are members of complex environments, and are under constant threat from a wide range of attackers, which includes organisms such as insect herbivores or plant pathogens. Plants have developed sophisticated defenses against these attackers, and include chemical responses such as the production of and emission of volatile compounds, which can be used by natural enemies of the herbivores to locate herbivore-infested plants. While the production and use of induced volatiles by foraging natural enemies has been well studied in a single attacker/natural enemy combination, in natural situations it is common for a plant to be challenged by multiple attackers. However there is little knowledge about what happens during multiple attack, especially when one of the secondary attackers is a plant pathogen. The aim of this thesis was to explore how dual attack modifies plant chemistry, and how changes in the emitted volatile blends then affect the behavior of foraging parasitoid, with a strong focus on the effects of non-host herbivore density and plant pathogen challenge. This study focused on the system consisting of black mustard plants (Brassica nigra), the large cabbage white butterfly, Pieris brassicae, and its larval parasitoid, Cotesia glomerata. Butterfly eggs, Brevicoryne brassicae aphids and the plant pathogen Xanthomonas campestris were used as secondary attackers. The results presented in the thesis showed that C. glomerata wasps are attracted to host-infested plants, irrespective of the presence and identity of the non-host attacker. However, when the responses of several parasitoid species to volatiles of plants infested with Pieris and/or aphids were compared, the wasp species were not equally affected and aphid infestation altered, in a density-dependent manner, the foraging behavior of all three species. In terms of volatiles, while differences in induced blends could be seen between individual attackers, these effects disappeared when plants were subjected to dual attack with caterpillars, and in the case of infestation with different aphid densities, non-linear volatile responses were revealed. Furthermore, the effects of dual attack by aphids and caterpillars were present at the level of leaf chemistry. Single and dual herbivory, as well as aphid density, led to metabolome-wide effects, driven mainly by changes in glucosinolates, sugars and antioxidant-related metabolites. The effect of pathogen challenge was further assessed by comparing the effects of virulent and avirulent pathovars, and results showed that both virulence and disease severity strongly affected the induced plant volatile blend. Remarkably, C.glomerata wasps were strongly attracted to volatiles of all the pathogen-challenged treatments, even in the absence of hosts. The data presented in this thesis contribute to our understanding of how dual attack affects the chemistry of B. nigra plants, and modifies plant interactions with the natural enemies of attacking herbivores. This work reveals that plant pathogen challenge can affect volatile-mediated tritrophic interactions, and shows that focusing only on general species-specific effects of dual attack is too simplistic of an approach. The outcomes of this thesis contribute to our understanding of how plants modulate their defense responses against multiple attackers.