Plants need to defend themselves against their natural enemies without compromising their interactions with beneficial organisms. Chemical mechanisms underpin many of these interactions and changes in plant metabolism are critical to both robust defences against antagonists and effective signals to mutualists. Further, such plant responses can be systemic, so mediating interactions between spatially separated organisms above and below ground. This thesis aimed to characterise the changes in the ragwort (Senecio jacobaea L.) metabolome caused by two different belowground organisms, an antagonistic herbivorous nematode (Pratylenchus penetrans (Cobb, 1917) Filipjev & Schuurmans Stekhoven, 1941) and a mutualist arbuscular mycorrhizal fungus (AMF) (Glomus intraradices Smith & Schenck). Initially, vegetative and reproductive stage ragwort plants were sampled in the field and the chemical composition of leaf and flower tissues was assessed using a metabolomic approach. Techniques for the identification of key ragwort secondary metabolites were trialled and results demonstrated that plants of different ages differed in their allocation of within plant defences such as flavanoids, pyrrolizidine alkaloids (PAs) and chlorogenic acids. Subsequent experiments with nematodes and AMF focussed on the analysis of leaf tissues from vegetative stage plants. Feeding by the nematode species P. penetrans resulted in increased concentrations of metabolites associated with plant defence, including the main class of ragwort defence compounds PAs. In contrast, colonisation of root material by AMF caused increases in the concentrations of metabolites associated with the maintenance of the beneficial interaction between plant and fungi, such as a number of apocarotenoids known as blumenols. The findings of both experimental studies detected unexpected and previously unreported changes in plant metabolism, highlighting the importance of an untargeted approach when examining the chemical ecology of plant interactions.