Although often investigated within the context of plant growth and development and/or seed composition, plant lipids have roles in responses to environment. To dissect changes in lipid and fatty acid composition linked to drought tolerance responses in oats, we performed a detailed profiling of (>90) different lipids classes during a time course of water stress. We used two oat cultivars, Flega and Patones previously characterized as susceptible and tolerant to drought, respectively. Significant differences in lipid classes (mono, di and triacylglycerols; [respectively MAG, DAG, and TAG] and free fatty acids [FFA]) and in their fatty acid (FA) composition was observed between cultivars upon drought stress. In Flega there was an increase of saturated FAs, in particular 16:0 in the DAG and TAG fractions. This led to significant lower values of the double bond index and polyunsaturated/saturated ratio in Flega compared with Patones. By contrast, Patones was characterized by the early induction of signaling-related lipids and fatty acids, such as DAGs and linolenic acid. Since the latter is a precursor of jasmonates, we investigated further changes of this signaling molecule. Targeted measurements of jasmonic acid (JA) and Ile-JA indicated early increases in the concentrations of these molecules in Patones upon drought stress whereas no changes were observed in Flega. Altogether, these data suggest a role for jasmonates and specific fatty acids in different lipid classes in coping with drought stress in oat.