The mustard leaf beetle, Phaedon cochleariae (F.) (Coleoptera: Chrysomelidae), is specialized to feed and develop on various species within the Brassicaceae. In this study, we investigated the acceptance of several host plant species (Brassica rapa L. and Sinapis alba L.), commonly used by the beetle (familiar plants), and of various unfamiliar plants, including systematically and chemically related [Bunias orientalis L. (Brassicaceae) and Tropaeolum majus L. (Tropaeolaceae), both Brassicales], as well as unrelated non-host plant species [Plantago lanceolata L. (Plantaginaceae); Lamiales]. Emphasis was laid on the acceptance of the neophyte B. orientalis, and on underlying cues responsible for the acceptance of the various species. Behavioural responses to plant volatiles were studied using a static four-chamber olfactometer. Stimulants and deterrents were investigated by bioassay-guided solid phase extraction and semi-preparative high performance liquid chromatography. A difference in acceptance of plant species was found: odours and polar compounds of all Brassicales evoked attraction and feeding stimulation, respectively, in Ph. cochleariae. Glucosinolates and their volatile hydrolysis products could be the main compounds that are involved in attraction of the beetles. In contrast, Ph. cochleariae did not respond to odours of the non-host P. lanceolata, and some fractions of this plant had feeding-deterrent effects, due to the presence of iridoid glycosides, among others. Although adult females accepted the neophyte B. orientalis for oviposition, neonate larvae did not survive on it. The flavonoid-containing fraction of this plant was deterrent, whereas a similar fraction had been shown to cause some feeding stimulation when derived from S. alba. Differences in qualitative and quantitative composition of related metabolites lead to differentiated plant acceptance, proving the complexity of plant cues and of insect responses that determine host acceptance behaviour. The possibility of a diet breadth enlargement to B. orientalis and the role of Ph. cochleariae as a putative native biocontrol agent of this invasive plant are discussed.