Arboreta serve as effective tools for identifying alien insect pests and novel trophic associations. In this study, we used an arboretum in Slovenia to survey woody plants and identify both alien and native leaf miners. The leaves and twigs of 50 woody plant species and their cultivars were examined for characteristic damage. We used an integrative approach that combined identification based on leaf mines and DNA barcoding of the larvae and pupae found in the mines. In total, 62 leaf-mining species were identified, including eight alien species, of which the heliozelid Coptodisca lucifluella (Clemens, 1860) and the agromyzid Cerodontha unisetiorbita Zlobin, 1992 were documented for Slovenia for the first time. Additionally, three presumably native Gracillariidae moths Phyllocnistis labyrinthella (Bjerkander, 1790), P. ramulicola Langmaid & Corley, 2007 and P. saligna (Zeller, 1839) represented the first record for Slovenia. Furthermore, we documented 23 novel-to-science trophic associations, 20 of which involved native insects and alien woody plants, primarily from Asia. This study highlights the importance of arboreta and botanical gardens for the interception of invasive alien insects and the early detection of trophic shifts of native insects to alien plants, which can aid in predicting their potential spread.