AbstractAnt diversity in Australia is exceptionally high compared to other regions of the world, with semiarid regions being the most species‐rich. Ants perform numerous ecological functions and have been used in various biomonitoring programs. The role of ants in vineyards is unclear although other studies have focussed on controlling the invasive Argentine ant, Linepithema humile (Mayr), which significantly promotes mealybug and scale insect densities. While ants have been surveyed in a range of habitats throughout the continent, little is known about the diversity of ants in Australian vineyards. In this study, 147 native ant species were identified with pitfall trapping from 50 vineyards distributed in five regions in Victoria and South Australia. Species richness, compositional similarities and assemblage structure varied within and across regions although the ant fauna in semiarid vineyards was the most diverse. Linepithema humile was only detected in seven vineyards in one region. High ant diversity in vineyards could potentially contribute to ecosystem services, including biological control, and provide insurance to ecosystem functioning. Vineyards and other perennial agroecosystems could be important in maintaining invertebrate biodiversity particularly in a structurally diverse agricultural landscape.