As remnant vegetation covers <15% of the Australian sheep-wheat belt, it is important to identify conservation strategies suitable for use in agricultural landscapes. Tree lines are widespread ecological structures in rural areas, and are now the subject of government subsidy schemes in New South Wales. However, the contribution of tree lines to biodiversity conservation is poorly understood. To identify the conservation value of tree lines, the bird communities in 36 tree lines in Cowra Shire, New South Wales, were surveyed 4 times each. The results demonstrated that tree lines were used by a large number of species, six of which were threatened. Different taxa were associated with different physical tree line attributes, with tree line age an important predictor of species occurrence. While mature tree lines tended to support more species, as was reflected in higher Shannon Diversity Index scores, they also harboured relatively more introduced species and nest predators, while young tree lines provided the best habitat for threatened species. However, these tree lines will mature, and as they do so they will provide increasingly suitable habitat for the hyperaggressive native honeyeater, the Noisy Miner (Manorina melanocephala), and introduced species. Therefore, tree lines in agricultural landscapes may only be serving an important role for conservation on short time scales, and the suite of threatened species young tree lines currently support appear likely to decline further in the future.