Natural flow regimes are important for sustaining riverine vegetation. The regulation of river flows to provide water for agriculture often results in changes to flow timing. This study assesses the impact of altered seasonal flow patterns on riverine flora. Within temperate Australia, we surveyed the vegetation of five lowland rivers, three of which have large dams that alter their seasonal flow patterns; the other two are unregulated. From four to six sites were selected on each river, and these were classified into three levels of regulation based on the extent to which the timing of their seasonal flow patterns were altered. Sites were surveyed in winter and the following summer. Permanent quadrats were also established at a number of the surveyed sites and resurveyed every 3months. Of the 267 plant taxa identified, 145 were exotic (non-native). More exotic taxa and fewer native taxa were associated with increasing level of seasonal flow inversion (regulation). In particular, greater numbers of short-lived exotic terrestrial taxa and fewer native woody taxa were associated with increasing level of regulation. Some exotic woody species (e.g. willows) were more common in the unregulated rivers and may have life-history traits favoured by the natural seasonal flow patterns of study area. Multivariate analyses showed that level of regulation had a significant effect on the overall composition of the riverine vegetation. Our results provide support for the hypotheses that flow regulation adversely affects native species diversity and increases the vulnerability of riparian zones to invasion by exotic species; however, these effects are dependent on plant species' life-history strategies. Our study highlights the importance of natural seasonal flow patterns for sustaining native riverine plant communities. Flow management aimed at maintaining or restoring ecological values should consider seasonal flow patterns. Winter/spring flow peaks may be particularly important for the recruitment of native riverine plants, especially trees and shrubs, and reducing the extent of exotic annuals and grasses. © 2012 John Wiley Sons, Ltd.