The world's most iconic forests are under threat from climate change. Climate-firevegetation feedback mechanisms are altering the usual successional trajectories of forests. Many obligate seeder forests across the globe are experiencing regeneration failures and subsequent alterations to their recovery trajectories. For example, the persistence of Eucalyptus regnans F. Muell. forests in southeast Australia is highly vulnerable to the effects of climate-driven increases in wildfire frequency. Shortening of the wildfire return interval from >100 years to < 20 years would inhibit or entirely stop regeneration of E. regnans, leading to replacement with understorey species such as Acacia dealbata Link. In this study, it is hypothesised that following such replacement, forest overstorey structure and transpiration will diverge. An experiment was designed to test this hypothesis by measuring and comparing overstorey transpiration and structural properties, including sapwood area and leaf area, between E. regnans and A. dealbata over a chronosequence (10-, 20-, 35-and 75-/80-year-old forests). We found that overstorey structure significantly diverged between the two forest types throughout the life cycle of A. dealbata after age 20.The study revealed strikingly different temporal patterns of water use, indicating a highly significant eco-hydrologic change as a result of this species replacement.Overall, the results provide a strong indication that after age 20, overstorey transpiration in Acacia-dominated forests is substantially lower than in the E. regnans forests they replace. This difference may lead to divergence in water yield from forested catchments where this species replacement is widespread.