Australia’s southeastern subalpine regions were severely impacted by the 2019-2020 bushfires. Many of the plant species most affected were poorly known and had no conservation management plan. Ex situ conservation approaches provide insurance against plant species’ extinction and germplasm for associated research. Through integrated horticultural and seed science research, the Australian National Botanic Gardens (ANBG) aimed to secure ex situ germplasm of 12 fire-affected subalpine species. Plant material was collected across 40+ locations in Namadgi National Park (ACT) and Kosciuszko National Park (NSW) between November 2021 and January 2024. At the National Seed Bank, experiments investigated seed viability and germination response to various seed pre-treatments including smoke chemicals, light/dark, gibberellic acid, scarification and warm/cold stratification. At the ANBG Nursery, a range of propagation treatments were trialled to determine the optimal treatment(s) for each species. These included different environments, hormones, watering regimes and propagation mixes. Despite extensive fire damage, cuttings for all primary target species were successfully sourced, and 11/12 species now have at least one seed collection in long-term storage. However, seeds of most species were difficult to obtain, low in quality and/or difficult to germinate, raising concerns about genetic representation and collection utility in the future. Here, we present case studies detailing how issues were tackled through propagation and cultivation techniques, tissue culture and rigorous seed biology and ecology research. This work demonstrates the valuable role that botanic gardens can play in preventing local extinctions of wild plant populations.