With large wildfires becoming more frequent^1,2, we must rapidly learn how megafires impact biodiversity to prioritize mitigation and improve policy. A key challenge is to discover how interactions among fire-regime components, drought and land tenure shape wildfire impacts. The globally unprecedented^3,4 2019–2020 Australian megafires burnt more than 10 million hectares⁵, prompting major investment in biodiversity monitoring. Collated data include responses of more than 2,000 taxa, providing an unparalleled opportunity to quantify how megafires affect biodiversity. We reveal that the largest effects on plants and animals were in areas with frequent or recent past fires and within extensively burnt areas. Areas burnt at high severity, outside protected areas or under extreme drought also had larger effects. The effects included declines and increases after fire, with the largest responses in rainforests and by mammals. Our results implicate species interactions, dispersal and extent of in situ survival as mechanisms underlying fire responses. Building wildfire resilience into these ecosystems depends on reducing fire recurrence, including with rapid wildfire suppression in areas frequently burnt. Defending wet ecosystems, expanding protected areas and considering localized drought could also contribute. While these countermeasures can help mitigate the impacts of more frequent megafires, reversing anthropogenic climate change remains the urgent broad-scale solution.

Post-photosynthetic carbon isotope fractionation might alter the isotopic signal imprinted on organic matter (OM) during primary carbon fixation by Rubisco. To characterise the influence of post-photosynthetic processes, we investigated the effect of starch storage and remobilisation on the stable carbon isotope signature (d13C) of different carbon pools in the Eucalyptus delegatensis R. T. Baker leaf and the potential carbon isotope fractionation associated with phloem transport and respiration. Twig phloem exudate and leaf water-soluble OM showed diel variations in d13C of up to 2.5 and 2‰, respectively, with 13C enrichment during the night and depletion during the day. Damped diel variation was also evident in bulk lipids of the leaf and in the leaf wax fraction. d13C of nocturnal phloem exudate OM corresponded with the d13C of carbon released from starch. There was no change in d13C of phloem carbon along the trunk. CO2 emitted from trunks and roots was 13C enriched compared with the potential organic substrate, and depleted compared with soil-emitted CO2. The results are consistent with transitory starch accumulation and remobilisation governing the diel rhythm of d13C in phloem-transported OM and fragmentation fractionation occurring during respiration. When using d13C of OM or CO2 for assessing ecosystem processes or plant reactions towards environmental constraints, post-photosynthetic discrimination should be considered.