Early warning indicators for heat-induced mortality in temperate tree saplings

{middle dot} Globally, forest ecosystems face widespread mortality events. However, the independent impacts of distinct stressors, such as heat stress vs edaphic drought, remain poorly understood and physiological early warning indicators for tree mortality are urgently required. {middle dot} We exposed well-watered saplings of Fagus sylvatica, Pseudotsuga menziesii and Picea abies to summer heat waves and subsequent natural winter-desiccation. Physiological parameters (e.g. gas exchange, water uptake velocity via 2H labelling, and volatile organic compound emissions) were monitored throughout the growing season and survival was assessed regularly until subsequent spring to capture immediate and delayed mortality as a consequence of legacy effects. {middle dot} Heat exposure without soil water deficit, followed by winter desiccation, triggered species-specific mortality rates (51.8% F. sylvatica, 48.2% P. abies, 16.9% P. menziesii), with P. abies exhibiting significantly faster mortality response than the other species. Reduced water uptake, lower stomatal conductance, impaired photosynthetic efficiency, and altered VOC emissions distinguished non-surviving from surviving saplings months before visible damage in all three species. {middle dot} Heat stress drives mortality independent of edaphic drought, with sub-lethal physiological indicators detectable up to 10 months before visual signs. These early warning indicators could enable damage detection before lethal thresholds are crossed, offering new strategies for mitigating climate change-driven forest decline.