Photoprotective compounds as early markers to predict holm oak crown defoliation in declining Mediterranean savannahs.

Dehesas, human-shaped savannah-like ecosystems, where the overstorey is mainly dominated by the evergreen holm oak (Quercus ilex L. subsp. ballota (Desf.) Samp.), are classified as a global conservation priority. Despite being Q. ilex a species adapted to the harsh Mediterranean environmental conditions, recent decades have witnessed worrisome trends of climate-change-induced holm oak mortality. Holm oak decline is evidenced by tree vigour loss, gradual defoliation and ultimately, death. However, before losing leaves, trees undergo leaf-level physiological adjustments in response to stress that may represent a promising field to develop biochemical early markers of holm oak decline. This study explored holm oak photoprotective responses (pigments, tocopherols and photosynthetic performance) in 144 mature holm oak trees with different health statuses (i.e., crown defoliation percentages) from healthy to first-stage declining individuals. Our results indicate differential photochemical performance and photoprotective compounds concentration depending on the trees' health status. Declining trees showed higher energy dissipation yield, lower photochemical efficiency and enhanced photoprotective compounds. In the case of total violaxanthin cycle pigments (VAZ) and tocopherols, shifts in leaf contents were significant at very early stages of crown defoliation, even before visual symptoms of decline were evident, supporting the value of these biochemical compounds as early stress markers. Linear mixed-effects models results showed an acute response, both in the photosynthesis performance index and in the concentration of foliar tocopherols, during the onset of tree decline, whereas VAZ showed a more gradual response along the defoliation gradient of the crown. These results collectively demonstrate that once a certain threshold of leaf physiological damage is surpassed, that leaf cannot counteract oxidative stress and progressive loss of leaves occurs. Therefore, the use of both photosynthesis performance indexes and the leaf tocopherols concentration as early diagnostic tools might predict declining trends, facilitating the implementation of preventive measures to counteract crown defoliation.