Strawberry tree (Arbutus unedo) is a small resilient species with a circum-Mediterranean distribution, high ecological relevance in southern European forests and with several economical applications. As most orchards are usually installed on marginal lands where plants usually face severe drought, selecting plants that can better cope with water restriction is critical, and a better understanding of the tolerance mechanisms is required. Strawberry tree plants under drought follow a typical isohydric strategy, by limiting transpiration through stomata closure. However, the contribution of genotype and its bio-geographic origin on plant performance needs clarification, as well as the involvement of a specific metabolic reactions associated with the mechanical response. To test this hypothesis, several eco-physiological and biochemical parameters were assessed on different genotypes, and the metabolic profiles studied, including important stress-related phytohormones, on plants under different water regimes (plants watered to 70% and 18% field capacity) and a recovery assay. A contrasting drought tolerance was found in plants from different genotypes, associated with physiological and metabolic responses. Metabolomics revealed more than 500 metabolic features were differentially accumulated, including abscisic and salicylic acids, for the genotype with better performance under drought (A4). This genotype also recovered faster when the imposed stress was interrupted, thus indicating the relevance of metabolic adaptation under water deficit conditions. By correlating carbon assimilation with identified metabolites, some proved to be satisfactory predictors of plant performance under drought and might be used for marker assisted breeding. Therefore, our study proves the importance of genotype as a major selection criterion of resistant plants to drought and provides empirical knowledge of the metabolic response involved. We also hypothesized the involvement of phenolics on response mechanisms under drought, which is worth to be explored to shed light on the metabolic pathways involved in plant response to water stress.

Arbutus unedo L. is a small Ericaceae tree with a circum-Mediterranean distribution. It has a huge ecological impact on southern Europe forests and a great economic importance as a source of phytochemicals with bioactive properties and for fruit production. On the foreseen climate change context, breeding toward drought tolerance is necessary in order to ameliorate plant performance. Therefore, the aim of this work was to study the reproduction mechanisms of the strawberry tree, obtain new genetic combinations by hybridization, and select genotypes more tolerant to drought stress. A morphological analysis of flowers and pollen was carried out, and controlled pollinations were performed both in vitro and ex vitro. The very first approach on strawberry tree breeding by means of hybridization is also presented. Several physiological parameters were evaluated on 26 genotypes submitted to a water-deficit regime. Plant behavior under drought greatly varied among genotypes, which showed high phenotype plasticity. Three genotypes that were able to cope with water restriction without compromising net CO₂ assimilation were identified as highly tolerant to drought stress. The results obtained elucidate the reproduction mechanisms of the strawberry tree and open the way for a long-term breeding program based on the selection of drought-tolerant plants.

This study investigated the effect of drought stress on the expression of chloroplast genes in two different genotypes (A1 and A4) of strawberry tree plants with contrasting performances. Two-year-old plants were subjected to drought (20 days at 18% field capacity), and the photosynthetic activity, chlorophyll content, and expression levels of 16 chloroplast genes involved in photosynthesis and metabolism-related enzymes were analyzed. Genotype-specific responses were prominent, with A1 displaying wilting and leaf curling, contrasting with the mild symptoms observed in A4. Quantification of damage using the net CO₂ assimilation rates and chlorophyll content unveiled a significant reduction in A1, while A4 maintained stability. Gene expression analysis revealed substantial downregulation of A1 (15 out of 16 genes) and upregulation of A4 (14 out of 16 genes). Notably, psbC was downregulated in A1, while it was prominently upregulated in A4. Principal Component Analysis (PCA) highlighted genotype-specific clusters, emphasizing distinct responses under stress, whereas a correlation analysis elucidated intricate relationships between gene expression, net CO₂ assimilation, and chlorophyll content. Particularly, a positive correlation with psaB, whereas a negative correlation with psbC was found in genotype A1. Regression analysis identified potential predictors for net CO₂ assimilation, in particular psaB. These findings contribute valuable insights for future strategies targeting crop enhancement and stress resilience, highlighting the central role of chloroplasts in orchestrating plant responses to environmental stressors, and may contribute to the development of drought-tolerant plant varieties, which are essential for sustaining agriculture in regions affected by water scarcity.