Physiological and biochemical adaptive traits support the specific breeding of alfalfa (Medicago sativa) for severely drought-stressed or moisture-favourable environments

The adaptation of alfalfa to moisture-favourable and drought-prone environments and its underlying mechanisms have not been thoroughly investigated despite its crucial importance to maximize the crop breeding progress. The main aim of this study was to support breeding strategies by exploring the relationship of leaf morphophysiological traits assessed in three growth chamber experiments with the adaptive response of five cultivars (Mamuntanas; Demnat 203; Erfoud 1; Prosementi; SARDI 10) that showed large genotype x environment interaction (GEI) across 10 drought-prone to moisture-favourable Mediterranean environments. Changes in shoot and root metabolite profiles across moisture-contrasting conditions were assessed for two cultivars with contrasting adaptation pattern. The cultivar specifically adapted to drought-prone environments (Mamuntanas), compared with that specifically adapted to moisture-favourable environments (Demnat 203), displayed: (i) lower leaf wilting under drought stress; (ii) delayed leaf senescence under stress, assessed as limited chlorophyll reduction either in vivo from upper to central leaves or in vitro from control to polyethylene glycol-treated leaflets, (iii) greater leaf tolerance to desiccation, measured in vitro as less pronounced reduction of leaf dry weight from control to polyethylene glycol-treated leaflets, (iv) smaller leaves, (v) lower early root and shoot growth and (vi) lower leaf stomata density. The other cultivars exhibited different combinations and/or intermediate values of these traits. The metabolite profiles of Mamuntanas and Demnat 203 differed more in shoots than in roots. Mamuntanas accumulated more amino acids than Demnat 203 under moisture-favourable and drought conditions. The distinct and partly incompatible combination of traits featuring the cultivars that maximized the crop yield in favourable or drought-prone conditions (Mamuntanas and Demnat 203) improves our understanding of traits contributing to large GEI across moisture-contrasting environments and support the breeding for specific adaptation to either condition. We identified a few inexpensive morphophysiological traits that could be used for early selection targeting drought-prone environments.