In the literature, frost hardiness (FH) studies in trees have often been restricted to one organ (buds, leaves, needles or twigs). To extend our knowledge and gain a unified view, FH differences between organs and tissues or throughout the life of the tree have to be characterized in relation to physiological changes. In this study, different organs and tissues of young potted and mature orchard walnut trees (Juglans regia L.) were compared for seasonal changes in FH during different years. FH was assessed using the electrolyte leakage method. Physiological parameters were concomitantly monitored focusing on two significant traits: water content (WC) and carbohydrate content (glucose + fructose + sucrose, GFS). No seasonal variation in FH was observed in the root system, but acclimation and deacclimation were observed aboveground. Among organs and tissues, cold sensitivity levels were different in deep winter, with buds most sensitive and bark most resistant, but acclimation/deacclimation dynamics followed similar patterns. Physiological variation was also similar among organs: FH increased when WC decreased and/or soluble carbohydrates increased. Based on these results, relations between soluble carbohydrate content, WC and FH were calculated independently or in interaction. The key results were that: (i) the relationship between FH and physiological parameters (GFS and WC), which had previously been shown for branches only, could be generalized to all aboveground organs; (ii) lower WC increased the cryoprotective effect of GFS, showing a synergic effect of the two factors; (iii) the best fit was a non-linear function of WC and GFS, yielding a predictive model with an root mean square error of 5.07 °C on an independent dataset and 2.59 °C for the most sensitive stages; and (iv) the same parameters used for all organs yielded a unified model of FH depending on physiology, although the variability of GFS or WC was wide. The model should be of value for predicting FH in walnut independently of previous growing conditions (i.e., after sublethal stress accumulation).

Frost damages develop when exposure overtakes frost vulnerability. Frost risk assessment therefore needs dynamic simulation of frost hardiness using temperature and photoperiod in interaction with developmental stage. Two models, including or not the effect of photoperiod, were calibrated using five years of frost hardiness monitoring (2007-2012), in two locations (low and high elevation) for three walnut genotypes with contrasted phenology and maximum hardiness (Juglans regia cv Franquette, J. regia × nigra 'Early' and 'Late'). The photothermal model predicted more accurate values for all genotypes (efficiency = 0.879; Root Mean Standard Error Predicted (RMSEP) = 2.55 °C) than the thermal model (efficiency = 0.801; RMSEP = 3.24 °C). Predicted frost damages were strongly correlated to minimum temperature of the freezing events (ρ = -0.983) rather than actual frost hardiness (ρ = -0.515), or ratio of phenological stage completion (ρ = 0.336). Higher frost risks are consequently predicted during winter, at high elevation, whereas spring is only risky at low elevation in early genotypes exhibiting faster dehardening rate. However, early frost damages, although of lower value, may negatively affect fruit production the subsequent year (R = 0.381, P = 0.057). These results highlight the interacting pattern between frost exposure and vulnerability at different scales and the necessity of intra-organ studies to understand the time course of frost vulnerability in flower buds along the winter.

Predicting tree frost tolerance is critical to select adapted species according to both the current and predicted future climate. The relative change in water to carbohydrate ratio is a relevant trait to predict frost acclimation in branches from many tree species. The objective of this study is to demonstrate the interspecific genericity of this approach across nine tree species. In the studied angiosperm species, frost hardiness dynamics were best correlated to a decrease in water content at the early stage of acclimation (summer and early autumn). Subsequently, frost hardiness dynamics were more tightly correlated to soluble carbohydrate contents until spring growth resumption. Based on different model formalisms, we predicted frost hardiness at different clade levels (angiosperms, family, genus and species) with high to moderate accuracy (1.5–6.0°C RMSE) and robustness (2.8–6.1°C RMSEP). The TOT model, taking all soluble carbohydrate and polyols into account, was more effective and adapted for large scale studies aiming to explore frost hardiness across a wide range of species. The ISC model taking the individual contribution of each soluble carbohydrate molecule into account was more efficient at finer scale such as family or species. The ISC model performance also suggests that the role of solutes cannot be reduced to a 'bulk' osmotic effect as could be computed if all of them were located in a single, common, compartment. This study provides sets of parameters to predict frost hardiness in a wide range of species, and clues in targeting specific carbohydrate molecules to improve frost hardiness.