Tree-ring Δ13C reveals the impact of past forest management on water-use efficiency in a Mediterranean oak coppice in Tuscany (Italy)

? The effects of thinning and heavy stand density reduction was investigated in Turkey oak (Quercus cerris L.) forests of central Italy, to evaluate the physiological responses and the growth status of trees that survived a past coppice cut and thinning to convert the stand to high-forest.

? The working hypothesis was that a strong decrease in stand density would cause a decreasing in canopy-intrinsic water-use efficiency (measured as the ratio of CO₂ assimilation to stomatal conductance, A/g), thus an increase in tree-ring carbon isotopic discrimination (Δ¹³C).

? The tree-ring Δ¹³C of the remaining trees (“survivors”) was found to have significantly (P < 0.05) raised between year two and year seven since the coppice stand was thinned (high-forest conversion thinnings). This effect was mostly caused by a large decrease in tree-rings Δ¹³C at control site which was characterized by high density and competition by trees. An increase in survivors tree-rings Δ¹³C probably indicates an improved water availability, possibly induced by a decrease in competition and in stand density or a decrease in the precipitation intercepted by the canopy (i.e., a stronger increase in g over A since a decrease in A is highly unlikely). A change in foliar nitrogen, foliar Δ¹³C and content in chlorophylls was also recorded seven years after thinning.

? Thinnings carried out to convert old abandoned coppices into high-forest stands induce short-term stimulation of Turkey oak growth by increasing light and water availability. We were able to make a detailed reconstruction of the impact of past silvicultural treatment on the stand using a tree-ring wood Δ¹³C time-series.

     

Intraspecific variation in drought response of Populus cathayana grown under ambient and enhanced UV-B radiation

? The effects of drought, enhanced UV-B radiation and their combination on plant growth and physiological traits were investigated in a greenhouse experiment in two populations of Populus cathayana Rehder originating from high and low altitude in south-west China.

? In both populations, drought significantly decreased biomass accumulation and gas exchange parameters, including net CO₂ assimilation rate (A), stomatal conductance (g _s ), transpiration rate (E) and photosynthetic nitrogen use efficiency (PNUE). However, instantaneous water use efficiency (WUE _i ), transpiration efficiency (WUE _T ), carbon isotope composition (δ¹³C) and nitrogen (N) content, as well as the accumulation of soluble protein, UV-absorbing compounds and abscisic acid (ABA) significantly increased in response to drought. On the other hand, cuttings from both populations, when kept under enhanced UV-B radiation, showed very similar changes, as under drought, in all above-mentioned parameters.

? Compared with the low altitude population, the high altitude population was more tolerant to drought and enhanced UV-B, as indicated by the higher level of biomass accumulation, gas exchange, water-use efficiency, ABA concentration and UV-absorbing compounds.

? After one growing season of exposure to different UV-B levels and watering regimes, the decrease in biomass accumulation and gas exchange, induced by drought, was more pronounced under the combination of UV-B and drought. Significant interactions between drought and UV-B were observed in WUE _i , WUE_T, δ¹³C, soluble protein, UV-absorbing compounds, ABA and in the leaf and stem N, as well as in the leaf and stem C:N ratio.

? Our results showed that UV-B acts as an important signal allowing P. cathayana seedlings to respond to drought and that the combination of drought and UV-B may cause synergistically detrimental effects on plant growth.

     

Genetic variation for carbon isotope composition in Juglans regia L.: relationships with growth, phenology and climate of origin

? Among the traits of breeding interest for the common walnut tree Juglans regia L., characteristics such as timing of budbreak and leaf fall, water-use efficiency and growth performance are regarded as being of utmost relevance in Mediterranean conditions.

? The authors evaluated intraspecific variation in δ¹³C (carbon isotope composition, surrogate of intrinsic water-use efficiency, WUE_i) for 22 J. regia families grown in a progeny test under supplementary irrigation, and investigated whether such variation correlated with climatic indicators of native habitats. The genetic relationships between δ¹³C, growth and phenology were also assessed during two consecutive years.

? Overall, the most water-use-efficient families (i.e. with higher δ¹³C), which originated mainly from drought-prone provenance regions which have a high vapour pressure deficit and low rainfall, exhibited less height growth and smaller DBH. Using a stepwise regression procedure, δ¹³C was included as the main explanatory variable of genotypic variation in growth traits, together with growing season duration (for DBH in both years) and flushing (for height in 2007).

? It was concluded that WUE_i is largely unconnected to phenology effects in the explanation of growth performance for J. regia, therefore suggesting the opportunity of simultaneously selecting for low WUE_i and extended growing period to maximise productivity in non-water-limited environments.

     

Genetic variation in productivity, leaf traits and carbon isotope discrimination in hybrid poplars cultivated on contrasting sites

? We examined the relationships between productivity, leaf traits and carbon isotope discrimination in bulk leaf matter (Δ₁) and in phloem sap (Δ_s) from more than 5-year-old trees belonging to Populus deltoides × P. nigra and Populus trichocarpa × P. deltoides; trees were grown in alluvial and non alluvial sites in a commercial poplar plantation.

? On both sites, a large genetic variability was evidenced for all variables. The genotypic ranking remained stable between years for all variables, while it differed between sites. Δ₁ scaled positively with Δ_s and neither Δ₁ nor Δ_s were correlated with productivity. A significant genotype by site interaction was evident for all variables. The non alluvial site resulted in lower productivity, and in thicker/denser leaves with lower nitrogen and carbon contents. Noteworthy, the genotypic ranking for Δ₁ measured at the alluvial site was similar to that previously established in a glasshouse.

? As observed in previous studies from younger trees, there is a potential to select genotypes, combining high productivity and high water-use efficiency, for growth in moderately drought-prone areas.

     

Mesophyll conductance to CO2 and leaf morphological characteristics under drought stress during Quercus ilex L. resprouting

? Quercus ilex L., the dominant species in Mediterranean forests and one with a great capacity for resprouting after disturbances, is threatened by the expected increase in fire frequency and drought associated with climate change.

? The aim of this study was to determine the contribution of photosynthesis limitants, especially mesophyll conductance (g _mes) during this species’ resprouting and under summer drought.

? Resprouts showed 5.3-fold increased g _mes and 3.8-fold increased stomatal conductance (g _s) at mid-day with respect to leaves of undisturbed individuals. With increased drought, structural changes (decreased density and increased thickness) in resprouts contributed to the observed higher photosynthesis and increased g _mes. However, g _mes only partially depended on leaf structure, and was also under physiological control. Resprouts also showed lower non-stomatal limitations (around 50% higher carboxylation velocity (V _c,max) and capacity for ribulose-1,5-bisphosphate regeneration (J _max)). A significant contribution of g _mes to leaf carbon isotope discrimination values was observed.

? g _mes exhibits a dominant role in photosynthesis limitation in Q. ilex and is regulated by factors other than morphology. During resprouting after disturbances, greater capacity to withstand drought, as evidenced by higher g _mes, g _s and lower non-stomatal limitants, enables increased photosynthesis and rapid growth.

     

Current uptake of 15N-labeled ammonium and nitrate in flooded and non-flooded black spruce and tamarack seedlings

? We investigated the effects of flooding for three weeks on physiological responses and uptake of NH ₄ ⁺ and NO ₃ ^? by black spruce (Picea mariana (Mill.) BSP.) and tamarack (Larix laricina (Du Roi) K. Koch) seedlings fertilized with labeled (¹⁵NH₄)₂SO₄ or K¹⁵NO₃ in a growth chamber experiment.

? Flooding reduced photosynthesis (A), transpiration (E), water use efficiency (WUE), and current uptake of NH ₄ ⁺ and NO ₃ ^? in both species.

? Under flooding, there were no significant differences between the two species in uptake of either NH ₄ ⁺ or NO ₃ ^? at the whole-plant level but black spruce had higher translocation of NH ₄ ⁺ to the shoots than did tamarack.

? Under non-flooded conditions, black spruce seedlings exhibited higher uptake of both NH ₄ ⁺ and NO ₃ ^? than did tamarack and demonstrated preferential uptake of NH ₄ ⁺ (19. 7 mg g^?1dw) over NO ₃ ^? (12.3 mg g^?1dw after three weeks). In contrast, non-flooded tamarack seedlings had equal uptake of NH ₄ ⁺ (4.96 mg g^?1dw) and NO ₃ ^? (4.97 mg g^?1dw).

? We hypothesize that the ability of tamarack to equally exploit both ¹⁵NH ₄ ⁺ and ¹⁵NO ₃ ^? would confer an advantage over black spruce, when faced with limitations in the availability of different forms of soil nitrogen.

     

Sap flow measurement by a single thermal dissipation probe: exploring the transient regime

? In thermal dissipation method the stationary regime is used to estimate sap flow density from the temperature difference between a heating probe and a non-heating one.

? A new approach, based on the non-stationary regime of a single heating probe, is investigated on an olive tree branch. Probe temperature is monitored for different flow densities and for heating and cooling modes. Measurement sequences of 10 min using 5 s time steps are made just after the heating is switched on or off.

? Results show that temperature variation has a quasi-exponential phase whose duration decreases for increasing flow densities. Thermal indices using temperature of the probe measured at initial, final, and intermediate times t _i were developed. Relationships between sap flow density and thermal indices were found to be linear for t_i = 10 s?60 s with better coefficients of determination in cooling mode (R ² = 0.94–0.96) than in heating (R ² = 0.83–0.93). Validation tests confirmed the robustness of the single probe method with lowest error obtained for t _i = 20 s.

? Single probe method seems to be a promising technique to measure tree sap flow. Its application could be considered under cooling mode using t _i = 20 s and relatively short measurement sequences e.g. 5–10 min.

     

Why and where do adult trees die in a young secondary temperate forest? The role of neighbourhood

? The density and identity of tree neighbourhood is a key factor to explain tree mortality in forests, especially during the stem exclusion phase.

? To understand this process, we built a logistic model for mortality in a spatially explicit context, including tree and neighbourhood predictors. Additionally, we used this model to build mortality risk frequency distributions. Finally, we tested this model against a random mortality model to predict the spatial pattern of the forest.

? Annual mortality rate was high for pedunculate oak (Quercus robur, 6.99%), moderate for birch (Betula celtiberica, 2.19%) and Pyrenean oak (Q. pyrenaica, 1.58%) and low for beech (Fagus sylvatica, 0.26%). Mortality risk models for pedunculate oak and birch included stem diameter, tree height, canopy position and neighbourhood. Mortality was affected by the specific nature of the neighbourhood showing a clear competitive hierarchy: beech > pedunculate oak > birch. Models based on random mortality and logistic regression model were able to predict the spatial pattern of survivors although logistic regression predictions were more accurate.

? Our study highlights how simple models such as the random mortality one may obscure much more complex spatial interactions.

     

Leaf water status and stem xylem flux in relation to soil drought in five temperate broad-leaved tree species with contrasting water use strategies

? Five temperate broad-leaved tree species were compared with respect to their water consumption strategies under ample and restricted water supply. We measured synchronously leaf conductance (g _L) in the sun canopy, xylem sap flux (J _s) and leaf water potential (predawn, ψ_pd and noon, ψ_noon) in adult trees in a mixed stand and related them to the fluctuations in vapor pressure deficit (D) and soil moisture.

? Maximum g _L was particularly high in F. excelsior, C. betulus and T. cordata and revealed a higher D sensitivity. ψ_pd remained constantly high in A. pseudoplatanus, C. betulus and F. excelsior, but decreased in T. cordata and F. sylvatica with decreasing soil moisture.

? J _sddecreased linearly with decreasing soil matrix potential in all species except for F. excelsior. Apparent hydraulic conductance in the soil-to-leaf flow path (L _c) was higher in A. pseudoplatanus than in the other species.

? F. sylvatica maintained a low maximum g _L and reduced J _sd markedly upon drought, but faced severe decreases in ψ_pd and ψ_noon. F. excelsior represents an opposite strategy with high maximum g _L and stable ψ_pd.

? The species drought sensitivity increases in the sequence F. excelsior < C. betulus < T. cordata < A. pseudoplatanus < F. sylvatica.

     

Genetic structure of Tunisian natural carob tree (Ceratonia siliqua L.) populations inferred from RAPD markers

? Seven RAPD markers were used to assess the genetic diversity and structure of ten Tunisian natural Ceratonia siliqua L. populations from different geographic and bioclimatic zones.

? The species maintain a high diversity within population as estimated by the percentage of polymorphic loci and Shannon’s index (P% = 76.31, $\bar H'_{pop} = 0.569$ ). The range of variation between populations was large. Populations from the upper semi-arid bioclimates, with more continuous distribution area showed the highest level of variation.

? A high genetic differentiation among populations (Φ_ST = 0.250 and $\bar G_{ST} = 0.347$ ), as a result of population isolation was revealed. Nevertheless, the genetic structure is in accordance with bioclimate indicating that ecological factors also should influence differentiation. Populations from the sub-humid, upper semi-arid and mean semi-arid zones clustered together and were distinct from those of the lower semi-arid ones.

? Conservation strategy should be made according to the level of polymorphism within population and bioclimate.

     

Drought susceptibility and recovery of transplanted Quercus rubra seedlings in relation to root system morphology

? Transplant shock, implicated by depressed seedling physiological status associated with moisture stress immediately following planting, limits early plantation establishment. Large root volume (Rv) has potential to alleviate transplant shock because of higher root growth potential and greater access to soil water.

? We investigated impacts of drought and transplant Rv on photosynthetic assimilation (A), transpiration (E), stomatal conductance (g _s ), predawn leaf xylem water potential (Ψ_L), and growth of northern red oak (Quercus rubra L.) seedlings to explain mechanisms associated with susceptibility to transplant shock. One year-old barerooot seedlings were graded into four Rv categories and either well watered or subjected to drought consisting of low, medium, or high moisture stress by discontinuing irrigation at 22-day intervals for 3 months. Thereafter, all treatments were re-watered to examine recovery.

? Transplant shock was signified by reduced A, E, g _s, and Ψ_L, which generally increased with increasing moisture stress and Rv. Physiological status improved during recovery, though stress was still evident in seedlings exposed to medium or high moisture stress and in larger Rv seedlings. Growth declined with increasing moisture stress but was generally similar among Rv treatments, likely reflecting greater A at the whole plant level and/or reliance upon stored reserves in large Rv seedlings.

? The most effective drought avoidance mechanisms were root growth, stomatal regulation, reduced leaf area, and higher growth allocation to roots relative to shoots. Our results suggest that large initial Rv does not enhance drought avoidance during the first season after transplant in northern red oak seedlings.

     

Cold hardiness and transplant response of Juglans nigra seedlings subjected to alternative storage regimes

? Effects of overwinter storage regimes on seedling cold hardiness and physiological vigor are relatively unexplored, particularly for temperate deciduous forest tree species.

? We evaluated influence of storage duration (0, 66, 119, or 175 d) on electrolyte leakage of stem and root collar tissues following exposure to a series of freeze-test temperatures in black walnut (Juglans nigra L.) seedlings sampled from cold (3 °C) or freezer (?2 °C) storage. Seedlings were subsequently transplanted into a controlled growth chamber environment for two months.

? Regardless of storage temperature, mean LT₅₀ was lowest for seedlings stored for 66 d (≤ ?34 °C) and increased dramatically after 119 d (≥ ?13 °C).

? Root collar tissue had lower LT₅₀ than stem tissue after 119 d for cold-stored seedlings, reflecting importance of evaluative tissue type. Days to bud break shortened with increasing storage duration up to 119 d and stabilized thereafter for both storage regimes. Root growth potential was maximized after 119 d of storage, and subsequently declined for cold-stored seedlings. Height growth increased following storage, regardless of duration.

? To promote stress resistance and transplant growth response, we recommend that black walnut seedlings from this genetic source be outplanted after approximately 66–119 d of storage.

     

Extending sapwood — Leaf area relationships from stems to roots in Coast Douglas-fir

? Studies of allometric relationships between leaf area and the cross-sectional area (CSA) of sapwood in the stem have shed light on the structural and functional relationships between water-conducting and photosynthetic tissues.

? The purpose of this study was to test whether sapwood-leaf area relationships could be extended from stems to roots in coast Douglas-fir (Pseudotsuga menziesii var. menziesii (Mirb.) Franco). Twelve trees were felled, their stumps were excavated, and the CSA of sapwood and heartwood were estimated for individual roots, entire root systems, and stem section.

? Root sapwood CSA was greater than sapwood CSA throughout the stem, and the ratio of leaf area to sapwood CSA (A _l :A _s ) was accordingly lower for root sapwood. The relationship between sapwood CSA and leaf area was more variable in roots and at groundline compared to crown base. Root A _l :A _s decreased with relative tree height (tree height/mean stand height).

? The strong allometric relationship between leaf area and the CSA of sapwood in the stem generally holds when extended to roots. The greater CSA of sapwood in roots versus stems may reflect differences in their roles in supporting the tree.

     

Saprotrophic versus symbiotic strategy during truffle ascocarp development under holm oak. A response based on 13C and 15N natural abundance

? The development of truffles in the soil is not well understood. It is not known if a direct transfer of carbohydrates takes place between the host tree and the developing ascocarps through ectomycorrhizal structures or whether sporophores become independent from their hosts after several weeks or months and are able to use dead host tissues or soil organic matter as carbon (C) and nitrogen (N) sources.

? To study saprophytic or symbiotic capacities of truffle ascocarps the natural abundance of ¹⁵N and ¹³C in foliage, wood, fine roots, mycorrhizae, fungal sporophores and soil were determined in a truffle orchard.

? The processes of carbon and nitrogen allocation remained unchanged during the entire period of ascocarp development of Tuber melanosporum. From ¹³C and ¹⁵N natural abundance measurements, T. melanosporum, T. brumale and T. rufum did not exhibit saprotophic strategy during ascocarp development, which is contradictory to common statements found in handbooks regarding truffle cultivation.

     

Genetic diversity in Tunisian Crataegus azarolus L. var. aronia L. populations assessed using RAPD markers

? The genetic diversity of nine wild Tunisian Crataegus azarolus var. aronia L. populations from different bioclimates was assessed using RAPD markers.

? Eight selected primers generated a total of 105 bands, 81 of which were polymorphic. Shannon’s index (H′) ranged from 0.222 to 0.278 according to a population with an average of 0.245. The genetic variation within the species (H _SP = 0.423) was relatively low. A high differentiation (G _ST = 0.421) among populations coupled with a low level of gene flow (N _m = 0.472) were observed. The analysis of molecular variance (AMOVA) revealed also significant differentiation among populations (Φ_ST = 0.371), even at a low scale space. The majority of variation occurred within populations (63.31%). The Mantel test performed on genetic (Φ_ST) and geographic distance matrices among population pairs did not reveal an isolation by distance.

? Interpretation of Neighbour-joining tree based on Nei’s and Li’s genetic distance among individuals showed distinct population groupings. The UPGMA dendrogram based on Φ_ST values revealed two population sub-clusters, each including populations from different bioclimates and/or geographic regions.

? The low level of genetic diversity and the high genetic structure of populations resulted from genetic drift caused both by habitat fragmentation and the low size of populations.

? The high differentiation among populations and the similar low level of diversity within populations suggest that in situ conservation should interest all populations. The ex situ conservation should be based on the collection of seeds rather within than among populations because of the maximum of variation was revealed within populations.

     

Fall fertilization of Pinus resinosa seedlings: nutrient uptake, cold hardiness, and morphological development

? Fall fertilization may increase plant nutrient reserves, yet associated impacts on seedling cold hardiness are relatively unexplored.

? Bareroot red pine (Pinus resinosa Ait.) seedlings in north-central Minnesota, USA were fall fertilized at the end of the first growing season with ammonium nitrate (NH₄NO₃) at 0, 11, 22, 44, or 89 kg N ha^?1. Seedling morphology and cold hardiness [assessed by freeze induced electrolyte leakage (FIEL)] were evaluated six weeks after fertilization and following the second growing season.

? Seedling height and number of needle primordia increased with fertilizer rate for both sampling years. Seedlings fertilized with 44 and 89 kg N ha^?1 attained target height (15 cm) after the second growing season. Shoot and root N concentration increased after the first growing season in fall fertilized seedlings compared to controls. Fall fertilized seedlings had lower FIEL (i.e., increased cold hardiness) compared to controls when tested at ?40 °C after the first growing season, but no significant differences in FIEL of control and fertilized seedlings were observed after the second growing season.

? Results suggest that fall fertilization of red pine seedlings can help render desired target height in the nursery, while maintaining or increasing cold hardiness levels.

     

Variations in water use by a mature mangrove of Avicennia germinans, French Guiana

? In the tropical intertidal zones, little is known on water uptake by mangroves. Transpiration rates are generally measured at leaf level, but few studies exist on water use at tree or stand levels.

? The objective of this study was to measure sap flow in trees of different sizes to appreciate the range of variation in water use that may exist in a site dominated by 80% mature Avicennia germinans.

? The results showed that from the dry to the wet season the mean water use increased from 3.2 to 5.3 dm³ d^?1 in small trees (DBH ～ 13 cm), from 11.5 to 30.8 dm³ d^?1 in medium trees (～24 cm) and from 40.8 to 64.1 dm³ d^?1 in large ones (～45 cm).

? Sapwood remained active up to a depth of 8 cm with radial variations within the stem. Weak correlations were obtained with VPD and net radiation.

? This study confirmed that transpiration was larger under low levels of salinity. Water use at stand level (～1900 living stems ha^?1) was estimated to be in the range of 5.8 to 11.8 m³ ha?1 d^?1 according to the season.

     

What sampling is needed for reliable estimations of genetic diversity in Fraxinus excelsior L. (Oleaceae)?

? Sample size is a critical issue for genetic diversity studies and conservation programs. However, sample size evaluation requires previous knowledge of allele frequencies estimated with precision and this is not often the case.

? Here, we evaluated sample size requirements for accurate genetic diversity in adult trees and family arrays in a 12 ha plot of Fraxinus excelsior L. (Oleaceae) in a community forest in central France. Data consisted of 579 adult trees and 480 offspring from 24 families genotyped at four nuclear microsatellites.

? Mean square errors (MSE) estimates performed on Monte Carlo simulations of resampled data indicated that several adult individuals (> 300) are necessary for accurate measures of allele richness. However, expected heterozygosity requires smaller samples (< 30). Seeds captured about 90% of adult allelic diversity requiring a sampling effort roughly 50% larger than that of adult trees (480 seeds vs. 300 adults) suggesting that seed sampling is heavily penalized for allele counts. Nevertheless, gene diversity of seeds was essentially identical to that of the adult population.

? Extrapolation of these results to other ash tree populations appears feasible because of similar levels of diversity reported in the literature but it is not granted for species with significant selfing or high genetic structure.

     

Effects of organic residue management and legume cover on growth of pine seedlings, nutrient leaching and soil properties

? The short-term effect of organic residue management on the growth and nutrition of Pinus pinaster Ait. seedlings, and on nutrient leaching and chemical properties of an acid soil was assessed through a lysimeter experiment. Treatments included absence, placement on the soil surface, and incorporation into the soil (with and without legume cover cropping) of organic residues (forest floor litter or forest floor litter plus harvest residues).

? Residues placed on the soil surface enhanced seedling growth. Organic residues reduced nutrient losses (NO ₃ ^? , Ca and Mg) and resulted in nutrient accumulation in the soil. Harvest residues positively affected K seedling nutrition status and enhanced K soil accumulation.

? Legume cover cropping reduced soil nutrient losses (N, Ca, Mg and K) during the early stage of seedling growth; it also improved seedling nutrition status (N and P), but without any effect on growth.

? Harvest residues plus forest floor litter placed on the soil surface was the most appropriate management to both reduce nutrient losses through leaching and increase height of seedlings at the end of the experimental period (two years).

     

Evidence of altitudinal increase in photosynthetic capacity: gas exchange measurements at ambient and constant CO2 partial pressures

? Because all microclimatic variables change with elevation, it is difficult to compare plant performance and especially photosynthetic capacity at different elevations. Indeed, most previous studies investigated photosynthetic capacity of low- and high-elevation plants using constant temperature, humidity and light but varying CO₂ partial pressures (P _{CO 2}).

? Using gas exchange measurements, we compared here maximum assimilation rates (A _max) at ambient and constant-low-elevation P _{CO 2}for two temperate tree species along an altitudinal gradient (100 to 1600 m) in the Pyrénées mountains.

? Significant differences in A _max were observed between the CO₂ partial pressure treatments for elevations above 600 m, the between-treatment differences increasing with elevation up to 4 μmol m^?2 s^?1. We found an increase in A _max with increasing elevation at constant-low-elevation P _{CO 2} but not at ambient P _{CO 2} for both species. Given a 10% change in P _{CO 2}, a proportionally higher shift in maximum assimilation rate was found for both species.

? Our results showed that high elevation populations had higher photosynthetic capacity and therefore demonstrated that trees coped with extreme environmental conditions by a combination of adaptation (genetic evolution) and of acclimation. Our study also highlighted the importance of using constant CO₂ partial pressure to assess plant adaptation at different elevations.