Genomic and phenotypic divergence unveil microgeographic adaptation in the Amazonian hyperdominant tree Eperua falcata Aubl. (Fabaceae).

Plant populations can undergo very localized adaptation, allowing widely distributed populations to adapt to divergent habitats in spite of recurrent gene flow. Neotropical trees-whose large and undisturbed populations often span a variety of environmental conditions and local habitats-are particularly good models to study this process. Here, we explore patterns of adaptive divergence from large (i.e., regional) to small (i.e., microgeographic) spatial scales in the hyperdominant Amazonian tree Eperua falcata Aubl. (Fabaceae) under a replicated design involving two microhabitats (~300 m apart) in two study sites (~300 km apart). A three-year reciprocal transplant illustrates that, beyond strong maternal effects and phenotypic plasticity, genetically driven divergence in seedling growth and leaf traits was detected both between seedlings originating from different regions, and between seedlings from different microhabitats. In parallel, a complementary genome scan for selection was carried out through whole-genome sequencing of tree population pools. A set of 290 divergence outlier SNPs was detected at the regional scale (between study sites), while 185 SNPs located in the vicinity of 106 protein-coding genes were detected as replicated outliers between microhabitats within regions. Outlier-surrounding genomic regions are involved in a variety of physiological processes, including plant responses to stress (e.g., oxidative stress, hypoxia and metal toxicity) and biotic interactions. Together with evidence of microgeographic divergence in functional traits, the discovery of genomic candidates for microgeographic adaptive divergence represents a promising advance in our understanding of local adaptation, which probably operates across multiple spatial scales and underpins divergence and diversification in Neotropical trees.

Acclimation and adaptation components of the temperature dependence of plant photosynthesis at the global scale.

The temperature response of photosynthesis is one of the key factors determining predicted responses to warming in global vegetation models (GVMs). The response may vary geographically, owing to genetic adaptation to climate, and temporally, as a result of acclimation to changes in ambient temperature. Our goal was to develop a robust quantitative global model representing acclimation and adaptation of photosynthetic temperature responses. We quantified and modelled key mechanisms responsible for photosynthetic temperature acclimation and adaptation using a global dataset of photosynthetic CO2 response curves, including data from 141 C3 species from tropical rainforest to Arctic tundra. We separated temperature acclimation and adaptation processes by considering seasonal and common-garden datasets, respectively. The observed global variation in the temperature optimum of photosynthesis was primarily explained by biochemical limitations to photosynthesis, rather than stomatal conductance or respiration. We found acclimation to growth temperature to be a stronger driver of this variation than adaptation to temperature at climate of origin. We developed a summary model to represent photosynthetic temperature responses and showed that it predicted the observed global variation in optimal temperatures with high accuracy. This novel algorithm should enable improved prediction of the function of global ecosystems in a warming climate.

Increased light-use efficiency sustains net primary productivity of shaded coffee plants in agroforestry system.

In agroforestry systems, shade trees strongly affect the physiology of the undergrown crop. However, a major paradigm is that the reduction in absorbed photosynthetically active radiation is, to a certain extent, compensated by an increase in light-use efficiency, thereby reducing the difference in net primary productivity between shaded and non-shaded plants. Due to the large spatial heterogeneity in agroforestry systems and the lack of appropriate tools, the combined effects of such variables have seldom been analysed, even though they may help understand physiological processes underlying yield dynamics. In this study, we monitored net primary productivity, during two years, on scales ranging from individual coffee plants to the entire plot. Absorbed radiation was mapped with a 3D model (MAESPA). Light-use efficiency and net assimilation rate were derived for each coffee plant individually. We found that although irradiance was reduced by 60% below crowns of shade trees, coffee light-use efficiency increased by 50%, leaving net primary productivity fairly stable across all shade levels. Variability of aboveground net primary productivity of coffee plants was caused primarily by the age of the plants and by intraspecific competition among them (drivers usually overlooked in the agroforestry literature) rather than by the presence of shade trees.

Vapour pressure deficit during growth has little impact on genotypic differences of transpiration efficiency at leaf and whole-plant level: an example from Populus nigra†L.

Poplar genotypes differ in transpiration efficiency (TE) at leaf and whole-plant level under similar conditions. We tested whether atmospheric vapour pressure deficit (VPD) affected TE to the same extent across genotypes. Six Populus nigra genotypes were grown under two VPD. We recorded (1) (13)C content in soluble sugars; (2) (18)O enrichment in leaf water; (3) leaf-level gas exchange; and (4) whole-plant biomass accumulation and water use. Whole-plant and intrinsic leaf TE and (13)C content in soluble sugars differed significantly among genotypes. Stomatal conductance contributed more to these differences than net CO2 assimilation rate. VPD increased water use and reduced whole-plant TE. It increased intrinsic leaf-level TE due to a decline in stomatal conductance. It also promoted higher (18)O enrichment in leaf water. VPD had no genotype-specific effect. We detected a deviation in the relationship between (13)C in leaf sugars and (13)C predicted from gas exchange and the standard discrimination model. This may be partly due to genotypic differences in mesophyll conductance, and to its lack of sensitivity to VPD. Leaf-level (13)C discrimination was a powerful predictor of the genetic variability of whole-plant TE irrespective of VPD during growth.

Performance of a new dynamic model for predicting diurnal time courses of stomatal conductance at the leaf level.

Under natural conditions, plants are subjected to continuous changes of irradiance that drive variations of stomatal conductance to water vapour (g(s)). We propose a dynamic model to predict the temporal response of g(s) at the leaf level using an asymmetric sigmoid function with a unique parameter describing time constants for increasing and decreasing g(s). The model parameters were adjusted to observed data using Approximate Bayesian Computation. We tested the model performance for (1) instant changes of irradiance; or (2) continuous and controlled variations of irradiance simulating diurnal time courses. Compared with the two mostly used steady-state models, our dynamic model described daily time courses of g(s) with a higher accuracy. In particular, it was able to describe the hysteresis of g(s) responses to increasing/decreasing irradiance and the resulting rapid variations of intrinsic water-use efficiency. Compared to the mechanistic model of temporal responses of g(s) by Kirschbaum, Gross & Pearcy, for which time constants were estimated with a large variance, our model estimated time constants with a higher precision. It is expected to improve predictions of water loss and water-use efficiency in higher scale models by using a small number of parameters.

Genotype differences in 13C discrimination between atmosphere and leaf matter match differences in transpiration efficiency at leaf and whole-plant levels in hybrid Populus deltoides x nigra.

(13) C discrimination between atmosphere and bulk leaf matter (Δ(13) C(lb) ) is frequently used as a proxy for transpiration efficiency (TE). Nevertheless, its relevance is challenged due to: (1) potential deviations from the theoretical discrimination model, and (2) complex time integration and upscaling from leaf to whole plant. Six hybrid genotypes of Populus deltoides×nigra genotypes were grown in climate chambers and tested for whole-plant TE (i.e. accumulated biomass/water transpired). Net CO(2) assimilation rates (A) and stomatal conductance (g(s) ) were recorded in parallel to: (1) (13) C in leaf bulk material (δ(13) C(lb) ) and in soluble sugars (δ(13) C(ss) ) and (2) (18) O in leaf water and bulk leaf material. Genotypic means of δ(13) C(lb) and δ(13) C(ss) were tightly correlated. Discrimination between atmosphere and soluble sugars was correlated with daily intrinsic TE at leaf level (daily mean A/g(s) ), and with whole-plant TE. Finally, g(s) was positively correlated to (18) O enrichment of bulk matter or water of leaves at individual level, but not at genotype level. We conclude that Δ(13) C(lb) captures efficiently the genetic variability of whole-plant TE in poplar. Nevertheless, scaling from leaf level to whole-plant TE requires to take into account water losses and respiration independent of photosynthesis, which remain poorly documented.

Mesophyll diffusion conductance to CO2: an unappreciated central player in photosynthesis.

Mesophyll diffusion conductance to CO(2) is a key photosynthetic trait that has been studied intensively in the past years. The intention of the present review is to update knowledge of g(m), and highlight the important unknown and controversial aspects that require future work. The photosynthetic limitation imposed by mesophyll conductance is large, and under certain conditions can be the most significant photosynthetic limitation. New evidence shows that anatomical traits, such as cell wall thickness and chloroplast distribution are amongst the stronger determinants of mesophyll conductance, although rapid variations in response to environmental changes might be regulated by other factors such as aquaporin conductance. Gaps in knowledge that should be research priorities for the near future include: how different is mesophyll conductance among phylogenetically distant groups and how has it evolved? Can mesophyll conductance be uncoupled from regulation of the water path? What are the main drivers of mesophyll conductance? The need for mechanistic and phenomenological models of mesophyll conductance and its incorporation in process-based photosynthesis models is also highlighted.

Tree age-related effects on sun acclimated leaves in a chronosequence of beech (Fagus sylvatica) stands.

The assessment of the effect of tree age on leaves is usually limited by the difficulty of sampling sun leaves from tall ageing trees. In this study, we investigated tree age-related effects on sun leaves in a chronosequence of beech (Fagus sylvatica L.) stands. The effects of stand age on leaf mass to area ratio (LMA), chlorophyll (Chl), epidermal polyphenols (EPhen), nitrogen and carbon contents in sun leaves were investigated in 17 even-aged stands distributed into six age classes (14-175 years old). Chl and EPhen were assessed in vivo with SPAD and Dualex portable leaf-clips respectively. Leaves were sampled by shooting and sun leaves were identified based on criteria obtained from a vertical profile of the ratio abaxial vs adaxial EPhen across the canopy. Sun leaves were characterised by a high and similar adaxial and abaxial EPhen contents, high LMA value and low mass-based Chl content. These sun leaf characteristics, together with leaf nitrogen and carbon contents, were not significantly affected by stand age. Along the chronosequence, beech trees invested a stable fraction of leaf mass into nitrogen, carbon, Chl and EPhen with decreasing leaf size, i.e. dry mass and area.

Is mesophyll conductance to CO2 in leaves of three Eucalyptus species sensitive to short-term changes of irradiance under ambient as well as low O2?

Mesophyll conductance to CO2 (g m) limits the diffusion of CO2 to the sites of carboxylation, and may respond rapidly (within minutes) to abiotic factors. Using three Eucalyptus species, we tested the rapid response of g m to irradiance under 21% and 1% O2. We used simultaneous measurements of leaf gas exchange and discrimination against 13CO2 with a tuneable diode laser absorption spectrometer. Measurements under 1% O2 were used to limit uncertainties due to 13C-12C fractionation occurring during photorespiration. Switching irradiance from 600 to 200µmolm-2s-1 led to a ≈60% decrease of g m within minutes in all species under both 21% O2 and 1% O2. The g m response to irradiance is unlikely to be a computation artefact since using different values for the parameters of the discrimination model changed the absolute values of g m but did not affect the relative response to irradiance. Simulations showed that possible rapid changes of any parameter were unable to explain the observed variations of g m with irradiance, except for13C-12C fractionation during carboxylation (b), which, in turn, is dependent on the fraction of leaf C assimilated by phospho-enol pyruvate carboxylase (PEPc) (ß). g m apparently increased by ≈30% when O2 was switched from 21% to 1% O2. Again, possible changes of ß with O2 could explain this apparent g m response to O2. Nevertheless, large irradiance or O2-induced changes in ß would be required to fully explain the observed changes in g m, reinforcing the hypothesis that g m is responsive to irradiance and possibly also to O2.

Time course of δ1¹³C in poplar wood: genotype ranking remains stable over the life cycle in plantations despite some differences between cellulose and bulk wood.

Genetic differences in δ¹³C (isotopic composition of dry matter carbon) have been evidenced among poplar genotypes at juvenile stages. To check whether such differences were maintained with age in trees growing in plantations, we investigated the time course of δ¹³C as recorded in annual tree rings from different genotypes growing at three sites in southwestern France and felled at ∼15-17 years. Wood cores were cut from tree discs to record the time course of annual basal area increment (BAI). The isotopic ratio δ¹³C was recorded in bulk wood and in extracted cellulose from the annual rings corresponding to the period 1996-2005. Discrimination against ¹³C between atmosphere and tissues (Δ¹³C) was computed by taking into account the inter-annual time course of δ¹³C in the atmosphere. Annual BAI increased steadily and stabilized at about 8 years. An offset in δ¹³C of ∼1‰ was recorded between extracted cellulose and bulk wood. It was relatively stable among genotypes within sites but varied among sites and increased slightly with age. Site effects as well as genotype differences were detected in Δ¹³C recorded from the cellulose fraction. Absolute values as well as the genotype ranking of Δ¹³C remained stable with age in the three sites. Genotype means of Δ¹³C were not correlated to annual BAI. We conclude that genotypic differences of Δ¹³C occur in older poplar trees in plantations, and that the differences as well as the genotype ranking remain stable while trees age until harvest.

Variations in bulk leaf carbon isotope discrimination, growth and related leaf traits among three Populus nigra L. populations.

The ongoing global change could be an additional threat to the establishment and the long-term survival of Populus nigra L., an emblematic European riparian species. With the general aim of gaining insights into the adaptive potential of this species, we (i) quantified variations within and among three French P. nigra populations for key physiological attributes, i.e., water-use efficiency (assessed from bulk leaf carbon isotope discrimination, Δ(13)C), growth performance and related leaf traits, (ii) examined genotype and population by environment interactions, and (iii) explored the relationship between Δ(13)C and growth. Thirty genotypes were sampled in each of three naturally established populations and grown in two different sites, Orléans (ORL) and Guémené-Penfao (GMN). In ORL, two similar plots were established and different watering regimes were applied in order to test for the drought response. Significant variations were observed for all traits within and among populations irrespective of site and watering. Trait variation was larger within than among populations. The effect of drought was neither genotype- nor population-dependent, contrary to the effect of site. The population ranking was maintained in all sites and watering regimes for the two most complex traits: Δ(13)C and growth. Moreover, these two traits were unrelated, which indicates that (i) water-use efficiency and growth are largely uncoupled in this species, and (ii) the environmental factors driving genetic structuration for Δ(13)C and growth act independently. The large variations found within populations combined with the consistent differences among populations suggest a large adaptive potential for P. nigra.

Mesophyll conductance to CO2, assessed from online TDL-AS records of ¹³CO2 discrimination, displays small but significant short-term responses to CO2 and irradiance in Eucalyptus seedlings.

Mesophyll conductance (g(m)) is now recognized as an important limiting process for photosynthesis, as it results in a significant decrease of CO(2) diffusion from substomatal cavities where water evaporation occurs, to chloroplast stroma. Over the past decade, an increasing number of studies proposed that g(m) can vary in the short term (e.g. minutes), but these variations are still controversial, especially those potentially induced by changing CO(2) and irradiance. In this study, g(m) data estimated with online (13)C discrimination recorded with a tunable diode laser absorption spectrometer (TDL-AS) during leaf gas exchange measurements, and based on the single point method, are presented. The data were obtained with three Eucalyptus species. A 50% decrease in g(m) was observed when the CO(2) mole fraction was increased from 300 µmol mol(-1) to 900 µmol mol(-1), and a 60% increase when irradiance was increased from 200 µmol mol(-1) to 1100 µmol mol(-1) photosynthetic photon flux density (PPFD). The relative contribution of respiration and photorespiration to overall (13)C discrimination was also estimated. Not taking this contribution into account may lead to a 50% underestimation of g(m) but had little effect on the CO(2)- and irradiance-induced changes. In conclusion, (i) the observed responses of g(m) to CO(2) and irradiance were not artefactual; (ii) the respiratory term is important to assess absolute values of g(m) but has no impact on the responses to CO(2) and PPFD; and (iii) increasing irradiance and reducing the CO(2) mole fraction results in rapid increases in g(m) in Eucalyptus seedlings.

Compound-specific differences in (13)C of soluble carbohydrates in leaves and phloem of 6-month-old Eucalyptus globulus (Labill).

Movement of photoassimilates from leaves to phloem is an important step for the flux of carbon through plants. Fractionation of carbon isotopes during this process may influence their abundance in heterotrophic tissues. We subjected Eucalyptus globulus to 20, 25 and 28 °C ambient growth temperatures and measured compound-specific δ(13)C of carbohydrates obtained from leaves and bled phloem sap. We compared δ(13)C of sucrose and raffinose obtained from leaf or phloem and of total leaf soluble carbon, with modelled values predicted by leaf gas exchange. Changes in δ(13)C of sucrose and raffinose obtained from either leaves or phloem sap were more tightly coupled to changes in c(i)/c(a) than was δ(13)C of leaf soluble carbon. At 25 and 28 °C, sucrose and raffinose were enriched in (13)C compared to leaf soluble carbon and predicted values - irrespective of tissue type. Phloem sucrose was depleted and raffinose enriched in (13)C compared to leaf extracts. Intermolecular and tissue-specific δ(13)C reveal that multiple systematic factors influence (13)C composition during export to phloem. Predicting sensitivity of these factors to changes in plant physiological status will improve our ability to infer plant function at a range of temporal and spatial scales.

A cost-benefit analysis of acclimation to low irradiance in tropical rainforest tree seedlings: leaf life span and payback time for leaf deployment.

The maintenance in the long run of a positive carbon balance under very low irradiance is a prerequisite for survival of tree seedlings below the canopy or in small gaps in a tropical rainforest. To provide a quantitative basis for this assumption, experiments were carried out to determine whether construction cost (CC) and payback time for leaves and support structures, as well as leaf life span (i) differ among species and (ii) display an irradiance-elicited plasticity. Experiments were also conducted to determine whether leaf life span correlates to CC and payback time and is close to the optimal longevity derived from an optimization model. Saplings from 13 tropical tree species were grown under three levels of irradiance. Specific-CC was computed, as well as CC scaled to leaf area at the metamer level. Photosynthesis was recorded over the leaf life span. Payback time was derived from CC and a simple photosynthesis model. Specific-CC displayed only little interspecific variability and irradiance-elicited plasticity, in contrast to CC scaled to leaf area. Leaf life span ranged from 4 months to >26 months among species, and was longest in seedlings grown under lowest irradiance. It was always much longer than payback time, even under the lowest irradiance. Leaves were shed when their photosynthesis had reached very low values, in contrast to what was predicted by an optimality model. The species ranking for the different traits was stable across irradiance treatments. The two pioneer species always displayed the smallest CC, leaf life span, and payback time. All species displayed a similar large irradiance-elicited plasticity.

Monitoring the regulation of gene expression in a growing organ using a fluid mechanics formalism.

BACKGROUND: Technological advances have enabled the accurate quantification of gene expression, even within single cell types. While transcriptome analyses are routinely performed, most experimental designs only provide snapshots of gene expression. Molecular mechanisms underlying cell fate or positional signalling have been revealed through these discontinuous datasets. However, in developing multicellular structures, temporal and spatial cues, known to directly influence transcriptional networks, get entangled as the cells are displaced and expand. Access to an unbiased view of the spatiotemporal regulation of gene expression occurring during development requires a specific framework that properly quantifies the rate of change of a property in a moving and expanding element, such as a cell or an organ segment. RESULTS: We show how the rate of change in gene expression can be quantified by combining kinematics and real-time polymerase chain reaction data in a mechanistic model which considers any organ as a continuum. This framework was applied in order to assess the developmental regulation of the two reference genes Actin11 and Elongation Factor 1-beta in the apex of poplar root. The growth field was determined by time-lapse photography and transcript density was obtained at high spatial resolution. The net accumulation rates of the transcripts of the two genes were found to display highly contrasted developmental profiles. Actin11 showed pulses of up and down regulation in the accelerating and decelerating parts of the growth zone while the dynamic of EF1beta were much slower. This framework provides key information about gene regulation in a developing organ, such as the location, the duration and the intensity of gene induction/repression. CONCLUSIONS: We demonstrated that gene expression patterns can be monitored using the continuity equation without using mutants or reporter constructions. Given the rise of imaging technologies, this framework in our view opens a new way to dissect the molecular basis of growth regulation, even in non-model species or complex structures.

Interactive effects of high irradiance and moderate heat on photosynthesis, pigments, and tocopherol in the tree-fern Dicksonia antarctica.

Effects of high irradiance and moderate heat on photosynthesis of the tree-fern Dicksonia antarctica (Labill., Dicksoniaceae) were examined in a climate chamber under two contrasting irradiance regimes (900 and 170 µmol photons m-2 s-1) and three sequential temperature treatments (15°C; 35°C; back to 15°C). High irradiance led to decline in predawn quantum yield of photochemistry, Fv/Fm (0.73), maximal Rubisco activity (Vcmax; from 37 to 29 µmol m-2s-1), and electron transport capacity (Jmax; from 115 to 67 µmol m-2 s-1). Temperature increase to 35°C resulted in further decreases in Fv/Fm (0.45) and in chlorophyll bleaching of high irradiance plants, while Vcmax and Jmax were not affected. Critical temperature for thylakoid stability (Tc) of D. antarctica was comparable with other higher plants (c. 47°C), and increases of Tc with air temperature were greater in high irradiance plants. Increased Tc was not associated with accumulation of osmotica or zeaxanthin formation. High irradiance increased the xanthophyll cycle pigment pool (V+A+Z, 91 v. 48 mmol mol-1 chlorophyll-1), de-epoxidation state (56% v. 4%), and α-tocopherol. Temperature increase to 35°C had no effect on V+A+Z and de-epoxidation state in both light regimes, while lutein, ß-carotene and α-tocopherols increased, potentially contributing to increased membrane stability under high irradiance.

Hydraulic properties of naturally regenerated beech saplings respond to canopy opening.

Enhanced sapling growth in advance regeneration requires gaps in the canopy, but is often delayed after canopy opening, because acclimation of saplings to the new environment is gradual and may last for several years. Canopy opening is expected to result in an increased transpiration because of a larger climatic demand and a higher stomatal conductance linked to the higher rates of photosynthesis. Therefore, we focused on the changes in water relations and the hydraulic properties of beech (Fagus sylvatica L.) saplings during 2 years after canopy opening. We tested the hypothesis that an increase in leaf-specific hydraulic conductance and a decrease in vulnerability to cavitation occur to sustain an enhanced transpiration. Hydraulic conductance of defoliated shoots, vulnerability to cavitation, size and density of xylem vessels as well as stomatal conductance were recorded on saplings growing in shade (S saplings) or in gaps created by opening the canopy (shade-to-light, SL saplings). Hydraulic conductance per unit cross-sectional area (K(AS)) did not differ in the shoots of S and SL saplings. But a higher ratio stem cross-sectional area/leaf area resulted in a higher leaf-specific hydraulic conductance of the shoots (K(AL)) of SL saplings. Contrary to expectations, vulnerability to cavitation increased transitorily in stems during the first year after canopy opening and no difference was observed between the two treatments in light-saturated stomatal conductance. During the second year, vulnerability to cavitation was similar in the S and SL saplings and light-saturated stomatal conductance increased in SL saplings. These results demonstrate a release of the hydraulic constraints after canopy opening with an adjustment of the ratio stem cross-sectional area/leaf area. But the larger vulnerability to cavitation during the first year could limit stomatal opening and therefore the ability of beech saplings to use the available light for photosynthesis and could therefore partly explain why the growth increase was delayed to the second growing season after canopy opening.

Seasonal time-course of gradients of photosynthetic capacity and mesophyll conductance to CO2 across a beech (Fagus sylvatica L.) canopy.

Leaf photosynthesis is known to acclimate to the actual irradiance received by the different layers of a canopy. This acclimation is usually described in terms of changes in leaf structure, and in photosynthetic capacity. Photosynthetic capacity is likely to be affected by mesophyll conductance to CO(2) which has seldom been assessed in tree species, and whose plasticity in response to local irradiance is still poorly known. Structural [N and chlorophyll content, leaf mass to area ratio (LMA)] and functional leaf traits [maximum carboxylation rate (V(cmax)), maximum light-driven electron flux (J(max)), and mesophyll conductance (g(i))] were assessed by measuring leaf response curves of net CO(2) assimilation versus intercellular CO(2) partial pressure, along a vertical profile across a beech canopy, and by fitting a version of the Farquhar model including g(i). The measurements were repeated five times during a growth season to catch potential seasonal variation. Irradiance gradients resulted in large decreasing gradients of LMA, g(i), V(cmax), and J(max). Relative allocation of leaf N to the different photosynthetic processes was only slightly affected by local irradiance. Seasonal changes after leaf expansion and before induction of leaf senescence were only minor. Structural equation modelling confirmed that LMA was the main driving force for changes in photosynthetic traits, with only a minor contribution of leaf Nitrogen content. In conclusion, mesophyll conductance to CO(2) displays a large plasticity that scales with photosynthetic capacity across a tree canopy, and that it is only moderately (if at all) affected by seasonal changes in the absence of significant soil water depletion.

The diversity of (13)C isotope discrimination in a Quercus robur full-sib family is associated with differences in intrinsic water use efficiency, transpiration efficiency, and stomatal conductance.

(13)C discrimination in organic matter with respect to atmospheric CO(2) (Delta(13)C) is under tight genetic control in many plant species, including the pedunculate oak (Quercus robur L.) full-sib progeny used in this study. Delta(13)C is expected to reflect intrinsic water use efficiency, but this assumption requires confirmation due to potential interferences with mesophyll conductance to CO(2), or post-photosynthetic discrimination. In order to dissect the observed Delta(13)C variability in this progeny, six genotypes that have previously been found to display extreme phenotypic values of Delta(13)C [either very high ('high Delta') or low ('low Delta') phenotype] were selected, and transpiration efficiency (TE; accumulated biomass/transpired water), net CO(2) assimilation rate (A), stomatal conductance for water vapour (g(s)), and intrinsic water use efficiency (W(i)=A/g(s)) were compared with Delta(13)C in bulk leaf matter, wood, and cellulose in wood. As expected, 'high Delta' displayed higher values of Delta(13)C not only in bulk leaf matter, but also in wood and cellulose. This confirmed the stability of the genotypic differences in Delta(13)C recorded earlier. 'High Delta' also displayed lower TE, lower W(i), and higher g(s). A small difference was detected in photosynthetic capacity but none in mesophyll conductance to CO(2). 'High Delta' and 'low Delta' displayed very similar leaf anatomy, except for higher stomatal density in 'high Delta'. Finally, diurnal courses of leaf gas exchange revealed a higher g(s) in 'high Delta' in the morning than in the afternoon when the difference decreased. The gene ERECTA, involved in the control of water use efficiency, leaf differentiation, and stomatal density, displayed higher expression levels in 'low Delta'. In this progeny, the variability of Delta(13)C correlated closely with that of W(i) and TE. Genetic differences of Delta(13)C and W(i) can be ascribed to differences in stomatal conductance and stomatal density but not in photosynthetic capacity.

Stomatal regulation of photosynthesis in apple leaves: evidence for different water-use strategies between two cultivars.

BACKGROUND AND AIMS: Leaf responses to environmental conditions have been frequently described in fruit trees, but differences among cultivars have received little attention. This study shows that parameters of Farquhar's photosynthesis and Jarvis' stomatal conductance models differed between two apple cultivars, and examines the consequences of these differences for leaf water use efficiency. METHODS: Leaf stomatal conductance (g(sw)), net CO2 assimilation rate (A(n)), respiration (R(d)) and transpiration (E) were measured during summer in 8-year-old 'Braeburn' and 'Fuji' apple trees under well-watered field conditions. Parameters of Farquhar's and Jarvis' models were estimated, evaluated and then compared between cultivars. Leaf carbon isotope discrimination (delta(13)C) was measured at the end of the growing season. KEY RESULTS: A single positive relationship was established between V(Cmax) (maximum carboxylation rate) and N(a) (leaf nitrogen concentration per unit area), and between J(max) (maximum light-driven electron transport rate) and N(a). A higher leaf R(d) was observed in 'Fuji'. The g(sw) responded similarly to increasing irradiance and leaf temperature in both cultivars. g(sw) responded to lower vapour pressure deficit in 'Fuji' than in 'Braeburn'. Maximal conductance (g(swmax)) was significantly smaller and A(n) was more limited by g(sw) in 'Braeburn' than 'Fuji'. Lower g(sw), E and higher intrinsic water use efficiency were shown in 'Braeburn' and confirmed by smaller leaf delta(13)C compared with 'Fuji' leaves. CONCLUSIONS: The use of functional model parameters allowed comparison of the two cultivars and provided evidence of different water use 'strategies': 'Braeburn' was more conservative in water use than 'Fuji', due to stomatal limitation of A(n), higher intrinsic water use efficiency and lower delta(13)C. These physiological traits need to be considered in relation to climate adaptation, breeding of new cultivars and horticultural practice.