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.

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.

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.

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.

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.

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.

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.

Intra- and interspecific diversity in the response to waterlogging of two co-occurring white oak species (Quercus robur and Q. petraea).

Quercus robur L. and Quercus petraea (Matt.) Liebl. are sympatric oak species with different ecological requirements. Quercus robur is more tolerant to waterlogging than Q. petraea. This ecological divergence may play a role in the maintenance of the two species despite the absence of an insurmountable reproductive barrier between them. We predicted that the genetic architecture of traits related to waterlogging tolerance differs between the species. To gain insight into this architecture in the absence of genetic markers for waterlogging tolerance, we compared populations of seedlings of each species for diversity in the expression of quantitative phenotypic traits induced by severe hypoxia. To determine the capacity for hypertrophied lenticel formation, we applied gas-impermeable mastic to stems of seedlings. Two months after application, the mastic treatment had induced the formation of 3 (+/- 2) cm(-2) hypertrophied lenticels in the absence of root hypoxia. Leaf epinasty during root hypoxia was an early predictor of seedling mortality. Four weeks of waterlogging resulted in greater epinasty in Q. petraea than in Q. robur, but fewer hypertrophied lenticels (16 +/- 6 versus 21 +/- 9 cm(-2)) and adventitious roots (2.7 +/- 4.7 versus 5.2 +/- 5.9). Differences between species in these traits were associated with differences in the frequencies of extreme phenotypes rather than with a generally higher tolerance to waterlogging in Q. robur seedlings.

In vivo and in situ rhizosphere respiration in Acer saccharum and Betula alleghaniensis seedlings grown in contrasting light regimes.

A perfusive method combined with an open-system carbon dioxide measurement system was used to assess rhizosphere respiration of Acer saccharum Marsh. (sugar maple) and Betula alleghaniensis Britton (yellow birch) seedlings grown in 8-l pots filled with coarse sand. We compared in vivo and in situ rhizosphere respiration between species, among light regimes (40, 17 and 6% of full daylight) and at different times during the day. To compute specific rhizosphere respiration, temperature corrections were made with either species-specific coefficients (Q10) based on the observed change in respiration rate between 15 and 21 degrees C or an arbitrarily assigned Q10 of 2. Estimated, species-specific Q10 values were 3.0 and 3.4 for A. saccharum and B. alleghaniensis, respectively, and did not vary with light regime. Using either method of temperature correction, specific rhizosphere respiration did not differ either between A. saccharum and B. alleghaniensis, or among light regimes except in A. saccharum at 6% of full daylight. At this irradiance, seedlings were smaller than in the other light treatments, with a larger fine root fraction of total root dry mass, resulting in higher respiration rates. Specific rhizosphere respiration was significantly higher during the afternoon than at other times of day when temperature-corrected on the basis of an arbitrary Q10 of 2, suggesting the possibility of diurnal variation in a temperature-independent component of rhizosphere respiration.

Irradiance-induced plasticity in the hydraulic properties of saplings of different temperate broad-leaved forest tree species.

We assessed the irradiance-related plasticity of hydraulic architecture in saplings of Betula pendula Roth., a pioneer species; Acer pseudoplatanus L., Fraxinus excelsior L. and Quercus robur L., which are post-pioneer light-requiring species; and Quercus petraea Matt. Liebl. and Fagus sylvatica L. Plants were grown in pots in 36%, 16% and 4% of full sunlight. Hydraulic conductance was measured with a high-pressure flow-meter in entire, in situ root systems and in excised shoots. Leaf-specific whole-plant conductance (LSC) increased with irradiance, due, in part, to an effect of irradiance on plant size. In addition, there was a size-independent effect of irradiance on LSC due, in part, to an increase in root hydraulic conductance paralleled by an increase in root biomass scaled to leaf area. Changes in shoot conductivity also contributed to the size-independent plasticity of LSC. Vulnerability to cavitation measured in current-year twigs was much larger in shade-grown plants. Betula pendula had the highest whole-plant, root and shoot conductances and also the greatest vulnerability to cavitation. The other species were similar in LSC, but showed some variation in root conductance scaled to biomass, with Q. robur, Q. petraea and F. sylvatica having the lowest root conductance and susceptibility to cavitation. All species showed a similar irradiance-related plasticity in LSC.

Analysis of carbohydrates in plants by high-performance anion-exchange chromatography coupled with electrospray mass spectrometry.

A mass spectrometer was coupled to high-performance anion-exchange chromatography (HPAEC) with the help of electrochemical neutralization of the eluent and post-column addition of lithium chloride for carbohydrate analysis. Parallel selective channels (single ion monitoring) were used to decrease the detection limits and separate unresolved peaks. The mass specific detection allowed the simultaneous analysis of a wide range of sugar alcohols, mono-, di- and oligosaccharides. Carbohydrates extracted from leaves of poplar submitted to drought stress were analyzed using pulsed amperometric detection (PAD), then mass spectrometry. It allowed the confirmation of peak attribution and the identification of salicin as a major compound in the extracts. Different responses to water deficit and re-hydration were obtained for several carbohydrates, suggesting different roles in osmoprotection processes.

Leaf photosynthetic traits of 14 tropical rain forest species in relation to leaf nitrogen concentration and shade tolerance.

Variability of leaf traits related to photosynthesis was assessed in seedlings from 14 tree species growing in the tropical rain forest of French Guiana. Leaf photosynthetic capacity (maximum rate of carboxylation and maximum rate of electron transport) was estimated by fitting a biochemical model of photosynthesis to response curves of net CO2 assimilation rate versus intercellular CO2 mole fraction. Leaf morphology described by leaf mass per unit leaf area (LMA), density and thickness, as well as area- and mass-based nitrogen (N) and carbon (C) concentrations, were recorded on the same leaves. Large interspecific variability was detected in photosynthetic capacity as well as in leaf structure and leaf N and C concentrations. No correlation was found between leaf thickness and density. The correlations between area- and mass-based leaf N concentration and photosynthetic capacity were poor. Conversely, the species differed greatly in relative N allocation to carboxylation and bioenergetics. Principal component analysis (PCA) revealed that, of the recorded traits, only the computed fraction of total leaf N invested in photosynthesis was tightly correlated to photosynthetic capacity. We also used PCA to test to what extent species with similar shade tolerances displayed converging leaf traits related to photosynthesis. No clear-cut ranking could be detected among the shade-tolerant groups, as confirmed by a one-way ANOVA. We conclude that the large interspecific diversity in photosynthetic capacity was mostly explained by differences in the relative allocation of N to photosynthesis and not by leaf N concentration, and that leaf traits related to photosynthetic capacity did not discriminate shade-tolerance ranking of these tropical tree species.

Diversity of leaf traits related to productivity in 31 Populus deltoides x Populus nigra clones.

To test if some leaf parameters are predictors of productivity in a range of Populus deltoides (Bartr.) Marsh. x P. nigra L. clones, we assessed leaf traits and productivity in 2-month-old rooted cuttings from 31 clones growing in 4-l pots in a greenhouse, under conditions of controlled temperature and optimal irrigation. We evaluated four groups of variables describing (1) productivity (total biomass), (2) leaf growth (total leaf number increment and total leaf area increment rate), (3) leaf structure (specific leaf area and nitrogen and carbon contents) and (4) carbon isotope discrimination (delta), which is negatively correlated with time-integrated water-use efficiency. High-yielding clones did not necessarily display high leaf growth rates, but they displayed a larger total leaf area, lower specific leaf area and lower leaf nitrogen concentration than clones with low productivity. Total leaf area was mainly controlled by maximal individual leaf area and total leaf area increment rate (r = 0.51 and 0.56, respectively). Carbon isotope discrimination did not correlate with total biomass, but it was associated with total number of leaves and total leaf area increment rate (r = 0.39 and 0.45, respectively). Therefore, leaf area and specific leaf area were better indicators of productivity than leaf growth traits. The observed independence of delta from biomass production provides opportunities for selecting poplar clones combining high productivity and high water-use efficiency.