Search

Recent research has shown that plant acclimation to diverse patterns of light intensity modifies the dynamics of their stomatal response. Therefore, whether plants are grown in controlled conditions or in the field may impact their stomatal dynamics. We analysed the stomatal dynamics of two Populus euramericana and two Populus nigra genotypes grown in the field under contrasting water availability. By comparing their stomatal dynamics with that of the same genotypes grown in a glasshouse, we were able to test whether differences between these growing conditions interacted with genotypic differences in affecting stomatal dynamics and responses to soil water deficit. We found that, despite higher stomatal density and smaller size, in the field stomatal dynamics were much slower than in the glasshouse. Overall, differences among genotypes and their response to soil water deficit were much less pronounced in the field compared with the glasshouse. These results indicate that stomatal dynamics are regulated by both genotype-specific and environmental factors. Moreover, having slower stomata may be advantageous under some conditions. While stomatal dynamics were linked with whole-plant transpiration per leaf area in both experiments, the contribution of stomatal morphology varies dependent on the environmental conditions.

Human-caused CO 2 emissions over the past century have caused the climate of the Earth to warm and have directly impacted on the functioning of terrestrial plants. We examine the global response of terrestrial gross primary production (GPP) to the historic change in atmospheric CO 2. The GPP of the terrestrial biosphere has increased steadily, keeping pace remarkably in proportion to the rise in atmospheric CO 2. Water-use efficiency, namely the ratio of CO 2 uptake by photosynthesis to water loss by transpiration, has increased as a direct leaf-level effect of rising CO 2. This has allowed an increase in global leaf area, which has conspired with stimulation of photosynthesis per unit leaf area to produce a maximal response of the terrestrial biosphere to rising atmospheric CO 2 and contemporary climate change. Rising Atmospheric CO 2 and Global Climate Change Emissions of CO 2 associated with human industrial activity and land-use change over the past century have significantly impacted on global climate, causing global warming of about 1.0°C [1]. The anthropogenic CO 2 emission rate is continuing to increase, and the future rise in atmospheric CO 2 will undoubtedly lead to more climate change, including increases in the frequency of extreme climate events such as heatwaves, droughts, and storms [2]. Global climate change has the potential to significantly stress terrestrial vegetation [3], for example with hot, dry air, soil moisture deficits, or flooding. This could lead to a carbon-climate feedback in which widespread tree mortality and forest decline contribute to accelerating accumulation of CO 2 in the atmosphere [4-6]. On the other hand, plants interact directly with atmospheric CO 2 , and they can potentially respond to rising atmospheric CO 2 concentrations by increasing photosynthetic rates and water-use efficiency (see Glossary) [7-10]. Water-use efficiency in this context is defined as the amount of CO 2 taken up by photosynthesis for a given amount of water lost to the atmosphere by transpiration (Box 1). Understanding emergent responses of the production of terrestrial vegetation to the potentially opposing impacts of global climate change and CO 2 fertilization is crucial for formulating effective mitigation and adaptation strategies [11]. At a global scale, there is currently an imbalance between the amount of CO 2 absorbed by the terrestrial biosphere through photosynthesis and the amount released back to the atmosphere through plant respiration, decomposition, fire, and emissions from land-use change [12]. This is commonly referred to as the land carbon sink. It is slowing the rate of increase in atmospheric CO 2 that would otherwise result from anthropogenic CO 2 emissions. Predicting the future behaviour of the land carbon sink is one of the most important challenges in carbon cycle science, given the potential that feedbacks could accelerate the rate of future climate change [13]. This requires a thorough understanding of the process through which the terrestrial biosphere captures CO 2-photosynthesis. Highlights Global climate change caused by CO 2 emissions can stress terrestrial vegetation , potentially decreasing production. On the other hand, CO 2 interacts directly with plants, stimulating leaf-level photo-synthesis and water-use efficiency. The rise in atmospheric CO 2 concentration over the past century presents an opportunity for gauging the strength of the terrestrial biosphere response to these potential impacts. Atmospheric proxy and model analysis both suggest that global terrestrial photosynthesis has increased in nearly constant proportion to the rise in atmospheric CO 2 concentration, a maximal response by the terrestrial biosphere. An accurate understanding of the impacts of climate change on terrestrial vegetation is essential for managing risks associated with human-caused climate change: gauging the historic response of terrestrial photosynthesis is an important step in this direction.

Ozone (O-3) and drought increase tree oxidative stress. To protect forest health, we need to improve risk assessment, using metric model such as the phytotoxic O-3 dose above a threshold of y nmol.m(-2).s(-1) (PODy), while taking into account detoxification mechanisms and interacting stresses. The impact of drought events on the effect of O-3 pollution deserves special attention. Water deficit may decrease O-3 entrance into the leaves by reducing stomatal opening; however, water deficit also induces changes in cell redox homeostasis. Besides, the behaviour of the cell antioxidative charge in case of stress combination (water deficit and O-3) still remains poorly investigated. To decipher the response of detoxification mechanisms relatively to the Halliwell-Asada-Foyer cycle (HAF), we exposed poplar saplings (Populus nigra x deltoides) composed of two genotypes (Carpaccio and Robusta), to various treatments for 17 days, i.e. i) mild water deficit, ii) 120 ppb O-3, and iii) a combination of these two treatments. Ozone similarly impacted the growth of the two genotypes, with an important leaf loss. Water deficit decreased growth by almost one third as compared to the control plants. As for the combined treatment, water deficit protected the saplings from leaf ozone injury, but with an inhibitory effect on growth. The pool of total ascorbate was not modified by the different treatments, while the pool of total glutathione increased with POD0. We noticed a few differences between the two genotypes, particularly concerning the activity of monodehydroascorbate reductase and glutathione reductase relatively to POD0. The expression profiles of genes coding for the dehydroascorbate reductase and glutathione reductase isoforms differed, probably in link with the putative localisation of ROS production in response to water deficit and ozone, respectively. Our result would argue for a major role of MDHAR, GR and glutathione in the preservation of the redox status. (C) 2018 Elsevier B.V. All rights reserved.

Element content and expression of genes of interest on single cell types, such as stomata, provide valuable insights into their specific physiology, improving our understanding of leaf gas exchange regulation. We investigated how far differences in stomatal conductance (g(s)) can be ascribed to changes in guard cells functioning in amphistomateous leaves. g(s) was measured during the day on both leaf sides, on well-watered and drought-stressed trees (two Populus euramericana Moench and two Populus nigra L. genotypes). In parallel, guard cells were dissected for element content and gene expressions analyses. Both were strongly arranged according to genotype, and drought had the lowest impact overall. Normalizing the data by genotype highlighted a structure on the basis of leaf sides and time of day both for element content and gene expression. Guard cells magnesium, phosphorus, and chlorine were the most abundant on the abaxial side in the morning, where g(s) was at the highest. In contrast, genes encoding H+-ATPase and aquaporins were usually more abundant in the afternoon, whereas genes encoding Ca2+-vacuolar antiporters, K+ channels, and ABA-related genes were in general more abundant on the adaxial side. Our work highlights the unique physiology of each leaf side and their analogous rhythmicity through the day.

The impact of ozone (O3) pollution events on the plant drought response needs special attention because spring O3 episodes are often followed by summer drought. By causing stomatal sluggishness, O3 could affect the stomatal dynamic during a subsequent drought event. In this context, we studied the impact of O3 exposure and water deficit (in the presence or in the absence of O3 episode) on the stomatal closure/opening mechanisms relative to irradiance or vapour pressure deficit (VPD) variation. Two genotypes of Populus nigra x deltoides were exposed to various treatments for 21 days. Saplings were exposed to 80 ppb/day O3 for 13 days, and then to moderate drought for 7 days. The curves of the stomatal response to irradiance and VPD changes were determined after 13 days of O3 exposure, and after 21 days in the case of subsequent water deficit, and then fitted using a sigmoidal model. The main responses under O3 exposure were stomatal closure and sluggishness, but the two genotypes showed contrasting responses. During stomatal closure induced by a change in irradiance, closure was slower for both genotypes. Nonetheless, the genotypes differed in stomatal opening under light. Carpaccio stomata opened more slowly than control stomata, whereas Robusta stomata tended to open faster. These effects could be of particular interest, as stomatal impairment was still present after O3 exposure and could result from imperfect recovery. Under water deficit alone, we observed slower stomatal closure in response to VPD and irradiance, but faster stomatal opening in response to irradiance, more marked in Carpaccio. Under the combined treatment, most of the parameters showed antagonistic responses. Our results highlight that it is important to take genotype-specific responses and interactive stress cross-talk into account to improve the prediction of stomatal conductance in response to various environmental modifications.

International audience; Recent findings were able to show significant variability of stomatal dynamics between species, but not much is known about factors influencing stomatal dynamics and its consequences on biomass production, transpiration and water-use efficiency (WUE). We assessed the dynamics of stomatal conductance (g(s)) to a change of irradiance or vapour-pressure deficit (VPD) in two Populus euramericana and two Populus nigra genotypes grown under control and drought conditions. Our objectives were to determine the diversity of stomatal dynamics among poplar genotypes, and if soil water deficit can alter it. Physiological and morphological factors were investigated to find their potential links with stomatal morphology, WUE and its components at the whole-plant level. We found significant genotypic variability of g(s) dynamics to both irradiance and VPD. Genotypes with faster stomatal dynamics were correlated with higher stomatal density and smaller stomata, and the implications of these correlations are discussed. Drought slowed g(s) dynamics, depending on genotype and especially during stomatal closing. This finding is contrary to previous research on more drought-tolerant species. Independently of the treatment, faster stomatal dynamics were negatively correlated with daily whole-plant transpiration, presenting new evidence of a previously hypothesized contribution of stomatal dynamics to whole-plant water use.

International audience; The adaptive capacity of long-lived organisms such as trees to the predicted climate changes, including severe and successive drought episodes, will depend on the presence of genetic diversity and phenotypic plasticity. Here, the involvement of epigenetic mechanisms in phenotypic plasticity toward soil water availability was examined in Populus×euramericana. This work aimed at characterizing (i) the transcriptome plasticity, (ii) the genome-wide plasticity of DNA methylation, and (iii) the function of genes affected by a drought-rewatering cycle in the shoot apical meristem. Using microarray chips, differentially expressed genes (DEGs) and differentially methylated regions (DMRs) were identified for each water regime. The rewatering condition was associated with the highest variations of both gene expression and DNA methylation. Changes in methylation were observed particularly in the body of expressed genes and to a lesser extent in transposable elements. Together, DEGs and DMRs were significantly enriched in genes related to phytohormone metabolism or signaling pathways. Altogether, shoot apical meristem responses to changes in water availability involved coordinated variations in DNA methylation, as well as in gene expression, with a specific targeting of genes involved in hormone pathways, a factor that may enable phenotypic plasticity.

Ozone induces stomatal sluggishness, which impacts photosynthesis and transpiration. Stomatal responses to variation of environmental parameters are slowed and reduced by ozone and may be linked to difference of ozone sensitivity. Here we determine the ozone effects on stomatal conductance of each leaf surface. Potential causes of this sluggish movement, such as ultrastructural or ionic fluxes modification, were studied independently on both leaf surfaces of three Euramerican poplar genotypes differing in ozone sensitivity and in stomatal behaviour. The element contents in guard cells were linked to the gene expression of ion channels and transporters involved in stomatal movements, directly in microdissected stomata. In response to ozone, we found a decrease in the stomatal conductance of the leaf adaxial surface correlated with high calcium content in guard cells compared with a slight decrease on the abaxial surface. No ultrastructural modifications of stomata were shown except an increase in the number of mitochondria. The expression of vacuolar H(+) /Ca(2+) -antiports (CAX1 and CAX3 homologs), β-carbonic anhydrases (βCA1 and βCA4) and proton H(+) -ATPase (AHA11) genes was strongly decreased under ozone treatment. The sensitive genotype characterized by constitutive slow stomatal response was also characterized by constitutive low expression of genes encoding vacuolar H(+) /Ca(2+) -antiports.

Wood is a renewable resource that can be employed for the production of second generation biofuels by enzymatic saccharification and subsequent fermentation. Knowledge on how the saccharification potential is affected by genotype-related variation of wood traits and drought is scarce. Here, we used three Populus nigra L. genotypes from habitats differing in water availability to (i) investigate the relationships between wood anatomy, lignin content and saccharification and (ii) identify genes and co-expressed gene clusters related to genotype and drought-induced variation in wood traits and saccharification potential. The three poplar genotypes differed in wood anatomy, lignin content and saccharification potential. Drought resulted in reduced cambial activity, decreased vessel and fiber lumina, and increased the saccharification potential. The saccharification potential was unrelated to lignin content as well as to most wood anatomical traits. RNA sequencing of the developing xylem revealed that 1.5% of the analyzed genes were differentially expressed in response to drought, while 67% differed among the genotypes. Weighted gene correlation network analysis identified modules of co-expressed genes correlated with saccharification potential. These modules were enriched in gene ontology terms related to cell wall polysaccharide biosynthesis and modification and vesicle transport, but not to lignin biosynthesis. Among the most strongly saccharification-correlated genes, those with regulatory functions, especially kinases, were prominent. We further identified transcription factors whose transcript abundances differed among genotypes, and which were co-regulated with genes for biosynthesis and modifications of hemicelluloses and pectin. Overall, our study suggests that the regulation of pectin and hemicellulose metabolism is a promising target for improving wood quality of second generation bioenergy crops. The causal relationship of the identified genes and pathways with saccharification potential needs to be validated in further experiments.

Particulate matter (PM) deposited on Platanus acerifolia tree leaves has been sampled in the urban areas of 28 European cities, over 20 countries, with the aim of testing leaf deposited particles as indicator of atmospheric PM concentration and composition. Leaves have been collected close to streets characterized by heavy traffic and within urban parks. Leaf surface density, dimensions, and elemental composition of leaf deposited particles have been compared with leaf magnetic content, and discussed in connection with air quality data. The PM quantity and size were mainly dependent on the regional background concentration of particles, while the percentage of iron-based particles emerged as a clear marker of traffic-related pollution in most of the sites. This indicates that Platanus acerifolia is highly suitable to be used in atmospheric PM monitoring studies and that morphological and elemental characteristics of leaf deposited particles, joined with the leaf magnetic content, may successfully allow urban PM source apportionment.

(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.

In an increasingly urbanized world, air pollution mitigation is considered one of most important issues in city planning. Urban trees help to improve air quality by facilitating widespread deposition of various gases and particles through the provision of large surface areas as well as through their influence on microclimate and air turbulence. However, many of these trees produce wind-dispersed pollen (a known allergen) and emit a range of gaseous substances that take part in photochemical reactions - all of which can negatively affect air quality. The degree to which these air-quality impacts are manifested depends on species-specific tree properties: that is, their traits. We summarize and discuss the current knowledge on how such traits affect urban air pollution. We also present aggregated traits of some of the most common tree species in Europe, which can be used as a decision-support tool for city planning and for improving urban air-quality models.

The responses of Populus euphratica Oliv. plants to soil water deficit were assessed by analyzing gene expression, protein profiles, and several plant performance criteria to understand the acclimation of plants to soil water deficit. Young, vegetatively propagated plants originating from an arid, saline field site were submitted to a gradually increasing water deficit for 4 weeks in a greenhouse and were allowed to recover for 10 d after full reirrigation. Time-dependent changes and intensity of the perturbations induced in shoot and root growth, xylem anatomy, gas exchange, and water status were recorded. The expression profiles of approximately 6,340 genes and of proteins and metabolites (pigments, soluble carbohydrates, and oxidative compounds) were also recorded in mature leaves and in roots (gene expression only) at four stress levels and after recovery. Drought successively induced shoot growth cessation, stomatal closure, moderate increases in oxidative stress-related compounds, loss of CO2 assimilation, and root growth reduction. These effects were almost fully reversible, indicating that acclimation was dominant over injury. The physiological responses were paralleled by fully reversible transcriptional changes, including only 1.5% of the genes on the array. Protein profiles displayed greater changes than transcript levels. Among the identified proteins for which expressed sequence tags were present on the array, no correlation was found between transcript and protein abundance. Acclimation to water deficit involves the regulation of different networks of genes in roots and shoots. Such diverse requirements for protecting and maintaining the function of different plant organs may render plant engineering or breeding toward improved drought tolerance more complex than previously anticipated.

International audience; Ozone exposure- and dose-response relationships based on photosynthetic leaf traits (CO2 assimilation, chlorophyll content, Rubisco and PEPc activities) were established for wheat, maize and poplar plants grown in identical controlled conditions, providing a comparison between crop and tree species, as well as between C3 and C4 plants. Intra-specific variability was addressed by comparing two wheat cultivars with contrasting ozone tolerance. Depending on plant models and ozone levels, first-order, second-order and segmented linear regression models were used to derive ozone response functions. Overall, flux-based functions appeared superior to exposure-based functions in describing the data, but the improvement remained modest. The best fit was obtained using the POD0.5 for maize and POD3 for poplar. The POD6 appeared relevant for wheat, although intervarietal differences were found. Our results suggest that taking into account the dynamics of leaf antioxidant capacity could improve current methods for ozone risk assessment for plants.

Stomata, the microscopic pores on the surface of the aerial parts of plants, are bordered by two specialized cells, known as guard cells, which control the stomatal aperture according to endogenous and environmental signals. Like most movements occurring in plants, the opening and closing of stomata are based on hydraulic forces. During opening, the activation of plasma membrane and tonoplast transporters results in solute accumulation in the guard cells. To re-establish the perturbed osmotic equilibrium, water follows the solutes into the cells, leading to their swelling. Numerous studies have contributed to the understanding of the mechanism and regulation of stomatal movements. However, despite the importance of transmembrane water flow during this process, only a few studies have provided evidence for the involvement of water channels, called aquaporins. Here, we microdissected Zea mays stomatal complexes and showed that members of the aquaporin plasma membrane intrinsic protein (PIP) subfamily are expressed in these complexes and that their mRNA expression generally follows a diurnal pattern. The substrate specificity of two of the expressed ZmPIPs, ZmPIP1;5 and ZmPIP1;6, was investigated by heterologous expression in Xenopus oocytes and yeast cells. Our data show that both isoforms facilitate transmembrane water diffusion in the presence of the ZmPIP2;1 isoform. In addition, both display CO2 permeability comparable to that of the CO2 diffusion facilitator NtAQP1. These data indicate that ZmPIPs may have various physiological roles in stomatal complexes.

Oxidative stress caused by ozone (O-3) affects plant development, but the roles of specific redox-homeostatic enzymes in O-3 responses are still unclear. While growth day length may affect oxidative stress outcomes, the potential influence of day length context on equal-time exposures to O-3 is not known. In ArabidopsisCol-0, day length affected the outcome of O-3 exposure. In short-days (SD), few lesions were elicited by treatments that caused extensive lesions in long days (LD). Lesion formation was not associated with significant perturbation of glutathione, ascorbate, NADP(H) or NAD(H). To investigate roles of two genes potentially underpinning this redox stability, O-3 responses of mutants for cytosolic NADP-isocitrate dehydrogenase (icdh) and glutathione reductase 1 (gr1) were analysed. Loss of ICDH function did not affect O-3-induced lesions, but slightly increased glutathione oxidation, induction of other cytosolic NADPH-producing enzymes and pathogenesis-related gene 1 (PR1). In gr1, O-3-triggered lesions, salicylic acid accumulation, and induction of PR1 were all decreased relative to Col-0 despite enhanced accumulation of glutathione. Thus, even at identical irradiance and equal-time exposures, day length strongly influences phenotypes triggered by oxidants of atmospheric origin, while in addition to its antioxidant function, the GR-glutathione system seems to play novel signalling roles during O-3 exposure.

SummaryPlant water loss and CO2 uptake are controlled by valve-like structures on the leaf surface known as stomata. Stomatal aperture is regulated by hormonal and environmental signals. We show here that stomatal sensitivity to the drought hormone abscisic acid (ABA) is acquired during leaf development by exposure to an increasingly dryer atmosphere in the rosette plant Arabidopsis. Young leaves, which develop in the center of the rosette, do not close in response to ABA. As the leaves increase in size, they are naturally exposed to increasingly dry air as a consequence of the spatial arrangement of the leaves, and this triggers the acquisition of ABA sensitivity. Interestingly, stomatal ABA sensitivity in young leaves is rapidly restored upon water stress. These findings shed new light on how plant architecture and stomatal physiology have coevolved to optimize carbon gain against water loss in stressing environments.

Cell capacity for cytosolic NADPH regeneration by NADP-dehydrogenases was investigated in the leaves of two hybrid poplar (Populus deltoides × Populus nigra) genotypes in response to ozone (O3 ) treatment (120 ppb for 17 days). Two genotypes with differential O3 sensitivity were selected, based on visual symptoms and fallen leaves: Robusta (sensitive) and Carpaccio (tolerant). The estimated O3 flux (POD0 ), that entered the leaves, was similar for the two genotypes throughout the treatment. In response to that foliar O3 flux, CO2 assimilation was inhibited to the same extent for the two genotypes, which could be explained by a decrease in Rubisco (EC 4.1.1.39) activity. Conversely, an increase in PEPC (EC 4.1.1.31) activity was observed, together with the activation of certain cytosolic NADP-dehydrogenases above their constitutive level, i.e. NADP-G6PDH (EC 1.1.1.49), NADP-ME (malic enzyme) (EC 1.1.1.40) and NADP-ICDH (NADP-isocitrate dehydrogenase) (EC1.1.1.42). However, the activity of non-phosphorylating NADP-GAPDH (EC 1.2.1.9) remained unchanged. From the 11th fumigation day, NADP-G6PDH and NADP-ME profiles made it possible to differentiate between the two genotypes, with a higher activity in Carpaccio than in Robusta. At the same time, Carpaccio was able to maintain high levels of NADPH in the cells, while NADPH levels decreased in Robusta O3 -treated leaves. All these results support the hypothesis that the capacity for cells to regenerate the reducing power, especially the cytosolic NADPH pool, contributes to improve tolerance to high ozone exposure.

Norway spruce and red oak trees were planted directly into the soil and enclosed in open-top chambers. For 2 years the trees were exposed to both ambient and elevated CO2 concentrations (700 μmol mol-1) and during this time variations in nutrient concentrations were studied. CO2-treated plants had decreases in global leaf concentrations of nitrogen, potassium, calcium and manganese for both species. When different areas of the foliage were analysed however, the response showed much variability between the respective sites and between species. Furthermore the nutrient concentrations changed differently as the plant material aged and this change showed inter-treatment differences. These results show how it may be important to analyse plant material of different ages and at different cell sites when studying nutrient levels.

SUMMARY Norway spruce and red oak trees were planted directly into the soil and exposed to 700 ,tmol mol' CO2 in opentop chambers. There were large inter-specific differences in response to naturally occurring drought during the second year of exposure to elevated CO Both species had decreased assimilation rates. CO2-treated red oak had no loss of photosynthetic enhancement when undroughted, whereas C02-treated Norway spruce showed a relative increase in assimilation rates only when droughted. The effect of CO2 on radial growth of both species was less marked in the second growing season, but this may have been a result of different biomass partitioning as Norway spruce shoot extension had a different pattern of growth in elevated CO2* Stomatal density and chlorophyll content were largely unaffected by the CO2 treatment. A precise method for measuring Norway spruce needle surface area was also developed.

absent

Tests were carried out to determine whether variations in the hydraulic architecture of eight Populus deltoides×Populus nigra genotypes could be related to variations in leaf function and growth performance. Measurements were performed in a coppice plantation on 1-year-old shoots under optimal irrigation. Hydraulic architecture was characterized through estimates of hydraulic efficiency (the ratio of conducting sapwood area to leaf area, A(X):A(L); leaf- and xylem-specific hydraulic conductance of defoliated shoots, k(SL) and k(SS), respectively; apparent whole-plant leaf-specific hydraulic conductance, k(plant)) and xylem safety (water potential inducing 50% loss in hydraulic conductance). The eight genotypes spanned a significant range of k(SL) from 2.63 kg s(-1) m(-2) MPa(-1) to 4.18 kg s(-1) m(-2) MPa(-1), variations being mostly driven by k(SS) rather than A(X):A(L). There was a strong trade-off between hydraulic efficiency and xylem safety. Values of k(SL) correlated positively with k(plant), indicating that high-pressure flowmeter (HPFM) measurements of stem hydraulic efficiency accurately reflected whole-plant water transport efficiency of field-grown plants at maximum transpiration rate. No clear relationship could be found between hydraulic efficiency and either net CO(2) assimilation rates, water-use efficiency estimates (intrinsic water-use efficiency and carbon isotope discrimination against (13)C), or stomatal characteristics (stomatal density and stomatal pore area index). Estimates of hydraulic efficiency were negatively associated with relative growth rate. This unusual pattern, combined with the trade-off observed between hydraulic efficiency and xylem safety, provides the rationale for the positive link already reported between relative growth rate and xylem safety among the same eight P. deltoides×P. nigra genotypes.

International audience; Genetic differences in delta C-13 (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 delta C-13 as recorded in annual tree rings from different genotypes growing at three sites in southwestern France and felled at similar to 15-17 years. Wood cores were cut from tree discs to record the time course of annual basal area increment (BAI). The isotopic ratio delta C-13 was recorded in bulk wood and in extracted cellulose from the annual rings corresponding to the period 1996-2005. Discrimination against C-13 between atmosphere and tissues (delta C-13) was computed by taking into account the inter-annual time course of delta C-13 in the atmosphere. Annual BAI increased steadily and stabilized at about 8 years. An offset in delta C-13 of similar to 1 parts per thousand 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 delta C-13 recorded from the cellulose fraction. Absolute values as well as the genotype ranking of delta C-13 remained stable with age in the three sites. Genotype means of delta C-13 were not correlated to annual BAI. We conclude that genotypic differences of delta C-13 occur in older poplar trees in plantations, and that the differences as well as the genotype ranking remain stable while trees age until harvest.

BackgroundComparative genomics has emerged as a promising means of unravelling the molecular networks underlying complex traits such as drought tolerance. Here we assess the genotype-dependent component of the drought-induced transcriptome response in two poplar genotypes differing in drought tolerance. Drought-induced responses were analysed in leaves and root apices and were compared with available transcriptome data from otherPopulusspecies.ResultsUsing a multi-species designed microarray, a genomic DNA-based selection of probesets provided an unambiguous between-genotype comparison. Analyses of functional group enrichment enabled the extraction of processes physiologically relevant to drought response. The drought-driven changes in gene expression occurring in root apices were consistent across treatments and genotypes. For mature leaves, the transcriptome response varied weakly but in accordance with the duration of water deficit. A differential clustering algorithm revealed similar and divergent gene co-expression patterns among the two genotypes. Since moderate stress levels induced similar physiological responses in both genotypes, the genotype-dependent transcriptional responses could be considered as intrinsic divergences in genome functioning. Our meta-analysis detected several candidate genes and processes that are differentially regulated in root and leaf, potentially under developmental control, and preferentially involved in early and long-term responses to drought.ConclusionsIn poplar, the well-known drought-induced activation of sensing and signalling cascades was specific to the early response in leaves but was found to be general in root apices. Comparing our results to what is known in arabidopsis, we found that transcriptional remodelling included signalling and a response to energy deficit in roots in parallel with transcriptional indices of hampered assimilation in leaves, particularly in the drought-sensitive poplar genotype.

International audience; Liming is a forest practice used to counteract forest decline induced by soil acidification. It consists of direct Ca and Mg input in forest soil and restores tree mineral nutrition, but also causes drastic changes in nutrient availability in soil. Ectomycorrhizal (ECM) fungi significantly contribute in nutrient uptake by trees, and can recover them through organic acid secretion or through enzymatic degradation of organic matter. The symbiotic fungi use their extraradical mycelium for nutrient uptake, and then store them into the ECM mantle. In this study we measured how liming influences element contents in the mantle of Lactarius subdulcis ECMs, an abundant and particularly active in oxalate and laccase secretion in beech stands. For this purpose we used SEM observation coupled with energy- (EDX) and wavelength-dispersive-X-ray microanalyses (WDX). Results showed that ECM mantles of this species presented significantly higher Ca, Mg, Mn, K, Si, Al and Fe contents in limed plots. The nutrient amounts of L. subdulcis ECMs were significantly different between individuals for all the elements, showing a differential storage ability between individuals. The storage role of the ECM mantle can be interpreted in two different ways: i) a detoxification role for Al or heavy metals and ii) an increased potential nutrient resource by the fungus, which can benefit the tree.

We examined the relationships between xylem resistance to cavitation and 16 structural and functional traits across eight unrelated Populus deltoides x Populus nigra genotypes grown under two contrasting water regimes. The xylem water potential inducing 50% loss of hydraulic conductance (Psi(50)) varied from -1.60 to -2.40 MPa. Drought-acclimated trees displayed a safer xylem, although the extent of the response was largely genotype dependant, with Psi(50) being decreased by as far as 0.60 MPa. At the tissue level, there was no clear relationship between xylem safety and either xylem water transport efficiency or xylem biomechanics; the only structural trait to be strongly associated with Psi(50) was the double vessel wall thickness, genotypes exhibiting a thicker double wall being more resistant. At the leaf level, increased cavitation resistance was associated with decreased stomatal conductance, while no relationship could be identified with traits associated with carbon uptake or bulk leaf carbon isotope discrimination, a surrogate of intrinsic water-use efficiency. At the whole-plant level, increased safety was associated with higher shoot growth potential under well-irrigated regime only. We conclude that common trade-offs between xylem resistance to cavitation and other physiological traits that are observed across species may not necessarily hold true at narrower scales.

Several reports on annual plants have already shown the involvement of epigenetic modifiers such as DNA methylation in their adaptation to abiotic stresses. Nevertheless, the genotypic variations of epigenetic modifiers, their possible correlations with morphological traits and the impact of water deficit have not been described for perennial plants. Six genotypes of Populus deltoides × P. nigra were subjected or not to a moderate water deficit treatment. Various morphological traits such as the height of the plants, their biomass and the total leaf area were measured to characterize the productivity in both conditions. Levels of DNA methylation, histone acetylation and the activities and isoform accumulation of the corresponding enzymes were measured at the shoot apex, the site of morphogenesis. Genotypic variation was observed for the morphological traits and the epigenetic variables and correlations were established among them. Genotypic variation for DNA methylation was detected in hybrid poplars. A positive correlation was demonstrated between DNA methylation percentage and productivity under well watered conditions. While there was a general decrease of growth for all genotypes in response to a moderate water deficit, genotypic dependant variations of DNA methylation were found suggesting different strategies among hybrids., Plusieurs études sur des plantes annuelles ont déjà montré l’implication des modifications épigénétiques telles que la méthylation de l’ADN dans la plasticité de leurs réponses aux contraintes abiotiques. Néanmoins, les variations génotypiques de ces modifications épigénétiques, leur possible corrélation avec des variables de croissance et l’impact d’un déficit hydrique n’ont pas été décrits sur une plante pérenne. Six génotypes de Populus deltoïdes × P. nigra ont été soumis ou non à un déficit hydrique modéré et plusieurs variables de croissance ont été mesurées afin de caractériser leur productivité. Les niveaux de méthylation de l’ADN, d’acétylation des histones, les activités enzymatiques et l’accumulation des isoformes correspondantes ont été mesurés sur des apex caulinaires, site de la morphogenèse. Des variations génotypiques ont été observées pour les variables de croissance et épigénétiques. Une corrélation positive a été mise en évidence entre la méthylation de l’ADN et la productivité en condition hydrique favorable. Bien qu’il y ait une diminution générale de la croissance de tous les génotypes en réponse à un déficit hydrique modéré, des variations génotype-dépendant de la méthylation de l’ADN ont été trouvées suggérant différentes stratégies entre hybrides.

Young poplar trees (Populus tremula Michx. x Populus alba L. clone INRA 717-1B4) were subjected to 120 ppb of ozone for 35 days in phytotronic chambers. Treated trees displayed precocious leaf senescence and visible symptoms of injury (dark brown/black upper surface stippling) exclusively observed on fully expanded leaves. In these leaves, ozone reduced parameters related to photochemistry (Chl content and maximum rate of photosynthetic electron transport) and photosynthetic CO 2 fixation [net CO 2 assimilation, Rubisco (ribulose-1,5-bisphosphate carboxylase oxygenase) activity and maximum velocity of Rubisco for carboxylation]. In fully expanded leaves, the rate of photorespiration as estimated from Chl fluorescence was markedly impaired by the ozone treatment together with the activity of photorespiratory enzymes (Rubisco and glycolate oxidase). Immunoblot analysis revealed a decrease in the content of serine hydroxymethyltransferase in treated mature leaves, while the content of the H subunit of the glycine decarboxylase complex was not modified. Leaves in the early period of expansion were exempt from visible symptoms of injury and remained unaffected as regards all measured parameters. Leaves reaching full expansion under ozone exposure showed potential responses of protection (stimulation of mitochondrial respiration and transitory stomatal closure). Our data underline the major role of leaf phenology in ozone sensitivity of photosynthetic processes and reveal a marked ozone-induced inhibition of photorespiration.

Genetic variation for intrinsic water use efficiency (W i) and related traits was estimated in a full-sib family of Quercus robur L. over 3 years. The genetic linkage map available for this F1 family was used to locate quantitative trait loci (QTL) for W i, as estimated by leaf carbon stable isotope composition (δ 13C) or the ratio of net CO2 assimilation rate (A) to stomatal conductance to water vapour (g w) and related leaf traits. Gas exchange measurements were used to standardize estimates of A and g w and to model the sensitivity of g w to leaf-to-air vapour pressure deficit (sgVPD). δ 13C varied by more than 3‰ among the siblings, which is equivalent to 40% variation of W i. Most of the studied traits exhibited high clonal mean repeatabilities (>50%; proportion of clonal mean variability in global variance). Repeatabilities for δ 13C, leaf mass per area (LMA) and leaf nitrogen content were higher than 70%. For δ 13C, ten QTLs were detected, one of which was detected repeatedly for all 3 years and consistently explained more than 20% of measured variance. Four genomic regions were found in which co-localizing traits linked variation in W i to variations in leaf chlorophyll and nitrogen content, LMA and sgVPD. A positive correlation using clonal means between δ 13C and A/g w, as well as a co-localisation of QTL detected for both traits, can be seen as validation of the theoretical model linking the genetic architecture of these two traits.

SEM and light-microscopical observations, supported by chemical microanalysis with an EDXA system, revealed that light-saturated pixels observed in X-ray negatives of sessile oak (Quercus petraea Liebl.) wood were caused by inorganic deposits present inside multiseriate ray and axial parenchyma cells. Calcium oxalate crystals, silica grains and amorphous granules with varied mineral compositions have been identified. The wood strips of three out of six sampled trees contained measurable amounts of mineral inclusions which were quantified using image analysis. Based on the variations of mineral content observed between trees and within and between annual rings of the same tree, some hypotheses were formulated concerning the factors involved in the formation of inorganic deposits in oak wood. Their occurrence varies depending on the mineral concerned and seems to be controlled largely by a tree effect. The time of formation appears to coincide with a shifting of the oak wood’s functions as a result of heartwood formation processes (inter-annual scale) or changes in leaf phenology and climate (intra-annual scale). In addition, the technical consequences of their presence as well as their effects on wood density measurements through microdensitometry are discussed, Composition, distribution et origine supposée d’inclusions minérales dans le bois de chêne sessile – conséquences pour l’analyse microdensitométrique. Des observations à l’aide de microscopes électronique à balayage et optique, appuyées par des analyses élémentaires au moyen d’un système EDXA, ont révélé que les points-images saturés en niveau de gris, constatés dans les négatifs de radiographies de bois de chêne sessile (Quercus petraea Liebl.), étaient dus à des dépôts inorganiques présents à l’intérieur des cellules parenchymateuses des rayons ligneux multisériés et du parenchyme axial. Des cristaux d’oxalate de calcium, des grains de silice et des granules amorphes ayant des compositions minérales variées ont été identifiés. Des barrettes de trois sur six arbres échantillonnés contenaient des quantités mesurables d’inclusions minérales qui ont été quantifiées par analyse d’images. En se basant sur les variations du contenu en dépôts minéraux observées entre arbres ainsi qu’à l’intérieur d’un arbre, aux niveaux intra- et intercerne, quelques hypothèses ont été avancées concernant les facteurs potentiellement responsables de la formation de dépôts minéraux chez le chêne. Leur présence dépend du minéral concerné et semble être contrôlée par un fort effet arbre. L’apparition des minéraux coïncide apparemment avec des changements de fonctions du bois relatifs aux processus de duraminisation (échelle interannuelle) ou correspondant à des évolutions phénologiques ou climatiques (échelle intra-annuelle). Enfin, les conséquences techniques ainsi que les effets de leur présence sur des mesures de la densité du bois par analyse microdensitométrique sont discutés

To understand key processes governing defense mechanisms in poplar (Populus spp.) upon infection with the rust fungus Melampsora larici-populina, we used combined histological and molecular techniques to describe the infection of Populus trichocarpa × Populus deltoides ‘Beaupré’ leaves by compatible and incompatible fungal strains. Striking differences in host-tissue infection were observed after 48-h postinoculation (hpi) between compatible and incompatible interactions. No reactive oxygen species production could be detected at infection sites, while a strong accumulation of monolignols occurred in the incompatible interaction after 48 hpi, indicating a late plant response once the fungus already penetrated host cells to form haustorial infection structures. P. trichocarpa whole-genome expression oligoarrays and sequencing of cDNAs were used to determine changes in gene expression in both interactions at 48 hpi. Temporal expression profiling of infection-regulated transcripts was further compared by cDNA arrays and reverse transcription-quantitative polymerase chain reaction. Among 1,730 significantly differentially expressed transcripts in the incompatible interaction, 150 showed an increase in concentration ≥3-fold, whereas 62 were decreased by ≥3-fold. Regulated transcripts corresponded to known genes targeted by R genes in plant pathosystems, such as inositol-3-P synthase, glutathione S-transferases, and pathogenesis-related proteins. However, the transcript showing the highest rust-induced up-regulation encodes a putative secreted protein with no known function. In contrast, only a few transcripts showed an altered expression in the compatible interaction, suggesting a delay in defense response between incompatible and compatible interactions in poplar. This comprehensive analysis of early molecular responses of poplar to M. larici-populina infection identified key genes that likely contain the fungus proliferation in planta.

International audience; • We examined the relationships among roductivity, water use efficiency (WUE) and drought tolerance in 29 genotypes of Populus × euramericana (Populus deltoides × Populus nigra), and investigated whether some leaf traits could be used as predictors for productivity, WUE and drought tolerance. • At Orléans, France, drought was induced on one field plot by withholding water, while a second plot remained irrigated and was used as a control. Recorded variables included stem traits (e.g. biomass) and leaf structural (e.g. leaf area) and functional traits [e.g. intrinsic water use efficiency (Wi) and carbon isotope discrimination (∆)]. • Productivity and ∆ displayed large genotypic variability and were not correlated. ∆ scaled negatively with Wi and positively with stomatal conductance under moderate drought, suggesting that the diversity for ∆ was mainly driven by stomatal conductance. • Most of the productive genotypes displayed a low level of drought tolerance (i.e. a large reduction of biomass), while the less productive genotypes presented a large range of drought tolerance. The ability to increase WUE in response to water deficit was necessary but not sufficient to explain the genotypic diversity of drought tolerance.

http://www.elsevier.com/locate/issn/09819428 The effects of ambient and elevated ozone (O3) levels on photosynthesis, growth, pigment, biomass and element contents of Aleppo pine (Pinus halepensis Mill.) were studied for two growing seasons (1997, 1998). Two-year-old seedlings were exposed to elevated O3 in open-top chambers. The treatments were charcoal-filtered air and non-filtered air + 50 nl l–1 O3 (24 h per day, 7 days per week). In summer 1998, half of the seedlings were drought-stressed (leaf water potential down to approximately –2 MPa), while the other half were kept well-watered. At the beginning of the season (1998), current (c) and previous-year (c + 1) needles under O3 stress showed an increase in stomatal conductance and net photosynthesis. During the drought period, only stomatal conductance increased in both needle age-classes, whereas the net photosynthesis decreased. At the end of the measuring period, both parameters were reduced in the O3 treatment. Both O3 and drought decreased chlorophyll a and b concentrations, growth and biomass.A carry-over effect of O3 on pigments was also observed. Needle K content was increased in the O3 treatment. Drought protected Aleppo pine against O3 (less chlorotic mottle and less decrease of stem and branch biomass).

Pinus cembra (L.) trees growing at an elevation of 1,800 m in the Mercantour Alps National Park in southern France have shown various degrees of damage (yellowing and needle loss) since the early 1980s. Two stands were selected, one populated by healthy and the other by damaged trees, and 1-year-old needles were analysed. The needles of the declining trees displayed chlorotic mottled patterns associated with a significantly lower level of total chlorophyll compared to healthy trees. Measurements of gas exchange showed a lower photosynthetic rate linked to a lower activity of ribulose bisphosphate carboxylase and a higher stomatal conductance in needles of declining trees compared to healthy trees. In contrast, phosphoenolpyruvate carboxylase activity was found to be higher in declining tree needles, reaching twice the level measured in healthy tree needles. A significant oxidative stress was also observed in the needles which presented a 68% higher malondialdehyde content than healthy tree needles. The atmospheric concentration of ozone was quite high (40 nl·l–1, monthly mean in summer) in both stands but preliminary results based on stomatal conductance and ozone concentration measurements suggested a higher ozone flux in needles of declining trees. Ozone is thus suspected as a contributing factor in the observed decline of Pinus cembra trees.

International audience; The effects of 700 μmol mol−1 CO2 and 200 nmol mol−1 ozone on photosynthesis in Pinus halepensis seedlings and on N translocation from its mycorrhizal symbiont, Paxillus involutus, were studied under nutrient-poor conditions. After 79 days of exposure, ozone reduced and elevated CO2 increased net assimilation rate. However, the effect was dependent on daily accumulated exposure. No statistically significant differences in total plant mass accumulation were observed, although ozone-treated plants tended to be smaller. Changes in atmospheric gas concentrations induced changes in allocation of resources: under elevated ozone, shoots showed high priority over roots and had significantly elevated N concentrations. As a result of different shoot N concentration and net carbon assimilation rates, photosynthetic N use efficiency was significantly increased under elevated CO2 and decreased under ozone. The differences in photosynthesis were mirrored in the growth of the fungus in symbiosis with the pine seedlings. However, exposure to CO2 and ozone both reduced the symbiosis-mediated N uptake. The results suggest an increased carbon cost of symbiosis-mediated N uptake under elevated CO2, while under ozone, plant N acquisition is preferentially shifted towards increased root uptake.

Nine-year-old Norway spruce from two clones and beech grown in open-top chambers were exposed to both drought and elevated (ambient +0.05 μl l −1 ) ozone. Beech was less damaged from the effects of ozone by drought. Compared to trees exposed to elevated ozone and well watered, beech trees grown under elevated ozone and exposed to drought had lower CO 2 compensation points and were not affected by foliar necroses observed in well watered trees. Both Norway spruce clones were more resistant than beech to ozone alone, but the Istebna clone grown in drought and elevated ozone showed an increased stomatal conductance. Therefore these trees have significantly more negative pre-dawn leaf water potentials than droughted trees grown in filtered air (without ozone). The increased stomatal conductance implies a greater ozone flux to the needles. In association with this increased ozone uptake, 1-year-old needles became discoloured and were prematurely shed.

summary Two clones of eight-year old trees of Norway spruce and beech, planted directly into the soil and in open-top chambers, were exposed to elevated ozone concentrations and subjected to mild soil drought. When ambient ozone was increased by 50 ppb, the shoot extension of both Norway spruce clones was significantly reduced. The mild drought stress was begun in July 1993 and following a soil-drying associated with a spell of fine weather, the drought remained relatively constant for over a month. During this period, gas exchange parameters were regularly monitored between 7.00 and 9.00 GMT and on two occasions followed throughout the day. Both clones responded to the mild drought by reducing stomatal conductance and photosynthesis. There were no apparent interactions between drought and ozone. The reaction of beech was more complicated. Ozone caused a significant decrease in stomatal conductance and photosynthesis in well-watered treatments. All the ozone-drought stress treated trees had greater stomatal conductance than the non-drought stressed equivalents in the morning, but the situation was reversed as the day went on. As peak ozone concentrations occur in the afternoon, it is hypothesized that over a period of time the drought stressed trees are less affected by ozone as they receive a lower cumulative dose. Coupled with the low values of vapour pressure deficit occurring in the morning, the ozone-treated, drought-stressed trees can support relatively higher rates of photosynthesis than the well-watered equivalents. The chlorophyll contents of beech lent support to this hypothesis, as ozone associated reductions were less marked in drought stressed trees.

International audience; Previously, a large intra-specific diversity and a tight genetic control have been shown for Δ13C (carbon isotope discrimination) in a pedunculate oak (Quercus robur L.) family, which is an estimator for intrinsic water use efficiency (Wi), a complex trait defined as the ratio of net CO2 assimilation rate (A) to stomatal conductance for water vapour (gs).* In the present study, twelve genotypes with extreme phenotypic values of Δ13C were selected within this family to (i) asses the stability of genotype differences across contrasting environments and for different measures Wi; (ii) quantify the relationship between Δ13C and Wi within this family; (iii) identify which leaf traits drive the diversity in Wi observed in this family. * Genetic variability of Δ13C and Wi was largely independent from different temporal integration scales and their correlation was found to be strong (R2 = 88% for leaf sugars) within this family.* Weak correlations between measures of Wi with estimators of photosynthetic capacity, suggest a minor role of the latter in the diversity of Wi.* However, the tight correlation between gs and Δ13C as well as Wi, and the related genotypic variation in stomatal density, suggest that the genotypic diversity in Wi within this pedunculate oak family might be due to differences in gs.; Une large diversité intra-spécifique et un fort contrôle génétique ont été mis en évidence pour Δ13C (discrimination isotopique du carbone) dans une famille de chêne pédonculé (Quercus robur L.). Δ13C est un estimateur de l'efficience intrinsèque d'utilisation de l'eau (Wi), un caractère complexe défini comme le rapport entre l'assimilation nette de CO2 (A) et la conductance stomatique pour la vapeur d'eau (gs).* Douze génotypes présentant des valeurs phénotypiques extrêmes de Δ13C ont été sélectionnés dans cette famille pour (i) évaluer la stabilité des différences génotypiques dans des environnements contrastés et pour différents estimateurs de l'efficience d'utilisation de l'eau ; (ii) quantifier la relation entre Δ13C et Wi dans cette famille; (iii) identifier quels caractères foliaires sont impliqués dans la diversité de Wi observée dans cette famille.* La variabilité génétique de Δ13C et Wi était largement indépendante des différentes échelles d'intégration temporelles et leur corrélation était forte dans cette famille (R2 = 88% lorsque Δ13C était mesuré dans les sucres foliaires).* Les faibles corrélations entre Wi (ou Δ13C) et des estimateurs de la capacité photosynthétique suggèrent un rôle mineur de celle-ci dans la diversité de Wi.* Par contre, la corrélation étroite entre gs et Δ13C ainsi que Wi et la variation génétique de la densité stomatique, suggèrent que la diversité génétique de Wi dans cette famille de chêne pédonculé est liée à des différences de conductance stomatique gs.