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Fagus sylvatica L. (Fagaceae), European beech, is one of the most important deciduous tree species in Central Europe and the most common broadleaved tree species in Germany. We investigated the leaves of six individual Fagus sylvatica trees growing in a beech forest in Kiel, Schleswig-Holstein, Germany, for seasonal variations in the content of phenolic natural products over three consecutive growing seasons. The investigated compound classes comprised hydroxycinnamic acid and flavonoid derivatives. The content of phenolic compounds showed clear trends in all years. A sharp decline in the total content of phenolic substances was observed from mid-April to the end of May. During the summer months, the content of phenolic compounds remained stable with only slight fluctuations until fall. The values for individual trees deviated more pronouncedly from one another in spring, but converged during the course of the growing period. These trends, despite differences in absolute values, were identical in three consecutive growing seasons (2016-2018). Our results contribute to a better understanding of the dynamics of plant natural products of deciduous trees in temperate climates caused by seasonal variations.

Temperate trees could cope with climate change through phenotypic plasticity of phenological key events or adaptation in situ via selection on genetic variation. However, the relative contribution of local adaptation and phenotypic plasticity to phenological change is unclear for many ecologically important tree species. Here, we analyzed the leaf-out data of European beech (Fagus sylvatica L.) from 50 provenances planted in 7 trial sites. We first constructed a function between chilling accumulation (CA) and photoperiod-associated heat requirement (PHR) of leaf-out date for each provenance and quantified the relationship between parameters of the CA-PHR function and climatic variables at provenance origins by using the random forest model. Furthermore, we used the provenance-specific CA-PHR function to simulate future leaf-out dates under two climate change scenarios (RCP 4.5 and 8.5) and two assumptions (no adaptation and adaptation). The results showed that both CA, provenance, and their interactions affected the PHR of leaf-out. The provenances from southeastern Europe exhibited a stronger response of PHR to CA and thus flushed earlier than northwestern provenances. The parameters of the CA-PHR function were connected with climatic variables (e.g., mean diurnal temperature range, temperature seasonality) at the originating sites of each provenance. If only considering the phenotypic plasticity, the leaf-out date of European beech in 2070-2099 will advance by 6.8 and 9.0 days on average relative to 1951-2020 under RCP 4.5 and RCP 8.5, respectively. However, if F. sylvatica adapts to future climate change by adopting the current strategy, the advance of the leaf-out date will weaken by 1.4 and 3.4 days under RCP 4.5 and RCP 8.5, respectively. Our results suggest that the European beech could slow down its spring phenological advances and reduce its spring frost risk if it adopts the current strategy to adapt to future climate change.

Woody debris is found in all habitats of the land-sea continuum. While isolated experimental studies of wood degradation exist, few programs have observed the dynamics of wood degradation, in situ across this gradient. Since 2014, we have been conducting a series of long-term observations of wood decay in three characteristic areas of a Mediterranean Sea coastal watershed: forest leaf litter ('Forest'), river bed ('River') and the near-shore marine environment ('Sea'). The study sites are within the Massane River watershed (France) whose headwaters are in a protected beech tree (Fagus sylvatica) dominated forest. Branch sections from a recently fallen beech tree and standardized blocks of Norway spruce (Picea abies) were installed in all three environments. The proportion of remaining mass and volumetric mass of the individual wood samples were determined periodically over 4.2 years. Regardless of wood type, there were marked differences in the decay dynamics. Mass losses at the Forest and River sites were well-described by continuous negative exponential models. At the Sea site, there was a short latency period followed by rapid degradation for the wood fraction exploited by shipworms; in this case, a Weibull-type function was fitted to the data. Integrated mass loss rates at the coastal location were 6 to 20 times faster than in the other two environments. Our study suggests that the early dynamics of wood degradation in a land-sea meta-ecosystem are dominated by the marine invertebrate community. This means woody debris, once it reaches the sea, is likely to break down rapidly within near shore coastal habitats. These results highlight the need to quantify the mass transport dynamics between local ecosystems.

The species trait-environment relationships in understory vegetation were carried out on metalliferous sites created by historical Zn-Pb mining (S Poland), on which natural revegetation took place. The study sites were 31 small heaps of waste rock covered by an 80-120 year old beech forest. The sites were described in terms of plant coverage, species richness and composition, and the composition of plant traits. Three types of understory communities that were identified were compared for environmental variables and plant parameters. Despite a high concentration of heavy metals in soils, namely Cd, Pb, and Zn, the degree of shading by the tree canopy was the main factor determining variation in understory communities. The understories that developed in low and strong shading were similar to some extent: They had high number of species, and they were characterized by a high proportion of the ancient forest species. The species composition changed substantially with increasing coverage of trees; mixed strategy plants withdrew, while the stress tolerant species became more abundant. The other predictor of community structure was the ratio of Cd to Ca, which was responsible for the decrease of woody plants and the endangered species. This study proved that, in the case of highly complex and productive ecosystems, even with a high degree of contamination with heavy metals, the biotic factors play a primary role.

The primary objective of this study was to describe parameters related to the leaf biochemical assimilation capacity of photosynthesis, such as the maximum rates of carboxylation (V-cmax) and electron transport (J(max)), as a function of the leaf nitrogen content throughout a canopy of Siebold's beech and Japanese oak grown under elevated ozone (O-3) conditions during a growing season. To this end, we investigated the relationship between photosynthetic traits and leaf nitrogen content in various canopy positions of two tree species under free-air O-3 exposure (60 nmol mol(-1), during daylight hours) in June, August, and October 2012. We observed O-3-induced reduction in V-cmax and J(max) without reduction of leaf nitrogen content in both tree species. In Siebold's beech, V-cmax and J(max) in leaves with higher N-area were largely decreased by O-3 from August, while little effect of O-3 was observed in leaves with lower N-area. On the other hand, there was no difference in the extent of O-3-induced reduction in V-cmax and J(max) across the range of N-area in leaves of Japanese oak. Reduction of leaf nitrogen content under elevated O-3 conditions was observed only in Siebold's beech in October. These results indicated that the decrease in the efficiency of photosynthetic nitrogen use is in an earlier step in O-3-induced decline of photosynthesis in Siebold's beech and Japanese oak. Based on these results, we emphasize the importance of integration of O-3 effects into the conventional estimation of V-cmax and J(max) from leaf nitrogen content for evaluating canopy photosynthesis under current and future elevated O-3 conditions. (C) 2018 Published by Elsevier B.V.

This study aimed to simulate oak and beech forest growth under various scenarios of climate change and to evaluate how the forest response depends on site properties and particularly on stand characteristics using the individual process-based model HETEROFOR. First, this model was evaluated on a wide range of site conditions. We used data from 36 long-term forest monitoring plots to initialize, calibrate, and evaluate HETEROFOR. This evaluation showed that HETEROFOR predicts individual tree radial growth and height increment reasonably well under different growing conditions when evaluated on independent sites. In our simulations under constant CO2 concentration ([CO2]cst) for the 2071-2100 period, climate change induced a moderate net primary production (NPP) gain in continental and mountainous zones and no change in the oceanic zone. The NPP changes were negatively affected by air temperature during the vegetation period and by the annual rainfall decrease. To a lower extent, they were influenced by soil extractable water reserve and stand characteristics. These NPP changes were positively affected by longer vegetation periods and negatively by drought for beech and larger autotrophic respiration costs for oak. For both species, the NPP gain was much larger with rising CO2 concentration ([CO2]var) mainly due to the CO2 fertilisation effect. Even if the species composition and structure had a limited influence on the forest response to climate change, they explained a large part of the NPP variability (44% and 34% for [CO2]cst and [CO2]var, respectively) compared to the climate change scenario (5% and 29%) and the inter-annual climate variability (20% and 16%). This gives the forester the possibility to act on the productivity of broadleaved forests and prepare them for possible adverse effects of climate change by reinforcing their resilience.

This study investigates the functionality of a Mediterranean-mountain beech forest in Central Italy using simultaneous determinations of optical measurements, carbon (C) fluxes, leaf eco-physiological and biochemical traits during two growing seasons (2014-2015). Meteorological variables showed significant differences between the two growing seasons, highlighting a heat stress coupled with a reduced water availability in mid-summer 2015. As a result, a different C sink capacity of the forest was observed between the two years of study, due to the differences in stressful conditions and the related plant physiological status. Spectral indices related to vegetation (VIs, classified in structural, chlorophyll and carotenoid indices) were computed at top canopy level and used to track CO2 fluxes and physiological changes. Optical indices related to structure (EVI 2, RDVI, DVI and MCARI 1) were found to better track Net Ecosystem Exchange (NEE) variations for 2014, while indices related to chlorophylls (SR red edge, CL red edge, MTCI and DR) provided better results for 2015. This suggests that when environmental conditions are not limiting for forest sink capacity, structural parameters are more strictly connected to C uptake, while under stress conditions indices related to functional features (e.g., chlorophyll content) become more relevant. Chlorophyll indices calculated with red edge bands (SR red edge, NDVI red edge, DR, CL red edge) resulted to be highly correlated with leaf nitrogen content (R2 > 0.70), while weaker, although significant, correlations were found with chlorophyll content. Carotenoid indices (PRI and PSRI) were strongly correlated with both chlorophylls and carotenoids content, suggesting that these indices are good proxies of the shifting pigment composition related to changes in soil moisture, heat stress and senescence. Our work suggests the importance of integrating different methods as a successful approach to understand how changing climatic conditions in the Mediterranean mountain region will impact on forest conditions and functionality.

In this study we analysed a novel tree-growth dataset, inferred from annual ring-width measurements, of 7 forest tree species from 12 mountain regions in Greece, in order to identify tree growth - climate relationships. The tree species of interest were: Abies cephalonica, Abies borisii-regis, Picea abies, Pinus nigra, Pinus sylvestris, Fagus sylvatica and Quercus frainetto growing across a gradient of climate conditions with mean annual temperature ranging from 5.7 to 12.6°C and total annual precipitation from 500 to 950mm. In total, 344 tree cores (one per tree) were analysed across a network of 20 study sites. We found that water availability during the summer period (May-August) was a strong predictor of interannual variation in tree growth for all study species. Across species and sites, annual tree growth was positively related to summer season precipitation (PSP). The responsiveness of annual growth to PSP was tightly related to species and site specific measurements of instantaneous photosynthetic water use efficiency (WUE), suggesting that the growth of species with efficient water use is more responsive to variations in precipitation during the dry months of the year. Our findings support the importance of water availability for the growth of mountainous Mediterranean tree species and highlight that future reductions in precipitation are likely to lead to reduced tree-growth under climate change conditions.

Phosphorus is often the least available macronutrient in soil. Lack in phosphorus has detrimental effect on growth and biomass production of European Fagus sylvatica L., a major trees species in temperate forests. In contrast to leaf tissues, few studies have examined changes in the root system and no study has ever investigated the proteomic changes affected in beech roots by a lack in available phosphate (P). Here, we studied roots of young Fagus sylvatica L. trees in their native soils from two forests sites with contrasting availability of P: one P rich and P poor soil. To understand also the response to P fertilization, the trees were fertilized with triple superphosphate and the proteome of fine roots of all conditions was compared. Gel-free mass-spectrometry-based shotgun proteomics revealed that the proteome was differentially affected by diverging P availabilities. The proteomic changes that took place as the result of P fertilization were dependent on the supply level of P before the fertilization. When P was supplied to the P-rich soil proteins related to cell biogenesis exhibited increased abundances. Addition of P to soil that was strongly limited in P resulted in increased abundance of proteins associated with amino acid metabolism and transport. Biological significance Beech ( Fagus sylvatica L.) forests have a huge ecological and economic value across Europe. In recent years, however, these forest sites increasingly suffer under phosphorus (P) deficiency. As the consequence, growth and vitality of beech forests is impaired. For this reason, this study was conducted with the aim to identify and understand proteomic impairments and adjustments that evolve in the fine roots under both, a P deficiency and an amelioration thereof. For this, we analyzed (1) the fine root proteome of young beech trees grown (2) at two soil sites that contrast in their degree of availability P (low vs. high) in dependency (3) to a fertilization with P. This experiment revealed fundamental differences with respect to proteomic changes in dependency on the severity of P limitation and helped to identify processes that take place after amelioration of the deficiency. This information is useful to understand which physiological processes are impaired under P deficiency and, thus, impair growth. The fertilization experiment enabled to identify developmental processes that take place in fine roots when concentration of available P was increased. They are “cellular component organization and biogenesis” in the P rich soil and “synthesis of organonitrogen-containing compounds” in the P poor soil.

A physicochemical characteristic of the cold-pressed oil obtained from seeds of common beech (Fagus sylvatica L.) has been presented. This plant may be considered as unconventional oilseeds crops because of relatively high content of fat (27.25%). The analyzed beech seeds oil has been classified as oleic-linoleic acids oil with more than 76% percentage share of those species. Beech seeds oil contains 4.2% of gamma-linolenic acid (GLA). Unique characteristic is the high content of γ-tocopherol (75.4mg/100g) and δ-tocopherol (34.05mg/100g). γ-Tocopherol is effective scavengers of reactive nitrogen species and prevents DNA bases nitration, what makes beech seeds oil interesting raw material in the production of cosmetics. Additionally the content of carotenoids, very effective photooxidation inhibitors, is at high level in comparison with other cold-pressed oils. It was demonstrated that PCA analysis may help to determine the authenticity of oil obtained from beech seeds.

Electron beam irradiation (EBI) is an alternative treatment for intrinsic viscosity (IV) control in cellulose pulps, but has never been integrated in full bleaching sequences for comparison to conventional methods. Both eucalyptus kraft (EK) paper pulp and beech sulfite (BS) dissolving pulp were subjected to totally chlorine free (TCF) bleaching sequences comprising either EBI, ozone, or both for IV control. Additionally, effects of EBI on hexenuronic acid (HexA) and xylan were investigated. IV was adjusted to 450–500 mL g−1 and properties including carbonyl content, kappa, brightness, alkali-resistance and sugar composition were compared. Pulps produced with EBI had a higher alkali-resistance, uniformity and less cellulose oxidation. However, the degree of bleaching (DoB) was low without the use of ozone. HexA content in a birch pulp was halved by EBI. Isolated xylans were more resistant to irradiation than cellulose with little decrease of molar masses and moderate oxidation.

Climate change is increasing the frequency of extreme climate events, causing profound impacts on forest function and composition. Late frost defoliation (LFD) events, the loss of photosynthetic tissues due to low temperatures at the start of the growing season, might become more recurrent under future climate scenarios. Therefore, the detection of changes in late-frost risk in response to global change emerges as a high-priority research topic. Here, we used a tree-ring network from southern European beech (Fagus sylvatica L.) forests comprising Spain, Italy and the Austrian Alps, to assess the incidence of LFD events in the last seven decades. We fitted linear-mixed models of basal area increment using different LFD indicators considering warm spring temperatures and late-spring frosts as fixed factors. We reconstructed major LFD events since 1950, matching extreme values of LFD climatic indicators with sharp tree-ring growth reductions. The last LFD events were validated using remote sensing. Lastly, reconstructed LFD events were climatically and spatially characterized. Warm temperatures before the late-spring frost, defined by high values of growing-degree days, influenced beech growth negatively, particularly in the southernmost populations. The number of LFD events increased towards beech southern distribution edge. Spanish and the southernmost Italian beech forests experienced higher frequency of LFD events since the 1990s. Until then, LFD events were circumscribed to local scales, but since that decade, LFD events became widespread, largely affecting the whole beech southwestern distribution area. Our study, based on in-situ evidence, sheds light on the climatic factors driving LFD occurrence and illustrates how increased occurrence and spatial extension of late-spring frosts might constrain future southern European beech forests' growth and functionality. Observed alterations in the climate-phenology interactions in response to climate change represent a potential threat for temperate deciduous forests persistence in their drier/southern distribution edge.

Wood rotting basidiomycetes produce extracellular mucilaginous sheaths interfacing fungal hyphae and plant biomass. While the versatility of these fungal sheaths has been addressed, sheaths generated by selective white-rot fungi remain poorly understood. To fill this gap, the sheath produced by the basidiomycete Ceriporiopsis subvermispora, which degrades lignin while inflicting limited cellulose damage, was analyzed in this study. Fluorescence and transmission electron microscopy revealed that the sheath formed three days after inoculation into a beech wood slice on an agar plate and was embedded at the interface between fungal hyphae and wood cell walls. The sheath's chemical structure was evaluated from fungus cultures in a liquid medium containing [U-13C6]-d-glucose and beech wood slices. Compositional analysis, methylation analysis, and 13C NMR demonstrated that the sheath mainly consisted of a comb-like β-1,6-glucopyranose residue-branched β-1,3-glucan, which is advantageous to retain water and extracellular secondary metabolites.

A lactic acid (LA)-producing strain of the hyper-lignin-degrading fungus Phanerochaete sordida YK-624 with the lactate dehydrogenase-encoding gene from Bifidobacterium longum (Blldh) was constructed. When the endogenous pyruvate decarboxylase gene-knocked down and Blldh-expressing transformant was cultured with beech wood meal, the transformant was able to successively delignify and ferment the substrate. Supplementation of calcium carbonate into the culture medium, significantly increased the level of LA accumulation. Direct LA production (at 0.29g/l) from wood was confirmed, and additional inclusion of exogenous cellulase in this fermentation yielded significant further improvement in LA accumulation (up to 1.44g/l). This study provides the first report of direct production of LA by fermentation from woody biomass by a single microorganism, and indicates that transgenic white-rot fungi have a potential use for development of simple/easy applications for wood biorefinery.

The aim of this study was to analyze four conceptual beech wood based biorefineries generated during process design in terms of environmental and economic criteria. Biorefinery 1 annually converts 400,000 dry metric tons of beech wood into the primary products 41,600t/yr polymer-grade ethylene and 58,520tDM/yr organosolv lignin and the fuels 90,800tDM/yr hydrolysis lignin and 38,400t/yr biomethane. Biorefinery 2 is extended by the product of 58,400t/yr liquid "food-grade" carbon dioxide. Biorefinery 3 produces 69,600t/yr anhydrous ethanol instead of ethylene. Compared to biorefinery 3, biorefinery 4 additionally provides carbon dioxide as product. Biorefinery 3 and 4 seem most promising, since under basic assumptions both criteria, (i) economic effectiveness and (ii) reduction of potential environmental impacts, can be fulfilled. All four alternatives may reduce potential environmental impacts compared to reference systems using the ReCiPe methodology. Economic feasibilities of the analyzed biorefineries are highly sensitive.

Tension wood that is an abnormal part formed in angiosperms has been barely used for wood industry. In this study, to utilize the tension wood effectively by means of liquefaction using ionic liquid, we performed morphological and topochemical determination of the changes in tension wood of Japanese beech (Fagus crenata) during ionic liquid treatment at the cellular level using light microscopy, scanning electron microscopy and confocal Raman microscopy. Ionic liquid treatment induced cell wall swelling in tension wood. Changes in the tissue morphology treated with ionic liquids were different between normal wood and tension wood, moreover the types of ionic liquids. The ionic liquid 1-ethyl-3-methylimidazolium chloride liquefied gelatinous layers rapidly, whereas 1-ethylpyridinium bromide liquefied slowly but delignified selectively. These novel insights into the deconstruction behavior of tension wood cell walls during ionic liquid treatment provide better understanding of the liquefaction mechanism. The obtained knowledge will contribute to development of an effective chemical processing of tension wood using ionic liquids and lead to efficient use of wood resources.

Herpes simplex virus belongs to Herpesviridae family and causes infection of humans from ancient times. 4OMe-glucuronoxylans as the renewable biopolymers can be promising glycomaterials for various applications in pharmacy. Control enzymatic degradation of the native 4OMe-glucuronoxylan (GX1) followed by targeted sulfation procedure afforded a range of 4OMe-glucuronoxylan sulfates differed in the degree of sulfation (10-16%) and molecular mass (21,000-5000 g/mol; GXS1 > GXS2 > GXS3 > GXS4). Antiviral activity tests on GXS1-4 against herpes simplex virus (HSV) types 1 and 2 revealed the positive effect of all compounds against strains of herpes virus. Of them, the compounds GXS1 and GXS4 were shown to be the most active for both HSV serotypes. The antiviral activity of GXS1 and GXS4 was similar to those of heparin or dextran sulfate, used as reference compounds. It was found that GXS1 and GXS4 were active as well against Polio and dengue viruses, however, on a smaller scale. The mode of antiviral action of 4OMe-glucuronoxylan sulfates is due to inhibition of the virus binding to the cell receptors. Fil: Pujol, Carlos Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Damonte, Elsa Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina Fil: Turjan, J. Slovak Academy of Sciences; Eslovaquia Fil: Yanbo, K. Z.. University of Hong Kong; Hong Kong Fil: Capek, P.. Slovak Academy of Sciences; Eslovaquia

In spring 2019, forecasts of the incidence of tick-borne encephalitis (TBE) for the next two years, i.e. 2019 and 2020, were made for the first time. For this purpose, negative binomial regression models with 4–5 predictors were fitted to the time series of annual human TBE incidences from Austria, Germany and Switzerland. The most important predictor for TBE incidences is the fructification index of the European beech (Fagus sylvatica) 2 years prior as a proxi for the intensity of the TBE virus transmission cycle. These forecasts were repeated in spring 2020 after the updated predictors and the confirmed TBE cases for 2019 became available. Forecasting TBE incidences for 2020 and 2021 results in 156±19 and 131±23 TBE cases for Austria, 663±95 and 543±112 TBE cases for Germany as well as 472±56 and 350±62 TBE cases for Switzerland. The newly implemented operational TBE forecasts will be verified every year with confirmed TBE cases. An initial verification for 2019 demonstrates the high reliability of the forecasts.

Drought responses of mature trees are still poorly understood making it difficult to predict species distributions under a warmer climate. Using mature European beech (Fagus sylvatica L.), a widespread and economically important tree species in Europe, we aimed at developing an empirical stress-level scheme to describe its physiological response to drought. We analysed effects of decreasing soil and leaf water potential on soil water uptake, stem radius, native embolism, early defoliation and crown dieback with comprehensive measurements from overall nine hydrologically distinct beech stands across Switzerland, including records from the exceptional 2018 drought and the 2019/2020 post-drought period. Based on the observed responses to decreasing water potential we derived the following five stress levels: I (predawn leaf water potential-0.4 MPa): no detectable hydraulic limitations; II (-0.4 to -1.3): persistent stem shrinkage begins and growth ceases; III (-1.3 to -2.1): onset of native embolism and defoliation; IV (-2.1 to -2.8): onset of crown dieback; V (-2.8): transpiration ceases and crown dieback is20%. Our scheme provides, for the first time, quantitative thresholds regarding the physiological downregulation of mature European beech trees under drought and therefore synthesises relevant and fundamental information for process-based species distribution models. Moreover, our study revealed that European beech is drought vulnerable, because it still transpires considerably at high levels of embolism and because defoliation occurs rather as a result of embolism than preventing embolism. During the 2018 drought, an exposure to the stress levels III-V of only one month was long enough to trigger substantial crown dieback in beech trees on shallow soils. On deep soils with a high water holding capacity, in contrast, water reserves in deep soil layers prevented drought stress in beech trees. This emphasises the importance to include local data on soil water availability when predicting the future distribution of European beech.

The pyrolysis kinetics of a typical biomass energy feedstock, beech, was investigated based on thermogravimetric analysis over a wide heating rate range from 5K/min to 80K/min. A three-component (corresponding to hemicellulose, cellulose and lignin) parallel decomposition reaction scheme was applied to describe the experimental data. The resulting kinetic reaction model was coupled to an evolutionary optimization algorithm (Shuffled Complex Evolution, SCE) to obtain model parameters. To the authors' knowledge, this is the first study in which SCE has been used in the context of thermogravimetry. The kinetic parameters were simultaneously optimized against data for 10, 20 and 60K/min heating rates, providing excellent fits to experimental data. Furthermore, it was shown that the optimized parameters were applicable to heating rates (5 and 80K/min) beyond those used to generate them. Finally, the predicted results based on optimized parameters were contrasted with those based on the literature.

Southern forests are at the highest ozone (O3) risk in Europe where ground-level O3 is a pressing sanitary problem for ecosystem health. Exposure-based standards for protecting vegetation are not representative of actual field conditions. A biologically-sound stomatal flux-based standard has been proposed, although critical levels for protection still need to be validated. This innovative epidemiological assessment of forest responses to O3 was carried out in 54 plots in Southeastern France and Northwestern Italy in 2012 and 2013. Three O3 indices, namely the accumulated exposure AOT40, and the accumulated stomatal flux with and without an hourly threshold of uptake (POD1 and POD0) were compared. Stomatal O3 fluxes were modeled (DO3SE) and correlated to measured forest-response indicators, i.e. crown defoliation, crown discoloration and visible foliar O3 injury. Soil water content, a key variable affecting the severity of visible foliar O3 injury, was included in DO3SE. Based on flux-effect relationships, we developed species-specific flux-based critical levels (CLef) for forest protection against visible O3 injury. For O3 sensitive conifers, CLef of 19mmolm-2 for Pinus cembra (high O3 sensitivity) and 32mmolm-2 for Pinus halepensis (moderate O3 sensitivity) were calculated. For broadleaved species, we obtained a CLef of 25mmolm-2 for Fagus sylvatica (moderate O3 sensitivity) and of 19mmolm-2 for Fraxinus excelsior (high O3 sensitivity). We showed that an assessment based on PODY and on real plant symptoms is more appropriated than the concentration-based method. Indeed, POD0 was better correlated with visible foliar O3 injury than AOT40, whereas AOT40 was better correlated with crown discoloration and defoliation (aspecific indicators). To avoid an underestimation of the real O3 uptake, we recommend the use of POD0 calculated for hours with a non-null global radiation over the 24-h O3 accumulation window. © 2015 Elsevier B.V.

Lignocellulose biorefineries are distinguished by an explicitly integrative, multi-functional concept that transforms biomass into multiple products, using a variety of conversion and separation processes. This study focuses on the technical design and economic evaluation of a lignocellulose biorefinery, that converts 400,000tDM/a (≙250MW) of beech wood into chemicals and fuel. A model was simulated with Aspen Plus® including the process steps pre-treatment, enzymatic hydrolysis, alcoholic fermentation, dehydration and biogas generation and upgrading. Mass and energy balances showed that 61,600t/a polymer-grade ethylene, 58,520tDM/a organosolv lignin, 38,400t/a biomethane and 90,800tDM/a hydrolysis lignin can be produced with a total energy efficiency of 87.1%. A discounted cash flow analysis indicated that the heat integrated biorefinery concept is not yet profitable. However, the economic results are greatly sensitive regarding various assumptions, in particular in terms of the prices for beech wood, ethylene and organosolv lignin.

We present a coupled experimental and theoretical framework for quantifying the kinetics of two-phase enzymatic hydrolysis of hemicellulose. For xylan loading of 1-5mg/ml, the nature of inhibition by the product xylose (non-competitive), the kinetic constants (Km=3.93 mg/ml, Vmax=0.0252 mg/ml/min) and the xylose inhibition constant (Kx=0.122 mg/ml) are experimentally determined. Our multi-step two-phase kinetic model incorporating enzyme adsorption to the solid substrate and non-competitive product inhibition is simulated using our kinetic data and validated against our experimentally measured temporal dynamics of xylose and reducing sugars. Further experiments show that higher substrate loading reduces the specific adsorption of the endoxylanase to the solid xylan and the enzyme's solid-liquid distribution ratio, which decelerates the solid hydrolysis and accelerates the liquid phase reactions. Thus, the xylose yield increases with substrate loading, which increases product inhibition and decreases reducing sugar yields. An operating cost analysis gives 3mg/ml as the optimal substrate loading.

A strong increase in the level of dehydrin/response ABA transcripts expression reported from the 14th week after flowering coincident with the accumulation of 26 and 44 kDa dehydrins in the embryonic axes of developing beech (Fagus sylvatica L.) seeds. Both transcript and protein levels were strongly correlated with maturation drying. These results suggest that the 44-kDa dehydrin protein is a putative dimer of dehydrin/response ABA protein migrating as a 26-kDa protein. Dehydrins and dehydrin-like proteins form large oligomeric complexes under native conditions and are shown as several spots differing in pI through isoelectrofocusing analyses. Detailed prediction of specific sites accessible for various post-translational modifications (PTMs) in the dehydrin/response ABA protein sequence revealed sites specific to acetylation, amidation, glycosylation, methylation, myristoylation, nitrosylation, O-linked β-N-acetylglucosamination and Yin-O-Yang modification, palmitoylation, phosphorylation, sumoylation, sulfation, and ubiquitination. Thus, these results suggest that specific PTMs might play a role in switching dehydrin function or activity, water binding ability, protein-membrane interactions, transport and subcellular localization, interactions with targeted molecules, and protein stability. Despite the ability of two Cys residues to form a disulfide bond, -SH groups are likely not involved in dimer arrangement. His-rich regions and/or polyQ-tracts are potential candidates as spatial organization modulators. Dehydrin/response ABA protein is an intrinsically disordered protein containing low complexity regions. The lack of a fixed structure and exposition of amino acids on the surface of the protein structure enhances the accessibility to 40 predicted PTM sites, thereby facilitating dehydrin multifunctionality, which is discussed in the present study.

The long-term storage of seeds generally reduces their viability and vigour. The aim of this work was to evaluate the effect of long-term storage on beech (Fagus sylvatica L.) seeds at optimal conditions, over 9 years, on the total and soluble protein levels and activity of proteolytic enzymes, including endopeptidases, carboxypeptidases and aminopeptidases, as well as free amino acid levels and protein synthesis, in dry seeds, after imbibition and during cold stratification leading to dormancy release and germination. The same analyses were conducted in parallel on seeds gathered from the same tree in the running growing season and stored under the same conditions for only 3 months. The results showed that germination capacity decreased from 100% in freshly harvested seeds to 75% in seeds stored for 9 years. The levels of total and soluble proteins were highest in freshly harvested seeds and decreased significantly during storage, these proportions were retained during cold stratification and germination of seeds. Significant differences between freshly harvested and stored seeds were observed in the activities of proteolytic enzymes, including endopeptidases, aminopeptidases and carboxypeptidases, and in the levels of free amino acids. The neosynthesis of proteins during dormancy release and in the early stage of seed germination was significantly weaker in stored seeds. These results confirm the importance of protein metabolism for seed viability and the consequences of its reduction during seed ageing.

The aim of the present work was the high-performance liquid chromatographic separation and multistage mass spectrometric characterization of the polyphenolic compounds of beech bark, as well as the extraction optimization of the identified compounds. Beech is a common and widely used material in the wood industry, yet its bark is regarded as a by-product. Using appropriate extraction methods these compounds could be extracted and utilized in the future. Different extraction methods (stirring, sonication, microwave assisted extraction) using different solvents (water, methanol:water 80:20 v/v, ethanol:water 80:20 v/v) and time/temperature schedules have been compared basing on total phenol contents (Folin-Ciocâlteu) and MRM peak areas of the identified compounds to investigate optimum extraction efficiency. Altogether 37 compounds, including (+)-catechin, (-)-epicatechin, quercetin-O-hexoside, taxifolin-O-hexosides (3), taxifolin-O-pentosides (4), B-type (6) and C-type (6) procyanidins, syringic acid- and coumaric acid-di-O-glycosides, coniferyl alcohol- and sinapyl alcohol-glycosides, as well as other unknown compounds with defined [M-H](-) m/z values and MS/MS spectra have been tentatively identified. The choice of the method, solvent system and time/temperature parameters favors the extraction of different types of compounds. Pure water can extract compounds as efficiently as mixtures containing organic solvents under high-pressure and high temperature conditions. This supports the implementation of green extraction methods in the future. Extraction times that are too long and high temperatures can result in the decrease of the concentrations. Future investigations will focus on the evaluation of the antioxidant capacity and utilization possibilities of the prepared extracts.

The common beech (Fagus sylvatica L.) is propagated by seeds, but the seed set is irregular with five to ten years in between crops. It is therefore necessary to store the seeds. However, beech seeds lose germinability during long-term storage. In this study, beech seeds were stored at −10 °C under controlled conditions for 2, 5, 8, 11 and 13 years. Our results show that beech seeds lose germinability during storage in proportion to the duration of storage. The decrease in germinability correlated with increased electrolyte leakage and accumulation of superoxide anion radicals, hydrogen peroxide and hydroxyl radicals. Furthermore, a strong positive correlation was observed among the releases of superoxide anion radicals, hydrogen peroxide and hydroxyl radicals. In situ localization showed that superoxide anion radicals and hydrogen peroxide were first detectable in root cap cells. When the seed storage time was extended, the reactive oxygen species fluorescence expanded to more areas of the radicle, reaching the root apical meristem. A storage time-dependent decrease in catalase activity, observed in both embryonic axes and cotyledons, was also positively correlated with germinability. DNA fragmentation was observed in beech seeds during storage and occurred predominantly in embryonic axes stored for 5 years and more. Altogether, these results suggest that the loss of germinability in beech seeds during long-term storage depends on several factors, including strong of reactive oxygen species accumulation accompanied by reduced catalase activity as well as membrane injury and DNA alternations, which may be aging-related and ROS-derived. We suggest that the accumulating reactive oxygen species that spread to the root apical meristem are key factors that affect seed germinability after long-term storage.

The effect of long-term exposure of twice-ambient O(3) (2 × O(3)) on whole-tree nitrogen (N) uptake and partitioning of adult beech and spruce was studied in a mixed forest stand, SE-Germany. N uptake as (15)N tracer and N pools were calculated using N concentrations and biomass of tree compartments. Whole-tree N uptake tended to be lower under 2 × O(3) in both species compared to trees under ambient O(3) (1 × O(3)). Internal partitioning in beech showed significantly higher allocation of new N to roots, with mycorrhizal root tips and fine roots together receiving about 17% of new N (2 × O(3)) versus 7% (1 × O(3)). Conversely, in spruce, N allocation to roots was decreased under 2 × O(3). These contrasting effects on belowground N partitioning and pool sizes, being largely consistent with the pattern of N concentrations, suggest enhanced N demand and consumption of stored N with higher relevance for tree-internal N cycling in beech than in spruce.

sous-presse; International audience; Karst environments are unusual because their dry, stony and shallow soils seem to be unfavorable to vegetation, and yet they are often covered with forests. How can trees survive in these environments ? Where do they find the water that allows them to survive? This study uses midday and predawn water potentials and xylem water isotopes of branches to assess tree water status and the origin of transpired water. Monitoring was conducted during the summers of 2014 and 2015 in two dissimilar plots of Mediterranean forest located in karst environments. The results show that the three monitored tree species (Abies alba Mill, Fagus sylvatica L, and Quercus ilex L.) use deep water resources present in the karst vadose zone (unsaturated zone) more intensively during drier years. Quercus ilex, a species welladapted to water stress, which grows at the drier site, uses the deep water resource very early in the summer season. Conversely, the two other species exploit the deep water resource only during severe drought. These results open up new perspectives to a better understanding of ecohydrological equilibrium and to improved water balance modeling in karst forest settings.

Cuticle micromorphology of all eight species of Fagus and an outgroup were examined in the present study. The genus Trigonobalanus was selected as the outgroup. Thirteen characteristics of the inner surface and five of the outer surface of the cuticle were described. Some characteristics, such as the subsidiary cell shape, size of stomata, arrangement of subsidiary cell, shape of anticlinal and periclinal cell walls, texture of periclinal cell wall, development epicuticular wax, and presence of papillae, were considered important for infrageneric classification. The topology was obtained from the analysis using two major lineages: (1) Fagus engleriana + Fagus japonica + Fagus longipetiolata and (2) Fagus sylvatica + Fagus crenata + Fagus lucida + Fagus hayatae + Fagus grandifolia. The first clade supported a bootstrap value of 98% and the second clade a bootstrap value of 97%. Based on the cuticle morphology, our results support the previous study, by revealing F. engleriana, F. japonica, and F. longipetiolata with long peduncles in one group and the remaining extant species of short- to medium-length peduncles in another group. In addition, molecular phylogenetic study of Fagus based on ribosomal DNA ITS and chloroplast DNA sequences data supports this assemblage. This study shows that cuticle micromorphological characteristics provide useful and important information for analyzing the evolutionary aspects of Fagus.

The development of a cost-effective hydrolysis for crude cellulose is an essential part of biorefinery developments. To establish such high solid hydrolysis, a new solid state reactor with static mixing is used. However, concentrations >10% (w/w) cause a rate and yield reduction of enzymatic hydrolysis. By optimizing the synergetic activity of cellulolytic enzymes at solid concentrations of 9%, 17% and 23% (w/w) of crude Organosolv cellulose, glucose concentrations of 57, 113 and 152 g L(-1) are reached. However, the glucose yield decreases from 0.81 to 0.72 g g(-1) at 17% (w/w). Optimal conditions for hydrolysis scale-up under minimal enzyme addition are identified. As result, at 23% (w/w) crude cellulose the glucose yield increases from 0.29 to 0.49 g g(-1). As proof of its applicability, biobutanol, succinic and itaconic acid are produced with the crude hydrolysate. The potential of the substrate is proven e.g. by a high butanol yield of 0.33 g g(-1).

In the present study, we performed a large-scale protein analysis based on 2-DE DIGE to examine the effects of ozone on the leaves of juvenile European beech ( Fagus sylvatica L.), one of the most important deciduous tree species in Central Europe. To this end, beech trees were grown under field conditions and subjected to ambient and twice ambient ozone concentrations during the vegetation periods of four consecutive years. The twice ambient ozone concentration altered the abundance of 237 protein spots, which showed relative ratios higher than 30% compared to the ambient control trees. A total of 74 protein spots were subjected to mass spectrometry identification (LC–MS/MS), followed by homology-driven searches. The differentially expressed proteins participate in key biological processes including the Calvin cycle and photosynthesis, carbon metabolism, defense- and stress-related responses, detoxification mechanisms, protein folding and degradation, and mechanisms involved in senescence. The ozone-induced responses provide evidence of a changing carbon metabolism and counteraction against increased levels of reactive oxygen species. Biological significance This study provides useful information on how European beech, an economically and ecologically important tree species, reacts on the molecular level to increased ozone concentrations expected in the near future. The main emphasis in the present study was placed on identifying differentially abundant proteins after long-term ozone exposure under climatically realistic settings, rather than short-term responses or reactions under laboratory conditions. Additionally, using nursery-grown beech trees, we took into account the natural genotypic variation of this species. As such, the results presented here provide information on molecular responses to ozone in an experimental plant system at very close to natural conditions. Furthermore, this proteomic approach was supported by previous studies on the present experiment. Ultimately, the combination of this proteomic approach with several approaches including transcriptomics, analysis of non-structural carbohydrates, and morphological effects contributes to a more global picture of how beech trees react under increased ozone concentrations.

The estimate of growth losses by ozone exposure of forest trees is a significant part in current C sequestration calculations and will also be important in future modeling. It is therefore important to know if the relationship between ozone flux and growth reduction of young trees, used to derive a Critical Level for ozone, is also valid for mature trees. Epidemiological analysis of stem increment data from Fagus sylvatica L. and Picea abies Karst. observed in Swiss forest plots was used to test this hypothesis. The results confirm the validity of the flux-response relationship at least for beech and therefore enable estimating forest growth losses by ozone on a country-wide scale. For Switzerland, these estimates amount to 19.5% growth reduction for deciduous forests, 6.6% for coniferous forests and 11.0% for all forested areas based on annual ozone stomatal uptake during the time period 1991-2011.

The xylanase gene from Gloeophyllum trabeum was cloned and expressed in Pichia pastoris GS115. Xyl10g has a molecular weight of approximately 50 kDa, and exhibits maximum specific activity at 70 °C and a broad range of pH 4.0–7.0. Purified recombinant Xyl10g efficiently degraded popping-pretreated corn stover and newspaper waste at 50 °C and pH 4.0 after 24 h, and showed synergistic effects with Cel5B (endoglucanase) and BglB (β-glucosidase) to increase reduced sugar levels by about 1.71- to 1.88-fold and 2.26- to 2.48-fold, respectively. Although Xyl10g has low specific activity for beechwood xylan, as compared to XynA, Xyl10g more efficiently degraded corn stover than did XynA. According to immunogold labeling analysis, Xyl10g can attack highly substituted, unsubstituted, and low-substituted xylans, whereas XynA cannot efficiently attack highly substituted xylans, which is important for lignocellulose degradation. These results suggest that GH10 Xyl10g can be used for lignocellulose degradation.

The paper deals with the management problem how to decide on tree species suitability under changing environmental conditions. It presents an algorithm that classifies the output of a range shift model for major tree species in Europe into multiple classes that can be linked to qualities characterizing the ecological niche of the species. The classes: i) Core distribution area, ii) Extended distribution area, iii) Occasional occurrence area, and iv) No occurrence area are first theoretically developed and then statistically described. The classes are interpreted from an ecological point of view using criteria like population structure, competitive strength, site spectrum and vulnerability to biotic hazards. The functioning of the algorithm is demonstrated using the example of a generalized linear model that was fitted to a pan-European dataset of presence/absence of major tree species with downscaled climate data from a General Circulation Model (GCM). Applications of the algorithm to tree species suitability classification on a European and regional level are shown. The thresholds that are used by the algorithm are precision-based and include Cohen's Kappa. A validation of the algorithm using an independent dataset of the German National Forest Inventory shows good accordance of the statistically derived classes with ecological traits for Norway spruce, while the differentiation especially between core and extended distribution for European beech that is in the centre of its natural range in this area is less accurate. We hypothesize that for species in the core of their range regional factors like forest history superimpose climatic factors. Problems of uncertainty issued from potentially applying a multitude of modelling approaches and/or climate realizations within the range shift model are discussed and a way to deal with the uncertainty by revealing the underlying attitude towards risk of the decision maker is proposed.

A multiplicative stomatal conductance model was constructed to estimate stomatal O3 uptake of Fagus crenata exposed to O3 under different N loads to the soil. Our stomatal conductance model included environmental functions such as the stomatal responses of F. crenata to diurnal changes, chronic O3 stress (AOT0), acute O3 stress (O3 concentration), and nitrogen load to soil. The model could explain 62% of the variability in stomatal conductance. We suggest therefore that stomatal closure induced by O3 and N load-induced soil acidification must be taken into account in developing a stomatal conductance model for estimating stomatal O3 uptake for future risk assessment of O3 impact on Japanese forest tree species such as F. crenata.

To determine the effects of ozone (O 3 ) on the canopy carbon budget, we investigated photosynthesis and respiration of leaves of Siebold's beech saplings under free air O 3 exposure (60 nmol mol −1 , during daytime) in relation to the within-canopy light gradient; we then calculated the canopy-level photosynthetic carbon gain (PCG) and respiratory carbon loss (RCL) using a canopy photosynthesis model. Susceptibilities of photosynthesis and respiration to O 3 were greater in leaves of upper canopy than in the lower canopy. The canopy net carbon gain (NCG) was reduced by O 3 by 12.4% during one growing season. The increased RCL was the main factor for the O 3 -induced reduction in NCG in late summer, while contributions of the reduced PCG and the increased RCL to the NCG were almost the same in autumn. These results indicate contributions of changes in PCG and RCL under O 3 to NCG were different between seasons.

The saccharification of beech wood using alkaline polyol pulping (AlkaPolP) and enzymatic hydrolysis was investigated. It will be demonstrated that the AlkaPolP process yields high quality pulps which can easily be hydrolyzed by cellulases. In order to find optimum reaction conditions chips of Fagus sylvatica were pretreated by alkaline glycerol at temperatures between 190 and 230 °C for 15, 20, and 25 min. The impacts of temperature and time were expressed using a severity factor R0. The dependencies of the conversion during enzymatic hydrolysis on severity, pulp yield, delignification and pulp composition are shown. In further experiments it was investigated if the sugar yields can be increased by the application of ultrasound or surfactants before enzyme addition. Up to 95% of the initial cellulose in wood were converted into glucose using cellulases from Trichoderma reesei and β-glucosidase from Aspergillus niger.

Endo-1,4-β-xylanase A (XynA) from Schizophyllum commune was cloned into pPCZαA and expressed in Pichia pastoris GS115. The open reading frame of the xynA gene is composed of 684 bp, encoding 278 amino acids with a molecular weight of 26 kDa. Based on sequence similarity, XynA belongs to the CAZy glycoside hydrolase family 11. The optimal activity of XynA was at pH 5 and 50 °C on beechwood xylan. Under these conditions, the K(m), V(max) and specific activity of XynA were 5768 units mg(-1), 4 mg ml(-1) and 9000 μmol min(-1)mg(-1), respectively. XynA activity was enhanced in the presence of cations, such as K(+), Na(+), Li(2+), Cd(2+), and Co(2+). However, in the presence of EDTA, Hg(2+) and Fe(3+), xylanase activity was significantly inhibited. This enzyme effectively degraded approximately 45% of unsubstituted xylans in the cell wall from poplar stems. The high level of XynA activity might increase the yield of enzyme hydrolysis from biomass. Thus, XynA could be used as a major component of a lignocellulosic degrading enzyme cocktail.

An exact wavelength separation of the spectrum cannot be accomplished when utilizing common ultraviolet (UV) light sources, so the dependence of photodegradation on wavelength cannot be done directly. The photodegradation of wood resulted in by laser irradiation on various wavelengths can be studied by comparing the changes in the DRIFT spectra of various wood materials. The analysis of the difference spectrum is a widely used method to investigate the changes in the infrared spectrum. As this procedure cannot be used to compare the changes quantitatively, a new indicator was introduced, termed Band Damage Index (BDI). The BDI and the results obtained by its analysis are going to be discussed.

Triterpenoids, 1-3, 8 and 9, along with 24 known compounds were isolated from leaves and twigs of Fagus hayatae. Of these, compound 1, 1,10-seco-3β,10α,23-trihydroxyolean-12-ene-1,28-dioic acid 1,23-lactone, possesses a hitherto unknown 1,10-seco-oleanane skeleton. In addition, 2,3-seco-20(29)-lupene-2,3-dioic acid (16), previously described as a synthetic product, is now established as a plant natural product; the neolignan-9'-O-rhamnoside 19 is also characterized herein. Their structures were deduced mainly by 1D and 2D NMR spectroscopic analyses. Seven of these compounds possess moderate inhibitory activity against α-glucosidase type IV (Bacillus stearothermophilus).

The influence of Pleurotus ostreatus inoculation on wood degradation and on fungal community structure was studied. The experiments were performed on an organically poor fly ash deposit covered with a 10 cm layer of beech wood chips inoculated with P. ostreatus isolate ZIM76. Compared to non-inoculated wood chips, inoculation increased the temperatures and relative humidities and, in the first 6 months, accelerated Klason lignin degradation by 9% and also, after 17 months, increased iron translocation into wood chips by 30%. After 6 months, PCR-DGGE showed 22-28 and 13-21 fungal taxa in non-inoculated and P. ostreatus-inoculated beech chips, respectively. The differences in number of taxa and in the fungal community structure (based on Dice coefficient) between non-inoculated and inoculated wood chips diminished with time. The results indicate that the naturally occurring processes of wood degradation are as efficient as those occurring in sites inoculated with P. ostreatus.

Soil acidification was assessed in the Eastern Sudetes Mountains (Czech Republic) between 1941 and 2003, i.e. before and after the period of major industrial pollution (1950s–1990s). The twenty sites included in our study were distributed along a gradient of altitude ranging 1000 m. Values of pH have decreased in 80–90% of the pairs of samples after the six decades, on average by 0.7 for pH-H 2 O and 0.6 for pH-KCl. Organic matter increased in the topsoil, probably reflecting a change in decomposition conditions. The most important finding is that the acidification varies along the joint gradient of altitude/tree layer composition, and displays a changing pattern in three soil horizons (A, B and C). Contrary to expectations, most acidified were soils in beech forests at lower elevations.

Forests that are composed of two or more tree species with similar ecological strategies appear to contradict the competitive exclusion principle. Beech-maple communities are a well-known example of such a system. On a local scale, a number of mechanisms have been proposed to explain the coexistence of these two species. These are reciprocal replacement, external factors that favour alternatively one or the other species and demographic stochasticity. This paper presents and analyses a simple mathematical model that shows that external factors are not an essential requirement for coexistence. Rather, coexistence requires interspecific differences in light transmissivity through the crowns of adult trees. However, all the three mechanisms mentioned above can be interpreted within the framework of the model. Furthermore, many models of forest dynamics make use of shade tolerance as a key feature in describing successional dynamics. Despite its importance, however, shade tolerance does not have a commonly accepted quantitative definition. Here, a simple scheme is proposed where the relationship between shade tolerance, individual traits (growth and survival) and successional status is defined. This might have important implications in understanding the overall dynamics. Theoretical results have been compared with a number of studies carried out in North American forests. In particular, coexistence in beech-maple communities and the relation between shade tolerance and successional status in a beech-hemlock-birch community have been discussed.

For a quantitative estimate of the ozone effect on vegetation reliable models for ozone uptake through the stomata are needed. Because of the analogy of ozone uptake and transpiration it is possible to utilize measurements of water loss such as sap flow for quantification of ozone uptake. This technique was applied in three beech (Fagus sylvatica) stands in Switzerland. A canopy conductance was calculated from sap flow velocity and normalized to values between 0 and 1. It represents mainly stomatal conductance as the boundary layer resistance in forests is usually small. Based on this relative conductance, stomatal functions to describe the dependence on light, temperature, vapour pressure deficit and soil moisture were derived using multivariate nonlinear regression. These functions were validated by comparison with conductance values directly estimated from sap flow. The results corroborate the current flux parameterization for beech used in the DO3SE model.

Effects of elevated N deposition on forest aboveground biomass were evaluated using long-term data from N addition experiments and from forest observation plots in Switzerland. N addition experiments with saplings were established both on calcareous and on acidic soils, in 3 plots with Fagus sylvatica and in 4 plots with Picea abies. The treatments were conducted during 15 years and consisted of additions of dry NH4NO3 at rates of 0, 10, 20, 40, 80, and 160 kg N ha−1 yr−1. The same tree species were observed in permanent forest observation plots covering the time span between 1984 and 2007, at modeled N deposition rates of 12–46 kg N ha−1 yr−1. Experimental N addition resulted in either no change or in a decreased shoot growth and in a reduced phosphorus concentration in the foliage in all experimental plots. In the forest, a decrease of foliar P concentration was observed between 1984 and 2007, resulting in insufficient concentrations in 71% and 67% of the Fagus and Picea plots, respectively, and in an increasing N:P ratio in Fagus. Stem increment decreased during the observation period even if corrected for age. Forest observations suggest an increasing P limitation in Swiss forests especially in Fagus which is accompanied by a growth decrease whereas the N addition experiments support the hypothesis that elevated N deposition is an important cause for this development.

Tropospheric ozone causes severe oxidative stress in plants. To investigate the transcriptional responsiveness of adult trees to ozone, fully-expanded sun and shade leaves of mature beech trees were harvested at four time points over the entire vegetation period in 2005 and 2006. Microarray analyses were conducted on leaves from trees grown in the field under ambient and twice-ambient ozone concentrations at Kranzberger Forst (Bavaria). Beech trees changed their transcript levels in response to ozone. In the years 2005 and 2006 different transcription patterns were observed; this may have been a result of different weather conditions and ozone uptake. Furthermore, we obtained differences in mRNA expression patterns between shade and sun leaves. In the ozone-treated sun leaves of 2005, slightly up- and down-regulated transcript levels were detected, particularly in the spring and autumn, whereas shade leaves clearly exhibited reduced mRNA levels, particularly at the end of the vegetation period. In 2006, this pattern could not be confirmed, and in the autumn, four other transcripts were slightly up-regulated in ozone-treated shade leaves. In addition, two other transcripts were found to be influenced in sun leaves in the spring/summer. While we detected changes in the levels of only a few transcripts, the observed effects were not identical in both years. In conclusion, elevated ozone exhibited very small influence on the transcription levels of genes of mature beech trees.

In 2006, a controlled infection study was performed in the 'Kranzberger Forst' to address the following questions: (1) Will massive artificial inoculation with Apiognomonia errabunda override the previously observed inhibitory effect of chronic ozone? (2) Can biochemical or molecular markers be detected to account for the action of ozone? To this end six adult beech trees were chosen, three ozone fumigated (2x ozone) and three control trees (ambient = 1x ozone). Spore-sprayed branches of sun and shade crown positions of each of the trees, and uninoculated control branches, were enclosed in 100-L plastic bags for one night to facilitate infection initiation. Samples were taken within a five-week period after inoculation. A. errabunda infestation levels quantified by real-time PCR increased in leaves that were not fumigated with additional ozone. Cell wall components and ACC (ethylene precursor 1-amino cyclopropane-1-carboxylic acid) increased upon ozone fumigation and may in part lead to the repression of fungal infection.

The effects of experimentally elevated O(3) on soil respiration rates, standing fine-root biomass, fine-root production and delta(13)C signature of newly produced fine roots were investigated in an adult European beech/Norway spruce forest in Germany during two subsequent years with contrasting rainfall patterns. During humid 2002, soil respiration rate was enhanced under elevated O(3) under beech and spruce, and was related to O(3)-stimulated fine-root production only in beech. During dry 2003, the stimulating effect of O(3) on soil respiration rate vanished under spruce, which was correlated with decreased fine-root production in spruce under drought, irrespective of the O(3) regime. delta(13)C signature of newly formed fine-roots was consistent with the differing g(s) of beech and spruce, and indicated stomatal limitation by O(3) in beech and by drought in spruce. Our study showed that drought can override the stimulating O(3) effects on fine-root dynamics and soil respiration in mature beech and spruce forests.

In a 50- to 70-year-old mixed stand of Norway spruce (Picea abies (L.) Karst.) and European beech (Fagus sylvatica L.) in Germany, tree cohorts have been exposed to double ambient ozone (2xO(3)) from 2000 through 2007 and can be compared with trees in the same stand under the ambient ozone regime (1xO(3)). Annual diameter growth, allocation pattern, stem form, and stem volume were quantified at the individual tree and stand level. Ozone fumigation induced a shift in the resource allocation into height growth at the expense of diameter growth. This change in allometry leads to rather cone-shaped stem forms and reduced stem stability in the case of spruce, and even neiloidal stem shapes in the case of beech. Neglect of such ozone-induced changes in stem shape may lead to a flawed estimation of volume growth. On the stand level, 2xO(3) caused, on average, a decrease of 10.2 m(3) ha(-1) yr(-1) in European beech.