Long-term responses of Scots pine (Pinus sylvestris L.) and European beech (Fagus sylvatica L.) to the contamination of light soils with diesel oil.

Research into trees plays a very important role in evaluations of soil contamination with diesel oil. Trees are ideal for reclaiming contaminated soils because their large biomass renders them more resistant to higher concentrations of pollutants. In the literature, there is a general scarcity of long-term studies performed on trees, in particular European beeches. The aim of this study was to evaluate the responses of Scots pines and European beeches grown for 8 years on soil contaminated with diesel oil. Selected morphological and physiological parameters of trees were analyzed. The biomass yield of Scots pines was not significantly correlated with increasing concentrations of diesel oil, but it was more than 700% higher than in European beeches. Scots pines were taller and had a larger stem diameter than European beeches during the 8-year study. The diameter of trees grown on the most contaminated soil was reduced 1.5-fold in Scots pines and more than twofold in European beeches. The length of Scots pine needles from the most contaminated treatment decreased by 50% relative to control needles. The shortest needles were heaviest. The fluctuating asymmetry (FA) of needle length was highest in Scots pines grown on the most contaminated soil, whereas the reverse was noted in the FA of needle weight. Diesel oil decreased the concentrations of chlorophylls a and b, total chlorophyll, and carotenoids. The Fv/Fm ratio of needles and leaves was influenced by the tested concentrations of diesel oil. The results of the study indicate that the Scots pine better adapts (grows more rapidly and produces higher biomass) to long-term soil contamination with diesel oil than the European beech. In European beeches, growth inhibition and leaf discoloration (a decrease in chlorophyll content) were observed already after the first year of the experiment, which indicates that 1-year-old seedlings of European beech are robust bioindicators of soil contamination with diesel oil.

Ultraviolet radiation modulates C:N stoichiometry and biomass allocation in Fagus sylvatica saplings cultivated under elevated CO2 concentration.

Under the conditions of ongoing climate change, terrestrial ecosystems will be simultaneously exposed to a permanent rise in atmospheric CO2 concentration and increasing variability of such environmental factors as temperature, precipitation, and UV radiation. This will result in numerous interactions. The interactive effects caused by exposure to such multiple environmental factors are not yet well understood. We tested the hypotheses that enhanced UV radiation reduces the stimulatory effect of elevated CO2 concentration on plant biomass production and that it alters biomass allocation in broadleaved European beech (Fagus sylvatica L.) saplings. Our results after 2 years of exposure confirmed interactive effects of CO2 concentration and UV radiation on biomass production, and particularly on biomass allocation to roots and aboveground biomass. The strongest stimulatory effect of elevated CO2 on aboveground biomass and roots was found under ambient UV radiation, while both low and high UV doses reduced this stimulation. Nitrogen content in the roots and the distribution of nitrogen among leaves and roots were also significantly affected by interaction of CO2 concentration and UV radiation. The observed changes in leaf and root C:N stoichiometry were associated with altered morphological traits, and particularly with a change in the proportion of fine roots. As the biomass allocation and especially the proportion of fine roots can play an important role in effective water and nutrient use and acclimation to future climates, it is essential to obtain a deeper understanding of the links between C:N stoichiometry and biomass accumulation.

Interactive effects of ultraviolet radiation and elevated CO2 concentration on photosynthetic characteristics of European beech saplings during the vegetation season.

To test the hypothesis that ultraviolet radiation (UV) modulates photosynthetic responses to elevated CO2 concentration ([CO2]) in plants, saplings of European beech were grown for two vegetation seasons under ambient (400 ppm) and elevated (700 ppm) atmospheric [CO2]. From April to November the saplings were exposed to (i) ambient UV radiation, (ii) excluded and (iii) enhanced UV (150% of ambient). Gas-exchange and chlorophyll fluorescence techniques were used throughout the second vegetation season together with biochemical analyses of the amount and activity of the Rubisco enzyme. We found support for the hypothesis that an impact of elevated [CO2] on photosynthesis is substantially modulated by UV radiation. Moreover, we found that the [CO2] × UV interaction is changing along the vegetation season: an enhanced UV radiation stimulated a positive effect of elevated [CO2] on plant photosynthesis at the beginning of the vegetation season (short-term effect), whilst long-term cultivation reduced the stimulatory effect of elevated [CO2] (a clear down-regulation of photosynthesis). Down-regulation was, however, not found in plants grown under the conditions of excluded UV radiation. We found evidence that the down-regulation of photosynthesis is associated with a complex acclimation at different hierarchical and functional levels, including an acclimation of primary photochemical reactions, carboxylation activity of Rubisco enzyme, and stomatal conductance.

Photosynthetic activity in relation to a gradient of leaf nitrogen content within a canopy of Siebold's beech and Japanese oak saplings under elevated ozone.

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 (Vcmax) and electron transport (Jmax), as a function of the leaf nitrogen content throughout a canopy of Siebold's beech and Japanese oak grown under elevated ozone (O3) 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 O3 exposure (60 nmol mol-1, during daylight hours) in June, August, and October 2012. We observed O3-induced reduction in Vcmax and Jmax without reduction of leaf nitrogen content in both tree species. In Siebold's beech, Vcmax and Jmax in leaves with higher Narea were largely decreased by O3 from August, while little effect of O3 was observed in leaves with lower Narea. On the other hand, there was no difference in the extent of O3-induced reduction in Vcmax and Jmax across the range of Narea in leaves of Japanese oak. Reduction of leaf nitrogen content under elevated O3 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 O3-induced decline of photosynthesis in Siebold's beech and Japanese oak. Based on these results, we emphasize the importance of integration of O3 effects into the conventional estimation of Vcmax and Jmax from leaf nitrogen content for evaluating canopy photosynthesis under current and future elevated O3 conditions.

Behaviour of aluminium in forest soils with different lithology and herb vegetation cover.

The aim of this study was to determine the content, distribution and behaviour of Al in soils under beech forest with different parent rock, and to assess the role of herbaceous vegetation on soil Al behaviour. We hypothesize that the contents of elements in the soil sorption complex (Al etc.) are strongly influenced by vegetation cover. Also, low molecular mass organic acids (LMMOA) can be considered as an indicator of soil organic matter (SOM) decomposition and vegetation litter turnover. Speciation of LMMOA, nutrition content (PO43-, Ca2+, K+) and element composition in aqueous extracts were determined by means of ion chromatography and inductively coupled plasma - optical emission spectrometry (ICP-OES) respectively. Active and exchangeable pH, sorption characteristics and exchangeable Al (Alex) were determined in BaCl2 extracts by ICP-OES. Elemental composition of parent rocks was assessed by means of X-ray fluorescence spectroscopy. Herb-poor localities showed lower pH, less nutrients (PO43-, Ca2+, K+), less LMMOA, a larger stock of SOM and greater cation exchange capacity. There was also lower mobilisation of Al in organic horizons, which explains the larger pools of Al. Generally, we can conclude that LMMOA, and thus soil vegetation cover, play an important role in the Al soil cycle.

Anthropogenic nitrogen deposition alters growth responses of European beech (Fagus sylvativa L.) to climate change.

Global change affects the functioning of forest ecosystems and the services they provide, but little is known about the interactive effects of co-occurring global change drivers on important functions such as tree growth and vitality. In the present study we quantified the interactive (i.e. synergistic or antagonistic) effects of atmospheric nitrogen (N) deposition and climatic variables (temperature, precipitation) on tree growth (in terms of tree-ring width, TRW), taking forest ecosystems with European beech (Fagus sylvatica L.) as an example. We hypothesised that (i) N deposition and climatic variables can evoke non-additive responses of the radial increment of beech trees, and (ii) N loads have the potential to strengthen the trees' sensitivity to climate change. In young stands, we found a synergistic positive effect of N deposition and annual mean temperature on TRW, possibly linked to the alleviation of an N shortage in young stands. In mature stands, however, high N deposition significantly increased the trees' sensitivity to increasing annual mean temperatures (antagonistic effect on TRW), possibly due to increased fine root dieback, decreasing mycorrhizal colonization or shifts in biomass allocation patterns (aboveground vs. belowground). Accordingly, N deposition and climatic variables caused both synergistic and antagonistic effects on the radial increment of beech trees, depending on tree age and stand characteristics. Hence, the nature of interactions could mediate the long-term effects of global change drivers (including N deposition) on forest carbon sequestration. In conclusion, our findings illustrate that interaction processes between climatic variables and N deposition are complex and have the potential to impair growth and performance of European beech. This in turn emphasises the importance of multiple-factor studies to foster an integrated understanding and models aiming at improved projections of tree growth responses to co-occurring drivers of global change.

Five-year volume growth of European beech does not respond to ozone pollution in Italy.

A unique database of stand volume growth, estimated as periodic annual volume increment (in m3 ha-1 per year over the period 2001-2005) from 728 European beech (Fagus sylvatica L.) sites distributed across Italy, was used to assess the effects of ambient ozone (O3), expressed as annual average (M24), accumulated exposure above a 40 ppb hourly threshold (AOT40), and total stomatal ozone flux (POD0). Growth data were from the National forest inventory of Italy, while climate data and ozone concentrations were computed by the WRF and CHIMERE models, respectively. Results show that the growth increased with increasing solar radiation and air temperature and decreased with increasing number of cold days, while effects of soil water content and O3 were not significant. In contrast, the literature results suggest that European beech is sensitive to both drought and O3. Ozone levels resulted to be very high (48 ppb M24, 51,200 ppb h AOT40, 21.08 mmol m-2 POD0, on average) and thus able to potentially affect European beech growth. We hypothesize that the high-frequency signals of soil water and O3 got lost when averaged over 5 years and recommended finer time-resolution investigations and inclusion of other factors of variability, e.g., thinning, tree age, and size.

Photosynthetic responses to ozone of upper and lower canopy leaves of Fagus crenata Blume seedlings grown under different soil nutrient conditions.

We aimed to clarify the effects of ozone (O3) on photosynthetic ability of upper and lower canopy leaves of Fagus crenata Blume seedlings grown under different soil nutrient conditions. To accomplish this objective, we analyzed the response of photosynthetic parameters such as maximum carboxylation rate (Vcmax) to cumulative stomatal O3 uptake (ΣFst) and reduction rate of Vcmax per unit ΣFst as an index of detoxification capacity for O3. The seedlings of Fagus crenata were grown for two growing seasons (2014-2015) in nine treatments comprised of a combination of three levels of gas treatments (charcoal-filtered air or 1.0- or 1.5-times ambient O3 concentration) and three levels of soil nutrient treatments (non-fertilized or a supply of relatively low or high concentrations of compound fertilizer). The nutrient supply significantly increased the degree of O3-induced reduction in Vcmax in September. However, nutrient supply did not significantly increase ΣFst and reduce the detoxification capacity for O3. On the other hand, the degree of O3-induced reduction in Vcmax of upper canopy leaves was higher as compared with that of lower canopy leaves in August due to the higher ΣFst. However, the reduction rate of Vcmax per unit ΣFst in lower canopy leaves was higher than that in upper canopy leaves, indicating lower detoxification capacity for O3 in lower canopy leaves. Reduction rate of Vcmax per unit ΣFst over the threshold, which is assumed to be proportional to gross photosynthetic rate, was similar between upper and lower canopy leaves. Therefore, capacity of photosynthetic CO2 assimilation is likely to be associated with detoxification capacity for O3 in upper and lower canopy leaves of F. crenata seedlings grown under different soil nutrient conditions.

Beech wood Fagus sylvatica dilute-acid hydrolysate as a feedstock to support Chlorella sorokiniana biomass, fatty acid and pigment production.

This work evaluates the possibility of using beech wood (Fagus sylvatica) dilute-acid (H2SO4) hydrolysate as a feedstock for Chlorella sorokiniana growth, fatty acid and pigment production. Neutralized wood acid hydrolysate, containing organic and mineral compounds, was tested on Chlorella growth at different concentrations and compared to growth under phototrophic conditions. Chlorella growth was improved at lower loadings and inhibited at higher loadings. Based on these results, a 12% neutralized wood acid hydrolysate (Hyd12%) loading was selected to investigate its impact on Chlorella growth, fatty acid and pigment production. Hyd12% improved microalgal biomass, fatty acid and pigment productivities both in light and in dark, when compared to photoautotrophic control. Light intensity had substantial influence on fatty acid and pigment composition in Chlorella culture during Hyd12%-based growth. Moreover, heterotrophic Chlorella cultivation with Hyd12% also showed that wood hydrolysate can constitute an attractive feedstock for microalgae cultivation in case of lack of light.

Enhancement of biomass conversion in catalytic fast pyrolysis by microwave-assisted formic acid pretreatment.

This study investigated microwave-assisted formic acid (MW-FA) pretreatment as a possible way to improve aromatic production from catalytic fast pyrolysis (CFP) of lignocellulosic biomass. Results showed that short duration of MW-FA pretreatment (5-10min) could effectively disrupt the recalcitrant structure of beech wood and selectively remove its hemicellulose and lignin components. This increased the accessibility of cellulose component of biomass to subsequent thermal conversion in CFP. Consequently, the MW-FA pretreated beech wood produced 14.0-28.3% higher yields (26.4-29.8C%) for valuable aromatic products in CFP than the untreated control (23.2C%). In addition, the yields of undesired solid residue (char/coke) decreased from 33.1C% for the untreated control to 28.6-29.8C% for the MW-FA pretreated samples. These results demonstrate that MW-FA pretreatment can provide an effective way to improve the product distribution from CFP of lignocellulose.

Photosynthetic traits of Siebold's beech seedlings in changing light conditions by removal of shading trees under elevated CO2.

The purpose of this study was to obtain basic information on acclimation capacity of photosynthesis in Siebold's beech seedlings to increasing light intensity under future elevated CO2 conditions. We monitored leaf photosynthetic traits of these seedlings in changing light conditions (before removal of shade trees, the year after removal of shade trees and after acclimation to open conditions) in a 10-year free air CO2 enrichment experiment in northern Japan. Elevated CO2 did not affect photosynthetic traits such as leaf mass per area, nitrogen content and biochemical photosynthetic capacity of chloroplasts (i.e. maximum rate of carboxylation and maximum rate of electron transport) before removal of the shade trees and after acclimation to open conditions; in fact, a higher net photosynthetic rate was maintained under elevated CO2 . However, in the year after removal of the shade trees, there was no increase in photosynthesis rate under elevated CO2 conditions. This was not due to photoinhibition. In ambient CO2 conditions, leaf mass per area and nitrogen content were higher in the year after removal of shade trees than before, whereas there was no increase under elevated CO2 conditions. These results indicate that elevated CO2 delays the acclimation of photosynthetic traits of Siebold's beech seedlings to increasing light intensity.

Effects of rhizopheric nitric oxide (NO) on N uptake in Fagus sylvatica seedlings depend on soil CO2 concentration, soil N availability and N source.

Rhizospheric nitric oxide (NO) and carbon dioxide (CO2) are signalling compounds known to affect physiological processes in plants. Their joint influence on tree nitrogen (N) nutrition, however, is still unknown. Therefore, this study investigated, for the first time, the combined effect of rhizospheric NO and CO2 levels on N uptake and N pools in European beech (Fagus sylvatica L.) seedlings depending on N availability. For this purpose, roots of seedlings were exposed to one of the nine combinations (i.e., low, ambient, high NO plus CO2 concentration) at either low or high N availability. Our results indicate a significant effect of rhizospheric NO and/or CO2 concentration on organic and inorganic N uptake. However, this effect depends strongly on NO and CO2 concentration, N availability and N source. Similarly, allocation of N to different N pools in the fine roots of beech seedlings also shifted with varying rhizospheric gas concentrations and N availability.

Soil bacterial community structure responses to precipitation reduction and forest management in forest ecosystems across Germany.

Soil microbial communities play an important role in forest ecosystem functioning, but how climate change will affect the community composition and consequently bacterial functions is poorly understood. We assessed the effects of reduced precipitation with the aim of simulating realistic future drought conditions for one growing season on the bacterial community and its relation to soil properties and forest management. We manipulated precipitation in beech and conifer forest plots managed at different levels of intensity in three different regions across Germany. The precipitation reduction decreased soil water content across the growing season by between 2 to 8% depending on plot and region. T-RFLP analysis and pyrosequencing of the 16S rRNA gene were used to study the total soil bacterial community and its active members after six months of precipitation reduction. The effect of reduced precipitation on the total bacterial community structure was negligible while significant effects could be observed for the active bacteria. However, the effect was secondary to the stronger influence of specific soil characteristics across the three regions and management selection of overstorey tree species and their respective understorey vegetation. The impact of reduced precipitation differed between the studied plots; however, we could not determine the particular parameters being able to modify the response of the active bacterial community among plots. We conclude that the moderate drought induced by the precipitation manipulation treatment started to affect the active but not the total bacterial community, which points to an adequate resistance of the soil microbial system over one growing season.

Ozone induces stomatal narrowing in European and Siebold's beeches: a comparison between two experiments of free-air ozone exposure.

Stomata tend to narrow under ozone (O(3)) impact, leading to limitation of stomatal O(3) influx. Here, we review stomatal response under recently conducted free-air O(3) exposure experiments on two species of the same tree genus: Fagus sylvatica at Kranzberg Forest (Germany) and F. crenata at Sapporo Experimental Forest (Japan). Both beeches exhibited reduction in stomatal conductance (gs) by 10-20% under experimentally enhanced O(3) regimes throughout the summer relative to ambient-air controls. Stomatal narrowing occurred, in early summer, in the absence of reduced carboxylation capacity of Rubisco, although photosynthetic net CO(2) uptake rate temporarily reflected restriction to some minor extent. Observed stomatal narrowing was, however, diminished in autumn, suggesting gradual loss of stomatal regulation by O(3). Monotonic decline in gs with cumulative O(3) exposure or flux in current modeling concepts appear to be unrealistic in beech.

Contrasting carbon allocation responses of juvenile European beech (Fagus sylvatica) and Norway spruce (Picea abies) to competition and ozone.

Allocation of recent photoassimilates of juvenile beech and spruce in response to twice-ambient ozone (2 × O(3)) and plant competition (i.e. intra vs. inter-specific) was examined in a phytotron study. To this end, we employed continuous (13)CO(2)/(12)CO(2) labeling during late summer and pursued tracer kinetics in CO(2) released from stems. In beech, allocation of recent photoassimilates to stems was significantly lowered under 2 × O(3) and increased in spruce when grown in mixed culture. As total tree biomass was not yet affected by the treatments, C allocation reflected incipient tree responses providing the mechanistic basis for biomass partitioning as observed in longer experiments. Compartmental modeling characterized functional properties of substrate pools supplying respiratory C demand. Respiration of spruce appeared to be exclusively supplied by recent photoassimilates. In beech, older C, putatively located in stem parenchyma cells, was a major source of respiratory substrate, reflecting the fundamental anatomical disparity between angiosperm beech and gymnosperm spruce.

Whole-tree seasonal nitrogen uptake and partitioning in adult Fagus sylvatica L. and Picea abies L. [Karst.] trees exposed to elevated ground-level ozone.

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.

Modeling of stomatal conductance to estimate stomatal ozone uptake by Fagus crenata, Quercus serrata, Quercus mongolica var. crispula and Betula platyphylla.

To construct stomatal conductance models and estimate stomatal O3 uptake for Fagus crenata, Quercus serrata, Quercus mongolica var. crispula and Betula platyphylla, stomatal conductance (gs) was measured in seedlings of the four tree species. Better estimates of gs were made by incorporating the acute effects of O3 on gs into the models and the models could explain 34-52% of the variability in gs. Although the O3 concentration was relatively high in spring from April to May, COU of F. crenata, Q. serrata and Q. mongolica var. crispula were relatively low and the ratios of COU in spring to total COU in one year were 16.8% in all tree species because of low gs limited mainly by leaf pre-maturation and/or low temperature. The COU of B. platyphylla were relatively high mainly because of rapid leaf maturation and lower optimal temperature for stomatal opening.

Growth losses in Swiss forests caused by ozone: epidemiological data analysis of stem increment of Fagus sylvatica L. and Picea abies Karst.

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.

Stomatal and pavement cell density linked to leaf internal CO2 concentration.

BACKGROUND AND AIMS: Stomatal density (SD) generally decreases with rising atmospheric CO2 concentration, Ca. However, SD is also affected by light, air humidity and drought, all under systemic signalling from older leaves. This makes our understanding of how Ca controls SD incomplete. This study tested the hypotheses that SD is affected by the internal CO2 concentration of the leaf, Ci, rather than Ca, and that cotyledons, as the first plant assimilation organs, lack the systemic signal. METHODS: Sunflower (Helianthus annuus), beech (Fagus sylvatica), arabidopsis (Arabidopsis thaliana) and garden cress (Lepidium sativum) were grown under contrasting environmental conditions that affected Ci while Ca was kept constant. The SD, pavement cell density (PCD) and stomatal index (SI) responses to Ci in cotyledons and the first leaves of garden cress were compared. (13)C abundance (δ(13)C) in leaf dry matter was used to estimate the effective Ci during leaf development. The SD was estimated from leaf imprints. KEY RESULTS: SD correlated negatively with Ci in leaves of all four species and under three different treatments (irradiance, abscisic acid and osmotic stress). PCD in arabidopsis and garden cress responded similarly, so that SI was largely unaffected. However, SD and PCD of cotyledons were insensitive to Ci, indicating an essential role for systemic signalling. CONCLUSIONS: It is proposed that Ci or a Ci-linked factor plays an important role in modulating SD and PCD during epidermis development and leaf expansion. The absence of a Ci-SD relationship in the cotyledons of garden cress indicates the key role of lower-insertion CO2 assimilation organs in signal perception and its long-distance transport.

Phosphite protects Fagus sylvatica seedlings towards Phytophthora plurivora via local toxicity, priming and facilitation of pathogen recognition.

Phytophthora plurivora causes severe damage on Fagus sylvatica and is responsible for the extensive decline of European Beech throughout Europe. Unfortunately, no effective treatment against this disease is available. Phosphite (Phi) is known to protect plants against Phytophthora species; however, its mode of action towards P. plurivora is still unknown. To discover the effect of Phi on root infection, leaves were sprayed with Phi and roots were subsequently inoculated with P. plurivora zoospores. Seedling physiology, defense responses, colonization of root tissue by the pathogen and mortality were monitored. Additionally the Phi concentration in roots was quantified. Finally, the effect of Phi on mycelial growth and zoospore formation was recorded. Phi treatment was remarkably efficient in protecting beech against P. plurivora; all Phi treated plants survived infection. Phi treated and infected seedlings showed a strong up-regulation of several defense genes in jasmonate, salicylic acid and ethylene pathways. Moreover, all physiological parameters measured were comparable to control plants. The local Phi concentration detected in roots was high enough to inhibit pathogen growth. Phi treatment alone did not harm seedling physiology or induce defense responses. The up-regulation of defense genes could be explained either by priming or by facilitation of pathogen recognition of the host.