Seasonal ozone response of mature beech trees (Fagus sylvatica) at high altitude in the Bavarian forest (Germany) in comparison with young beech grown in the field and in phytotrons

Mature beech trees (Fagus sylvatica) grown at two different altitudes in the Bavarian forest were compared with young beech trees grown at nearby field sites or in phytotrons for their macroscopic and physiological responses to different ozone (O(3)) exposures. Cumulative O(3) exposure expressed as the sum of hourly mean concentrations above the canopy ranged between 100 and 150 microl l(-1) h, with the vertical O(3) profiles at the higher altitude site being enhanced by 30%. O(3) profiles at all sites were reduced by up to 20% with increasing depth within and beneath the canopy. The leaf discoloration that developed in the absence of premature leaf loss was similar in the sun foliage of mature and young trees (including plant grown in the phytotron). Injury became apparent at low O(3) exposures, expressed as accumulated hourly means over a threshold of 40 nl l(-1) (AOT40 <3.5 microl l(-1) h) at the lower site in both the mature trees and the young beech at the field site, but only occurred when AOT40 values reached 7 microl l(-1) h at the upper site, and 6 microl l(-1) h in the phytotrons. However, the association between injury and O(3) exposure was improved when cumulative ozone uptake to sun leaves was the ozone index, used with values of about 3 mmol m(-2) resulting in visible injury in both mature and young beech growing in phytotrons. Under high ozone exposure levels of inositol were lowered, whilst concentrations of lignin-like materials were enhanced in mature beech. Similar responses were observed in young beech grown in phytotrons. As the sun foliage was affected by only a small and variable extent each year, the seasonal O(3) impact at high altitude did not appear to pose an acute risk to mature beech trees.     

Ozone fumigation (twice ambient) reduces leaf infestation following natural and artificial inoculation by the endophytic fungus Apiognomonia errabunda of adult European 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 (2× ozone) and three control trees (ambient = 1× 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.     

Persistent stimulation of CO2 assimilation and stomatal conductance by summer ozone fumigation in Norway spruce

CO(2) assimilation rate, stomatal conductance and chlorophyll content of current and previous years' needles of Norway spruce were measured in May 1988, 205 days after the cessation of ozone fumigation during the summer of 1987. Rates of assimilation were consistently higher for both needle year age classes for ozone fumigated trees in comparison to control trees, although only statistically significant for part of the day for current year's needles. A 26% and 48% stimulation, overall, in mean daily rates of assimilation for current and previous years' needles of ozone fumigated trees was observed. This was due to an enhanced apparent quantum yield and light saturated rate of assimilation of ozone fumigated trees. The temperature response regression of assimilation versus temperature was also greater, such that at any given temperature, assimilation was higher for ozone treated trees than control trees. Stomatal conductance was greater for ozone fumigated trees than the controls, but this was only marginally statistically significant. Moreover, there was a consistent increase in chlorophyll content in both year classes in ozone-treated trees. These results are discussed in relation to a possible long term effect of ozone fumigation upon the processes of conifer winter hardening and spring de-hardening.     

Shoot growth of mature Fagus sylvatica and Picea abies in relation to ozone

Epidemiological analysis of sequential growth data may be a tool in assessing ozone sensitivity of mature trees. Annual shoot growth of mature Fagus sylvatica in 83 Swiss permanent forest observation plots and of Picea abies in 61 plots was evaluated for 11 and 8 consecutive years, respectively, using branches harvested every 4 years. The data were assessed as annual deviation from average growth and related to fructification, ozone, meteorological parameters, and modelled soil water content using a mixed linear model. In beech, a significant association between ozone and shoot growth was observed which corresponded to a 7.4% growth reduction between 0 and 10 ppm h AOT40 (accumulated ozone over threshold 40). This is in the same order of magnitude as the response observed in experiments with seedlings. No interaction was found between ozone and drought parameters. In Norway spruce, shoot growth was neither associated with ozone nor with drought.     

Growth and nutrition of Quercus rubra L. seedlings and mature trees after three seasons of ozone exposure

Seedling growth and nutritional status have been shown to be sensitive to ozone, but the influence of multi-season ozone exposure on mature tree growth and nutrition has not been examined. To determine if seedlings and mature trees were similarly affected by ozone exposure, growth and nutrient concentrations in northern red oak (Quercus rubra L.) 4-year-old seedlings and 32-year-old mature trees were examined after treatment with subambient, ambient and twice ambient concentrations of ozone for three growing seasons. SUM00 values summed over the three growing seasons were 147, 255 and 507 ppm-h, respectively, for the subambient, ambient and twice ambient exposures. For mature trees, no influence of ozone treatment on lower stem diameter growth, stem growth within the mid-canopy and foliar biomass was observed. Seedling height was increased by ozone, but biomass and diameter were unaffected. A reduction in the specific leaf weight of leaves in response to ozone coincident with the loss of recurrent flushing was observed in seedlings. Ozone exposure reduced foliar nitrogen concentrations and increased woody tissue nutrient concentrations in seedlings and mature trees at the end of the third growing season. These results suggest an influence of ozone on retranslocation processes in seedlings and mature trees.     

Differences in growth, leaf senescence and injury, and stomatal density in birch (Betula pendula Roth.) in relation to ambient levels of ozone in Finland

The differences in growth, leaf senescence, visible ozone injuries and stomatal density between one coastal site (natural ozone) and two inland sites (natural and elevated ozone) in Finland were determined for saplings of Betula pendula clones grown under open-field conditions during two growing seasons. Responses in growth, leaf senescence, visible injuries, and stomatal density were determined in relation to cumulative ozone exposure accumulated over the thresholds of 30, 40 and 50 ppb (10(9)) during the exposure period. In addition, the effects of the different ozone exposures on ultrastructure of chloroplasts were studied. Increasing ozone exposure resulted in reduced shoot dry weight, stimulated (first year) or reduced (second year) height growth, accelerated autumn yellowing of leaves, increased stomatal density, visible symptoms and chloroplast injuries, and increased number and size of plastoglobuli. Newly expanded mature leaves in midsummer were more sensitive to ozone episodes than younger developing leaves in the early growing season. In most parameters, the best correlation was achieved with the exposure index AOT30. Ozone risk for birch is highest in the southern coastal area of Finland, where background ozone concentrations are higher than in inland sites.     

Effects of ozone exposure in open-top chambers on poplar (Populus nigra) and beech (Fagus sylvatica): a comparison

Rooted cuttings of poplar (Populus nigra) and seedlings of beech (Fagus sylvatica) were exposed to ozone in open-top chambers for one growing season. Three treatments were applied: charcoal-filtered (CF), non-filtered (NF) and non-filtered air plus 30 ppb (nl l(-1)) ozone (NF+). Extra ozone was only added on clear days, from 09:00 until 17:00-20:00. The AOT40s (accumulated exposure over a threshold of 40 ppb), calculated from April to September were 4055 ppb.h for the NF and 8880 ppb.h for the NF+ treatments. For poplar ozone exposure caused highly significant reductions in growth rate, light-saturated net CO(2) assimilation rate, stomatal conductance, F(v)/F(m) and chlorophyll content. The largest effects were observed in August at which time ozone concentrations were elevated. A reduction was noticed in new leaf production, while accelerated ageing and visible damage to leaves caused high leaf losses. For beech the responses were similar but less pronounced: ozone exposure resulted in non-significant growth reductions, slight changes in light-saturated photosynthesis and accelerated leaf abscission. The chlorophyll content of beech leaves was not affected by the ozone treatments. The results confirmed previous observations that fast-growing tree species, such as most poplar species and hybrids, are more sensitive and responsive to tropospheric ozone than slower-growing species, such as beech. The growth reductions observed and reported here for beech were within the range of those reported in relationship to the AOT40 (accumulated exposure over a threshold of 40 ppb) critical level for ozone.     

Contents of free amino acids in needles of Norway Spruce trees in relation to novel forest decline. Studies on trees from a site in the northern Black Forest

The free amino acid content in needles of Norway spruce trees (about 45-year-old) was determined by means of HPLC. The studied trees have been growing at a forestry site in the Black Forest which is characterized by a high impact of ozone and magnesium deficiency. Measurements were carried out on visibly healthy and on damaged trees on several dates during two vegetation periods and during the course of a day. The amino acids occurring at the highest concentrations were glutamate, arginine, aspartate, and glutamine. The typical seasonal changes in the content of free amino acids, with their minimum during summer, were disturbed in the needles of the damaged trees. Particularly in summer and autumn the damaged trees showed increased levels of total amino acids and of most single amino acids. During the course of a day, the needles of the damaged trees showed a reduced amplitude in diurnal variations with the absolute level almost always higher in comparison to that of the undamaged trees. The results suggest that a shift in protein turnover towards enhanced degradation has taken place.     

Comparison of ozone uptake and sensitivity between a phytotron study with young beech and a field experiment with adult beech (Fagus sylvatica)

Chamber experiments on juvenile trees have resulted in severe injury and accelerated loss of leaves along with reduced biomass production under chronically enhanced O3 levels. In contrast, the few studies conducted on adult forest trees in the field have reported low O3 sensitivity. In the present study, young beech in phytotrons was more sensitive to O3 than adult beech in the field, although employed O3 regimes were similar. The hypotheses tested were that: (1) differences in O3 uptake were caused by the ontogenetically higher stomatal conductance of young compared to adult trees, (2) the experimental settings in the phytotrons enhanced O3 uptake compared to field conditions, and (3) a low detoxification capacity contributes to the higher O3 sensitivity of the young trees. The higher O3 sensitivity of juvenile beech in the phytotrons is demonstrated to relate to both the experimental conditions and the physiological responsiveness inherent to tree age.     

The influence of the season and the biotic factors on the cytosolic metal concentrations in the gills of the European chub (Leuciscus cephalus L.)

The influence of the season and the biotic factors (age and gill mass) on metal and protein levels in the gill cytosol of the young chubs (2 and 3 years old) was studied in Sava River in autumn 2005 and spring 2006. The obtained results are the first reported cytosolic concentrations of five metals and proteins for the chub gills. The average levels in autumn and spring, respectively, for total cytosolic proteins were 11.2 and 19.9 mg ml(-1), for Zn 6.3 and 10.3 microg ml(-1), for Fe 3.9 and 9.6 microg ml(-1), for Cu 68.4 and 79.0 ng ml(-1), for Mn 55.0 and 63.5 ng ml(-1), and for Cd 2.9 and 3.6 ng ml(-1). The influence of the gill mass on both the protein and the metal levels was observed, but it was seasonally dependent. In autumn, positive correlations were obtained between the gill mass and four parameters (total proteins, Mn, Zn and Fe), and negative with Cu. Contrary, in spring, even negative correlations of total proteins and some metals with the gill mass were observed. The proposed explanation for the different dependence of metal levels on the gill mass in autumn and spring was the seasonal difference in feeding intensity and metabolic rate, with presumably faster metabolism and water filtration through gills in spring. This hypothesis was further supported by the statistically significantly higher concentrations of the total proteins, Zn and Fe, as well as the Fulton condition indices in the spring period.     

Physiology, morphology, and ozone uptake of leaves of black cherry seedlings, saplings, and canopy trees

Patterns of ozone uptake were related to physiological, morphological, and phenological characteristics of different-sized black cherry trees (Prunus serotina Ehrh.) at a site in central Pennsylvania. Calculated ozone uptake differed among open-grown seedlings, forest gap saplings, and canopy trees and between leaves in the upper and lower crown of saplings and canopy trees. On an instantaneous basis, seedling leaves had the greatest ozone uptake rates of all tree size classes due to greater stomatal conductance and higher concentrations of ozone in their local environment. A pattern of higher stomatal conductance of seedlings was consistent with higher incident photosynthetically-active radiation, stomatal density, and predawn xylem water potentials for seedlings relative to larger trees. However, seedlings displayed an indeterminate pattern of shoot growth, with the majority of their leaves produced after shoot growth had ceased for canopy and sapling trees. Full leaf expansion occurred by mid-June for sapling and canopy trees. Because many of their leaves were exposed to ozone for only part of the growing season, seedlings had a lower relative exposure over the course of the growing season, and subsequently lower cumulative uptake, of ozone than canopy trees and a level of uptake similar to upper canopy leaves of saplings. Visible injury symptoms were not always correlated with patterns in ozone uptake. Visible symptoms were more apparent on seedling leaves in concurrence with their high instantaneous uptake rates. However, visible injury was more prevalent on leaves in the lower versus upper crown of canopy trees and saplings, even though lower crown leaves had less ozone uptake. Lower crown leaves may be more sensitive to ozone per unit uptake than upper crown leaves because of their morphology. In addition, the lower net carbon uptake of lower crown leaves may limit repair and anti-oxidant defense processes.     

California black oak response to nitrogen amendment at a high O3, nitrogen-saturated site

In a nitrogen (N) saturated forest downwind from Los Angeles, California, the cumulative response to long-term background-N and N-amendment on black oak (Quercus kelloggii) was described in a below-average and average precipitation year. Monthly measurements of leaf and branch growth, gas exchange, and canopy health attributes were conducted. The effects of both pollutant exposure and drought stress were complex due to whole tree and leaf level responses, and shade versus full sun leaf responses. N-amended trees had lower late summer carbon (C) gain and greater foliar chlorosis in the drought year. Leaf water use efficiency was lower in N-amended trees in midsummer of the average precipitation year, and there was evidence of poor stomatal control in full sun. In shade, N-amendment enhanced stomatal control. Small differences in instantaneous C uptake in full sun, lower foliar respiration, and greater C gain in low light contributed to the greater aboveground growth observed.     

Effects of simulated acid rain and ozone on foliar chemistry of field-grown Pinus ponderosa seedlings and mature trees

We investigated the additive and interactive effects of simulated acid rain and elevated ozone on C and N contents, and the C:N ratio of one-year-old and current-year foliage of field-grown mature trees and their half-sib seedlings of a stress tolerant genotype of ponderosa pine. Acid rain levels (pH 5.1 and 3.0) were applied weekly to foliage only (no soil acidification or N addition), from January to April, 1992. Plants were exposed to two ozone levels (ambient and twice-ambient) during the day from September 1991 to November 1992. The sequential application of acid rain and elevated ozone mimicked the natural conditions. Twice-ambient ozone significantly decreased foliar N content (by 12-14%) and increased the C:N ratio of both one-year-old and current-year foliage of seedlings. Although similar ozone effects were also observed on one-year-old foliage of mature trees, the only statistically significant effect was an increased C:N ratio when twice-ambient ozone combined with pH 3.0 rain (acid rain by ozone interaction). Enhancing the effect of twice-ambient ozone in increasing the C:N ratio of one-year-old foliage of mature trees in June was the only significant effect of acid rain.     

Effects of chronic elevated ozone exposure on gas exchange responses of adult beech trees (Fagus sylvatica) as related to the within-canopy light gradient

The effects of elevated O₃ on photosynthetic properties in adult beech trees (Fagus sylvatica) were investigated in relation to leaf mass per area as a measure of the gradually changing, within-canopy light availability. Leaves under elevated O₃ showed decreased stomatal conductance at unchanged carboxylation capacity of Rubisco, which was consistent with enhanced δ¹³C of leaf organic matter, regardless of the light environment during growth. In parallel, increased energy demand for O₃ detoxification and repair was suggested under elevated O₃ owing to enhanced dark respiration. Only in shade-grown leaves, light-limited photosynthesis was reduced under elevated O₃, this effect being accompanied by lowered F_v/F_m. These results suggest that chronic O₃ exposure primarily caused stomatal closure to adult beech trees in the field regardless of the within-canopy light gradient. However, light limitation apparently raised the O₃ sensitivity of photosynthesis and accelerated senescence in shade leaves.     

A comparison of two different approaches for mapping potential ozone damage to vegetation. A model study

Two very different types of approaches are currently in use today for indicating risk of ozone damage to vegetation in Europe. One approach is the so-called AOTX (accumulated exposure over threshold of Xppb) index, which is based upon ozone concentrations only. The second type of approach entails an estimate of the amount of ozone entering via the stomates of vegetation, the AFstY approach (accumulated stomatal flux over threshold of Y nmol m(-2) s(-1)). The EMEP chemical transport model is used to map these different indicators of ozone damage across Europe, for two illustrative vegetation types, wheat and beech forests. The results show that exceedences of critical levels for either type of indicator are widespread, but that the indicators give very different spatial patterns across Europe. Model simulations for year 2020 scenarios suggest reductions in risks of vegetation damage whichever indicator is used, but suggest that AOT40 is much more sensitive to emission control than AFstY values.     

Photosynthetic and stomatal conductance responses of Norway spruce and beech to ozone, acid mist and frost--a conceptual model

Two-year-old beech and Norway spruce seedlings were exposed to a combination of ozone and acid mist treatments in open-top chambers in Scotland during the months of July through to September 1988. Replicate pairs of chambers received charcoal-filtered air (control), ozone-enriched air (140 nl ozone litre(-1)) or 140 nl ozone litre(-1) plus a synthetic acid mist (pH 2.5) composed of ammonium nitrate and sulphuric acid. Field measurements of assimilation and stomatal conductance were made during August. In addition, measurements of assimilation and conductance were made during September in the laboratory. Light response curves of assimilation and conductance were determined using a GENSTAT nonrectangular hyperbolic model. During February 1988/9 the Norway spruce were subject to a four day warming period at 12 degrees C and the light response of assimilation determined. The same plants were then subject to a 3-h night-time frost of -10 degrees C. The following day the time-course of the recovery of assimilation was determined. It was found that ozone fumigation did not influence the light response of assimilation of beech trees in the field, although stomatal conductance was reduced in the ozone-fumigated trees. The rate of light-saturated assimilation of Norway spruce was increased by ozone fumigation when measured in the field. Measurements of assimilation of Norway spruce made during the winter showed that prior to rewarming there was no difference in the rate of light-saturated assimilation for control and ozone-fumigated trees. However, the ozone plus acid mist-treated trees exhibited a significantly higher rate. The 4-day period of warming to 12 degrees C increased the rate of light-saturated assimilation in all treatments but only the ozone plus acid mist-treated trees showed a significant increase. Following a 3-h frost to -10 degrees C the control trees exhibited a reduction in the rate of light-saturated assimilation (Amax) to 80% of the pre-frost value. In comparison, following the frost, the ozone-fumigated trees showed an Amax of 74% of the pre-frost value. The ozone plus acid mist-treated trees showed an Amax of 64% of the pre-frost trees. The time taken for Amax to attain 50% of the pre-frost value increased from 30 min (control) to 85 min for ozone-fumigated trees to 190 min (ozone plus acid mist). These results are discussed in relation to the impact of mild, short-term frosts, which are known to occur with greater frequency than extreme, more catastrophic frost events. A simple conceptual framework is proposed to explain the variable results obtained in the literature with respect to the impact of ozone upon tree physiology.     

Size-mediated foliar response to ozone in black cherry trees

Local ozone concentration and visible foliar injury were measured over the 1994 growing season on open-grown black cherry (Prunus serotina Ehrh.) trees of varying size (age) within forest stands and adjacent openings at a site in north-central Pennsylvania. Relationships were determined between visible ozone injury and ozone exposure, as well as calculated between injury and ozone uptake expressed as the product of stomatal conductance and ozone concentration. In addition, simultaneous measurements of visible symptoms and leaf gas exchange were also conducted to determine the correlation between visible and physiological injury and ozone exposure. By September, the amount of leaf area affected by visible foliar ozone injury was greatest for seedlings (46%), followed by canopy trees (20%) and saplings (15%). A large amount of variability in foliar ozone symptom expression was observed among trees within a size class. Sum40 and Sum60 (ozone concentration > 40 and > 60 nl liter(-1)) cumulative exposure statistics were the most meaningful indices for interpretation of foliar injury response. Seedlings were apparently more sensitive to ozone injury than larger trees because their higher rates of stomatal conductance resulted in higher rates of ozone uptake. Seedlings also had higher rates of early leaf abscission than larger trees with an average of nearly 30% of the leaves on a shoot abscised by 1 September compared to approximately 5% for larger trees. However, per unit ozone uptake into the leaf, larger trees exhibited larger amounts of foliar injury. The amount of visible foliar injury was negatively correlated (r(2) = 0.82) with net photosynthetic rates, but was not related to stomatal conductance. Net photosynthesis and stomatal conductance thus became uncoupled at high levels of visible foliar injury.     

Seasonal variations of monoterpene emissions from coniferous trees of different ages in Korea

Seasonal variations of emission rates and compositions from coniferous species were measured under controlled conditions using a vegetation enclosure method. Total emission rates and compositions of monoterpene compounds from young and adult trees in different seasons were compared.

It was found that the total emission rates and the components of monoterpene varied significantly with tree species, age, and season. Total emissions from C. japonica and P. koraiensis were higher for older trees than for younger trees; however, significantly higher emissions were found from younger trees for C. obtusa. Higher monoterpene emission rates from each plant were found in spring and summer compared with autumn and winter emissions.     

Use of sap flow measurements to validate stomatal functions for mature beech (Fagus sylvatica) in view of ozone uptake calculations

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.     

Beech (Fagus sylvatica) response to ozone exposure assessed with a chlorophyll a fluorescence performance index

This paper describes a relationship between ozone exposure, biomass, visual symptoms and a chlorophyll a fluorescence performance index for young beech trees (Fagus sylvatica). The plants were exposed to four levels of ozone in open-top fumigation chambers (50, 85, 100% of ambient, and 50% of ambient+30 nl l(-1) ozone) that fluctuated in parallel with ambient ozone during a single growing season. The trees were fumigated in the four treatments with ozone levels corresponding to an AOT40 (accumulated exposure above a threshold of 40 nl l(-1)) of 0.01, 3.35, 7.06 and 19.70 microl l(-1) h, respectively. Highly significant differences were found between the 50% of ambient+30 nl l(-1) ozone treatment and all other treatments, with a 70.5% reduction in primary photosynthetic performance, as measured with the PI index. The reduction of the PI values demonstrated a high correlation with visual symptom development (r(2)=0.98), and by the end of September with biomass loss (r(2)=0.99). A significant ozone exposure-response relationship was found between AOT40 and primary photochemistry (r(2)=0.97). Thus, analysis of PI provides an alternative method for regional monitoring of tree health within the context of the currently employed AOT40.