Psidium guajava 'Paluma' (the guava plant) as a new bio-indicator of ozone in the tropics

Psidium guajava 'Paluma' saplings were exposed to carbon filtered air (CF), ambient non-filtered air (NF), and ambient non-filtered air+40ppb ozone (NF+O(3)) 8h per day during two months. The AOT40 values at the end of the experiment were 48, 910 and 12 895ppbh(-1), respectively for the three treatments. After 5 days of exposure (AOT40=1497ppbh(-1)), interveinal red stippling appeared in plants in the NF+O(3) chamber. In the NF chamber, symptoms were observed only after 40 days of exposure (AOT40=880ppbh(-1)). After 60 days, injured leaves per plant corresponded to 86% in NF+O(3) and 25% in the NF treatment, and the average leaf area injured was 45% in NF+O(3) and 5% in the NF treatment. The extent of leaf area injured (leaf injury index) was explained mainly by the accumulated exposure of ozone (r(2)=0.91; p<0.05).     

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.     

Impact of ozone on the growth of birch (Betula pendula) saplings

Saplings of one half-sib family of birch, Betula pendula, were exposed to three levels of ozone in open-top chambers (OTCs) during two growing seasons 1997-1998. The ozone treatments were non-filtered air (NF, accumulated daylight AOT40 over the two growing seasons of 3.0 l l-1 h), non-filtered air with extra ozone (NF+, accumulated daylight AOT40 of 27.3 l l-1 h) and non-filtered air with additional extra ozone (NF++, accumulated daylight AOT40 of 120 l l-1 h). The birch saplings, including the roots, were harvested after the first and second growing seasons. After the first growing season, the NF++ treatment reduced the total wood biomass by 22%, relative to the NF treatment. There was no further reduction of the total wood biomass in the NF++ treatment after the second growing season. The root biomass was reduced by 30% after the first growing season. The shoot/root ratio, as well as the proportional biomass of leaves, were increased by ozone during both years. The ozone impact on the relative growth rate was estimated to -2% per 10 l l-1 h daylight AOT40 per growing season.     

Anthocyanins and tannins in ozone-fumigated guava trees

Psidium guajava “Paluma”, a tropical tree species, is known to be an efficient ozone indicator in tropical countries. When exposed to ozone, this species displays a characteristic leaf injury identified by inter-veinal red stippling on adaxial leaf surfaces. Following 30 days of three ozone treatments consisting of carbon filtered air (CF – AOT40 = 17 ppb h), ambient non-filtered air (NF – AOT40 = 542 ppb h) and ambient non-filtered air + 40 ppb ozone (NF + O₃ – AOT40 = 7802 ppb h), the amounts of residual anthocyanins and tannins present in 10 P. guajava (“Paluma”) saplings were quantified. Higher amounts of anthocyanins were found in the NF + O₃ treatment (1.6%) when compared to the CF (0.97%) and NF (1.30%) (p < 0.05), and of total tannins in the NF + O₃ treatment (0.16%) compared to the CF (0.14%). Condensed tannins showed the same tendency as enhanced amounts. Regression analyses using amounts of tannins and anthocyanins, AOT40 and the leaf injury index (LII), showed a correlation between the leaf injury index and quantities of anthocyanins and total tannins. These results are in accordance with the association between the incidence of red-stippled leaves and ozone polluted environments.     

Ozone exposure thresholds and foliar injury on forest plants in Switzerland

Canton Ticino in southern Switzerland is exposed to some of the highest concentrations of tropospheric ozone in Europe. During recent field surveys in Canton Ticino, foliar symptoms identical to those caused by ozone have been documented on native tree and shrub species. In Europe, the critical ozone level for forest trees has been defined at an AOT40 of 10 ppm.h O3 (10 ppm.h accumulated exposure of ozone over a threshold of 40 ppb) during daylight hours over a six-month growing season. The objective of this study was to determine the amount of ambient ozone required to induce visible foliar symptoms on various forest plant species in southern Switzerland. Species were grown within eight open-top chambers and four open plots at the Vivaio Lattecaldo Cantonal Forest Nursery in Ticino, Switzerland. Species differed significantly in terms of the ppb.h exposures needed to cause visible symptoms. The most to least symptomatic species grown within open-plots in this study rank as Prunus serotina, Salix viminalis, Vibrnum lantana, Rhamnus cathartica, Betula pendula, Rumex obtusifolius, Sambucus racemosa, Morus nigra, Prunus avium, Fraxinus excelsior, Rhamnus frangula, Alnus viridis, Fagus sylvatica and Acer pseudoplatanus. Similar rankings were obtained in the non-filtered chamber plots. The ranking of species sensitivity closely follows AOT values for the occurrence of initial symptoms and symptom progression across the remainder of the exposure season. Species that first showed evidence of foliar injury also demonstrated the most sensitivity throughout the growing season, with symptoms rapidly advancing over ca. 25-30% of the total plant leaf surfaces by the end of the observation period. Conversely, those species that developed symptoms later in the season had far less total injury to plant foliage by the end of the observation period (1.5 to < 5% total leaf area injured). The current European ambient ozone standard may be insufficient to protect native plant species from visible foliar injury, and many more native species may be sensitive to ozone-induced foliar injury than are currently known.     

Ozone exposure induces the activation of leaf senescence-related processes and morphological and growth changes in seedlings of Mediterranean tree species

Four Mediterranean tree taxa, Quercus ilex subsp. ilex, Quercus ilex subsp. ballota, Olea europaea cv. vulgaris and Ceratonia siliqua, were exposed to different ozone (O(3)) concentrations in open top chambers (OTCs) during 2 years. Three treatments were applied: charcoal-filtered air (CF), non-filtered air (NF) and non-filtered air plus 40 ppb(v) of O(3) (NF +). The photochemical maximal efficiency, Fv/Fm, decreased in NF + plants during the second year of exposure, especially during the most stressful Mediterranean seasons (winter and summer). An increase of delta(13)C was found in three of the four studied species during the first year of exposure. This finding was only maintained in C. siliqua during the second year. Decreases in the chlorophyll content were detected during the first year of fumigations in all the species studied, but not during the second year. The NF + treatment induced changes in foliar anatomical characteristics, especially in leaf mass per area (LMA) and spongy parenchyma thickness, which increased in some species. A reduction in N content and an increase in delta(15)N were found in all species during the second year when exposed in the NF + OTCs, suggesting a change in their retranslocation pattern linked to an acceleration of leaf senescence, as also indicated by the above mentioned biochemical and anatomical foliar changes. The two Q. ilex subspecies were the most sensitive species since the changes in N concentration, delta(15)N, chlorophyll, leaf area, LMA and biomass occurred at ambient O(3) concentrations. However, C. siliqua was the most responsive species (29% biomass reduction) when exposed to the NF + treatment, followed by the two Q. ilex subspecies (14-20%) and O. europaea (no significant reduction). Ozone resistance of the latter species was linked to some plant traits such as chlorophyll concentrations, or spongy parenchyma thickness.     

Ambient ozone and crop loss: establishing a cause-effect relationship

This paper provides the results of a retrospective mathematical analysis of the US NCLAN (National Crop Loss Assessment Network) open-top chamber data. Some 77% of the 73 crop harvests examined, showed no statistically significant yield differences between NF (non-filtered open-top chamber) and AA (chamberless, ambient air) treatments (no easily discernable chamber effects on yield). However, among these cases only seven acceptable examples showed statistically significant yield reductions in NF compared to the CF (charcoal filtered open-top chamber) treatment. An examination of the combined or cumulative hourly ambient O3 frequency distribution for cases with yield loss in NF compared to a similar match of cases without yield loss showed that the mean, median and the various percentiles were all higher (>/= 3 X) in the former in contrast to the latter scenario. The combined frequency distribution of hourly O3 concentrations for the cases with yield loss in NF were clearly separated from the corresponding distribution with no yield loss, at O3 concentrations > 49 ppb. Univariate linear regressions between various O3 exposure parameters and per cent yield losses in NF showed that the cumulative frequency of occurrence of O3 concentrations between 50 and 87 ppb was the best predictor (adjusted R2 = 0.712 and p = 0.011). This analysis also showed that the frequency distribution of hourly concentrations up to 87 ppb O3 represented a critical point, since the addition of the frequency distributions of > 87 ppb O3 did not improve the R2 values. In fact as the frequency of hourly O3 concentrations included in the regression approached 50-100 ppb, the R2 value decreased substantially and the p value increased inversely. Further, univariate linear regressions between the frequencies of occurrence of various O3 concentrations between 50 and 90 ppb and: (a) cases with no yield difference in NF and (b) cases with yield increase in NF compared to the CF treatment (positive effect) provided no meaningful statistical relationship (adjusted R2 = 0.000) in either category. These results support the basis that additional evaluation of the frequency of occurrence of hourly O3] concentrations between 50 and 87 ppb for cases with the yield reductions could provide a meaningful ambient O3 standard, objective or guideline for vegetation.     

Ozone exposure of field-grown winter wheat affects soil mesofauna in the rhizosphere

A 2-year open-top chamber experiment with field-grown winter wheat (Triticum aestivum L. cv. Astron) was conducted to examine the effects of ozone on plant growth and selected groups of soil mesofauna in the rhizosphere. From May through June in each year, plants were exposed to two levels of O₃: non-filtered (NF) ambient air or NF+ 40 ppb O₃ (NF+). During O₃ exposure, soil sampling was performed at two dates according to different plant growth stages. O₃ exposure reduced above- and below-ground plant biomass in the first year, but had little effect in the second year. The individual density of enchytraeids, collembolans and soil mites decreased significantly in the rhizosphere of plants exposed to NF+ in both years. Differences were highest around anthesis, i.e. when plants are physiologically most active. The results suggest that elevated O₃ concentrations may influence the dynamic of decomposition processes and the turnover of nutrients.     

Wheat-kernel growth characteristics during exposure to chronic ozone pollution

Chronic exposure to ozone (O(3)) air pollution can reduce yield in wheat; however, little is known concerning the effects of O(3) stress on kernel development. A field study was conducted to investigate the effects of chronic O(3) exposure on kernel-growth components of two soft red winter-wheat genotypes (Seven and MD5518308). Five air-quality treatments, including charcoal-filtered air (CF), non-filtered air (NF), NF + 20, and NF + 40 and NF + 80 nl O(3) liter(-1) air were applied 4 h d(-1), 5 d wk(-1) through maturity. In the case of the NF + treatments, O(3) was added to existing ambient O(3) levels. Spike samples were collected 16, 20, 24, 28, and 32 days after anthesis (DAA). Linear and quadratic equations were fitted to kernel-weight data to estimate kernel-growth rate (KGR) and kernel-fill duration (KFD). Effective filling period (EFP) and assimilate utilization (AU) were also determined. Rates of growth for individual kernels were 0.74 mg d(-1) and 1.07 mg d(-1) for the NF + 80 and CF treatments, respectively. The NF + 80 nL litter(-1) O(3) treatment significantly reduced KGR and AU compared with the CF treatment. Severn had a significantly loger KFD than MD5518308, but O(3) had no significant effect on KFD of either genotype. Each genotype had similar EFP values, and O(3) had no significant effect on EFP. Linear relationships between O(3) exposure and kernel weight suggests that O(3) effects on kernel weight begin soon after anthesis in MD5518308, but, in Severn, O(3) has a greater effect on kernel weight during the later stages of kernel development. These data suggest that decreased economic yield associated with chronic O(3) exposure is primarily the result of decreased KGR.     

Effects of ozone and soil water deficit on roots and shoots of field-grown soybeans

Water-stressed and well-watered soybean (Glycine max cvs. Williams and Corsoy) plants were exposed to increasing seasonal doses of ozone (O(3)) using open-top field chambers and ambient air plots. Chamber O(3) treatments included charcoal filtered (CF) air, non-filtered (NF) air, NF + 0.03, NF + 0.06 and NF + 0.09 microl litre(-1) O(3). Soil water potentials measured at 25 and 45 cm averaged -0.40 MPa and -0.05 MPa, respectively, for the plots in the water-stressed and well-watered series. Total root length/core, root length densities, and biomasses (dry weights) were determined. With Williams, a very popular cultivar in recent years, total root length for all O(3) treatments averaged 58% more under water-stress conditions than in well-watered plots, but the range was from 136% to 11% more for NF air and NF + 0.09 microl litre(-1) O(3), respectively. Increasing the O(3) exposure dose did not affect root lengths or weights in the well-watered series. With Corsoy, water stress did not significantly increase root development. In both soil moisture regimes, with both cultivars, there was a linear decrease in seed yield and top dry weight as the O(3) exposure dose increased.     

Contrasting effects of ozone under different water supplies in two Mediterranean tree species

The effects of ozone (O₃) exposure under different water availabilities were studied in two Mediterranean tree species: Quercus ilex and Ceratonia siliqua. Plants were exposed to different O₃ concentrations in open top chambers (charcoal-filtered air (CF), non-filtered air (NF)) and non-filtered air plus 40 ppb_v of O₃ ((7:00–17:00 solar time) (NF+)) during 2 years, and to different water regimes (IR, sample irrigation, and WS, reduced water dose to 50%) through the last of those 2 years. AOT40 in the NF+ treatment was 59265 ppb_v h (from March 1999 to August 1999) while in the NF treatment, the AOT40 was 6727 ppb_v h for the same period. AOT40 was always 0 in the CF treatment. WS plants presented lower stomatal conductances and net photosynthetic rates, and higher foliar N concentrations than IR plants in both species. The irrigation treatment did not change the response trends to ozone in Q. ilex, the most sensitive species to O₃ ambient concentrations, but it changed those of C. siliqua, the least sensitive species, since its ozone-fumigated WS plants did not decrease their net photosynthetic rates nor their biomass accumulation as it happened to its ozone-fumigated IR plants. These results show interspecific variations in O₃ sensitivity under different water availabilities.     

Yield loss assessments for cultivars of broccoli, lettuce, and onion exposed to ozone

The effects of the photochemical oxidant air pollutant ozone (O(3)) on growth and yield of three garden crops, broccoli (Brassica oleracea L.), lettuce (Lactuca sativa L.), and onion (Allium cepa L.) were studied in an open-top chamber experiment conducted in the field in southern California. Four cultivars each of leaf lettuce, broccoli, and globe onion were exposed to charcoal-filtered air (CF), non-filtered (NF) air, or NF plus 1.5 times ambient O(3) concentration from 4 weeks after germination in January or February until harvest. Exposures lasted 31 days for lettuce, 55 to 78 days for broccoli, and 105 days for onion. Results showed that despite severe O(3) injury to outer leaves, lettuce yields were not affected by O(3). Broccoli also was resistant to O(3) and no growth reduction was observed at ambient O(3) concentrations. Onions were more susceptible to O(3), but only one cv. 'Rio Bravo' had significant yield losses (ca. 5%) at ambient O(3) levels. These results suggest that, in general, concentrations of O(3) in the winter and spring may be below the threshold for adverse effects on yields of broccoli, lettuce and onion.     

From critical levels to critical loads for ozone: a discussion of a new experimental and modelling approach for establishing flux-response relationships for agricultural crops and native plant species

Present critical levels for ozone (O3) for protecting vegetation against adverse effects are based on exposure-response relationships mainly derived from open-top chamber experiments and are expressed as an Accumulated exposure Over a Threshold of 40 ppb (AOT40). In that context with a revision of the UN (United Nations)-ECE (Economic Commission for Europe) Gothenburg protocol, AOT40 values should be replaced by flux-oriented quantities, i.e. in the end by critical loads. At present, the database for the derivation of critical loads for O3 is extremely inadequate. Furthermore, the currently available flux-response relationships are also derived from open-top chamber experiments. The use of a relationship for spring wheat in a risk assessment for an agricultural site in Hesse, Germany, demonstrates in principle, the applicability of the critical load concept for O3. Comparisons of diurnal variation of stomatal uptake and AOT40 showed that a major part of toxicologically effective stomatal uptake occurred before noon whereas the AOT40 values were dominated by the O3 concentrations during afternoon. In other words, the AOT40 exposure index do not adequately address the O3 burden during hours when plants are sensitive to O3 uptake. However, due to the differences in radiation, air temperature and humidity between the chamber and the ambient microclimates, a derivation of flux-response relationships from chamber experiments is likely to be questionable, especially for species rich ecosystems: Here, without any changes in the pollution climate, significant modifications of species composition as well as an earlier beginning of the growing season has been previously observed. To overcome the problems associated with chamber-derived flux-response relationships, a new experimental and modelling concept, was developed. The approach, briefly described in this paper, combines methods in air pollution toxicology and micrometeorology. As an analogy to the free-air fumigation concept, O3 is released into the air by an injection system above the plant canopy. The assessment of dispersion and surface deposition of O3 released is based on Lagrangian trajectory modelling. Depending on wind direction and velocity, atmospheric stratification and surface roughness, without any disturbance of the microclimate and micrometeorology, several sub-areas can be identified around the source position with differing deposition rates above the ambient level. Taking into account the actual O3 background deposition, deposition rates and vegetation responses observed in these sub-areas can easily be used to derive flux-effect relationships under ambient conditions and more realistic limiting values to protect our environment.     

Ozone-induced foliar injury in saplings of Psidium guajava 'Paluma' in São Paulo, Brazil

Psidium guajava 'Paluma' was evaluated under field conditions as a tropical bioindicator species of ozone (O(3)). Three exposures of 90 days each were performed (June 21, 2004-March 19, 2005). In each one of them, saplings of 'Paluma' (n=30) were exposed to ambient air at a site in S?o Paulo (Brazil) with high ozone concentrations, and in a greenhouse with charcoal-filtered air. Ozone-like visible foliar injuries were observed during the winter, spring and summer exposures, when AOT40 reached 6166ppbh, 3504ppbh and 4828ppbh, respectively. No injuries were observed in the plants kept under filtered air. The injuries consisted in red stippling on adaxial leaf surfaces. They did not cover the veins and appeared first in older leaves, becoming more intense as the exposure period increased. Injury incidence, severity, and the cumulative exposure threshold at injury onset varied among the exposure periods. AOT40 explained partly the incidence, severity and leaf injury index LII (r(2)=0.52, 0.39, 0.38, respectively, p=0.002). The results confirm the potential of the species as an O(3)-sensitive bioindicator.     

Responses of spring wheat (Triticum aestivum L.) to ozone produced by either electric discharge and dry air or by UV-lamps and ambient air

The aim of the present study was to examine if ozone produced similar effects on spring wheat growth with and without small amounts of nitrogen oxides. Two methods were used to produce ozone: the first method consisted of dry pressurized air fed to an electric discharge generator generating the byproducts, N2O5 and N2O, the second method consisted of ambient air fed to UV-lamps. Two spring wheat cultivars (Triticum aestivum L. cvs Minaret and Eridano) were exposed in small open-top chambers to charcoal-filtered air, non-filtered ambient air, and non-filtered ambient air with the addition of ozone for 8 h (0900 to 1700 h) daily, for five weeks. Plants were harvested every week. The growth of Minaret was shown to be more sensitive to O3 than that of Eridano. Leaf senescence increased with increasing ozone level in both cultivars. The total above-ground biomass dry weight decreased with increasing ozone concentration in Minaret, but not in Eridano. The Minaret plants reacted with more damaged leaf dry weight and inhibition of growth when O3 was produced by UV-lamps than when O3 was produced by air fed to an electric discharge generator. This could be explained by more nitrogen content per plant but not by increased nitrogen concentration in plant tissue in plants exposed to increased O3 and small amounts of incidental nitrogen oxides.     

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.     

Growth and nutritive quality of Poa pratensis as influenced by ozone and competition

Interspecific plant competition has been hypothesized to alter effects of early-season ozone (O3) stress. A phytometer-based approach was utilized to investigate O3 effects on growth and nutritive quality of Poa pratensis grown in monoculture and in mixed cultures with four competitor-plant species (Anthoxanthum odoratum, Achillea millefolium, Rumex acetosa and Veronica chamaedrys). Mesocosms were exposed during April/May 2000-2002 to charcoal-filtered air+25 ppb O3 (control) or non-filtered air+50 ppb O3 (elevated O3). Biomass production was not affected by O3, but foliar injury symptoms were observed in May 2002. Early-season O3 exposure decreased relative food value of P. pratensis by an average of 8%, which is sufficient to have nutritional implications for its utilization by herbivores. However, forage quality response to O3 was not changed by interspecific competition. Lack of injury and nutritive quality response in P. pratensis harvested in September may reflect recovery from early-season O3 exposure.     

Ozone air pollution and foliar injury development on native plants of Switzerland

The objectives of this study were to examine the foliar sensitivity to ozone exposure of 12 tree, shrub, and herbaceous species native to southern Switzerland and determine the seasonal cumulative ozone exposures required to induce visible foliar injury. The study was conducted from the beginning of May through the end of August during 2000 and 2001 using an open-top chamber research facility located within the Lattecaldo Cantonal Forest Nursery in Canton Ticino, southern Switzerland (600 m asl). Plants were examined daily and dates of initial foliar injury were recorded in order to determine the cumulative AOT40 ppb h ozone exposure required to cause visible foliar injury. Plant responses to ozone varied significantly among species; 11 species exhibited visible symptoms typical of exposures to ambient ozone. The symptomatic species (from most to least sensitive) were Populus nigra, Viburnum lantana, Salix alba, Crataegus monogyna, Viburnum opulus, Tilia platyphyllos, Cornus alba, Prunus avium, Fraxinus excelsior, Ribes alpinum, and Tilia cordata; Clematis spp. did not show foliar symptoms. Of the 11 symptomatic species, five showed initial injury below the critical level AOT40 10 ppmh O3 in the 2001 season.     

Ozone exposure-response relationships for biomass and root/shoot ratio of beech (Fagus sylvatica), ash (Fraxinus excelsior), Norway spruce (Picea abies) and Scots pine (Pinus sylvestris)

Current-year seedlings of beech, ash, Norway spruce and Scots pine were exposed during one growing season to different, but moderate, ozone (O(3)) scenarios representative for Switzerland (50, 85, 100% ambient, 50% ambient+30 nl l(-1)) in open-top chambers (OTCs) and to ambient O(3) concentrations in the field. Biomass significantly decreased with increasing O(3) dose in all species except for spruce. Losses of 25.5% (ash), 17.4% (beech), 9.9% (Scots pine) were found per 10 microl l(-1) h accumulated O(3) exposure over a threshold concentration of 40 nl l(-1) during daylight hours (AOT40). Ratios of root/shoot biomass (RSR) also significantly decreased with increasing AOT40 levels in beech and ash, but not in Norway spruce and Scots pine. The data show that the deciduous species beech and ash were more susceptible to O(3) with respect to RSR and biomass than the coniferous species Norway spruce and Scots pine.