Impact of ozone and reduced water supply on the biomass accumulation of Norway spruce saplings

Norway spruce saplings [Picea abies (L.) Karst.] were exposed during four growing seasons to two different ozone treatments in open-top chambers: charcoal filtered air (CF), and non-filtered air with extra ozone (NF+, 1.4xambient concentrations). Within each ozone treatment the saplings were either kept well watered or treated with a 7-8 week period with reduced water supply each growing season. The total biomass of the trees was measured in April and September during each of the last three growing seasons. NF+ significantly reduced the total biomass accumulation of Norway spruce saplings during the fourth growing season. No interaction between ozone and reduced water supply could be detected. The magnitude of the ozone impact after 4 years of exposure was an 8% reduction of the total plant biomass and a 1.5% reduction of the RGR. The reduced water supply reduced the total biomass 29% and the RGR 12%.     

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.     

Effects of air pollutants on the growth and antioxidative system of Norway spruce exposed in open-top chambers

Grafted Norway spruce trees were subjected to exposure beginning in April 1988, to one of four different air treatments in open-top chambers: Charcoal filtered air (CF), non-filtered air (NF), non-filtered air with the addition of O(3) during summer (NFO), and SO(2) plus NO(2) during winter (NFOSN). CF trees were considered as the reference group. No effects on growth parameters were observed. Samples of the two youngest needle year classes were taken late in November 1989 for enzyme determinations. The activity of ascorbic acid peroxidase (A-POD) increased the same level in all treatments, and activities of catalase and dehydroascorbic acid reductase (DHA-R) increased only in NF and NFO treatments. A higher level of activity in the NFOSN treatment was observed only for glucose-6-phosphate-dehydrogenase (Glc-6-P-DH) and non-specific peroxidase (POD). Isoelectric focusing of POD showed a changed pattern in the NFOSN treatment. Neither activity nor isoelectric focusing of superoxidase dismutase (SOD) was changed in any of the treatments.     

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).     

Tibouchina pulchra (Cham.) Cogn., a native Atlantic Forest species, as a bio-indicator of ozone: visible injury

Tibouchina pulchra saplings were exposed to carbon filtered air (CF), ambient non-filtered air (NF) and ambient non-filtered air+40 ppb ozone (NF+O3) 8 h per day during two months. The AOT40 values at the end of the experiment were 48, 910 and 12,895 ppb h(-1), respectively, for the three treatments. After 25 days of exposure (AOT40=3871 ppb h(-1)), interveinal red stippling appeared in plants in the NF+O3 chamber. In the NF chamber, symptoms were observed only after 60 days of exposure (AOT40=910 ppb h(-1)). After 60 days, injured leaves per plant corresponded to 19% in NF+O3 and 1% in the NF treatment; and the average leaf area injured was 7% within the NF+O3 and 0.2% within the NF treatment. The extent of leaf area injured (leaf injury index) was mostly explained by the accumulated exposure of ozone (r2=0.89; p<0.05).     

Yield and grain quality of spring wheat (Triticum aestivum L., cv. Drabant) exposed to different concentrations of ozone in open-top chambers

Spring wheat (Triticum aestivum L., cv. Drabant) was exposed to different concentrations of ozone in open-top chambers for two growing seasons, 1987 and 1988, at a site located in south-west Sweden. The chambers were placed in a field of commercially grown spring wheat. The treatments were charcoal-filtered air (CF), non-filtered air (NF) and non-filtered air plus extra ozone (NF(+)). In 1988, one additional ozone concentration (NF(++)) was used. Grain yield was affected by the ozone concentration of the air. Air filtration resulted in an increase in grain yield of about 7% in both years, compared to NF. The addition of ozone (NF(+), NF(++)) reduced grain yield and increased the content of crude protein of the grain in both years. Filtration of the air had no significant effect on the content of crude protein, compared to NF. The results showed a strong positive chamber effect on grain yield in the cold and wet summer of 1987. In 1988, there was no net chamber effect on grain yield. The relative differences between the CF, NF and NF(+) treatments with respect to grain yield were of the same magnitude in the two years, despite the very different weather conditions.     

Does interspecific competition alter effects of early season ozone exposure on plants from wet grasslands? Results of a three-year experiment in open-top chambers

Chronic effects of ozone on wet grassland species early in the growing season might be altered by interspecific competition. Individual plants of Holcus lanatus, Lychnis flos-cuculi, Molinia caerulea and Plantago lanceolata were grown in monocultures and in mixed cultures with Agrostis capillaris. Mesocosms were exposed to charcoal-filtered air plus 25 nl l(-1) ozone (CF+25), non-filtered air (NF), non-filtered air plus 25 nl l(-1) ozone (NF+25) and non-filtered air plus 50 nl l(-1) ozone (NF+50) early in the growing seasons of 2000 through 2002. Ozone-enhanced senescence and visible foliar injury were recorded on some of the target plants in the first year only. Ozone effects on biomass production were minimal and plant response to ozone did not differ between monocultures and mixed cultures. After three years, above-ground biomass of the plants in mixed culture compared to monocultures was three times greater for H. lanatus and two to four times smaller for the other species.     

Simulations of stomatal conductance and ozone uptake to Norway spruce saplings in open-top chambers

A simulation model was developed to estimate the stomatal conductance and ozone flux to Norway spruce saplings in open-top chambers. The model was parameterized against needle conductance measurements that were made on 4-6-year-old spruce saplings, grown in open-top chambers, in July-September during three different seasons. The spruce saplings were either maintained well watered or subject to a 7-8 week drought period in July-September each year. The simulated conductance showed a good agreement with the measured conductance for the well-watered as well as the drought stress-treated saplings. The simulations were significantly improved when different vapour pressure deficit (VPD) functions were applied for well-watered and drought-stressed spruce saplings. The cumulated ozone uptake which was calculated from the conductance simulations showed less variation between years, compared to the cumulative ozone exposure index AOT40 (accumulated exposure over a threshold of 40 ppb or nl l(-1)) for the corresponding time periods. Measurements in May 1995 demonstrated the occurrence of long-term 'memory-effects' from the drought stress treatments on the conductance. Memory-effects need to be considered when simulation models for stomatal conductance are to be applied to long-lived forest trees under a multiple stress situation.     

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.     

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.     

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.     

Influence of ozone and ethylenediurea (EDU) on growth and yield of bean (Phaseolus vulgaris L.) in open-top field chambers

Greenhouse and ambient air experiments have shown ethylene diurea (EDU) to be a strong and specific protective suppressant of ozone injury in plants. To examine how EDU affects plant responses to various ozone (O(3)) levels under controlled field conditions, Phaseolus vulgaris L. cv. Lit was treated with 150 ppm EDU every 14 days and exposed in open-top chambers to charcoal-filtered air (CF), nonfiltered air (NF) or two cf treatments with ozone added. The ozone treatments were proportional additions of one (CF1) and two (CF2) times ambient ozone levels. The mean ozone concentrations in the CF, NF, CF1 and CF2 treatments were 0.98, 14.1, 14.98 and 31.56 nl litre(-1). A two-way split plot ANOVA revealed that shoot dry weight was significantly reduced by ozone. EDU treatment was highly significant for leaf dry weight, root dry weight and shoot dry weight, but not for pod dry weight; leading to a higher biomass of EDU-treated plants. Ozone/EDU interactions were significant for root weight only, indicating that EDU reduced growth suppression by ozone. These results show that EDU action on plant biomass could be interpreted as a delay in senescence since EDU-treated plants showed a significant decreased biomass loss even in the CF treatment.     

Effects of ozone on managed pasture: II. Yield, species composition, canopy structure, and forage quality

Managed pasture composed of grasses, clover and weeds was exposed in open-top chambers to different levels of ozone (O(3)) during two consecutive seasons to study changes in yield, species composition, canopy structure, and forage quality. The pasture was established in 1990 and exposed in 1991 and 1992. Ozone treatments included charcoal-filtered air (CF), non-filtered air (NF), and two treatments with O(3) added to NF air during periods with global radiation >/= 400 W m(-2) (NF(+), NF(++)). The ratio between the 2-year cumulative, radiation-weighted O(3) concentration in ambient air (= 365 microl litre(-1) h) and in the different treatments was 0.50 (CF), 0.85 (NF), 1.11 (NF(+)), and 1.64 (NF(++)). Plots were harvested four times in 1991, and five times in 1992. The total forage yield for both seasons was modified little by O(3). The yield reduction in NF(++) was only 10% as compared to the CF treatment. Also, only marginal changes were observed in forage quality (Ca, crude protein, crude fibre), and in leaf area index and fractional light penetration. Ozone strongly reduced the yield of clover (Trifolium repens L. and Trifolium pratense L.). The O(3)-effect on clover growth was small after the first harvest and increased with each growth period. In NF, the 2-year cumulative clover yield was reduced by 24% relative to CF. In NF(++), clover growth almost ceased near the end of the second season. The reduction in clover yield with increasing O(3) was associated with a slight increase in the yield of grasses (mainly Dacytlis glomerata L). The increase in the proportion of invading species (weeds or herbs) (Taraxacum officinale L.) during the experiment was not significantly affected by O(3). A second order polynomial function was fitted to the data to establish an exposure-response model for the cumulative clover yield and the cumulative, radiation-weighted O(3) dose, and linear models were developed for total forage mass, grass yield and yield of weeds. Reducing O(3) from elevated levels (NF(+) and NF(++)) during the first season to near-ambient levels (NF) during the second season resulted in a significant recovery of clover yield after two re-growth periods. It is concluded that continuous exposure to ambient levels of O(3) negatively affects the yield of clover in frequently cut, managed pasture, but because of the relatively small proportion of clover, the shift in species composition only marginally affects total forage yield and forage quality. It is emphasised, however, that limitations of the experimental system must be taken into account before extrapolations to real field situations can be made.     

Use of ethylenediurea (EDU) to ameliorate ozone effects on purple coneflower (Echinacea purpurea)

Purple coneflower plants (Echinacea purpurea) were placed into open-top chambers (OTCs) for 6 and 12 weeks in 2003 and 2004, respectively, and exposed to charcoal-filtered air (CF) or twice-ambient (2x) ozone (O3) in 2003, and to CF, 2x or non-filtered (NF), ambient air in 2004. Plants were treated with ethylenediurea (EDU) weekly as a foliar spray. Foliar symptoms were observed in >95% of the plants in 2x-treated OTCs in both years. Above-ground biomass was not affected by 2x treatments in 2003, but root and total-plant biomass decreased in 2004. As a result of higher concentrations of select cell wall constituents (% ADF, NDF and lignin) nutritive quality was lower for plants exposed to 2x-O3 in 2003 and 2004 (26% and 17%, respectively). Significant EDU x O3 interactions for concentrations of cell wall constituents in 2003 indicated that EDU ameliorated O3 effects on nutritive quality. Interactions observed in 2004 were inconsistent.     

Effects of air filtration on spring wheat grown in open-top field chambers at a rural site. II. Effects on mineral partitioning, sulphur and nitrogen metabolism and on grain quality

In 1988 the effect of ambient levels of air pollutants on the nutrients status and grain quality of spring wheat (Triticum aestivum cv. Pelican) was investigated by comparing plants grown in open-top chambers (OTC) ventilated with ambient air (NF treatments) and charcoal-filtered air (CF treatments) at a rural site (Tervuren, Belgium). Spring wheat cultivated in NF OTC showed only minor differences in the P, K, Ca, Mg, Mn and Na concentrations of the different plant parts at final harvest, as well as organic and inorganic S fractions, compared to those of the plants grown in CF air. The plants' total P content was reduced, as well as the P and K concentration of the flour. The total S concentration of the flour was increased by 4%. Effects on N concentrations and grain quality were much more pronounced. At final harvest the N concentrations of straw and flour of the NF air treated plants were much higher compared to CF air. However, the N content of the aerial biomass and the grain N yield were not significantly affected, implying a reduction of other structual compounds. Nitrogen harvest index (NHI) and the ratio of NHI over grain harvest index (GHI), indicated a significant reduction of N translocation from the above-ground biomass to the grain. Changes in the N status and partitioning of spring wheat had an effect on the baking quality of wheat flour. Several parameters that are commonly used as an indication of baking quality have been significantly increased in the NF treatment: total protein concentration, Zeleny value, dry and wet gluten concentration. A slightly increased Hagberg value indicated a reduced alpha-amylase activity. The possibility of foliar N uptake as an additional N source, especially after anthesis and implications of increased protein production instead of carbohydrate synthesis are discussed.     

Growth and yield responses of spring wheat (Triticum aestivum L. CV. Turbo) grown in open-top chambers to ozone and water stress

Spring wheat (Triticum aestivum L.) cv. Turbo was exposed to different levels of ozone and water supply in open-top chambers in 1991. The plants were grown either in charcoal filtered air (CF), not filtered air (NF), in charcoal filtered air with proportional addition of ambient ozone (CF1), or in charcoal filtered air with twice proportional addition of ambient ozone (CF2). The mean seasonal ozone concentrations (24 h mean) were 2.3, 20.6, 17.3, and 24.5 nl litre(-1) for CF, NF, CF1, and CF2 treatments, respectively. Ozone enhanced senescence and reduced growth and yield of the wheat plants. At final harvest, dry weight reductions were mainly due to reductions in ear weight. Grain yield loss by ozone mainly resulted from depressions of 1000 grain weight, whereas numbers of ears per plant and of grains per ear remained unchanged. Pollutants other than ozone did not alter the response to ozone, as was obvious from comparisons between CF1 and NF responses. Water stress alone did not enhance senescence, but also reduced growth and yield. However, yield loss mainly resulted from reductions in the number of ears per plant; 1000 grain weight was not influenced by water stress. No water supply by ozone treatment interactions were detected for any of the estimated parameters.     

Effects of ozone on 14C translocation velocity and growth of spring wheat (Triticum aestivum L.) exposed in open-top chambers

Growth and yield were reduced but (14)C translocation velocity was not affected by increasing levels of ozone in spring wheat exposed in open top chambers to the following treatments: charcoal filtered air (CF), non-filtered ambient air (NF), or NF with addition of 30 microl litre(-1) ozone, 8 h daily (NFO). Destructive harvests were performed at anthesis and at maturity. Parts of the flag leaf or the second leaf were exposed to (14)CO(2) in small cuvettes for 5 min before, during and after anthesis. The translocation velocity was followed by autoradiography and scintillation counting of the plants frozen and lyophilized at different times after labelling. The label was transported at the same velocity in all the treatments. Ozone induced changes in carbon allocation or partitioning should probably be explained as amounts of carbon transported (mg s(-1)), rather than as transportation velocity (mm s(-1)). The amount translocated may be governed by source conditions under O(3) stress: reduced healthy green biomass and photosynthesis, but perhaps also by impairment of phloem loading because of membrane damage.     

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.     

Interaction of ozone exposure and Fusarium subglutinans inoculation on growth and disease development of loblolly pine seedlings

Seedlings from ten half-sib families of loblolly pine (Pinus taeda) were exposed in open-top chambers to carbon-filtered air (CF), non-filtered air (NF), or air amended with ozone to 1.7 or 2.5 times ambient. After 105 days of exposure, half the seedlings within each family were wounded but not inoculated and half were wounded and inoculated with the pitch canker fungus, Fusarium subglutinans, to which five families were relatively resistant. After an additional 50 days of ozone treatment, seedling growth and canker development were recorded. Cankers were significantly (sigma < or = 0.05) smaller among resistant compared to susceptible families, and were significantly larger among seedlings receiving the highest (2.5) compared to the ambient (NF) ozone treatment. The wound scars of non-inoculated seedlings were also significantly larger among seedlings receiving the 2.5 compared to the NF treatment, but these dimensions did not differ significantly with seedling family or resistance. The weights of needles and large roots were significantly smaller at the 2.5 compared to the 1.7 ozone treatment for inoculated but not for non-inoculated seedlings; this resulted in a significant interaction for ozone and inoculation effects. Among resistant families, root weights were significantly smaller for inoculated seedlings. Diameter growth and dry weights of needles were significantly smaller among inoculated compared to non-inoculated seedlings, but did not differ between NF and 2.5 ozone treatments.     

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.