Damage to Eastern White Pine by Sulfur Dioxide,Semimature-Tissue Needle Blight,and Ozone

Eastern white pine (Pinus strobus L.) forests were severely damaged by atmospheric sulfur dioxide up to distances of 25 miles northeast of large smelters located in the Sudbury mining district of Ontario. Damage to white pine was measured in terms of foliage, bark, and biological injuries, radial and volume growth decrement, and tree mortality. The foliar symptoms of sulfur dioxide injury on white pine trees sometimes resembles those caused by a physiogenic disease, semimature-tissue needle blight (abbreviated to SNB). Studies on the nature and etiology of SNB were conducted in white pine forests in the Upper Ottawa Valley, which are remote from smelter operations which might pollute the atmosphere. These studies included the determination of the role that naturallyoccurring atmospheric ozone plays in the occurrence of SNB. Differences between the symptoms of sulfur dioxide injury, SNB, and ozone damage are outlined.     

Ozone-induced changes in the chloroplast structure of conifer needles, and their use in ozone diagnostics

Ozone induces characteristic symptoms in the chloroplasts of the needles of several coniferous species. Chloroplasts are (1) reduced in size and (2) the stroma is electron dense. Moreover, (3) these chloroplast alterations are more pronounced in the outer mesophyll cell layers and in the upper side of the needle compared to the inner layers and lower side. The syndrome, including the three symptoms (1)-(3), is found in the green needles of Scots pine and Norway spruce not only in the experimental fumigations, but also in mature trees in the field, and has potential for diagnosis of ozone stress. For sound ozone diagnostics all three symptoms must be present in the samples studied. The symptoms in relation to needle anatomy and physiology is discussed, and recommendations for sampling and analysis are given.     

Acid mist and ozone effects on the leaf chemistry of two western conifer species

Seedlings of Jeffrey pine (Pinus jeffreyi) and giant sequoia (Sequoiadendron gigantea) were more susceptible to leaf chemical changes following exposure to acid mist (pH 3.4-2.0) or acid mist/ozone combinations, than to ozone alone (0.1-0.2 microl/litre), when plants were exposed to alternating doses of these pollutants for 6-9 weeks. Under acid mist treatment, leaves exhibited higher levels of nitrogen and sulfur, two elements applied in acid mist. In addition, levels of foliar sodium, and, in the case of giant sequia, potassium, as well, increased under acid mist treatment. Iron and manganese were also mobilized, resulting in significant increases in these elements in pine, and decreases in manganese in giant sequoia foliage. The acid treatment also reduced chlorophyll b concentrations in pine, and, to a less significant extent, in giant sequoia. Calcium, magnesium, barium and strontium were differentially accumulated in giant sequoia compared to Jeffrey pine. Under acid mist treatment, all of these elements (except strontium) declined in concentration in giant sequoia, with calcium showing the most significant trend. The more extensive changes in leaf chemistry induced by acid mist are consistent with earlier observations of significant changes in spectral reflectance of these seedlings after 3 weeks of fumigation. Limited foliage samples collected from these two species in 1985 and 1986 in Sequoia/Kings Canyon National Parks in the southern Sierra Nevada do not in themselves indicate any clearcut or severe effects of ozone alone on leaf chemistry of these species, but a mild influence of nitrate-laden acid deposition, possibly in combination with ozone, is consistent with the rise in nitrogen and lignin levels in Jeffrey pine on sites observed to have moderate visible injury symptoms. No firm conclusions about effects of pollutants on leaf chemistry in these field sites is possible without further study.     

Ozone Suppression of White Pine Net Photosynthesis

Reduction in rates of net photosynthesis following exposure to ozone was observed in eastern white pine saplings prior to the development of visible injury. Branches exposed to consecutive doses of 50–80 pphm ozone for 4 hr and 80–90 pphm ozone for 3 hr appeared to have suppressed net photosynthetic rates but results were confounded by diurnal rate variation. Photosynthetic rates of treated branches returned to normal levels following this initial ozone exposure. A second ozonation involving dosages of 90–100 pphm for 5 and 3 hr was sufficient to reduce rates of net photosynthesis by approximately 80 percent.     

The use of light and electron microscopy to assess the impact of ozone on Norway spruce needles

Light (LM) and transmission electron (TEM) microscopy were used to study previously specified ozone symptoms in the foliage of Norway spruce. The three youngest green needle generations from twenty mature trees in two stands on sites of different soil fertility at Asa, southern Sweden, were sampled in 1999. The critical dose of ozone, expressed as AOT40, was 6,362 ppb.h. LM showed ozone-specific symptoms: decreased chloroplast size with electron dense stroma advancing gradually from the outer to the inner cell layers, being most severe in the needle side facing the sky. The symptoms were expressed as ozone syndrome indices at the needle generation, tree and stand levels. The index had higher values at the low fertility site. TEM was used to confirm the LM results. The study shows that LM can be used for diagnosis of the impact of ozone on conifers in the field.     

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.     

Summer Ozone Concentrations in Southern Ontario in Relation to Photochemical Aspects and Vegetation Damage

In the summers of 1960 and 1961, groups from the Canada Department of Agriculture, the Meteorological Service of Canada, and the Canada Department of National Health and Welfare conducted a joint study in a tobacco-growing area along the north shore of Lake Erie. The purpose of the study was to determine the causal agent for weather fleck damage to tobacco crops. A number of air pollutants were monitored and the results correlated with extensive observations of meteorological phenomena and effects on rate of growth and fleck damage to leaves of tobacco plants in experimental plots. Ozone concentrations followed a diurnal cycle, rising a few hours after sunrise, peaking in early afternoon at about 5 pphm, and dropping to a minimum of less than 1 pphm during the night. Other measurements indicated the presence of NO₂ in the order of 1 pphm, aldehydes about 0.2 pphm or lower, and negligible concentrations of SO₂. Cracking of stretched rubber strips followed the ozone values although, in general, the cracking index was greater than could be attributed to ozone (by oxidized KI) alone. The maximum ozone value recorded during the two growing seasons was IS pphm. A dosage of 20 pphm-hr was found sufficient to cause weather fleck or ozone damage to susceptible tobacco leaves. In addition meteorological data could be used to predict weather fleck attacks one to four days in advance.     

Seasonal differences and within-canopy variations of antioxidants in mature spruce (Picea abies) trees under elevated ozone in a free-air exposure system

The effect of free-air ozone fumigation and crown position on antioxidants were determined in old-growth spruce (Picea abies) trees in the seasonal course of two consecutive years (2003 and 2004). Levels of total ascorbate and its redox state in the apoplastic washing fluid (AWF) were increased under double ambient ozone concentrations (2xO3), whilst ascorbate concentrations in needle extracts were unchanged. Concentrations of apoplastic and symplastic ascorbate were significantly higher in 2003 compared to 2004 indicating a combined effect of the drought conditions in 2003 with enhanced ozone exposure. Elevated ozone had only weak effects on total glutathione levels in needle extracts, phloem exudates and xylem saps. Total and oxidised glutathione concentrations were higher in 2004 compared to 2003 and seemed to be more affected by enhanced ozone influx in the more humid year 2004 compared to the combined effect of elevated ozone and drought in 2003 as observed for ascorbate.     

An Evaluation of the Effects of Ozone Injury on Radial Growth of Ponderosa Pine (Pinus ponderosa) in the Southern Sierra Nevada

Growth of ponderosa pines with visible symptoms of ozone injury was compared with that of asymptomatic trees in the southern Sierra Nevada, California. Time series analysis indicated that there was no significant reduction in annual radial increment of symptomatic trees during recent years compared to past growth and growth of asymptomatic trees. First order autocorrelation and climatic variables accounted for a large proportion of the variance in growth index, and winter precipitation was positively correlated with growth for all size and age classes. Although ozone concentrations are high enough to cause chlorosis and premature needle senescence in ponderosa pine, there has been no significant change in growth associated with ozone injury.     

Quantifying ozone uptake at the canopy level of spruce,pine and larch trees at the alpine timberline: an approach based on sap flow measurement

Micro-climatic and ambient ozone data were combined with measurements of sap flow through tree trunks in order to estimate whole-tree ozone uptake of adult Norway spruce (Picea abies), cembran pine (Pinus cembra), and European larch (Larix decidua) trees. Sap flow was monitored by means of the heat balance approach in two trees of each species during the growing season of 1998. In trees making up the stand canopy, the ozone uptake by evergreen foliages was significantly higher than by deciduous ones, when scaled to the ground area. However, if expressed per unit of whole-tree foliage area, ozone flux through the stomata into the needle mesophyll was 1.09, 1.18 and 1.40 nmol m(-2) s(-1) in Picea abies, Pinus cembra and Larix decidua, respectively. These fluxes are consistent with findings from measurements of needle gas exchange, published from the same species at the study site. It is concluded that the sap flow-based approach offers an inexpensive, spatially and temporally integrating way for estimating ozone uptake at the whole-tree and stand level, intrinsicly covering the effect of boundary layers on ozone flux.     

Background ozone monitoring and phytodetection in the greater rural area of Corinth--Greece

Natural background ozone levels were monitored in three places within the greater rural area of Corinth, namely Bogdani Hill, Astronomical Observatory of Krionerion, and Kiato, and compared with ambient ozone monitored in the metropolitan area of Athens. Measurements were made sequentially, for a few weeks at each place, during the summer of 2000. In addition, ozone phytodetection, using tobacco (Nicotiana tabacum L.) plants of the Bel-W3 and Zichnomirodata varieties, was conducted in 12 places (the above included). Moreover, stomatal conductance was measured in the Bel-W3 plants, as well as in leaves of cultivated grape-vines (Vitis vinifera L.) and in needles of Aleppo pine (Pinus halepensis Mill.) trees and compared with the diurnal pattern of ozone concentrations.The 24 and 12 (08:00-20:00) hourly averages of ozone concentrations were high in Athens (37; 51 ppb), at Bogdani Hill (53; 56 ppb) and at the Astronomical Observatory (56; 55 ppb), but relatively low in Kiato (30; 34 ppb). Furthermore, the average daily AOT40 (accumulated exposure over 40 ppb for the daylight hours) (ppbh) was 193 in Athens, 212 at Bogdani Hill, 192 at the Astronomical Observatory and 47 in Kiato. Ozone concentrations exhibited the usual diurnal pattern in Athens (altitude 50 m), where they were maximum during midday and early afternoon hours, as well as at Bogdani Hill (300 m) and in Kiato (5 m) where, however, they were maximal 1-3 h later. At the Astronomical Observatory (altitude 920 m) ozone remained constant during both daylight and night hours. The differences in diurnal patterns are consistent with those in places of different elevation, reported elsewhere.The Bel-W3 plants were injured at all 12 places; Zichnomirodata plants exhibited lower injury and only in some of the places; probable ozone symptoms were also observed on vine plants and pine trees. The greatest injury was observed at the high altitude places of Astronomical Observatory and Mougostos. Stomatal conductance, in all three species, peaked during morning and early midday hours when ozone levels were higher in the high altitude, and lower in the low altitude, places.     

Effects of simultaneous ozone exposure and nitrogen loads on carbohydrate concentrations, biomass, and growth of young spruce trees (Picea abies)

Spruce saplings were grown under different nitrogen fertilization regimes in eight chamberless fumigation systems, which were fumigated with either charcoal-filtered (F) or ambient air (O3). After the third growing season trees were harvested for biomass and non-structural carbohydrate analysis. Nitrogen had an overall positive effect on the investigated plant parameters, resulting in increased shoot elongation, biomass production, fine root soluble carbohydrate concentrations, and also slightly increased starch concentrations of stems and roots. Only needle starch concentrations and fine root sugar alcohol concentrations were decreased. Ozone fumigation resulted in needle discolorations and affected most parameters negatively, including decreased shoot elongation and decreased starch concentrations in roots, stems, and needles. In fine roots, however, soluble carbohydrate concentrations remained unaffected or increased by ozone fumigation. The only significant interaction was an antagonistic effect on root starch concentrations, where higher nitrogen levels alleviated the negative impact of ozone.     

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.     

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.     

Analyses of enzyme activities and other metabolic criteria after five years of fumigation

Enzymatic activity (peroxidase, glutamate dehydrogenase, glutamine synthetase), foliage buffering capacity, soluble protein and nitrogen content were measured in current and previous year needles from young spruce (Picea abies) and fir (Abies alba). The trees were exposed to low levels of SO(2) and/or O(3) and simulated acidic precipitation (pH 4.0) in open-top chambers from 1983 through 1988. Needle samples were taken during March 1988 at the end of the five-year fumigation period. Exposure to SO(2) substantially increased sulphur content in both needle age classes of spruce and fir, and concomitantly reduced the foliage buffering capacity index (BCI), whereas the combined fumigation with SO(2) and O(3) had no effect on BCI. Peroxidase activity was markedly higher in year-old needles compared to current-year needles. However, trees from the SO(2) and SO(2) + O(3) treatments exhibited statistically significant stimulated peroxidase activities. Similarly, changes in the activities of the nitrogen-metabolizing enzymes indicated an altered cellular function of the trees after the long-term pollution stress. Levels of activity of both glutamate dehydrogenase and glutamine synthetase were increased by exposure to SO(2), especially in spruce. Although glutamate dehydrogenase in spruce was affected by all treatments, such changes in activity were found in fir only with the SO(2) treatment. The highest activity of glutamine synthetase, however, occurred in the older needles of trees exposed to SO(2) + O(3). Total nitrogen concentration was either unaffected by the pollutant treatments or decreased in spruce compared to the controls. No statistically significant changes due to the fumigation were found in soluble protein concentrations. Results indicated that chronic exposure to air pollutants lead to alterations in metabolic processes in conifer needles, detectable either by changes in typical stress indicating values or by increases in ammonium assimilation capacity.     

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.     

Variation in ozone sensitivity among clones of Betula pendula and Betula pubescens

Forty clones of Betula pendula and 6 clones of Betula pubescens, originating from southern and central Finland, were ranked in order of ozone sensitivity according to visible injuries, growth and leaf senescense under low ozone exposure. The plants were fumigated in natural climatic conditions using an open-air exposure system during two growing seasons. Control plants were grown under ambient air, and the elevated-ozone exposures were 1.6x the ambient in 1994 and 1.7x the ambient in 1995. The differences in ozone sensitivity among clones were large. Ozone tolerance was related to thicker leaves and higher stomatal density as compared to sensitive clones. Ultrastructural ozone-induced symptoms were found in chloroplasts of sensitive clones. Increased number of visibly injured leaves on fumigated plants was correlated with reduced leaf formation, foliage area, shoot dry wt and number of stomata, and increased yellowing of leaves. The results suggest that a considerable proportion of birch trees, showing high sensitivity to ozone, are at risk if ambient ozone exposures increase.     

Variation in CO2 assimilation rate induced by simulated dew waters with different sources of hydroxyl radical (*OH) on the needle surfaces of Japanese red pine (Pinus densifora Sieb. et Zucc.)

The hydroxyl radical (*OH) is generated in polluted dew on the needle surfaces of Japanese red pine (Pinus densiflora Sieb. et Zucc.). This free radical, which is a potent oxidant, is assumed to be a cause of ecophysiological disorders of declining trees on the urban-facing side of Mt. Gokurakuji, western Japan. Mists of *OH-generating N(III) (HNO2 and NO2-) and HOOH + Fe + oxalate solutions (50 and 100 microM, pH 5.1-5.4) simulating the dew water were applied to the foliage of pine seedlings grown in open-top chambers in the early morning. Needles treated with 100 microM N(III) tended to have a greater maximum CO2 assimilation rate (Amax), a greater stomatal conductance (g(s)) and a greater needle nitrogen content (Nneedle), suggesting that N(III) mist acts as a fertilizer rather than as a phytotoxin. On the other hand, needles treated with 100 microM HOOH + Fe + oxalate solution showed the smallest Amax, g(s), and Nneedle, suggesting that the combination of HOOH + Fe + oxalate caused a decrease in needle productivity. The effects of HOOH + Fe + oxalate mist on pine needles were very similar to the symptoms of declining trees at Mt. Gokurakuji.     

Seasonal changes in the tissue-metal (Cd, Zn and Pb) concentrations in two ecophysiologically dissimilar earthworm species: pollution-monitoring implications

During two consecutive growing seasons, the same potted individuals of European aspen (Populus tremula), grown from root cuttings of one clone, were fumigated with either ambient air or ozone concentrations of 0 (control), 0.05 or 0.1 microlitre litre(-1). Structure and biomass of the annually formed branches were analysed after excision at the end of each season. Only at 0.1 microlitre litre(-1) was branch weight reduced, and crooked axes occurred in each season. During the second season, branch length and leaf sizes were strongly reduced, while many leaves displayed yellowish deficiency symptoms and lowered cation concentrations. Such leaves contrasted to those showing characteristic O3-bronzing. Although foliage density was enhanced due to reduced branch length, the area of attached foliage was limited by the small leaf sizes, necrotic leaves and premature leaf loss. During mid-summer of the second fumigation period, photosynthetic capacity, carboxylation efficiency and water-use efficiency (WUE) declined in (attached) yellowish and bronze leaves at 0.1 microlitre litre(-1), whereas green leaves at 0.05 microlitre litre(-1) displayed accelerated senescence in late summer while maintaining WUE. It is concluded that the differences in branch growth between the two growing seasons were caused in part by internal changes in those plant organs (root and basal stem), which had experienced both fumigation periods.     

Ozone Dosage Response of Ponderosa Pine Seedlings

Photochemical oxidant injury to ponderosa pine (Pinus ponderosa Laws) is a severe problem in the southern California mountains. Three-year-old ponderosa pines fumigated in controlled environment chambers with ozone at 0.15, 0.30, or 0.45 ppm had apparent photosynthesis rates reduced by 10, 70, and 85%, respectively, after 30 days exposure. A fumigation with 0.30 ppm ozone for 33 days depressed the cold perchloric acid extracted polysaccharides of both current and one-year-old needles by 40%. The 80% ethanol soluble sugar concentration of current year, ozone-injured needles increased 16% and that of the one-year-old needles decreased slightly. Both carbohydrate fractions of control trees in carbon-filtered air increased moderately. Higher, endogenous concentrations of ascorbic acid in needles did not protect the tissue from ozone injury. Apparent photosynthesis rate was a sensitive index for ozone dosage response. Needle carbohydrate depletion probably induces premature abscission.