Spatial relationships between nitrogen status and pitch canker disease in slash pine planted adjacent to a poultry operation

Pitch canker disease (Fusarium circinatum Nirenberg & O'Donnell) causes serious shoot dieback, reduced growth and mortality in pines found in the southern and western USA, and has been linked to nutrient imbalances. Poultry houses with forced-air ventilation systems produce nitrogen (N) emissions. This study analyzed spatial correlations between pitch canker disease and foliar, forest floor, soil, and throughfall N in a slash pine (Pinus elliottii var. elliottii Engelm.) plantation adjacent to a poultry operation in north Florida, USA. Tissue and throughfall N concentrations were highest near the poultry houses and remained elevated for 400 m. Disease incidence ranged from 57-71% near the poultry houses and was spatially correlated with N levels. Similarly, stem mortality ranged from 41-53% in the most heavily impacted area, and declined to 0-9% at distances greater than 400 m. These results suggest that nutritional processes exacerbate changes in disease susceptibility and expression in slash pine.     

Sulphur dioxide adsorption in Scots pine canopies exposed to high ammonia emissions near a Cu-Ni smelter in SW Finland

Since 1994 the nickel-processing plant at the Cu-Ni smelter at Harjavalta, south-west Finland, has emitted considerable amounts of NH(3) into the atmosphere. The effects of NH(3) emissions on nitrogen and sulphur deposition in throughfall and the foliar nutrient status were investigated in a Scots pine stand at 0.5 km distance. Bulk deposition, stand throughfall and percolation water (20 cm depth) samples were collected at 4-week intervals during 1992-1998. pH and the Ca, Mg, K, NH(4) and SO(4) concentrations were determined on the samples. NH(3) emissions have strongly increased the scavenging of SO(2) from the air in the pine stand, and the increased levels of N and S deposition were clearly evident as increased foliar N and S concentrations and larger needle size. The increased input of SO(4) into the forest floor was not associated with an increase in the leaching of Ca and Mg from the surface soil layers.     

Limited compensation of ozone stress by potassium in Norway spruce

The effects of potassium fertilization and ozone stress were investigated in a clone of Picea abies (L.) Karst, by studies of the uptake of CO(2) by the crowns, the element content, on leaching of the youngest needles, and the longevity of the needles. All plants were exposed to 0.075 microl litre(-1) SO(2) from January to April 1986. The average ozone concentrations applied during the subsequent growing season (May-December) were 0, 0.027, 0.050 and 0.100 microl litre(-1). Half of the trees received liquid fertilizer applications from April to July 1986. CO(2) uptake by the crowns was significantly reduced in non-fertilized plants at ozone doses of 100-200 microl litre(-1) h, whereas similar reductions were recorded in fertilized plants only above an ozone dose of 300 microl litre(-1) h. Independent of the fertilization, however, the concentrations of calcium, magnesium and nitrogen in the needles increased in parallel with the ozone dose, whilst potassium, phosphorus and sulphur showed little response to ozone. In both nutrient regimes, the diffusive loss of elements from chloroform-washed needles was similar and tended to be reduced at the highest ozone concentration, when relating the leachate to the corresponding element content in the needles. Needles formed in the highest ozone treatment were significantly shed during the succeeding year, regardless of the nutrient supply. It appears that increased potassium supply has little compensating effect on ozone stress in spruce.     

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.     

Experimental work related to nitrogen deposition, nitrification and soil acidification--a case study

The leaching of major ions has been studied since August 1986 in two plots with different nitrogen fertilization levels and in a control plot in a 29-year-old stand of Norway spruce (Picea abies Karst.) in south-central Sweden. The fertilization started in 1967. The two fertilizer levels, both of which have caused a significant stimulation of the tree growth, correspond to an annual input of approximately 35 kg N ha(-1) and 75 kg N ha(-1) respectively, as NH4NO3. Phosphorus fertilizer is also applied. Field and laboratory incubations performed during 1986 showed that nitrification mainly occurs in the plot with the highest fertilization level, in accordance with the fact that nitrate could not be detected in the soil water in the other two plots. Fertilization has increased the ionic strength of the soil solution due mainly to sulphate in the phosphate fertilizer, but also nitrate at the highest fertilization level. This has caused an increase in total aluminium and a decline in pH. The preliminary data reported here are compared with results obtained at Swedish field sites with moderate to high levels of nitrogen deposition.     

A cumulative ozone uptake-response relationship for the growth of Norway spruce saplings

Norway spruce saplings [Picea abies (L.) Karst.] were exposed during four growing seasons to different ozone treatments in open-top chambers: charcoal filtered air (CF), non-filtered air (NF) and non-filtered air with extra ozone (NF+, 1.4xambient concentrations). The CF and NF+ ozone treatments were combined with phosphorous deficiency and drought stress treatments. The total biomass of the trees was harvested at different intervals during the experimental period. The ozone uptake to current-year needles of the Norway spruce saplings was estimated using a multiplicative stomatal conductance simulation model. There was a highly significant correlation between the reduction of total biomass and the estimated cumulative ozone uptake, which did not vary when different thresholds were applied for the rate of ozone uptake. The reduction of the total biomass was estimated to 1% per 10 mmol m(-2) cumulated ozone uptake, on a projected needle area basis.     

The effects of atmospheric nitrogen deposition and soil chemistry on the nutritional status of Pseudotsuga menziesii, Pinus nigra and Pinus sylvestris

The ammonium content and the base cation content, expressed relative to ammonium, are enhanced in the soil of Dutch forests, due to the extremely high deposition of ammonium to the forest floor. A nation-wide investigation was carried out to establish whether and how these changed nitrogen fluxes in deposition and soil affect the nutritional status of coniferous trees. The chemical composition of needles of Douglas fir, Scots pine and Corsican pine showed a regional trend similar to that of deposition and soil solution. Particularly nutrients, expressed relative to nitrogen, decreased from North to South. Of the macronutrients phosphorus was most often deficient and therefore probably limiting in the Douglas fir stands. Many pine trees suffered from relative magnesium shortages. In all stands, magnesium and, in Douglas stands, also phosphorus contents of the needles were negatively correlated with ammonium and ammonium/cation ratios in deposition. However, in contrast to pine trees, nutrient contents in needles of Douglas fir showed correlation with nitrate rather than with ammonium in the soil solution. Correlation analyses indicate that nitrogen fluxes in the soil, indirectly affect the nutritional status of coniferous trees.     

Does (K+Mg+Ca+P) fertilization lead to recovery of tree health in a nitrogen stressed Quercus rubra L. stand?

In The Netherlands atmospheric deposition of nitrogen compounds to forest ecosystems has been very high for some decades and has led to severe nutritional imbalances in soils as well as in trees. At this moment legislation is not fully in effect with respect to lowering emission/deposition fluxes, particularly of nitrogen. The trees suffer mainly from severe magnesium, potassium and calcium and sometimes phosphorus deficiencies. In this study it was investigated whether fertilization with potassium, magnesium, calcium and phosphorus could prevent a Quercus rubra stand from further decline by restoring the nutritional balance, though the detrimental input of nitrogen compounds still continues. Three fertilization treatments were applied: (i) a standard dose of 1250 kg.ha(-1) containing 60 kg P.ha(-1) (as P2O5), 100 kg K.ha(-1) (as K2O), 80 kg Mg.ha(-1) (as MgO) and 340 kg Ca.ha(-1) (as CaCO3); (ii) two times the standard dose; and (iii) three times the standard dose. Soil solution chemistry showed that the highest dose led to the most significant results: an improved nutrient balance and an increased availability of nutrients. After one growing season following fertilization, the trees looked much healthier and crown density had increased. This revitalization lasted for at least four years. For tree health the mid dose seemed appropriate. After the first growing season potassium is the most mobile nutrient, in the soil and also in the trees, but tended to decrease in the fourth year after fertilization. Magnesium and calcium reached normal values in the leaves after four growing seasons. No obvious effects of phosphorus were found.     

Impact of elevated CO(2) and nitrogen fertilization on foliar elemental composition in a short rotation poplar plantation

The experiment was carried out on a short rotation coppice culture of poplars (POP-EUROFACE, Central Italy), growing in a free air carbon dioxide enriched atmosphere (FACE). The specific objective of this work was to study whether elevated CO(2) and fertilization (two CO(2) treatments, elevated CO(2) and control, two N fertilization treatments, fertilized and unfertilized), as well as the interaction between treatments caused an unbalanced nutritional status of leaves in three poplar species (P. x euramericana, P. nigra and P. alba). Finally, we discuss the ecological implications of a possible change in foliar nutrients concentration. CO(2) enrichment reduced foliar nitrogen and increased the concentration of magnesium; whereas nitrogen fertilization had opposite effects on leaf nitrogen and magnesium concentrations. Moreover, the interaction between elevated CO(2) and N fertilization amplified some element unbalances such as the K/N-ratio.     

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%.     

Nitrate reductase activity of needles of Norway spruce fumigated with different mixtures of ozone, sulfur dioxide, and nitrogen dioxide

Four-year-old spruce clones (Picea abies (L.) Karst.) cultivated in sand and provided with a complete nutrient solution, or a solution deficient in magnesium and calcium, were exposed to the pollutant mixtures SO(2)/NO(2), O(3)/NO(2), and O(3)/SO(2)/NO(2), at realistic concentrations for 32 weeks. Fumigation caused a slight increase of total N contents in current year needles, whereas in one-year-old needles N concentrations did not change. The response of nitrate reductase activity to pollutant stress depended on needle age and nutrient supply, respectively. In one-year-old needles fumigation resulted in a significant inhibition of enzyme activity, particularly in Mg and Ca deficient trees. The combination of all three components proved to be most effective in causing a decrease by 60% compared to the control. In contrast, nitrate reductase activity was stimulated in current year needles, especially by O(3)/NO(2) and O(3)/SO(2)/NO(2). Changes in the activity of nitrate and nitrite reductases are considered as a factor contributing to the high phytotoxic potential of pollutant combinations with NO(2).     

Effects of elevated CO2, nitrogen supply and tropospheric ozone on spring wheat-II. Nutrients (N, P, K, S, Ca, Mg, Fe, Mn, Zn)

CO(2) enrichment is expected to alter leaf demand for nitrogen and phosphorus in plant species with C(3) carbon dioxide fixation pathway, thus possibly causing nutrient imbalances in the tissues and disturbance of distribution and redistribution patterns within the plants. To test the influence of CO(2) enrichment and elevated tropospheric ozone in combination with different nitrogen supply, spring wheat (Tritium aestivum L. cv. Minaret) was exposed to three levels of CO(2) (361, 523, and 639 microl litre(-1), 24 h mean from sowing to final harvest), two levels of ozone (28.4 and 51.3 nl litre(-1)) and two levels of nitrogen supply (150 and 270 kg ha(-1)) in a full-factorial design in open-top field chambers. Additional fertilization experiments (120, 210, and 330 kg N ha(-1)) were carried out at low and high CO(2) levels. Macronutrients (N, P, K, S, Ca, Mg) and three micronutrients (Mn, Fe, Zn) were analysed in samples obtained at three different developmental stages: beginning of shoot elongation, anthesis, and ripening. At each harvest, plant samples were separated into different organs (green and senescent leaves, stem sections, ears, grains). According to analyses of tissue concentrations at the beginning of shoot elongation, the plants were sufficiently equipped with nutrients. Elevated ozone levels neither affected tissue concentrations nor shoot uptake of the nutrients. CO(2) and nitrogen treatments affected nutrient uptake, distribution and redistribution in a complex manner. CO(2) enrichment increased nitrogen-use efficiency and caused a lower demand for nitrogen in green tissues which was reflected in a decrease of critical nitrogen concentrations, lower leaf nitrogen concentrations and lower nitrogen pools in the leaves. Since grain nitrogen uptake during grain filling depended completely on redistribution from vegetative pools in green tissues, grain nitrogen concentrations fell considerably with severe implications for grain quality. Ca, S, Mg and Zn in green tissues were influenced by CO(2) enrichment in a similar manner to nitrogen. Phosphorus concentrations in green tissues, on the other hand, were not, or only slightly, affected by elevated CO(2). In stems, 'dilution' of all nutrients except manganese was observed, caused by the huge accumulation of water soluble carbohydrates, mainly fructans, in these tissues under CO(2) enrichment. Whole shoot uptake was either remarkably increased (K, Mn, P, Mg), nearly unaffected (N, S, Fe, Zn) or decreased (Ca) under CO(2) enrichment. Thus, nutrient cycling in plant-soil systems is expected to be altered under CO(2) enrichment.     

Physiological and foliar symptom response in the crowns of Prunus serotina, Fraxinus americana and Acer rubrum canopy trees to ambient ozone under forest conditions

The crowns of five canopy dominant black cherry (Prunus serotina Ehrh.), five white ash (Fraxinus americana L.), and six red maple (Acer rubrum L.) trees on naturally differing environmental conditions were accessed with scaffold towers within a mixed hardwood forest stand in central Pennsylvania. Ambient ozone concentrations, meteorological parameters, leaf gas exchange and leaf water potential were measured at the sites during the growing seasons of 1998 and 1999. Visible ozone-induced foliar injury was assessed on leaves within the upper and lower crown branches of each tree. Ambient ozone exposures were sufficient to induce typical symptoms on cherry (0-5% total affected leaf area, LAA), whereas foliar injury was not observed on ash or maple. There was a positive correlation between increasing cumulative ozone uptake (U) and increasing percent of LAA for cherry grown under drier site conditions. The lower crown leaves of cherry showed more severe foliar injury than the upper crown leaves. No significant differences in predawn leaf water potential (psi(L)) were detected for all three species indicating no differing soil moisture conditions across the sites. Significant variation in stomatal conductance for water vapor (g(wv)) was found among species, soil moisture, time of day and sample date. When comparing cumulative ozone uptake and decreased photosynthetic activity (P(n)), red maple was the only species to show higher gas exchange under mesic vs. drier soil conditions (P < 0.05). The inconsistent differences in gas exchange response within the same crowns of ash and the uncoupling relationship between g(wv) and P(n) demonstrate the strong influence of heterogeneous environmental conditions within forest canopies.     

The effects of excess nitrogen deposition on young Norway spruce trees. Part II the vegetation

The effects of wet-deposited nitrogen on soil acidification and the health of Norway spruce were investigated in a pot experiment using an open-air spray/drip system. Nitrogen was applied as ammonium ((NH(4))(2)SO(4)) or nitrate (HNO(3)/NaNO(3)) in simulated rain to either the soil or the foliage. Symptoms of forest decline as observed in the field were not reproduced, and there was no evidence of direct toxicity. Treatments did, however, have significant effects on tree nutrition. Both NH(+)(4) and NO(-)(3) treatment applied to the foliage lowered foliar K concentrations. NH(+)(4) to a greater extent. Soil-applied NH(+)(4) reduced foliar Mg concentrations and increased foliar Al and Fe. Soil-applied NO(-)(3) significantly reduced foliar P concentrations, and at high doses prevented the alleviation of P deficiency by fertiliser. These effects could be important in some field situations. Ammonium deposition is predicted to be more damaging than nitrate deposition, although the latter may be critical for forests where P status is marginal, such as in parts of the British uplands.     

No general effect of ozone on foliar nutrient concentrations in mature scions of grafted Picea abies trees

The effect of ozone (< 10, 200, or 400 microg m(-3)), on foliar nutrient concentrations of Picea abies were determined by fumigating potted grafts from mature trees (> 55 and 125 years), representing six clones, in open-top chambers at two locations in Norway. The concentration of nutrients in needles of grafted plants were significantly affected by clone and location. Generally, the concentrations of nutrients were not affected by ozone, but a significant increase in the concentrations of potassium and iron in two of the clones were found. These two clones were the only ones injured (yellow needles) by the fumigation.     

Can the foliar nitrogen concentration of upland vegetation be used for predicting atmospheric nitrogen deposition? Evidence from field surveys

The deposition of atmospheric nitrogen can be enhanced at high altitude sites as a consequence of cloud droplet deposition and orographic enhancement of wet deposition on hills. The degree to which the increased deposition of nitrogen influences foliar nitrogen concentration in a range of upland plant species was studied in a series of field surveys in northern Britain. A range of upland plant species sampled along altitudinal transects at sites of known atmospheric nitrogen deposition showed marked increases in foliar nitrogen concentration with increasing nitrogen deposition and altitude (and hence with decreasing temperature). For Nardus stricta L., Deschampsia flexuosa (L.) Trin., Calluna vulgaris (L.) Hull, Erica cinerea L. and Hylocomium splendens (Hedw.) Br. Eur. on an unpolluted hill, foliar nitrogen increased by 0.07, 0.12, 0.15, 0.08 and 0.04% dry weight respectively for each 1 kg ha(-1) year(-1) increase in nitrogen deposition. Most species showed an approximately linear relationship between foliar nitrogen concentration and altitude but no trend with altitude for foliar phosphorus concentration. This provided evidence that the tissue nutrient status of upland plants reflects nutrient availability rather than the direct effects of climate on growth. However, differences in the relationship between foliar nitrogen concentration and atmospheric nitrogen deposition for N. stricta sampled on hills in contrasting pollution climates show that the possibility of temperature-mediated growth effects on foliar nitrogen concentration should not be ignored. Thus, there is potential to use upland plant species as biomonitors of nitrogen deposition, but the response of different species to nitrogen addition, in combination with climatic effects on growth, must be well characterised.     

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.     

Trichloroacetic acid cycling in Sitka spruce saplings and effects on sapling health following long term exposure

Trichloroacetic acid (TCA, CCl(3)COOH) has been associated with forest damage but the source of TCA to trees is poorly characterised. To investigate the routes and effects of TCA uptake in conifers, 120 Sitka spruce (Picea sitchensis (Bong.) Carr) saplings were exposed to control, 10 or 100 microg l(-1) solutions of TCA applied twice weekly to foliage only or soil only over two consecutive 5-month growing seasons. At the end of each growing season similar elevated TCA concentrations (approximate range 200-300 ng g(-1) dwt) were detected in both foliage and soil-dosed saplings exposed to 100 microg l(-1) TCA solutions showing that TCA uptake can occur from both exposure routes. Higher TCA concentrations in branchwood of foliage-dosed saplings suggest that atmospheric TCA in solution is taken up indirectly into conifer needles via branch and stemwood. TCA concentrations in needles declined slowly by only 25-30% over 6 months of winter without dosing. No effect of TCA exposure on sapling growth was measured during the experiment. However at the end of the first growing season needles of saplings exposed to 10 or 100 microg l(-1) foliage-applied TCA showed significantly more visible damage, higher activities of some detoxifying enzymes, lower protein contents and poorer water control than needles of saplings dosed with the same TCA concentrations to the soil. At the end of each growing season the combined TCA storage in needles, stemwood, branchwood and soil of each sapling was <6% of TCA applied. Even with an estimated half-life of tens of days for within-sapling elimination of TCA during the growing season, this indicates that TCA is eliminated rapidly before uptake or accumulates in another compartment. Although TCA stored in sapling needles accounted for only a small proportion of TCA stored in the sapling/soil system it appears to significantly affect some measures of sapling health.     

A high loading overland flow system: Impacts on soil characteristics, grass constituents, yields and nutrient removal

The objectives of this paper are to determine effects of different grass species and their harvests on pollutant removal, elucidate impacts on soil characteristics and grass constituents, observe grass yield and quantify nutrient uptake by vegetation in an overland flow system (OLFS). Polluted creek water was applied to eight channels in the OLFS, which were planted with Paragrass, Nilegrass, Cattail, and Vetiver, with each two channels being randomly planted with a given grass species. The grass in one channel was harvested while that in the other channel was not. At a high rate of 27.8 m d(-1) hydraulic loading, the removal efficiencies of conventional pollutants such as BOD, COD, suspended solids (SS), and total coliforms in wastewater are not affected by the type of the grasses species, but those of nitrogen and phosphorus are affected by different species. Overall average removal efficiencies of BOD, COD, SS, ammonia, total nitrogen, total phosphorus and total coliforms through the OLFS are 42%, 48%, 78%, 47%, 40%, 33% and 89%, respectively. The concentration of nitrate, however, increases due to nitrification. Soil characteristics in OLFS have been changed significantly; specific conductivity, organic matter, exchangeable magnesium, extractable copper and zinc in soils all increase with time while pHs decrease. During the winter season, there is a significant accumulation of nitrate in grass with the subsequent reduction during the active growing season (Spring). The contents of nitrate and phosphorus in grass tissue are higher than those of grass in general pastureland, probably due to nutrient luxury uptake by grass. The overall grass yield, growth rate and nutrient uptake are quantified and implication of such high rate OLFS discussed.     

Effects of ozone, acid mist and soil characteristics on clonal Norway spruce (Picea abies (L.) Karst.)--overall results and conclusions of the joint 14 month tree exposure experiment in closed chambers

This paper summarizes and evaluates the main findings of 14 preceding papers related to the joint 14-month tree-exposure experiment carried out by the 'Munich Working Party on Air Pollution' at the GSF, Munich, FRG, from July 1986 to September 1987. The experiment tested the hypothesis that an interaction of ozone/acid mist/soil/extreme climatic conditions is the cause of decline of Norway spruce (Picea abies (L.) Karst.) at higher altitudes of the Inner Bavarian Forest. The main findings of the individual studies are presented and their implications for the hypothesis are discussed. Clear effects of soil and genetic factors (differences between clones), for example on growth and frost resistance were found. Treatment with O(3)/acid mist was shown to have effects on plant biochemistry, physiology, histology/ cytology, and growth. The wide scattering of these effects, and the lack of a consistent pattern of response across all clones does not permits a firm conclusion on the validity of the experimental hypothesis. These effects were not confounded by the nutrient stresses imposed during the initial exposure period and were not found to be cumulative during repeated treatments, as was proposed by the hypothesis. It is concluded that the experimental evidence does not indicate that ozone/acid mist are major factors to explain the Norway spruce decline on acidic sites at higher altitudes of the Inner Bavarian Forest and probably similar forest areas.