Responses to Airborne Ozone and Soilborne Metal Pollution in Afforestation Plants with Different Life Forms.

With the current increases in environmental stress, understanding species-specific responses to multiple stress agents is needed. This science is especially important for managing ecosystems that are already confronted with considerable pollution. In this study, responses to ozone (O3, ambient daily course values + 20 ppb) and mixed metal contamination in soils (MC, cadmium/copper/lead/zinc = 25/1100/2500/1600 mg kg-1), separately and in combination, were evaluated for three plant species (Picea abies, Acer pseudoplatanus, Tanacetum vulgare) with different life forms and ecological strategies. The two treatments elicited similar stress reactions, as shown by leaf functional traits, gas exchange, tannin, and nutrient markers, irrespective of the plant species and life form, whereas the reactions to the treatments differed in magnitude. Visible and microscopic injuries at the organ or cell level appeared along the penetration route of ozone and metal contamination. At the whole plant level, the MC treatment caused more severe injuries than the O3 treatment and few interactions were observed between the two stress factors. Picea trees, with a slow-return strategy, showed the highest stress tolerance in apparent relation to an enhancement of conservative traits and an exclusion of stress agents. The ruderal and more acquisitive Tanacetum forbs translocated large amounts of contaminants above ground, which may be of concern in a phytostabilisation context. The deciduous Acer trees-also with an acquisitive strategy-were most sensitive to both stress factors. Hence, species with slow-return strategies may be of particular interest for managing metal-polluted sites in the current context of multiple stressors and for safely confining soil contaminants below ground.

Novel method to determine element concentrations in foliage of poplar and willow cuttings.

Measuring the uptake of the chemical elements by plants usually requires the destructive harvest of the plants. Analyzing individual leaves is unsatisfactory because their elemental concentration depends on their age and position on the branch or stem. We aimed to find an easy method to determine the elemental concentrations using a few suitable single leaves along the main shoot of poplar (Populus monviso) and willow (Salix viminalis) cuttings at the end of the first season. Using Ca, Cd, Mn, Fe, K, P, Pb, and Zn concentrations, measured in selected leaves along the main shoots of the cuttings, mathematical functions were derived, which described best their distribution. Elemental allocation patterns were independent of the soil characteristics and soil element concentrations. Based on these functions, three leaves from specific positions along the main shoot were selected, which could accurately describe the derived functions. The deviation of the calculated average concentration, based on the 3-leaves method, was ≤15% in approximately 65% of the cases compared to the measured concentration. This method could be used to calculate element concentrations and fluxes in phytomanagement, biomonitoring, or biomass productions projects using one-season poplar or willow cuttings.

Changes in the dynamics of foliar N metabolites in oak saplings by drought and air warming depend on species and soil type.

Climate change poses direct or indirect influences on physiological mechanisms in plants. In particular, long living plants like trees have to cope with the predicted climate changes (i.e. drought and air warming) during their life span. The present study aimed to quantify the consequences of simulated climate change for foliar N metabolites over a drought-rewetting-drought course. Saplings of three Central European oak species (i.e. Quercus robur, Q. petraea, Q. pubescens) were tested on two different soil types (i.e. acidic and calcareous). Consecutive drought periods increased foliar amino acid-N and soluble protein-N concentrations at the expense of structural N in all three oak species. In addition, transient effects on foliar metabolite dynamics were observed over the drought-rewetting-drought course. The lowest levels of foliar soluble protein-N, amino acid-N and potassium cation with a minor response to drought and air warming were found in the oak species originating from the driest/warmest habitat (Q. pubescens) compared to Q. robur and Q. petraea. Higher foliar osmolyte-N and potassium under drought and air warming were observed in all oak species when grown on calcareous versus acidic soil. These results indicate that species-specific differences in physiological mechanisms to compensate drought and elevated temperature are modified by soil acidity.

A phenological timetable of oak growth under experimental drought and air warming.

Climate change is expected to increase temperature and decrease summer precipitation in Central Europe. Little is known about how warming and drought will affect phenological patterns of oaks, which are considered to possess excellent adaptability to these climatic changes. Here, we investigated bud burst and intra-annual shoot growth of Quercus robur, Q. petraea and Q. pubescens grown on two different forest soils and exposed to air warming and drought. Phenological development was assessed over the course of three growing seasons. Warming advanced bud burst by 1-3 days °C⁻¹ and led to an earlier start of intra-annual shoot growth. Despite this phenological shift, total time span of annual growth and shoot biomass were not affected. Drought changed the frequency and intensity of intra-annual shoot growth and advanced bud burst in the subsequent spring of a severe summer drought by 1-2 days. After re-wetting, shoot growth recovered within a few days, demonstrating the superior drought tolerance of this tree genus. Our findings show that phenological patterns of oaks are modified by warming and drought but also suggest that ontogenetic factors and/or limitations of water and nutrients counteract warming effects on the biomass and the entire span of annual shoot growth.

Foliar symptoms triggered by ozone stress in irrigated holm oaks from the city of Madrid, Spain.

BACKGROUND: Despite abatement programs of precursors implemented in many industrialized countries, ozone remains the principal air pollutant throughout the northern hemisphere with background concentrations increasing as a consequence of economic development in former or still emerging countries and present climate change. Some of the highest ozone concentrations are measured in regions with a Mediterranean climate but the effect on the natural vegetation is alleviated by low stomatal uptake and frequent leaf xeromorphy in response to summer drought episodes characteristic of this climate. However, there is a lack of understanding of the respective role of the foliage physiology and leaf xeromorphy on the mechanistic effects of ozone in Mediterranean species. Particularly, evidence about morphological and structural changes in evergreens in response to ozone stress is missing. RESULTS: Our study was started after observing ozone -like injury in foliage of holm oak during the assessment of air pollution mitigation by urban trees throughout the Madrid conurbation. Our objectives were to confirm the diagnosis, investigate the extent of symptoms and analyze the ecological factors contributing to ozone injury, particularly, the site water supply. Symptoms consisted of adaxial and intercostal stippling increasing with leaf age. Underlying stippling, cells in the upper mesophyll showed HR-like reactions typical of ozone stress. The surrounding cells showed further oxidative stress markers. These morphological and micromorphological markers of ozone stress were similar to those recorded in deciduous broadleaved species. However, stippling became obvious already at an AOT40 of 21 ppm•h and was primarily found at irrigated sites. Subsequent analyses showed that irrigated trees had their stomatal conductance increased and leaf life -span reduced whereas the leaf xeromorphy remained unchanged. These findings suggest a central role of water availability versus leaf xeromorphy for ozone symptom expression by cell injury in holm oak.

Metal uptake and allocation in trees grown on contaminated land: implications for biomass production.

Phytostabilization aims to reduce environmental and health risks arising from contaminated soil. To be economically attractive, plants used for phytostabilization should produce valuable biomass. This study investigated the biomass production and metal allocation to foliage and wood of willow (Salix viminalis L.), poplar (Populus monviso), birch (Betula pendula), and oak (Quercus robur) on five different soils contaminated with trace elements (TE), with varying high concentrations of Cu, Zn, Cd, and Pb as well as an uncontaminated control soil. In the treatment soils, the biomass was reduced in all species except oak. There was a significant negative correlation between biomass and foliar Cd and Zn concentrations, reaching up to 15 mg Cd kg(-1) and 2000 mg Zn kg(-1) in willow leaves. Lead was the only TE with higher wood than foliage concentrations. The highest Pb accumulation occurred in birch with up to 135 mg kg(-1) in wood and 78 mg kg(-1) in foliage. Birch could be suitable for phytostabilization of soils with high Cd and Zn but low Pb concentrations, while poplars and willows could be used to stabilise soils with high Cu and Pb and low Zn and Cd concentrations.

Seasonal exposure to drought and air warming affects soil Collembola and mites.

Global environmental changes affect not only the aboveground but also the belowground components of ecosystems. The effects of seasonal drought and air warming on the genus level richness of Collembola, and on the abundance and biomass of the community of Collembola and mites were studied in an acidic and a calcareous forest soil in a model oak-ecosystem experiment (the Querco experiment) at the Swiss Federal Research Institute WSL in Birmensdorf. The experiment included four climate treatments: control, drought with a 60% reduction in rainfall, air warming with a seasonal temperature increase of 1.4 °C, and air warming + drought. Soil water content was greatly reduced by drought. Soil surface temperature was slightly increased by both the air warming and the drought treatment. Soil mesofauna samples were taken at the end of the first experimental year. Drought was found to increase the abundance of the microarthropod fauna, but reduce the biomass of the community. The percentage of small mites (body length ≤ 0.20 mm) increased, but the percentage of large mites (body length >0.40 mm) decreased under drought. Air warming had only minor effects on the fauna. All climate treatments significantly reduced the richness of Collembola and the biomass of Collembola and mites in acidic soil, but not in calcareous soil. Drought appeared to have a negative impact on soil microarthropod fauna, but the effects of climate change on soil fauna may vary with the soil type.

Boron accumulation and toxicity in hybrid poplar (Populus nigra × euramericana).

Poplars accumulate high B concentrations and are thus used for the phytomanagement of B contaminated soils. Here, we performed pot experiments in which Populus nigra × euramericana were grown on a substrate with B concentrations ranging from 13 to 280 mg kg(-1) as H(3)BO(3). Salix viminalis, Brassica juncea, and Lupinus albus were grown under some growing conditions for comparison. Poplar growth was unaffected at soil B treatment levels up to 93 mg kg(-1). Growth was progressively reduced at levels of 168 and 280 mg kg(-1). None of the other species survived at these substrate B levels. At leaf B concentrations <900 mg kg(-1) only <10% of the poplar leaf area showed signs of toxicity. Neutron radiography revealed that chlorotic leaf tissues had B concentrations of 1000-2000 mg kg(-1), while necrotic tissues had >2000 mg kg(-1). Average B concentrations of up to 3500 mg kg(-1) were found in leaves, while spots within leaves had concentrations >7000 mg kg(-1), showing that B accumulation in leaf tissue continued even after the onset of necrosis. The B accumulation ability of P. nigra × euramericana is associated with B hypertolerance in the living tissue and storage of B in dead leaf tissue.

Provenance-specific growth responses to drought and air warming in three European oak species (Quercus robur, Q. petraea and Q. pubescens).

Provenance-specific growth responses to experimentally applied drought and air warming were studied in saplings of three European oak species: Quercus robur, Quercus petraea and Quercus pubescens. Four provenances of each species were grown in large open-top chambers and subjected to four climates: control, periodic drought, air warming or their combination in 3 subsequent years. Overall growth responses were found among species and provenances, with drought reducing shoot height growth and stem diameter growth and air warming stimulating shoot height growth but reducing stem diameter growth and root length growth. Differential growth responses in shoots, stems and roots resulted in altered allometric growth relations. Root length growth to shoot height growth increased in response to drought but decreased in response to air warming. Stem diameter growth to shoot height growth decreased in response to air warming. The growth responses in shoots and stems were highly variable among provenances indicating provenance-specific sensitivity to drought and air warming, but this response variability did not reflect local adaptation to climate conditions of provenance origin. Shoot height growth was found to be more sensitive to drought in provenances from northern latitudes than in provenances from southern latitudes, suggesting that genetic factors related to the postglacial immigration history of European oaks might have interfered with selective pressure at provenance origins.

Compartmentation of metals in foliage of Populus tremula grown on soils with mixed contamination. I. From the tree crown to leaf cell level.

In order to achieve efficient phytoextraction of heavy metals using trees, the metal allocation to aboveground tissues needs to be characterised. In his study, the distribution of heavy metals, macro- and micronutrients and the metal micro-localisation as a function of the leaf position and heavy metal treatment were analysed in poplars grown on soil with mixed metal contamination. Zinc was the most abundant contaminant in both soil and foliage and, together with cadmium, was preferentially accumulated in older foliage whereas excess copper and lead were not translocated. Changes in other element concentrations indicated an acceleration in aging as a consequence of the metal treatment. Excess zinc was irregularly accumulated inside leaf tissues, tended to saturate the veins and was more frequently stored in cell symplast than apoplast. Storage compartments including metabolically safe and sensitive subcellular sites resulted in sizable metal accumulation as well as stress reactions.

Compartmentation of metals in foliage of Populus tremula grown on soils with mixed contamination. II. Zinc binding inside leaf cell organelles.

The phytoextraction potential of plants for removing heavy metals from polluted soils is determined by their capacity to store contaminants in aboveground organs and complex them safely. In this study, the metal compartmentation, elemental composition of zinc deposits and zinc complexation within leaves from poplars grown on soil with mixed metal contamination was analysed combining several histochemical and microanalytical approaches. Zinc was the only heavy metal detected and was stored in several organelles in the form of globoid deposits showing ß-metachromasy. It was associated to oxygen anions and different cations, noteworthy phosphorous. The deposit structure, elemental composition and element ratios indicated that zinc was chelated by phytic acid ligands. Maturation processes in vacuolar vs. cytoplasmic deposits were suggested by differences in size and amounts of complexed zinc. Hence, zinc complexation by phytate contributed to metal detoxification and accumulation in foliage but could not prevent toxicity reactions therein.

Erratum to "Structural and physiological responses to ozone in Manna ash (Fraxinus ornus L.) leaves of seedlings and mature trees under controlled and ambient conditions".

Leaf-level microscopical symptom structure and physiological responses were investigated in seedlings experimentally exposed to ozone (O3) in indoor chambers (150 ppb, 8 hd(-1) per 7 weeks), and field trees of Manna ash (Fraxinus ornus) exposed to ambient O3 (max 93 ppb per one growing season). Ozone-induced leaf injury, including leaf reddening and stippling, was observed in both seedlings and mature trees, but the morphology of injury in the stipples differed, being hypersensitive-like (HR-like) in the chamber seedlings and accelerated cell senescence (ACS) in the field trees. In both exposure conditions, the main structural impact of O3 was on the mesophyll and especially the upper assimilating cell layers. The main physiological impact was on carbon assimilation and on stomatal sluggishness. These effects were not due to stomatal structural injury and were more severe in juvenile compared to mature trees because of environmental (water availability, light) and constitutional (gas exchange capacity) factors and differences in the cell physiology processes (HR-like vs. ACS) triggered by ozone stress. Given the plasticity of plant responses to ozone stress, dose/response relationships for tree seedlings in the indoor chambers cannot be extrapolated to mature trees unless ambient conditions are closely simulated.

Structural and physiological responses to ozone in Manna ash (Fraxinus ornus L.) leaves of seedlings and mature trees under controlled and ambient conditions.

Leaf-level microscopical symptom structure and physiological responses were investigated in seedlings experimentally exposed to ozone (O3) in indoor chambers (150 ppb, 8 h d(-1)/7 weeks), and field trees of Manna ash (Fraxinus ornus) exposed to ambient O3 (max 93 ppb/one growing season). Ozone-induced leaf injury, including leaf reddening and stippling, was observed in both seedlings and mature trees, but the morphology of injury in the stipples differed, being hypersensitive-like (HR-like) in the chamber seedlings and accelerated cell senescence (ACS) in the field trees. In both exposure conditions, the main structural impact of O3 was on the mesophyll and especially the upper assimilating cell layers. The main physiological impact was on carbon assimilation and on stomatal sluggishness. These effects were not due to stomatal structural injury and were more severe in juvenile compared to mature trees because of environmental (water availability, light) and constitutional (gas exchange capacity) factors and differences in the cell physiology processes (HR-like vs. ACS) triggered by ozone stress. Given the plasticity of plant responses to ozone stress, dose/response relationships for tree seedlings in the indoor chambers cannot be extrapolated to mature trees unless ambient conditions are closely simulated.

Heavy metal accumulation and phytostabilisation potential of tree fine roots in a contaminated soil.

Root systems of Norway spruce (Picea abies) and poplar (Populus tremula) were long-term exposed to metal-contaminated soils in open-top chambers to investigate the accumulation of the heavy metals in the fine roots and to assess the plants suitability for phytostabilisation. The heavy metals from the contaminated soil accumulated in the fine roots about 10-20 times more than in the controls. The capacity to bind heavy metals already reached its maximum after the first vegetation period. Fine roots of spruce tend to accumulate more heavy metals than poplar. Copper and Zinc were mainly detected in the cell walls with larger values in the epidermis than in the cortex. The heavy metals accumulated in the fine roots made up 0.03-0.2% of the total amount in the soils. We conclude that tree fine roots adapt well to conditions with heavy metal contamination, but their phytostabilisation capabilities seem to be very low.

Leaf responsiveness of Populus tremula and Salix viminalis to soil contaminated with heavy metals and acidic rainwater.

Fast-growing trees such as Salix viminalis L. and Populus tremula L. are well suited to phytoremediate heavy metal contaminated soils. However, information on tree performance, particularly leaf function, under conditions of heavy metal contamination is scarce. We used yearly coppiced saplings of S. viminalis and P. tremula growing in model ecosytems to test four hypotheses: (1) heavy metal contamination impairs photosynthesis by injuring leaf structure; (2) the effects of heavy metal contamination are enhanced by acidified rainwater and low soil pH; (3) heavy metal contamination increases dark respiration and, thus, repair processes; and (4) heavy metal contamination is tolerated and remediated better by S. viminalis than by P. tremula. We investigated heavy metal accumulation, tissue injury and gas exchange in leaves of plants subjected to controlled soil contamination with heavy metal dust. Additional treatments included acidic and calcareous natural forest subsoils in combination with irrigation with rainwater at pH 5.5 or 3.5. In both provenances of P. tremula that were studied, but not in S. viminalis, heavy metal treatment reduced photosynthesis and transpiration by varying amounts, except in the hot and dry summer of 2003, but had no effect on dark respiration. At light saturation, net CO(2) uptake and water-use efficiency were reduced by heavy metal contamination, whereas the CO(2) concentration in the leaf intercellular air space was increased. Rainwater pH and subsoil pH only slightly modified the effects of the heavy metal treatment on P. tremula. Gas exchange responses of P. tremula to heavy metals were attributed to leaf structural and ultrastructural changes resulting from hypersensitive-response-like processes and accelerated mesophyll cell senescence and necroses in the lower epidermis, especially along the transport pathways of heavy metals in the leaf lamina. Overall, the effects of heavy metals on P. tremula corroborated Hypothesis 1, but refuted Hypotheses 2 and 3, and were inconclusive for Hypothesis 4. Both P. tremula and S. viminalis showed appreciable potential for storing heavy metals in aging foliage.

Impacts of air pollution and climate change on forest ecosystems--emerging research needs.

Outcomes from the 22nd meeting for Specialists in Air Pollution Effects on Forest Ecosystems "Forests under Anthropogenic Pressure--Effects of Air Pollution, Climate Change and Urban Development", September 10-16, 2006, Riverside, CA, are summarized. Tropospheric or ground-level ozone (O3) is still the phytotoxic air pollutant of major interest. Challenging issues are how to make O3 standards or critical levels more biologically based and at the same time practical for wide use; quantification of plant detoxification processes in flux modeling; inclusion of multiple environmental stresses in critical load determinations; new concept development for nitrogen saturation; interactions between air pollution, climate, and forest pests; effects of forest fire on air quality; the capacity of forests to sequester carbon under changing climatic conditions and coexposure to elevated levels of air pollutants; enhanced linkage between molecular biology, biochemistry, physiology, and morphological traits.

Linking stress with macroscopic and microscopic leaf response in trees: new diagnostic perspectives.

Visible symptoms in tree foliage can be used for stress diagnosis once validated with microscopical analyses. This paper reviews and illustrates macroscopical and microscopical markers of stress with a biotic (bacteria, fungi, insects) or abiotic (frost, drought, mineral deficiency, heavy metal pollution in the soil, acidic deposition and ozone) origin helpful for the validation of symptoms in broadleaved and conifer trees. Differentiation of changes in the leaf or needle physiology, through ageing, senescence, accelerated cell senescence, programmed cell death and oxidative stress, provides additional clues raising diagnosis efficiency, especially in combination with information about the target of the stress agent at the tree, leaf/needle, tissue, cell and ultrastructural level. Given the increasing stress in a changing environment, this review discusses how integrated diagnostic approaches lead to better causal analysis to be applied for specific monitoring of stress factors affecting forest ecosystems.

Diagnosis of abiotic and biotic stress factors using the visible symptoms in foliage.

Visible symptoms in the foliage of trees are recorded to monitor the effects of abiotic and biotic stress. Difficulties are reported in diagnosing the origin of stress. The present paper discusses several diagnostic criteria which are usable in different species for a better determination of the stress factor type. A new diagnosis scheme to differentiate between classes of abiotic and biotic stress factors is supplied. Abiotic stress generates gradients of symptoms. The symptom specificity is determined by the degree of interaction between the stress factor and plant defense system. Symptoms caused by abiotic stress and natural autumnal senescence can be morphologically different or undistinguishable according to the stress and plant species. With biotic stress, the class of parasitic is generally recognizable on the basis of the visible symptoms. Structurally and physiologically based explanations of the symptom morphology are still missing for many stress factors.

Effects of metal-contaminated soil on the performance of young trees growing in model ecosystems under field conditions.

Young Populus tremula, Salix viminalis, Betula pendula and Picea abies trees were grown together in large open-top chambers. The treatments were: without or with (Cu/Zn/Cd/Pb=640/3000/10/90 mg kg-1) metal contamination in the topsoil, irrigation pH 3.5 or 5.5, and acidic or calcareous subsoil. Growth, metal allocation to foliage and wood, as well as leaf gas exchange were measured. Biomass was reduced in P. tremula and B. pendula by the metal-contaminated topsoil relative to uncontaminated topsoil, whereas in P. tremula photosynthesis and transpiration were decreased. These effects were related to the elevated foliar Zn accumulation in P. tremula. S. viminalis showed a significant reduction in growth and an increased Zn and Cd accumulation on acidic vs. calcareous subsoil. Acidic irrigation produced only a few significant effects. P. abies showed the lowest metal uptake and no growth response to metal contamination.

Diagnosis of abiotic and biotic stress factors using the visible symptoms in foliage.

Visible symptoms in the foliage of trees are recorded to monitor the effects of abiotic and biotic stress. Difficulties are reported in diagnosing the origin of stress. The present paper discusses several diagnostic criteria which are usable in different species for a better determination of the stress factor type. A new diagnosis scheme to differentiate between classes of abiotic and biotic stress factors is supplied. Abiotic stress generates gradients of symptoms. The symptom specificity is determined by the degree of interaction between the stress factor and plant defense system. Symptoms caused by abiotic stress and natural autumnal senescence can be morphologically different or undistinguishable according to the stress and plant species. With biotic stress, the class of parasitic is generally recognizable on the basis of the visible symptoms. Structurally and physiologically based explanations of the symptom morphology are still missing for many stress factors.