Comparative analysis of abscisic acid levels and expression of abscisic acid-related genes in Scots pine and Norway spruce seedlings under water deficit.

Abscisic acid (ABA) is one of the main participants in the regulation of plant responses to water deficiency. Knowledge of the ABA signal transduction pathways in gymnosperms is rather limited, especially in comparison with those in angiosperms. Seedlings of Scots pine and Norway spruce are known for their contrasting behaviour strategies under water deficit. To characterize the possible role of ABA in these differences, ABA dynamics were investigated under conditions of water deficit in seedlings of these two species. The content of ABA and its catabolites was followed in the roots and needles of seedlings of Pinus sylvestris and Picea abies under conditions of polyethylene glycol (PEG)-induced water deficiency (-0.15 and -0.5 MPa) for 10 days. The expression of the main genes for ABA-biosynthetic enzymes was also analysed. ABA showed more pronounced stress-dependent dynamics in pine roots than in spruce roots, whereas in needles, the response was greater for spruce than pine. The ABA increase during drought was mainly due to de novo synthesis and the shift in the balance between ABA synthesis and catabolism towards synthesis. The ABA-glucosyl ester did not serve as a reserve for the release of free ABA under water deficiency. The expression levels of the main ABA biosynthetic genes showed a weak or no correlation with changes in ABA content under water stress, i.e., the ABA content in the seedlings of both species was not directly linked to the transcript levels of the main ABA biosynthetic genes. Less-pronounced stress-induced changes in ABA in pine needles than in spruce needles may be related to pine seedlings having a less conservative strategy of growth and maintenance of water balance under water deficit.

The effects of structurally different siderophores on the organelles of Pinus sylvestris root cells.

MAIN CONCLUSION: Siderophores are a driver of Pinus sylvestris root responses to metabolites secreted by pathogenic and mycorrhizal fungi. Structurally different siderophores regulate the uptake of Fe by microorganisms and may play a key role in the colonization of plants by beneficial or pathogenic fungi. Siderophore action, however, may be dependent on the distribution of Fe within cells. Here, the involvement of siderophores in determining the changes of organelle morphology and element composition of some cellular fractions of root cells in Pinus sylvestris to trophically diverse fungi was investigated. Changes in the morphology and concentrations of different elements within organelles of root cells in response to three structurally different siderophores were examined by transmission electron microscopy combined with energy-dispersive X-ray spectroscopy. Weak development of mitochondrial cristae and the deposition of backup materials in plastids occurred in the absence of Fe in the structures of triacetylfusarinine C and ferricrocin. In response to metabolites of both pathogenic and mycorrhizal fungi, Fe accumulated mainly in the cell walls and cytoplasm. Fe counts increased in all of the analyzed organelles in response to applications of ferricrocin and triacetylfusarinine C. Chelation of Fe within the structure of siderophores prevents the binding of exogenous Fe, decreasing the abundance of Fe in the cell wall and cytoplasm. The concentrations of N, P, K, Ca, Mn, Cu, Mg, and Zn also increased in cells after applications of ferricrocin and triacetylfusarinine C, while the levels of these elements decreased in the cell wall and cytoplasm when Fe was present within the structure of the siderophores. These results provide insight into the siderophore-driven response of plants to various symbionts.

Long-term responses of Scots pine (Pinus sylvestris L.) and European beech (Fagus sylvatica L.) to the contamination of light soils with diesel oil.

Research into trees plays a very important role in evaluations of soil contamination with diesel oil. Trees are ideal for reclaiming contaminated soils because their large biomass renders them more resistant to higher concentrations of pollutants. In the literature, there is a general scarcity of long-term studies performed on trees, in particular European beeches. The aim of this study was to evaluate the responses of Scots pines and European beeches grown for 8 years on soil contaminated with diesel oil. Selected morphological and physiological parameters of trees were analyzed. The biomass yield of Scots pines was not significantly correlated with increasing concentrations of diesel oil, but it was more than 700% higher than in European beeches. Scots pines were taller and had a larger stem diameter than European beeches during the 8-year study. The diameter of trees grown on the most contaminated soil was reduced 1.5-fold in Scots pines and more than twofold in European beeches. The length of Scots pine needles from the most contaminated treatment decreased by 50% relative to control needles. The shortest needles were heaviest. The fluctuating asymmetry (FA) of needle length was highest in Scots pines grown on the most contaminated soil, whereas the reverse was noted in the FA of needle weight. Diesel oil decreased the concentrations of chlorophylls a and b, total chlorophyll, and carotenoids. The Fv/Fm ratio of needles and leaves was influenced by the tested concentrations of diesel oil. The results of the study indicate that the Scots pine better adapts (grows more rapidly and produces higher biomass) to long-term soil contamination with diesel oil than the European beech. In European beeches, growth inhibition and leaf discoloration (a decrease in chlorophyll content) were observed already after the first year of the experiment, which indicates that 1-year-old seedlings of European beech are robust bioindicators of soil contamination with diesel oil.

Accumulative response of Scots pine (Pinus sylvestris L.) and silver birch (Betula pendula Roth) to heavy metals enhanced by Pb-Zn ore mining and processing plants: Explicitly spatial considerations of ordinary kriging based on a GIS approach.

Plants have an accumulative response to heavy metals present in soils or deposited from airborne sources of emissions. Therefore, their tissues are very often used in studies of heavy metal contamination originating from different sources as a bioindicator of environmental pollution. This research was undertaken to examine accumulation capacities of Pb, Zn, Cd, Cu and Cr in washed and unwashed needles of Scots pine (Pinus sylvestris L.) and leaves of silver birch (Betula pendula Roth) growing in a contaminated area. We collected needles of Scots pine and leaves of silver birch in an area around a sedimentation pond and metallurgic plant processing Pb and Zn ores near Olkusz, Poland. Concentrations of heavy metals, which have been linked with exposure to emissions, were determined from foliar samples collected at 33 sites. These sites were established at various distances (0.5-3.6 km) from the pond and metallurgic plant so as to identify the predominant accumulative response of plants. Spatial gradients for Pb and Zn were calculated using an ordinary kriging interpolation algorithm. A spatial pattern was identified by a GIS method to visualize maps over the Pb-Zn ore mining area. The accumulation of Zn (R2 = 0.74, p < 0.05) and Pb (R2 = 0.85, p < 0.01) in plant tissues correlated with soil concentrations. This tendency was not found in the case of Cu, Cd and Cr.

Trace element contamination differentiates the natural population of Scots pine: evidence from DNA microsatellites and needle morphology.

The Scots pine is often used in the biomonitoring of forests. Studies on the chemical composition plus variability of its needles morphological structure allow for an assessment of the state of environmental pollution. However, in their natural populations, the response of individual trees to stress differs. This study reports on the influence of long-term soil contamination with trace elements on the morphology of the needles, its possible relation to the differentiation of the genetic pool, and their implications for biomonitoring. In the natural and self-renewable pine stand growing near the point polluter (zinc smelter, Upper Silesia, Poland), two categories of trees are observed with respect to their health status: pollution-tolerant (T) and pollution-sensitive (S). A detailed analysis of the trace element content of the needles reveals that the concentration of Cd, Zn, Pb, and Cu in the needles is significantly higher in S as compared to T individuals. The metal accumulation pattern decidedly follows the sequence Pb > Cd > Cu > Zn. An analysis of the fluctuating asymmetry (FA) of the needles reveals that sensitive trees showed an FA index ten times higher in comparison to tolerant ones. Moreover, the high differences between these S and T tree groups are also observed in the basic genetic diversity parameters investigated by an analysis of DNA simple sequence repeats (SSR). The concentration of trace elements in pine needles, distinct in sensitive and tolerant trees and in connection with their morphological and genetic characteristics, may reflect an adaptation process. The level of Mg and Fe content in the needles could be a physiological-toxicological index for evaluating trace element "lethality" expressed as Mg and Fe mineral-survival strategies. The example of differences described in this Scots pine population should be taken into consideration in ecotoxicological research to better interpret the obtained results.

Effects of copper deficiency and copper toxicity on organogenesis and some physiological and biochemical responses of Scots pine (Pinus sylvestris L.) seedlings grown in hydroculture.

The morphological, physiological, and biochemical parameters of 6-week-old seedlings of Scots pine (Pinus sylvestris L.) were studied under deficiency (1.2 nM) and chronic exposure to copper (0.32, 1, 2.5, 5, and 10 µM CuSO4) in hydroculture. The deposit of copper in the seed allowed the seedlings to develop under copper deficiency without visible disruption of growth. The high sensitivity of Scots pine to the toxic effects of copper was shown, which manifested as a significant inhibition of growth and development. The loss of dominance of the main root and a strong inhibition of lateral root development pointed to a lack of adaptive reorganization of the root system architecture under copper excess. A preferential accumulation of copper in the root and a minor translocation in aerial organs confirmed that Scots pine belongs to a group of plants that exclude copper. Selective impairment in the absorption of manganese was discovered, under both deficiency and excess of copper in the nutrient solution, which was independent of the degree of development of the root system. Following 10 µM CuSO4 exposure, the absorption of manganese and iron from the nutrient solution was completely suppressed, and the development of seedlings was secured by the stock of these micronutrients in the seed. The absence of signs of oxidative stress in the seedling organs was shown under deficiency and excess of copper, as evidenced by the steady content of malondialdehyde and 4-hydroxyalkenals. Against this background, no changes in total superoxide dismutase activity in the organs of seedlings were revealed, and the increased content of low-molecular-weight antioxidants was observed in the roots under 1 µM and in the needles under 5 µM CuSO4 exposures.

Accumulation of heavy metals and antioxidant responses in Pinus sylvestris L. needles in polluted and non-polluted sites.

The purpose of this study was to determine the concentrations of heavy metals (cadmium, iron, manganese, lead and zinc) in current-year, 1-year old and 2-year old needles of Pinus sylvestris L. Trees were from three heavily polluted (immediate vicinity of zinc smelter, iron smelter and power plant) and three relatively clean sites (nature reserve, ecologically clean site and unprotected natural forest community) in southern Poland. Analysis also concerned the antioxidant response and contents of protein, proline, total glutathione, non-protein thiols and activity of guaiacol peroxidase (GPX) in the needles. Generally, in pine needles from the polluted sites, the concentrations of the metals were higher and increased with the age of needles, and in most cases, antioxidant responses also were elevated. The highest levels of Cd, Pb and Zn were found in 2-year old pine needles collected near the polluted zinc smelter (respectively: 6.15, 256.49, 393.5 mg kg(-1)), Fe in 2-year old pine needles in the vicinity of the iron smelter (206.82 mg kg(-1)) and Mn in 2-year old needles at the ecologically clean site (180.32 mg kg(-1)). Positive correlations were found between Fe, Mn and Pb and the content of proteins and NPTs, between Cd and non-protein -SH groups, and between Zn and proline levels. The activity of GPX increased under the influence of Mn, while glutathione levels tended to decrease as Mn levels rose. The data obtained show that the levels of protein and non-protein -SH groups may be useful in biological monitoring, and that these ecophysiological parameters seem to be good evidence of elevated oxidative stress caused by heavy metals.

Growth responses of Scots pine to climatic factors on reclaimed oil shale mined land.

Afforestation on reclaimed mining areas has high ecological and economic importance. However, ecosystems established on post-mining substrate can become vulnerable due to climate variability. We used tree-ring data and dendrochronological techniques to study the relationship between climate variables and annual growth of Scots pine (Pinus sylvestris L.) growing on reclaimed open cast oil shale mining areas in Northeast Estonia. Chronologies for trees of different age classes (50, 40, 30) were developed. Pearson's correlation analysis between radial growth indices and monthly climate variables revealed that precipitation in June-July and higher mean temperatures in spring season enhanced radial growth of pine plantations, while higher than average temperatures in summer months inhibited wood production. Sensitivity of radial increment to climatic factors on post-mining soils was not homogenous among the studied populations. Older trees growing on more developed soils were more sensitive to precipitation deficit in summer, while growth indices of two other stand groups (young and middle-aged) were highly correlated to temperature. High mean temperatures in August were negatively related to annual wood production in all trees, while trees in the youngest stands benefited from warmer temperatures in January. As a response to thinning, mean annual basal area increment increased up to 50 %. By managing tree competition in the closed-canopy stands, through the thinning activities, tree sensitivity and response to climate could be manipulated.

A cerato-platanin-like protein HaCPL2 from Heterobasidion annosum sensu stricto induces cell death in Nicotiana tabacum and Pinus sylvestris.

The cerato-platanin family is a group of small secreted cysteine-rich proteins exclusive for filamentous fungi. They have been shown to be involved in the interactions between fungi and plants. Functional characterization of members from this family has been performed mainly in Ascomycota, except Moniliophthora perniciosa. Our previous phylogenetic analysis revealed that recent gene duplication of cerato-platanins has occurred in Basidiomycota but not in Ascomycota, suggesting higher functional diversification of this protein family in Basidiomycota than in Ascomycota. In this study, we identified three cerato-platanin homologues from the basidiomycete conifer pathogen Heterobasidion annosum sensu stricto. Expression of the homologues under various conditions as well as their roles in the H. annosum s.s.-Pinus sylvestris (Scots pine) pathosystem was investigated. Results showed that HaCPL2 (cerato-platanin-like protein 2) had the highest sequence similarity to cerato-platanin from Ceratocystis platani and hacpl2 was significantly induced during nutrient starvation and necrotrophic growth. The treatment with recombinant HaCPL2 induced cell death, phytoalexin production and defense gene expression in Nicotiana tabacum. Eliciting and cell death-inducing ability accompanied by retardation of apical root growth was also demonstrated in Scots pine seedlings. Our results suggest that HaCPL2 might contribute to the virulence of H. annosum s.s. by promoting plant cell death.

Production and recovery of monosaccharides from lignocellulose hot water extracts in a pulp mill biorefinery.

Processing of hemicelluloses obtained with pressurized hot water extraction (PHWE) from Scots pine to monosaccharides and other chemicals was investigated experimentally. A process scheme consisting of ultrafiltration, acid hydrolysis, and chromatographic separation was proposed and evaluated. A two-stage ultrafiltration was found necessary for efficient fractionation of the wood extract. It was shown that the monosaccharides can be released from a concentrated hemicellulose fraction with sulfuric acid hydrolysis without a significant loss of yield due to decomposition of monosaccharides. Acid hydrolysate was successfully fractionated with ion exchange chromatography and the hydrolysis acid was recovered for reuse. The product fractions obtained include polyphenols and high molar mass hemicelluloses (from UF stage 1), arabinose (from UF stage 2), as well as acetic acid and a mixture of monosaccharides (xylose, galactose, mannose, glucose) from chromatography.

Rhizospheric NO affects N uptake and metabolism in Scots pine (Pinus sylvestris L.) seedlings depending on soil N availability and N source.

We investigated the interaction of rhizospheric nitric oxide (NO) concentration (i.e. low, ambient or high) and soil nitrogen (N) availability (i.e. low or high) with organic and inorganic N uptake by fine roots of Pinus sylvestris L. seedlings by (15) N feeding experiments under controlled conditions. N metabolites in fine roots were analysed to link N uptake to N nutrition. NO affected N uptake depending on N source and soil N availability. The suppression of nitrate uptake in the presence of ammonium and glutamine was overruled by high NO. The effects of NO on N uptake with increasing N availability showed different patterns: (1) increasing N uptake regardless of NO concentration (i.e. ammonium); (2) increasing N uptake only with high NO concentration (i.e. nitrate and arginine); and (3) decreasing N uptake (i.e. glutamine). At low N availability and high NO nitrate accumulated in the roots indicating insufficient substrates for nitrate reduction or its storage in root vacuoles. Individual amino acid concentrations were negatively affected with increasing NO (i.e. asparagine and glutamine with low N availability, serine and proline with high N availability). In conclusion, this study provides first evidence that NO affects N uptake and metabolism in a conifer.

Arboreal insects associated with herbicide-stressed Pinus resinosa and Pinus sylvestris used as Sirex noctilio trap trees in New York.

In September of 2004, Sirex noctilio F. (Hymenoptera: Siricidae) was detected in New York State and later found to be established over a larger area, including parts of southeastern Canada and the northeastern United States. A key component of S. noctilio detection and management plans in other parts of the world where S. noctilio has become established are chemically girdled trap trees. Trap tree usage in North America is confounded by the presence of diverse communities of organisms that inhabit dead and dying trees. We trapped a portion of the arboreal insect community arriving at Pinus resinosa Ait. and Pinus sylvestris L., trap trees girdled 3 mo before (April), one month before (June), and at S. noctilio flight (July) in central New York. Multiple-funnel traps attached to trap trees captured 30,031 individuals from 109 species of Scolytinae, Cerambycidae, and Siricidae. Ips pini (Say) and Ips grandicollis (Eichhoff) accounted for almost 50% of the scolytines captured at trap trees and were present on all girdling dates. Significantly more scolytines and cerambycids were captured on P. sylvestris compared with P. resinosa, but species richness of captured insects did not differ between the two trees. More total and conifer-inhabiting scolytines and cerambycids were captured in traps on trees girdled in April and June and higher observed species richness was found on trees girdled in April and controls. Results from this study suggest a large community of arboreal insects and associated organisms are attracted to chemically girdled trap trees and likely interact with S. noctilio.

Cavitation induced by a surfactant leads to a transient release of water stress and subsequent 'run away' embolism in Scots pine (Pinus sylvestris) seedlings.

Cavitation decreases the hydraulic conductance of the xylem and has, therefore, detrimental effects on plant water balance. However, cavitation is also hypothesized to relieve water stress temporarily by releasing water from embolizing conduits to the transpiration stream. Stomatal closure in response to decreasing water potentials in order to avoid excessive cavitation has been well documented in numerous previous studies. However, it has remained unclear whether the stomata sense cavitation events themselves or whether they act in response to a decrease in leaf water potential to a level at which cavitation is initiated. The effects of massive cavitation on leaf water potential, transpiration, and stomatal behaviour were studied by feeding a surfactant into the transpiration stream of Scots pine (Pinus sylvestris) seedlings. The stomatal response to cavitation in connection with the capacitive effect was also studied. A major transient increase in leaf water potential was found due to cavitation in the seedlings. As cavitation was induced by lowering the surface tension, the two mechanisms could be uncoupled, as the usual relation between xylem water potential and the onset of cavitation did not hold. Our results indicate that the seedlings responded more to leaf water potential and less to cavitation itself, as stomatal closure was insufficient to prevent the seedlings from being driven to 'run-away' cavitation in a manner of hours.

Manipulation of VOC emissions with methyl jasmonate and carrageenan in the evergreen conifer Pinus sylvestris and evergreen broadleaf Quercus ilex.

Plant defence can be induced by exposing plants to the plant hormone jasmonic acid (JA) or its volatile ester, methyl jasmonate (MeJA). Carrageenans (Carr) - sulphated D-galactans extracted from red algae - can also induce plant defences. In this study, the effects of exogenous MeJA and Carr application (concentration 300 and 12.7 µmol, respectively) on volatile emissions from two widespread evergreen woody species, Pinus sylvestris (nine Turkish and one Finnish provenance) and Quercus ilex (Italian provenance) were investigated. We collected headspace samples from seedlings and analysed the quality and quantity of volatile compounds emitted by treated and control plants. In total, 19 monoterpenes, 10 sesquiterpenes, 10 green leaf volatiles (GLVs) and two aromatic compounds were emitted by P. sylvestris from all the provenances studied. Foliar MeJA application clearly affected the volatile profiles of trees from all the provenances. Effects of Carr were genotype specific. In Q. ilex, emissions of sesquiterpenes, GLVs and the homoterpene (E)-DMNT were all induced by MeJA application. However, emissions of most constitutively emitted monoterpenes were significantly reduced. Carr application also led to a significant reduction in monoterpene emissions, but without corresponding increases in other emissions. Our results indicate that exogenously applied MeJA and Carr can both significantly modify the volatile profiles of P. sylvestris and Q. ilex, but also that there are important provenance- and species-specific differences in the overall degree of elicitation and compositions of elicited compounds.

Embryogenic potential and expression of embryogenesis-related genes in conifers are affected by treatment with a histone deacetylase inhibitor.

Somatic embryogenesis is used for vegetative propagation of conifers. Embryogenic cultures can be established from zygotic embryos; however, the embryogenic potential decreases during germination. In Arabidopsis, LEAFY COTYLEDON (LEC) genes are expressed during the embryonic stage, and must be repressed to allow germination. Treatment with the histone deacetylase inhibitor trichostatin A (TSA) causes de-repression of LEC genes. ABSCISIC ACID3 (ABI3) and its Zea mays ortholog VIVIPAROUS1 (VP1) act together with the LEC genes to promote embryo maturation. In this study, we have asked the question whether TSA treatment in a conifer affects the embryogenic potential and the expression of embryogenesis-related genes. We isolated two conifer LEC1-type HAP3 genes, HAP3A and HAP3B, from Picea abies and Pinus sylvestris. A comparative phylogenetic analysis of plant HAP3 genes suggests that HAP3A and HAP3B are paralogous genes originating from a duplication event in the conifer lineage. The expression of HAP3A is high, in both somatic and zygotic embryos, during early embryo development, but decreases during late embryogeny. In contrast, the expression of VP1 is initially low but increases during late embryogeny. After exposure to TSA, germinating somatic embryos of P. abies maintain the competence to differentiate embryogenic tissue, and simultaneously the germination progression is partially inhibited. Furthermore, when embryogenic cultures of P. abies are exposed to TSA during embryo maturation, the maturation process is arrested and the expression levels of PaHAP3A and PaVP1 are maintained, suggesting a possible link between chromatin structure and expression of embryogenesis-related genes in conifers.

Recovery of ectomycorrhiza after 'nitrogen saturation' of a conifer forest.

Trees reduce their carbon (C) allocation to roots and mycorrhizal fungi in response to high nitrogen (N) additions, which should reduce the N retention capacity of forests. The time needed for recovery of mycorrhizas after termination of N loading remains unknown. Here, we report the long-term impact of N loading and the recovery of ectomycorrhiza after high N loading on a Pinus sylvestris forest. We analysed the N% and abundance of the stable isotope (15) N in tree needles and soil, soil microbial fatty acid biomarkers and fungal DNA. Needles in N-loaded plots became enriched in (15) N, reflecting decreased N retention by mycorrhizal fungi and isotopic discrimination against (15) N during loss of N. Meanwhile, needles in N-limited (control) plots became depleted in (15) N, reflecting high retention of (15) N by mycorrhizal fungi. N loading was terminated after 20yr. The δ(15) N and N% of the needles decreased 6yr after N loading had been terminated, and approached values in control plots after 15yr. This decrease, and the larger contributions compared with N-loaded plots of a fungal fatty acid biomarker and ectomycorrhizal sequences, suggest recovery of ectomycorrhiza. High N loading rapidly decreased the functional role of ectomycorrhiza in the forest N cycle, but significant recovery occurred within 6-15yr after termination of N loading.

Effect of petrol fuel contamination on the growth of mature Scots pine, Pinus sylvestris L., trees.

The radial increment and crown status of mature Scots pine trees growing in polluted and unpolluted sites were compared. In 1996, as a result of some malfunction, unleaded petrol penetrated into the soil next to a plantation. Detailed geological and hydrological studies revealed the route of the spread of contamination and extent of the pollution. The trees growing in polluted sites revealed strong depletion of radial growth starting immediately after pollution. Such depletion lasted 2-3 years before the ring widths stabilised at a low level. After a few years the radial increment increased, and now do not differ from the increment of trees in the unpolluted sites.

[Effect of the complex of technogenic and recreational loads on development of trunk tissues of Scotch pine in the Krasnoyarsk forest-steppe].

The morphometric parameters of trunk tissues of Scotch pine in suburban pine forests of Krasnoyarsk are studied, which grow under the long-term impact of technogenic and recreational loads in comparison with the background. Technogenic and recreational loads decrease the intensity of work of the cambium, especially towards the xylem, its duration, the cell size, and the structure of storage tissues. In pine stands of the Krasnoyarsk forest-steppe, the effect of the complex of anthropogenic stressors is weakening and degradation of forest stands and inhibition of production process at the cell and tissue levels are taking place.

Northern plants and ozone.

Forests in northern Fennoscandia are mainly composed of the O3-sensitive species--Scots pine and downy, mountain, and silver birches. Seminatural vegetation also contributes to biodiversity, carbon cycling, and ecosystem services as a part of forests, mires, meadows, and road verges. Fumigation experiments show that current O3 concentrations of 30-50 ppb reduce plant biomass production and reproduction. Visible foliar injury is attributable to peak O3 concentrations and relates to fast phenological development and high growth rate. Trees can acclimate to O3-induced water stress by producing more xeromorphic leaves or needles. The direct effects of O3 on grassland vegetation also translate to changes in the structure and size of the soil microbial community, and ecosystem N cycling. It is necessary to reduce the emission of O3 precursors and maintain high biodiversity to protect northern ecosystems. Regular, systematic, countrywide monitoring and validation as well as quantification of the effects of O3 on plants in the Nordic countries are also necessary.

Impact of experimentally elevated ozone on seed germination and growth of Russian pine (Pinus sylvestris) and spruce (Picea spp.) provenances.

The impact of elevated ozone concentrations on early ontogenetic stages of pine (Pinus sylvestris) and spruce (Picea abies, Picea obovata, P. abies x P. obovata) seedlings originating from different provenances in Russia were studied in the open-field ozone fumigation system located in Kuopio, Finland, over a span of 2 y. The AOT40 value (accumulated ozone dose over the threshold 40 ppb during daylight hours) was 11 ppm hr per growing season, which was 1.4 times higher than the ambient air concentration. The plants were measured for germination rate; shoot increment; needle length; and dry mass of needles, shoots, and roots. Significant differences between pine and spruce provenance response to ozone were found in all parameters. Ozone stress immediately reduced the germination rate of Northern pine provenances, whereas biomass reductions became evident during the second year of the exposure in all pine provenances. Spruce species were more tolerant to elevated ozone concentrations. Our results indicate that seedling development is vulnerable to increasing ozone concentrations and that attention must be paid to the provenance selection.