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Monitoring environmental pollution in Erzurum by chemical analysis of Scots pine (Pinus sylvestris L.) needles.

Pollution distribution maps of copper (Cu), zinc (Zn) and lead (Pb) for Erzurum province were developed on the basis of chemical analysis of Scots pine (Pinus sylvestris L.) needles collected from randomly selected sampling points during 2 years. The maps show deposition zones for the studied elements and could help in identification of sources and directions of air pollution dispersion. This study indicated that vegetation in Erzurum was greatly endangered by sulphur dioxide (SO(2)), whereas Zn does not pose an immediate threat to vegetation in most of the country's territory. However, in the city center, main and secondary road agglomeration, very high pollution with Pb and Cu could limit growth. Higher levels of SO(2) measured in some areas of Erzurum might be harmful for human and animal health. Results of this study indicated that Scots pine needles were not contaminated by Zn.

Cities↗

Urban polluted forest soils induce elevated root peroxidase activity in Scots pine (Pinus sylvestris L.) seedlings.

Plant biomass. mycorrhizal status and root peroxidase activity were measured in ectomycorrhizal Scots pine (Pinus sylvestris L.) seedlings grown in urban polluted and native, non-polluted forest soils with added ammonium or potassium sulphates simulating N and S deposition of urban areas. Peroxidase activity in the fine roots of seedlings planted in polluted forest soils was higher than in those planted in non-polluted soils and correlated positively with the activities measured in an earlier study in the roots of mature Scots pines growing at the sites from where the soils were collected. Growth of seedlings and mycorrhizal status were not affected by the origin of soil. Exposing the seedlings to winter acclimation conditions for 6 weeks elevated peroxidase activity in the roots. The addition of ammonium or potassium sulphate to non-polluted soils did not induce elevated root peroxidase activity, although at the levels of 0.5 and 1.0 g of ammonium sulphate a slight increasing trend was observed. We suggest, that indirect biotic factors, i.e. changes in the community structure of soil fungi, early stages of recognition, and defence reactions of pine roots against saprophytic and pathogenic fungi may be participating in the elicitation of peroxidase (POD) activity, although the possible role of heavy metals cannot be excluded.

Adaptation, Physiological↗

Ozone-induced free polyamine response in scots pine in northern Finland.

The study aimed to assess the ozone-induced response in Scots pine (Pinus sylvestris L.) needles by measuring free polyamine concentrations. An open-top chamber experiment with realistically elevated ozone concentrations was carried out in northern Finland. A carry-over effect was detected: the concentrations of free polyamines, especially putrescine, were decreased at the beginning of the next growing season in the ozone-exposed trees. This indicates that the free polyamine pathway was not activated by ozone stress in Scots pines in northern conditions.

Air Pollutants↗

Shift in ectomycorrhizal community composition in Scots pine (Pinus sylvestris L.) seedling roots as a response to nickel deposition and removal of lichen cover.

Scots pine seedlings were exposed to wet-deposited nickel (Ni) and removal of lichen cover in a dry heath Scots pine forest. Ni deposition affected the colonization of roots by indigenous ectomycorrhizal fungi in contrasting ways in intact and skimmed quadrats. Highest frequencies of tubercle morphotypes of ectomycorrhiza were found in quadrats exposed to 100 mg m(-2) year(-1) Ni in lichen covered treatment, while in skimmed quadrats these peaked after the treatment with 10 mg Ni m(-2) year(-1). Removal of the lichen layer increased the value of diversity index (H') of ectomycorrhizal fungal community, probably due to the increase in the evenness of the morphotype distribution. Lichen removal seemed also to improve the condition of the short roots, as the frequencies of poor and senescent short roots were decreased by the removal.

Analysis of Variance↗

Fluid self-diffusion in Scots pine sapwood tracheid cells.

The self-diffusion coefficients of water and toluene in Scots pine sapwood was measured using low field pulsed field gradient nuclear magnetic resonance (PFG-NMR). Wood chips of 8 mm diameter were saturated with the respective liquids, and liquid self-diffusion was then traced in one dimension orthogonal to the tracheid cell walls in the wood's radial direction. The experimental echo attenuation curves were exponential, and characteristic self-diffusion coefficients were produced for diffusion times spanning from very short times to times on the order of magnitude of seconds. Observed self-diffusion coefficients were decaying asymptotically as a function of diffusion time, an effect which was ascribed to the cell walls' restriction on confined liquid diffusion. The observed self-diffusion behavior in Scots pine sapwood was compared to self-diffusion coefficients obtained from simulations of diffusion in a square. Principles of molecular displacements in confined geometries were used for elucidating the wood's cellular structure from the observed diffusion coefficients. The results were compared with a mathematical model for diffusion between parallel planes.

Cell Wall↗

Expression analysis of a cytosolic glutamine synthetase gene in cotyledons of Scots pine seedlings: developmental, light regulation and spatial distribution of specific transcripts.

The expression of a cytosolic glutamine synthetase (GS1; EC 6.3.1.2) gene was examined in cotyledons of Scots pine seedlings. Light strongly stimulated GS1 mRNA accumulation during development. Similarly, steady-state levels of GS1 transcripts increased in dark-grown seedlings transferred to light and decreased in dark-adapted seedlings. Light/dark adaptation affected rbcS and lhcb2 mRNA levels and chlorophyll contents in the same manner. Light-grown seedlings in the presence of the herbicide norflurazon showed a drastic decrease in mRNA for GS and photosynthetic proteins, whereas the effect of the herbicide on mitochondrial beta-ATP synthase mRNA was limited. These results indicate that factors associated with developing chloroplasts could be required for maximal GS1 gene expression during seedling development. The level of GS polypeptide, determined by immunoblot, was up-regulated during seedling development in the light or dark. However, the levels of the polypeptide detected were unaltered by the light/dark adaptation treatments. The analysis of GS1 mRNA association with polysomes indicated that the discrepancies between GS protein and mRNA levels are not a result of a differential translational rate of the transcript in darkness relative to light. Two GS isoproteins with different isoelectric point were resolved by two-dimensional PAGE in light- and dark-germinated plants. The relative abundance of one of them was markedly affected by light and correlated with the observed changes in GS mRNA, suggesting that the other form is not a product derived from the detected transcript. In situ hybridization of cotyledon sections showed the presence of GS1 mRNAs in mesophyll and phloem cells confirming gene expression in photosynthetic tissues. High levels of transcript were detected also in meristematic cells of apical primordia. These data suggest a dual role for the GS1 gene associated with chloroplast development/activity and glutamine biosynthesis for nitrogen mobilization during early growth of Scots pine.

Chloroplasts↗

The production of transgenic Scots pine (Pinus sylvestris L.) via the application of transformed pollen in controlled crossings.

The study demonstrates the production of a transgenic Scots pine (Pinus sylvestris L.) seedling through the application of transformed pollen in controlled crossings. The pollen lots were transformed by particle bombardment, resulting in transient transformation frequencies varying from 15 to 49% of the germinated pollen grains, and bombarded pollen was used to pollinate megasporangiate strobili. Progeny was screened by histochemical, GUS assays, and selected seedlings were further analysed by PCR. PCR amplification revealed the presence of both the nptII and gusA genes in one seedling (23/237). Results were confirmed by Southern blot analysis. The morphology and growth of this transgenic seedling was normal. Although the transformation frequency of recovered plants was very low (1/14999), the present protocol suggests that production of transgenic Scots pine is possible without the use of any tissue culture methods or the involvement of marker genes, for selection of transformants.

Crosses, Genetic↗

Defoliation-induced responses in peroxidases, phenolics, and polyamines in scots pine (Pinus sylvestris L.) needles.

Effects of artificial defoliation on defensive needle chemistry in Scots pine (Pitus sylvestris L.) were evaluated with particular emphasis on peroxidases, phenolic compounds, soluble sugars, polyamines, and foliar nitrogen levels. The study was carried out on a nutrient-poor Scots pine stand with 8- to 25-year-old trees. Defoliation treatment consisted of repeated defoliation in two successive years with respective control trees. Defoliation was done before needle flushing by removing all mature needles. Guaiacol peroxidase activity increased in the needles after the first defoliation. The difference between treatments diminished towards autumn, and disappeared before the second defoliation in the next summer. After the second defoliation, the activities showed a similar trend. Apparently, peroxidases are involved in inducible chemical changes and recovery reactions that occur in the intact needles shortly after defoliation. After the second defoliation, total nitrogen concentration in the current year needles was about 20% lower, and free putrescine (a polyamine) concentration was 40% lower in the defoliated trees than in control needles. These changes indicate a loss of nitrogen due to defoliation. Specific phenolic compounds such as quercitrin, (+)-catechin, and two catechin derivatives increased in current year needles in response to defoliation. Accumulation of starch and sucrose in the current year needles of repeatedly defoliated trees may imply decreased assimilate transport. The results are indicative that changes in needle phytochemistry in response to defoliation accompany changes in needle nitrogen metabolism.

Adaptation, Physiological↗

Genetic variation within and between populations of Pinus sylvestris L. (Scots pine) for susceptibility to Melampsora pinitorqua Rostr. (pine twist rust).

The genetic variability of Scots pine (Pinus sylvestris L.) for twist rust susceptibility and the consequences for genetic improvement of the species were evaluated after artificial inoculation of 1-year-old seedlings in greenhouse tests. Wind-pollinated progenies, factorial and incomplete diallel mating designs were used to compare two natural populations of Scots pine (Haguenau, France, and Taborz, Poland) and their hybrids (Haguenau x Taborz). Families from Taborz were significantly (P < 0.001) less susceptible to twist rust than those from Haguenau. Inter-population hybrids were as susceptible as the Haguenau population. No clear relationship between severity of infection, shoot length and phenological stage at time of inoculation was observed in intra- and interpopulation crosses. Differences observed between the two natural populations for their response to twist rust are suggested to be the consequence of local adaptation simultaneously for climatic conditions and pathogen pressure. Within each of the two populations, the effects of general combining abilities (GCA) for rust susceptibility were predominant, although estimation of individual heritabilities remained at a moderate level (0.30-0.54). In interpopulation crosses, GCA effects of Haguenau and Taborz populations were still predominant. Moreover, hybrid performance could be predicted with confidence based on the intrapopulation GCA values of parents. Genetic gain on twist rust resistance could be achieved quickly through intrapopulation mass or backward selection based on wind-pollinated progeny tests.

Basidiomycota↗

Tolerance and biodegradation of m-toluate by Scots pine, a mycorrhizal fungus and fluorescent pseudomonads individually and under associative conditions.

The tolerance to, and degradation of m-toluate by Scots pine (Pinus sylvestris), a symbiotic mycorrhizal fungus (Suillus bovinus) and Pseudomonas fluorescens strains, with or without m-toluate-degrading capacity, was determined individually and in all symbiotic/associative plant-microbe combinations. Fungal survival on medium with m-toluate was increased in co-culture with the degradative bacterial strains on agar plates (up to 0.02%, w/v). When fungi were grown in mycorrhizal association with Scots pine seedlings in test-tube microcosms containing expanded clay pellets and growth media, the fungus was able to withstand m-toluate concentrations up to 2.0%, w/v in all treatments. The seedling tolerance remained unaltered regardless of the presence or absence of mycorrhizal fungi or biodegradative bacteria. Reduction in m-toluate levels was only detected in treatments inoculated with bacterial strains harbouring TOL catabolic plasmids. The plant and fungus, alone or in mycorrhizal symbiosis, were unable to cleave m-toluate. The presence of easily available plant-derived carbon sources did not impede m-toluate degradation by the bacteria in the mycorrhizosphere.

Basidiomycota↗

Identification of Salmonella typhi genes expressed within macrophages by selective capture of transcribed sequences (SCOTS).

Salmonella enterica serovar Typhi (S. typhi) is a human-restricted pathogen which causes typhoid fever. Relatively little is known about S. typhi host interaction as animal models of this disease are severely limited by the lack of virulence of S. typhi in other hosts. The virulence of other Salmonella serovars in animal models is dependent on the abilities of these bacteria to survive within host macrophages. We have used selective capture of transcribed sequences (SCOTS) to identify S. typhi genes expressed during growth in human macrophages. This positive cDNA selection technique identified 28 distinct clones representing previously identified as well as novel, uncharacterized and hypothetical gene sequences that are expressed intracellularly. Transcripts for the Vi capsular antigen and genes whose products are involved in stress responses and nutrient acquisition were obtained from intracellular bacteria using SCOTS. Most of these clones are present in the S. typhimurium genome and are also expressed in murine macrophages. Nineteen of these gene sequences were disrupted insertionally in S. typhi, and most of the resulting mutants exhibited a lower level of survival within macrophages compared with the wild-type parent strain. Mutant strains, transformed with a copy of a wild-type gene, exhibited a macrophage survival level similar to that of the parental strain.

Bacterial Proteins↗

Diversity of fungi in organic soils under a moorland--Scots pine (Pinus sylvestris L.) gradient.

The conservation and regeneration of native Scots pine (Pinus sylvestris L.) woodlands is being actively encouraged by conservation agencies in the UK because of their high biodiversity value. In the present study, the consequences of regeneration on terrestrial fungal communities was determined in three parallel transects running from open moorland, through an intermediate zone showing seedling colonization, into a mature Scots pine forest at Abernethy Forest, Cairngorm, Scotland. Soil cores were taken at 18 m intervals along each 180 m transect, and the diversity of the soil fungal community was investigated by DGGE and sequence analysis of ITS fragments PCR-amplified from DNA extracted from soil. Analysis of DGGE profiles generated for two of the three transects indicates a clear shift in the community from the moorland region of the transects to the forest region. Whereas a few bands were present at all sampling points across the transects, the majority of bands were unique to either the moorland or forest samples. FASTA database searches of ITS sequence data generated from excised DGGE bands revealed the closest species match for each band. In some cases, the similarity of ITS sequences to database sequences was poor, but the remaining sequences were most closely related to ITS sequences of both mycorrhizal and non-mycorrhizal fungi. The data are discussed in relation to the effect of native pine woodland expansion on the soil fungal community.

Biodiversity↗

Seasonal and annual stem respiration of Scots pine trees under boreal conditions.

BACKGROUND AND AIMS: Stem respiration of trees is a major, but poorly assessed component of the carbon balance of forests, and important for geo-chemistry. Measurements are required under naturally changing seasonal conditions in different years. Therefore, intra- and inter-annual carbon fluxes of stems in forests were measured continuously from April to November in three consecutive years. METHODS: Stem respiratory CO2 fluxes of 50-year-old Scots pine (Pinus sylvestris) trees were continuously measured with a CO2 analyser, and, concomitantly, stem circumference, stem and air temperature and other environmental factors and photosynthesis, were also measured automatically. KEY RESULTS: There were diurnal, seasonal and inter-annual changes in stem respiration, which peaked at 1600 h during the day and was highest in July. The temperature coefficient of stem respiration (Q10) was greater during the growing season than when growth was slow or had stopped, and more sensitive to temperature in the growing season. The annual Q10 remained relatively constant at about 2 over the three years, while respiration at a reference temperature of 15 degrees C (R15) was higher in the growing than in the non-growing season (1.09 compared with 0.78 micromol m(-2) stem surface s(-1)), but was similar between the years. Maintenance respiration was 76 %, 82 % and 80 % of the total respiration of 17.46, 17.26 and 19.35 mol m2 stem surface in 2001, 2002 and 2003, respectively. The annual total stem respiration of the stand per unit ground area was 75.97 gC m(-2) in 2001 and 74.28 gC m(-2) in 2002. CONCLUSIONS: Stem respiration is an important component in the annual carbon balance of a Scots pine stand, contributing 9 % to total carbon loss from the ecosystem and consuming about 8 % of the carbon of the ecosystem gross primary production. Stem (or air) temperature was the most important predictor of stem carbon flux. The magnitude of stem respiration is modified by photosynthesis and tree growth. Solar radiation indirectly affects stem respiration through its effect on photosynthesis.

Carbon↗

Free amino acids and protein in Scots pine seedlings cultivated at different nutrient availabilities.

Scots pine (Pinus sylvestris L.) seedlings of a provenance from northern Sweden were cultivated hydroponically for 7 weeks in a climate chamber. The nutrient solution contained either 2.5 (low-N) or 50 (high-N) mg N l(-1) with other essential elements added in a fixed optimal proportion to the nitrogen. After 5 and 7 weeks, the seedlings were analyzed for growth, total nitrogen and other essential nutrients, protein and free amino acids. Low-N seedlings grew more slowly and had higher root/shoot ratios than high-N seedlings. With respect to total nitrogen, the effect of the lower nutrient supply was mainly on the nitrogen content of the whole plant and the allocation of nitrogen among tissues, not on tissue nitrogen concentration. This was also the case for potassium, phosphorus, calcium and magnesium. The proportions by weight among these macronutrients in the whole seedlings were similar in both nutrient regimes. The proportion and concentration of sulfur were significantly lower in low-N seedlings than in high-N seedlings, because of a lower net uptake of sulfur than of other macronutrients. The shoot, needles and stem of low-N seedlings had higher concentrations of free amino acids and lower concentrations of protein than the shoot, needles and stem of high-N seedlings. Arginine dominated the pool of free amino acids in the low-N seedlings, whereas glutamine predominated in the high-N seedlings. We conclude that Scots pine seedlings accumulated soluble nitrogen as arginine when net protein synthesis was limited by factors other than nitrogen availability. Nutritional imbalance, as revealed by growth characteristics and a suboptimal proportion and concentration of sulfur in the seedlings, probably affected synthesis of S-amino acids, resulting in the diversion of assimilated nitrogen to arginine instead of protein.

Journal Article↗

Performance of the LAI-2000 plant canopy analyzer in estimating leaf area index of some Scots pine stands.

The LAI-2000 plant canopy analyzer (Li-Cor, Inc., Lincoln, NE) was tested at six experimental plots of Scots pine (Pinus sylvestris L.) in central Sweden at peak leaf area in August and after litterfall in October 1990. An independent estimate of leaf area index for August 1990 was obtained based on an empirically derived regression of needle area on stem sapwood area, and the decrease in leaf area between the two measurements was estimated from measurements of litterfall. A strong linear relationship was found between estimates by the LAI-2000 (L(Li-Cor)) and the indirect estimates of leaf area index (taken as half of total surface area) (L). The finding that L(Li-Cor) was considerably smaller than L was explained theoretically. It was shown that if shoots, instead of individual needles, are randomly distributed in the canopy, L(Li-Cor) corresponds to L multiplied by a factor (beta) characterizing the mutual shading of needles on the shoot. The shading factor, beta, was equal to the ratio of spherically projected shoot area to spherically projected needle area, where the spherically projected area is defined as the average projection (silhouette) area taken over all directions in space. The quantity betaL was defined as the shoot silhouette area index (SSAI), and an equation for the relationship between SSAI and the mean silhouette to total area ratio (mean STAR) of shoots was derived. Measured values of mean STAR for Scots pine indicated that L(Li-Cor) corresponds to SSAI rather than L. However, the decrease in leaf area index due to litterfall occurring between August and October was only partly detected by the LAI-2000, possibly because SSAI did not change to the same degree as L, i.e., there was an increase in the factor beta. This hypothesis is supported by data showing a large increase in mean STAR with shoot age.

Journal Article↗

Climate influences the leaf area/sapwood area ratio in Scots pine.

We tested the hypothesis that the leaf area/sapwood area ratio in Scots pine (Pinus sylvestris L.) is influenced by site differences in water vapor pressure deficit of the air (D). Two stands of the same provenance were selected, one in western Scotland and one in eastern England, so that effects resulting from age, genetic variability, density and fertility were minimized. Compared with the Scots pine trees at the cooler and wetter site in Scotland, the trees at the warmer and drier site in England produced less leaf area per unit of conducting sapwood area both at a stem height of 1.3 m and at the base of the live crown, whereas stem permeability was similar at both sites. Also, trees at the drier site had less leaf area per unit branch cross-sectional area at the branch base than trees at the wetter site. For each site, the average values for leaf area, sapwood area and permeability were used, together with values of transpiration rates at different D, to calculate average stem water potential gradients. Changes in the leaf area/sapwood area ratio acted to maintain a similar water potential gradient in the stems of trees at both sites despite climatic differences between the sites.

Journal Article↗

Seasonal changes in the transient expression of a 35S CaMV-GUS gene construct introduced into Scots pine buds.

Seasonal changes in the transient expression of Beta-glucuronidase gene (GUS) driven by a constitutive 35S CaMV-promoter in Scots pine (Pinus sylvestris L.) buds were studied by the microprojectile DNA-delivery method. Buds were collected from 5-, 15- and 50-year-old trees. In buds from all age groups the amount of transient expression was dependent on the season; the highest values were found in March, and values were lowest both at the beginning and at the end of the growing season. Pretreatment with growth regulators increased both the amount of transient GUS expression and arginine decarboxylase (ADC) activity in buds indicating an increase in metabolic activity. These results confirm that the genetic transformation technique can be used to study seasonally dependent regulation in mature Scots pine tissues.

Journal Article↗

Photosynthetic responses to needle water potentials in Scots pine after a four-year exposure to elevated CO(2) and temperature.

Effects of needle water potential (Psi(l)) on gas exchange of Scots pine (Pinus sylvestris L.) grown for 4 years in open-top chambers with elevated temperature (ET), elevated CO(2) (EC) or a combination of elevated temperature and CO(2) (EC + ET) were examined at a high photon flux density (PPFD), saturated leaf to air water vapor pressure deficit (VPD) and optimal temperature (T). We used the Farquhar model of photosynthesis to estimate the separate effects of Psi(l) and the treatments on maximum carboxylation efficiency (V(c,max)), ribulose-1,5-bisphosphate regeneration capacity (J), rate of respiration in the light (R(d)), intercellular partial pressure of CO(2) (C(i)) and stomatal conductance (G(s)). Depression of CO(2) assimilation rate at low Psi(l) was the result of both stomatal and non-stomatal limitations on photosynthetic processes; however, stomatal limitations dominated during short-term water stress (Psi(l) < -1.2 MPa), whereas non-stomatal limitations dominated during severe water stress. Among the nonstomatal components, the decrease in J contributed more to the decline in photosynthesis than the decrease in V(c,max). Long-term elevation of CO(2) and temperature led to differences in the maximum values of the parameters, the threshold values of Psi(l) and the sensitivity of the parameters to decreasing Psi(l). The CO(2) treatment decreased the maximum values of V(c,max), J and R(d) but significantly increased the sensitivity of V(c,max), J and R(d) to decreasing Psi(l) (P < 0.05). The effects of the ET and EC + ET treatments on V(c,max), J and R(d) were opposite to the effects of the EC treatment on these parameters. The values of G(s), which were measured simultaneously with maximum net rate of assimilation (A(max)), declined in a curvilinear fashion as Psi(l) decreased. Both the EC + ET and ET treatments significantly decreased the sensitivity of G(s) to decreasing Psi(l). We conclude that, in the future, acclimation to increased atmospheric CO(2) and temperature could increase the tolerance of Scots pine to water stress.

Journal Article↗