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Cambial history of Scots pine trees (Pinus Sylvestris) prior and after the Chernobyl accident as encoded in the xylem.

Studies were carried out on wood samples collected in October 1997 from breast height of Scots pine trees (Pinus sylvestris) from site located 5 km south from the Chernobyl nuclear power plant. The radioactive contamination at the site was 3.7x10(5) kBq m(-2). These samples of secondary wood were used as an archive of information about the dynamics of a meristematic tissue cambium affected by ionising radiation from the Chernobyl reactor accident. The results show that frequency of the cambial cells events like anticlinal divisions, intrusive growth and cells elimination, was after the Chernobyl accident, about three times higher in comparison to preceding years. The most interesting finding was that after irradiation the length of tracheids increased. This increase is interpreted as an effect of intracambial competition among cells in the initial layer.

Journal Article↗

Gabaculine does not inhibit cytokinin-stimulated biosynthesis of chlorophyll in Pinus nigra seedlings in the dark.

Chlorophyll (Chl) accumulation was monitored during black pine (Pinus nigra L.) seed germination for 14 days in the light and in the dark in the presence of gabaculine (GAB) and cytokinin in order to elucidate the regulation of gymnosperm seedling greening in the dark, primarily at the level of aminolevulinic acid formation. In the light, GAB inhibited chlorophyll accumulation in a manner dependent on concentration and developmental stage, and in the dark it showed no effect. Cytokinin, 10(-5) M benzyl adenine (BA) partly overcame GAB-induced inhibition in the light, mainly during earlier developmental stages. In the seedlings grown in the dark, an equal quantity of Chl accumulated in the presence of cytokinin with and without GAB and it was approximately 20-40% higher than in the control seedlings or in the seedlings grown only in the presence of GAB. 5-Amino-levulinic acid (ALA) synthesis was equal in the light and in the dark in seedlings of the same age and seedlings treated with GAB grown in the dark. In the light, GAB inhibited ALA synthetic activity. The results indicate that ALA synthesis is not a rate-limiting step within Chl biosynthesis in pine seedlings grown in the dark.

Journal Article↗

Role of climate, crown position, tree age and altitude in calculated ozone flux into needles of Picea abies and Pinus cembra: a synthesis.

Ozone (O(3)) flux into Norway spruce (Picea abies) and cembran pine (Pinus cembra) needles was estimated under ambient conditions at six rural sites between 580 and 1950 m a.s.l. We also assessed age-related differences in O(3) flux by examining changes in leaf conductance across the life span of Norway spruce. At the leaf level O(3) flux into the needles was effectively controlled by stomatal conductance and, hence by factors such as temperature, irradiance and humidity, which control stomatal conductance. Seasonal variations in O(3) flux were mainly attributed to the course of the prevailing temperature. During the growing season, however, data have emphasised leaf-air vapour pressure difference as the environmental factor most likely to control stomatal conductance and O(3) flux into the needles. In the sun crown stomatal conductance averaged over the growing season decreased with increasing tree age from 42.0+/-3.5 mmol O(3) m(-2) s(-1) in 17-year-old trees to 7.1+/-1.0 mmol O(3) m(-2) s(-1) in 216-year-old trees, indicating that O(3) concentration in the substomatal cavities is higher in young than in old trees. Independent from tree age stomatal conductance and O(3) flux were approximately 50% lower in shade needles as compared to sun-exposed needles. Stomatal conductance was also greater in the current flush (24+/-5.6 mmol O(3) m(-2) s(-1)) and in 1-year old needles (16+/-4 mmol O(3) m(-2) s(-1)) than in older needle age classes (12+/-1 mmol O(3) m(-2) s(-1), averaged across the four older needle age classes). In trees similar in age (60-65 years old) average O(3) flux into sun needles increased from 0.55+/-0.36 nmol m(-2) s(-1) at the valley floor to 0.9 nmol m(-2) s(-1) in 1950 m a.s.l. Cumulative O(3) uptake during the vegetation period increased from 11.4+/-1.7 mol m(-2) in the valley to 14 mol m(-2) at the alpine timberline. Although stomatal conductance provides the principal limiting factor for O(3) flux, additional field research is necessary in order to improve our understanding concerning the quantitative 'physiological threshold dose' which internally can be active and can have adverse effects of O(3) on forest trees.

Journal Article↗

Impact of increased springtime O3 exposure on Scots pine (Pinus sylvestris) seedlings in central Finland.

Three-year-old Scots pine (Pinus sylvestris L.) seedlings were exposed to ambient or elevated ozone (O(3)) concentrations in open-air exposure fields in central Finland in 1995-97. Three different treatments were applied in 1996 and 1997: ambient air, elevated O(3) (1.3-1.5xambient) during the growing season (June-September) and elevated O(3) in March-September, i.e. the growing season including the springtime O(3) exposure. The ambient mean O(3) concentrations were 40% higher in springtime (March-May) compared to the concentrations during the growing seasons. Maximum O(3) concentrations were measured in April or early May, whereas a clear increase in the stomatal activity of the seedlings was observed by the middle of May. This suggests a low intake of O(3) by conifers despite the higher O(3) concentrations in spring. Stomatal conductance, and contents of chlorophyll and ribulosebisphosphate carboxylase/oxygenase (Rubisco) in current-year needles were not significantly affected by any O(3) treatment. Only a slight decrease in current-year shoot growth, slight increase in the abscission of 2-year-old needles and increased electron density of chloroplast stroma by springtime O(3) exposure suggest a rather small contribution of elevated springtime O(3) concentrations to total O(3) damage under current climatic conditions in Finland. However, the increases in springtime O(3) concentrations may enhance the cumulative effects of O(3) during long-term O(3) exposures.

Journal Article↗

Measurement of ethylene emission from Japanese red pine (Pinus densiflora) under field conditions in NOx-polluted areas.

Emission of ethylene from the needles of Japanese red pine, Pinus densiflora, was measured in air-polluted areas in Hiroshima, Japan. We applied a suitable protocol to determine the rate of ethylene emission from the excised needles. The influence of excision of needles on ethylene emission was not detected during the first 4 h of incubation at 20 degrees C. Ethylene emissions were low in the unpolluted (clean) areas regardless of the altitude or season. The emission of stress ethylene increased with the atmospheric NO2 concentration, suggesting that atmospheric NOx or related substances induced the higher ethylene emission in the polluted areas (near urban and industrial areas). In all cases, 1-year-old needles emitted significantly larger amounts of ethylene than the current needles. Ethylene emission did not increase evenly in the polluted areas, but the frequency of trees emitting high ethylene increased. Therefore, threshold rates for the baseline ethylene emission were proposed.

Air Pollutants↗

Anthropogenic impacts on natural nitrogen isotope variations in Pinus sylvestris stands in an industrially polluted area.

Natural variations of the nitrogen isotopes 15N/14N (delta15N values) and the N concentrations of one-year-old needles from 7-12-year-old pine trees (Pinus sylvestris L.) were determined on 27 sites in the heavily polluted Leipzig-Halle region (former GDR). At three selected sites measurements were repeated over a period of 2 years. N concentrations and delta15N values in different needle age classes were compared at the three sites. The delta15N values of the N in the humus layer and the potential plant available N in the A(h) horizon of the local soil were determined. The 15N/14N isotope ratios (delta15N values) of one-year-old pine needles in the region of Leipzig-Halle were found to vary depending on their specific location by a factor of up to one order of magnitude (-9.6 per thousand to + 0.4 per thousand ). N concentrations in one-year-old pine needles varied between 0.71 and 1.38 mmol eq N g dw(-1). Pine stands with positive or slightly negative delta15N values and high N concentrations in one-year-old needles were concentrated around the cities of Leipzig and Halle and in the industrial areas. More negative delta15N values and lower N concentrations in one-year-old pine needles were found on sites at greater distances from the industrial agglomerations, mainly in the NE forested part. Site specific differences in the delta15N values of the N in the humus layer from three selected sites were similar to those found for the needles. No site specific differences, however, were found for the delta15N values of the water soluble nitrogen fraction from the mineral soil horizons of the same sites.

Journal Article↗

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

Maps of the distribution of environmental pollution by sulfur (S), zinc (Zn), cadmium (Cd), lead (Pb), copper (Cu), and arsenic (As) for the territory of Poland and the Warsaw (Warszawa) district were developed on the basis of chemical analysis of Scots pine (Pinus sylvestris L.) needles collected from randomly selected sampling points during 1983-1985. The maps show deposition zones for the studied elements and can help in identification of sources and directions of air pollution dispersion. This study indicated that vegetation in Poland is greatly endangered by sulfur dioxide (SO(2)) and other sulfurous air pollutants, whereas Zn, Cd, Pb, and As do not pose an immediate threat to vegetation in most of the country's territory. However, in the urban-industrial agglomeration of Katowice-Cracow, very high pollution with Z, Cd, Pb and As could limit growth and development of some sensitive plant species. Higher than normal levels of As in some areas of Poland (Upper Silesia, Glogow-Lubin Copper Region, and areas close to the Russian border near Braniewo) might affect the health of humans and animals. Results of this study indicated that Poland's environment was not contaminated with Cu.

Journal Article↗

Inter-population differences in the essential oils of Pinus caribaea needles.

Differences in volatile terpene content of Pinus caribaea needles and soil qualities between two pine plantations (Uverito and Sartenejas) in Venezuela were analyzed. Soils in the Uverito pine plantation were sandy, low in nutritional quality, and deficient in available nitrogen and phosphorus content. Pines grown on these soils presented in their needles higher concentrations of alpha-pinene, camphene, sabinene, myrcene+alpha-phellandrene mixture, beta-phellandrene, beta-caryophyllene, alpha-humulene, and a higher total monoterpene content than pines of the Sartenejas plantation, where soils were nutritionally richer and higher in nitrogen and phosphorus content. The hypothesis of the carbon/nutrient balance could explain these results. Alternatively, continuous stress on the pines of the Uverito plantation due to herbivory by the leafcutter ant Atta laevigata may induce, in part, the differences observed between these pines and those of the Sartenejas plantation.

Journal Article↗

Ferulic acid excretion as a marker of consumption of a French maritime pine (Pinus maritima) bark extract.

French maritime pine (Pinus maritima) bark extract (PBE) is a polyphenol-rich food supplement patented under the name of Pycnogenol and known to have strong antioxidant activity and different beneficial effects on human health. Although its biological properties have begun to be extensively studied both in vitro, in laboratory animals and more recently in humans, little is known about its bioavailability. The present study investigated the urinary excretion of free and conjugated ferulic acid, present in quantitatively detectable amounts in PBE, after oral PBE administration to human subjects. Eleven healthy adult subjects (4 women and 7men) consumed either a single dose (200 mg PBE) or two doses of PBE (100 and 200 mg, respectively) within a 48-h interval. Two days before the oral administration of PBE and during the urine sample collection period volunteers adhered to a diet low in polyphenols. Aliquots of all urine production were collected over 24 h. Free and conjugated ferulic acid was assessed in urine by HPLC using diode array detection. A close association between the dietary intake of PBE and the urinary excretion of ferulic acid was detected. Moreover, the results indicate that a considerable proportion of ferulic acid is excreted as glucuronide or sulfate after PBE consumption, varying over the range 2 to 20% between individuals. The kinetics of excretion associated with the administration of 100 mg PBE was quite similar to that obtained after 200 mg PBE. A a biphasic trend was evident in a number of subjects. All subjects studied here displayed a significant, although variable level of excretion of ferulic acid after supplementation with PBE, Thus, the data provide evidence that at least a part of the phenolic components of PBE are absorbed, metabolized, and eliminated by humans.

Administration, Oral↗

Procyanidins extracted from Pinus maritima (Pycnogenol): scavengers of free radical species and modulators of nitrogen monoxide metabolism in activated murine RAW 264.7 macrophages.

Nitrogen monoxide (NO) has diverse physiological roles and also contributes to the immune defense against viruses, bacteria, and other parasites. However, excess production of NO is associated with various diseases such arthritis, diabetes, stroke, septic shock, autoimmune, chronic inflammatory diseases, and atheriosclerosis. Cells respond to activating or depressing stimuli by enhancing or inhibiting the expression of the enzymatic machinery that produce NO. Thus, maintenance of a tight regulation of NO production is important for human health. Phytochemicals have been traditionally utilized in ways to treat a family of pathologies that have in common the disregulation of NO production. Here we report the scavenging activity of Pycnogenol (the polyphenols containing extract of the bark from Pinus maritima) against reactive oxygen and nitrogen species, and its effects on NO metabolism in the murine macrophages cell line RAW 264.7. Macrophages were activated by the bacterial wall components lipopolysaccharide (LPS) and interferon (IFN-gamma), which induces the expression of large amounts of the enzyme nitric oxide synthase (iNOS). Preincubation of cells with physiological concentrations of Pycnogenol significantly decreased NO generation. It was found that this effect was due to the combination of several different biological activities, i.e., its ROS and NO scavenging activity, inhibition of iNOS activity, and inhibition of iNOS-mRNA expression. These data begin to provide the basis for the conceptual understanding of the biological activity of Pycnogenol and possibly other polyphenolic compounds as therapeutic agents in various human disorders.

Animals↗

Antioxidant activity and biologic properties of a procyanidin-rich extract from pine (Pinus maritima) bark, pycnogenol.

There is growing interest in the biologic activities of plant extracts such as that obtained from the bark of the French maritime pine Pinus maritima, Pycnogenol. Pycnogenol (PYC) is a standardized extract composed of a mixture of flavonoids, mainly procyandins and phenolic acids. Studies indicate that PYC components are highly bioavailable. Uniquely PYC displays greater biologic effects as a mixture than its purified components do individually indicating that the components interact synergistically. PYC has been reported to have cardiovascular benefits, such as a vasorelaxant activity, angiotensin-converting enzyme (ACE) inhibiting activity, and the ability to enhance the microcirculation by increasing capillary permeability. Investigations of the cellular mechanisms of these therapeutic effects have demonstrated that PYC has strong free radical-scavenging activity against reactive oxygen and nitrogen species. The oligomeric components of PYC contribute significantly to the ESR free radical signal. PYC also participates in the cellular antioxidant network as indicated by its ability to regenerate the ascorbyl radical and to protect endogenous vitamin E and glutathione from oxidative stress. PYC modulates NO metabolism in activated macrophages by quenching the NO radical and inhibiting both iNOS mRNA expression and iNOS activity. The spectrum of different effects of NO in the circulation and the nervous system suggest the potential applications of PYC in immune and circulatory disorders as well as in neurodegenerative disease. PYC can bind to proteins, altering their structure and thereby modulating the activity of key enzymes and proteins involved in metabolic pathways. PYC effects redox-sensitive signal transduction pathways and alters gene expression. Aspects of PYC's activity are presented and discussed together with possible future implications and directions in the field of flavonoid research.

Antioxidants↗

Allometry and biomass of Korean pine (Pinus koraiensis) in central Korea.

Aboveground tree biomass of Korean pine (Pinus koraiensis Sieb. et Zucc.) was determined for a natural forest of Korean pine and mixed deciduous trees and seven age classes of plantation forests in central Korea. Regression analyses of the dry weights of stem wood, stem bark, branches, and needles versus diameter at breast height were used to calculate regression equations of the form of log Y = a + b log X. Biomass of Korean pine in the mixed forest was 118 Mg ha(-1), and biomass in the plantations was linearly related to stand age, ranging from 52.3 Mg ha(-1) in 11 to 20-year-old stands to 317.9 Mg ha(-1) in 71 to 80-year-old stands. The proportions of stem wood and stem bark in the total aboveground biomass decreased with stand age while those of branch and needle increased. Specific leaf area of Korean pine ranging from 35.2 to 52.1 cm2 g(-1) was significantly different among crown positions and needle ages; in general, lower crown position and current needles had the greatest surface area per unit dry weight.

Biomass↗

Preliminary studies on TCF bleaching of Pinus pinaster acetosolv pulps.

Oxygen pre-treatment of Pinus pinaster acetosolv pulps has been studied as a first step towards TCF bleaching. Using a 2(3) factorial design, the influence of temperature (80-120 degrees C), time (1-2 h) and NaOH concentration (1.5-3%) on pulp yield in the oxygen stage, chemical composition and physical properties of the pulps obtained was studied. Pulps pre-bleached with oxygen in the conditions selected as optimal (80 degrees C, 1 h with 2.25% NaOH) have been bleached with TCF sequences which included stages with hydrogen peroxide or hydrogen peroxide-oxygen under pressure. Even if high degrees of delignification were reached, with a reduction in Kappa number up to 95% and without important loss of viscosity, the carbohydrates degradation, especially hemicelluloses in the acetic acid delignification, reduces the strength potential of the pulps.

Acetic Acid↗

Chemical composition of essential oils from needles and twigs of balkan pine (Pinus peuce grisebach) grown in Northern Greece.

The composition of essential oils from twigs and needles of Balkan pine (Pinus peuce Gris.) grown in northern Greece was investigated. The compounds were identified by using GC-MS analysis. The twig oil was rich in alpha-pinene (7.38%), beta-pinene (12.46%), beta-phellandrene (26.93%), beta-caryophyllene (4.48%), and citronellol (12.48%), and the needle oil was rich in alpha-pinene (23.07%), camphene (5.52%), beta-pinene (22.00%), beta-phellandrene (6.78%), bornyl acetate (9.76%), beta-caryophyllene (3.05%), and citronellol (13.42%). The mean oil yield was 2.85% for twigs and 0. 57% for needles.

Cycadopsida↗

Four new trisnorlanostene-type triterpenoids from the stem bark of Pinus luchuensis.

Four new 25,26,27-trisnorlanostene-type triterpenoids were isolated from the stem bark of Pinus luchuensis, together with five known compounds. These new compounds were characterized as 25,26, 27-trisnor-3 alpha-hydroxy-lanost-9(11)-en-24-oic acid (1), 25,26, 27-trisnor-3 alpha-methoxy-lanost-9(11)-en-24-oic acid (2), 25,26, 27-trisnor-3 beta-methoxy-lanost-9(11)-en-24-oic acid (3), and 25,26, 27-trisnor-3-oxo-lanost-9(11)-en-24-oic acid (4) on the basis of MS and NMR evidence.

Chromatography, Gel↗

Isolation and structure elucidation of new PKCalpha inhibitors from Pinus flexilis.

Bioassay-guided fractionation of the CH2Cl2-MeOH extract of Pinus flexilis using an assay for protein kinase C (PKC) inhibitory activity led to the isolation of the two new bioactive diarylheptanoids (3R)-1,7-bis(3, 4-dihydroxyphenyl)-3-(beta-D-glucopyranosyl)heptan-3-ol (1) and its aglycon (3R)-1,7-bis(3,4-dihydroxyphenyl)heptan-3-ol (2), together with the three known bioactive compounds, hirsutenone (3), oregonin (4), and hirsutanonol (5). The IC50 values of compounds 1-5 in the PKC assay were 1.4, 1.6, 1.4, 8.6, and 4.6 microg/mL, respectively.

Circular Dichroism↗

Characterization of a germin-like protein gene expressed in somatic and zygotic embryos of pine (Pinus caribaea Morelet).

Germin-like proteins (GLPs) ionically bound to the walls of preglobular somatic embryos of Pinus caribaea Morelet are markers of this early developmental stage. In order to reveal the physiological implications of such markers during early embryo development, we isolated a cDNA clone from somatic embryos predicted to encode a protein with sequence similarity to GLPs. PcGER1 has an open reading frame corresponding to a 220 amino acid polypeptide with a putative N-glycosylation site on Asn-69. The presence of a 24 amino acid putative signal peptide supports the hypothesis of an apoplastic location. The N-terminal 20 amino acid sequence of the predicted mature protein is identical to the amino terminal sequence of GP111, one of the extracellular pine GLPs previously identified. Southern blot hybridizations indicate that PcGER1 is probably unique in the pine genome. Transcripts homologous to PcGER1 are abundant in all embryogenic lines, absent from nonembryogenic lines, and present in quiescent zygotic embryos but not in the female gametophyte, the haploid storage tissue of conifers. Their abundance sharply decreases during germination. Isolation of gf-0.8, a genomic fragment identical to PcGER1 cDNA sequence, confirms that no introns disrupt the coding region as it has been already described for wheat gf-2.8 and gf-3.8 genomic clones. Recombinant PcGER1, produced in Escherichia coli, is recognized by antibodies raised against the GP111 N-terminal nonapeptide and the unglycosylated wheat germin monomer. The implications of GLPs in pine embryogenesis are discussed.

Amino Acid Sequence↗