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Effects of temperature and nutrient availability on plasma membrane lipid composition in Scots pine roots during growth initiation.

We studied the effects of root zone temperature (RZT) and nutrient availability on free sterols and phospholipids in the plasma membrane (PM) and on PM-ATPase activity in roots of 1-year-old Scots pine (Pinus sylvestris L.) seedlings during growth initiation in the spring. Seedlings were grown for 6 weeks in hydroponic cultures with low (0.5 mM N; LN) or high (3 mM N; HN) nutrient availability. The root zone was subjected to slow warming (SW) and fast warming (FW) treatments while maintaining similar air temperatures in both treatments. Decreases in the amount of phospholipids and in the phospholipid/free sterol ratio, an increase in the degree of saturation of phospholipid fatty acids and changes in free sterol composition were observed during root growth initiation. Changes in lipid composition of the PM associated with the cold deacclimation process were detected at RZTs above 9 degrees C. Nutrient availability affected the lipid composition of the PM only when RZT was increased slowly. When RZT increased from 4 to 6 degrees C in the SW treatment, the degree of saturation of phospholipid fatty acids decreased, especially in HN seedlings. The sitosterol/stigmasterol ratio remained higher in HN seedlings than in LN seedlings. After an RZT of 9 degrees C had been reached in the SW treatment, HN caused increases in the saturation of phospholipid fatty acids and root PM-ATPase activity, and a decrease in the phospholipid/free sterol ratio. Possible effects of changes in PM lipid composition on root growth and PM-ATPase activity are discussed.

Adenosine Triphosphatases↗

Light sources with different spectra affect root and mycorrhiza formation in Scots pine in vitro.

We studied the effects of broad-spectrum light quality on the interaction between the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch and Scots pine (Pinus sylvestris L.) seedlings and hypocotyl cuttings cultured in vitro. The light sources were cool white (CW), warm white (WW) and red-rich daylight (RD) fluorescent lamps. Inoculation with P. tinctorius enhanced adventitious root formation of the cuttings in all light treatments. Rooting of the inoculated cuttings was highest in WW light (89%), followed by CW (73%) and RD light (66%). During 6 weeks of in vitro culture, rooted cuttings formed only a few lateral roots. The fungus grew over lateral roots, but the Hartig net was absent in all light treatments. In non-inoculated cuttings, neither root formation nor subsequent root growth was affected by light quality. In the seedling experiment, inoculation in the WW treatment resulted in a significantly (P < 0.05) greater number of lateral roots than inoculation in the RD treatment. The percentage of lateral roots covered with fungal hyphae was also highest in WW light (62%), followed by CW (50%) and RD (27%) light. A similar pattern was observed in the intensity of Hartig net formation. We conclude that effects of broad-spectrum light quality on the ectomycorrhizal fungus-root interaction are dependent on the developmental stage of the root.

Basidiomycota↗

Carbon budget for Scots pine trees: effects of size, competition and site fertility on growth allocation and production.

Time series of carbon fluxes in individual Scots pine (Pinus sylvestris L.) trees were constructed based on biomass measurements and information about component-specific turnover and respiration rates. Foliage, branch, stem sapwood, heartwood and bark components of aboveground biomass were measured in 117 trees sampled from 17 stands varying in age, density and site fertility. A subsample of 32 trees was measured for belowground biomass excluding fine roots. Biomass of fine roots was estimated from the results of an earlier study. Statistical models were constructed to predict dry mass (DW) of components from tree height and basal area, and time derivatives of these models were used to estimate biomass increments from height growth and basal area growth. Biomass growth (G) was estimated by adding estimated biomass turnover rates to increments, and gross photosynthetic production (P) was estimated by adding estimated component respiration rates to growth. The method, which predicts the time course of G, P and biomass increment in individual trees as functions of height growth and basal area growth, was applied to eight example trees representing different dominance positions and site fertilities. Estimated G and P of the example trees varied with competition, site fertility and tree height, reaching maximum values of 22 and 43 kg(DW) year(-1), respectively. The site types did not show marked differences in productivity of trees of the same height, although height growth was greater on the fertile site. The G:P ratio decreased with tree height from 65 to 45%. Growth allocation to needles and branches increased with increasing dominance, whereas growth allocation to the stem decreased. Growth allocation to branches decreased and growth allocation to coarse roots increased with increasing tree size. Trees at the poor site allocated 49% more to fine roots than trees at the fertile site. The belowground parts accounted for 25 to 55% of annual G, increasing with tree size and decreasing with site fertility. Annual G and P per unit needle mass varied over the ranges 1.9-2.4 and 3.5-4.0 kg(DW) kg(-1), respectively. The relationship between P and needle mass in the example trees was linear and relatively independent of competition, site fertility and age.

Biomass↗

Respiratory responses of Scots pine stems to 5 years of exposure to elevated CO2 concentration and temperature.

Stem respiration in 20-year-old Scots pine (Pinus sylvestris L.) trees was examined following 5 years of exposure to ambient conditions (CON), elevated atmospheric carbon dioxide concentration ([CO2]) (ambient + 350 micromol mol(-1), (EC)), elevated temperature (ambient + 2-6 degrees C, (ET)) or a combination of elevated [CO2] and elevated temperature (ECT). Stem respiration varied seasonally regardless of the treatment and displayed a similar trend to temperature, with maximum rates occurring around Day 190 in summer and minimum rates in winter. Respiration normalized to 15 degrees C (R15) was higher in the growing season than in the non-growing season, whereas the temperature coefficient (Q10) was lower in the growing season. Annually averaged R15 was 0.36, 0.43, 0.40 and 0.44 micromol m(-2) s(-1) under CON, EC, ET and ECT conditions, respectively, whereas the corresponding values for total stem respiration were 6.55, 7.69, 7.50 and 7.90 mol m(-2) year(-1). The EC, ET and ECT treatments increased R15 by 18, 11 and 22%, respectively, relative to CON, and increased the modeled annual total stem respiration by 18, 15 and 21%. The increase in modeled annual stem respiration under EC and ECT conditions was caused mainly by higher maintenance respiration (22 and 25%, respectively, whereas the increase in growth respiration was 9 and 12%). Growth respiration was unaltered by ET. The treatments did not significantly affect the respiratory response to stem temperature; the mean Q10 value was 2.04, 2.10, 1.99 and 2.12 in the CON, EC, ET and ECT treatments, respectively. It is suggested that the increase in stem respiration was partly a result of the increased growth rate. We conclude that elevated [CO2] increased the maintenance component of respiration more than the growth component.

Carbon Dioxide↗

Scots pine responses to elevated temperature and carbon dioxide concentration: growth and wood properties.

Growth and wood properties of 20-year-old Scots pine (Pinus sylvestris L.) trees were studied for 6 years in 16 closed chambers providing a factorial combination of two temperature regimes (ambient and elevated) and two carbon dioxide concentrations ([CO2]) (ambient and twice ambient). The elevation of temperature corresponded to the predicted effect at the site of a doubling in atmospheric [CO2]. Annual height and radial growth and wood properties were analyzed during 1997-2002. Physical wood properties analyzed included early- and latewood widths and their proportions, intra-ring wood densities, early- and latewood density and mean fiber length. Chemical wood properties analyzed included concentrations of acetone-soluble extractives, lignin, cellulose and hemicellulose. There were no significant treatment effects on height growth during the 6-year study. Elevated [CO2] increased ring width by 66 and 47% at ambient and elevated temperatures, respectively. At ambient [CO2], elevated temperature increased ring width by 19%. Increased ring width in response to elevated [CO2] resulted from increases in both early- and latewood width; however, there was no effect of the treatments on early- and latewood proportions. Mean wood density, earlywood density and fiber length increased in response to elevated temperature. The chemical composition of wood was affected by elevated [CO2], which reduced the cellulose concentration, and by elevated temperature, which reduced the concentration of acetone-soluble extractives. Thus, over the 6-year period, radial growth was significantly increased by elevated [CO2], and some wood properties were significantly affected by elevated temperature or elevated [CO2], or both, indicating that climate change may affect the material properties of wood.

Carbon Dioxide↗

Estimating foliage biomass in Scots pine (Pinus sylvestris) and Norway spruce (Picea abies) plots.

Dynamic decomposition models are needed to estimate changes in the carbon stock of boreal soil because these changes are difficult to measure directly. An important aboveground carbon flux to the soil is foliage litterfall. To estimate this flux, both the amount and the turnover rate of the foliage biomass component must be known. Several methods for estimating foliage biomass of Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) Karst.), including biomass equations and biomass expansion factors (BEFs), were compared with predicted foliage biomass based on forest inventory plot-level measurements. Measured foliage biomass was up-scaled from the branch-level to the plot-level by combining forest inventory variables (diameter, height, height at the crown base and crown base diameter) based on the assumptions of pipe model theory. Combining the foliage biomass: cross-sectional area ratio with the forest inventory variables provided accurate estimates of foliage biomass at the plot-level for plots in southern Finland. The results emphasize the need to test biomass equations with independent data, especially when the equations applied are based on neighboring regions.

Biomass↗

Effects of timing of soil frost thawing on Scots pine.

Effects of the timing of soil thawing in the spring on Scots pine (Pinus sylvestris L.) were studied under controlled laboratory conditions. Sixteen 6-year-old saplings were lifted from the field, replanted in containers and placed in four treatments in controlled environment (CE) chambers with four replicate saplings per chamber. The saplings were held in the CE chambers during one simulated winter and one simulated growing season. The soil was frozen to -2 degrees C during a second simulated winter in the CE chambers, and the soil thawing treatments began at the end of the second simulated winter. Soil thawing began at various times before (no delay in thawing) and after (delay in thawing) chamber air conditions were changed from simulated winter to simulated summer. Delayed soil thawing subjected saplings to stress, with the severity of stress depending on the length of the delay in thawing. If there was no delay or only a short delay in soil thawing, stress was minor and reversible. A 2-week delay in soil thawing led to death of the saplings. Stress was apparent as decreases in the variable to maximal chlorophyll fluorescence ratio (Fv/Fm), chlorophyll a/b ratio and needle water potential. In needles of stressed saplings, apoplastic electrical resistance first decreased and then increased and there were anomalies in the electrical impedance spectra of the stems. Stress from the soil thawing treatments affected both root and shoot growth.

Freezing↗

Plant size, not age, regulates growth and gas exchange in grafted Scots pine trees.

We studied the effect of scion donor-tree age on the physiology and growth of 6- to 7-year-old grafted Scots pine (Pinus sylvestris L.) trees (4 and 5 years after grafting). Physiological measurements included photosynthethetic rate, stomatal conductance, transpiration, whole plant hydraulic conductance, needle nitrogen concentration and carbon isotope composition. Growth measurements included total and component biomasses, relative growth rates and growth efficiency. Scion donor trees ranged in age from 36 to 269 years at the time of grafting. Hydraulic conductance was measured gravimetrically, applying the Ohm's law analogy, and directly, with a high-pressure flow meter. We found no effect of scion donor-tree age on any of the variables measured. There was, however, great variation within scion donor-tree age groups, which was related to the size of the grafted trees. Differences in size may have been caused by variable initial grafting success, but there was no indication that grafting success and age were related. At the stem level, hydraulic conductance scaled with total leaf area so that total conductance per unit leaf area did not vary with crown size. However, leaf specific hydraulic conductance (gravimetric), transpiration, photosynthesis and stomatal conductance declined with increasing total tree leaf area and needle width. We hypothesize that needle width is inversely related to mesophyll conductance. We conclude that canopy and needle size and not scion donor-tree age determined gas exchange in our grafted trees.

Pinus sylvestris↗

Indole 3-acetic acid concentration in the leading shoot and living stem bark of Scots pine: seasonal variation and effects of pruning.

Seasonal changes in the concentration of indole-3-acetic acid (IAA) in the leading shoot and in the live tissue external to the xylem (living bark) of 2-3 m tall Scots pine (Pinus sylvestris L.) trees were observed over a period of three years. Both the leading shoot and the living bark showed a consistent pattern of seasonal variation in IAA content, with maxima of about 150 and 350 ng g(-1) (fresh weight), respectively. The IAA concentration in both tissues increased in late June at the time of shoot growth termination, decreased from late August, and by October reached a low value, which was maintained throughout the winter. Excision of current year shoots on the upper branches decreased the annual stem diameter increment without affecting the concentration of IAA in the living bark. However, excising the leading shoot, or girdling the mainstem beneath the leading shoot, caused a substantial reduction in the IAA content of the living stem bark.

Journal Article↗

Effects of low night temperature and light on chlorophyll fluorescence of field-grown seedlings of Scots pine (Pinus sylvestris L.).

In vivo chlorophyll fluorescence kinetics was observed in naturally regenerated and planted Scots pine (Pinus sylvestris L.) seedlings growing at two adjacent sites in northern Sweden. Some seedlings were shaded from direct solar radiation and some were protected from frost at night. Air temperature and solar radiation were monitored. In vivo chlorophyll fluorescence kinetics of current-year needles was measured at both room temperature and 77 K. On clear days during August, variable fluorescence of photosystem II of needles from exposed, naturally regenerated seedlings was reduced by night frosts, whereas frost caused little change in fluorescence of needles from shaded seedlings. Overnight frost-protection reduced the inhibition of photosystem II by direct sunlight. Recovery from photoinhibitory damage occurred on cloudy days. In September, the decline of variable fluorescence of photosystem II was more pronounced in unshaded than in shaded seedlings, and coincided with frosts at night. In addition, the reduction in variable fluorescence was larger for planted seedlings than for naturally regenerated seedlings. It is suggested that the inhibition of photosystem II activity was caused by an interaction between low temperatures and light. Natural night frosts are proposed to increase the susceptibility to photoinhibition in the following day(s).

Journal Article↗

Nitrate metabolism in Scots pine seedlings during their first growing season.

Fluctuations in nitrate reductase activity (NRA), nitrate, nitrite, protein and total nitrogen content of bare-root Scots pine seedlings (Pinus sylvestris L.) raised outdoors were investigated during the first growing season. Nitrate reductase activity was higher in roots than in needles, whereas NO(3) (-) content was higher in needles than in roots and increased in both from June to October. Nitrate reductase activity in roots correlated more closely with NO(3) (-) N in the soil than did NO(3) (-) in the roots. In autumn, there was a closer correlation between foliar NRA and NO(3) (-) in the needles than with NO(3) (-)-N in the soil. Nitrite was not detected in the seedlings during the growing season. Total nitrogen content decreased toward the autumn, whereas protein content initially decreased but increased again in autumn. Acrylic netting placed above the seedlings increased both air and soil temperatures and apparently accelerated the use of nitrate.

Journal Article↗

Seasonal changes in amino acids, protein and total nitrogen in needles of fertilized Scots pine trees.

Seasonal changes in amino acids, protein and total nitrogen in needles of 30-year-old, fertilized Scots pine (Pinus sylvestris L.) trees growing in Northern Sweden were investigated over two years in field experiments. The studied plots had been fertilized annually for 17 years with (i) a high level of N, (ii) a medium level of N, or (iii) a medium level of N, P and K. Trees growing on unfertilized plots served as controls. In control trees, glutamine, glutamic acid, gamma-aminobutyric acid, aspartic acid and proline represented 50-70% of the total free amino acids determined. Arginine was present only in low concentrations in control trees throughout the year, but it was usually the most abundant amino acid in fertilized trees. Glutamine concentrations were high during the spring and summer in both years of study, whereas proline concentrations were high in the spring but otherwise low throughout the year. In the first year of study, glutamic acid concentrations were high during the spring and summer, whereas gamma-aminobutyric acid was present in high concentrations during the winter months. This pattern was less pronounced in the second year of investigation. The concentrations of most amino acids, except glutamic acid, increased in response to fertilization. Nitrogen fertilization increased the foliar concentration of arginine from < 1 micromol g(dw) (-1) in control trees to a maximum of 110 micromol g(dw) (-1). Trees fertilized with nitrogen, phosphorus and potassium had significantly lower arginine concentrations than trees fertilized with the same amount of nitrogen only. Protein concentrations were similar in all fertilized trees but higher than those in control trees. For all treatments, protein concentrations were high in winter and at a minimum in early spring. In summer, the protein concentration remained almost constant except for a temporary decrease which coincided with the expansion of new shoots. Apart from arginine, the amino acid composition of proteins was similar in all treatments.

Journal Article↗

A case of invoking a Scot's economy or seven-in-a-stroke.

The principle of invoking a Scot's economy or "never throw anything away" is useful for nearly every reconstructive procedure. A case of unilateral brow ptosis and temporal hollowness is presented here whereby the application of this principle allowed a single flap to perform multiple functions in the course of correcting the deformity.

Aged↗

Gene-pool variation in caledonian and European Scots pine (Pinus sylvestris L.) revealed by chloroplast simple-sequence repeats.

We have used polymorphic chloroplast simple-sequence repeats to analyse levels of genetic variation within and between seven native Scottish and eight mainland European populations of Scots pine (Pinus sylvestris L.). Diversity levels for the Scottish populations based on haplotype frequency were far in excess of those previously obtained using monoterpenes and isozymes and confirmed lower levels of genetic variation within the derelict population at Glen Falloch. The diversity levels were higher than those reported in similar studies in other Pinus species. An analysis of molecular variance (AMOVA) showed that small (3.24-8.81%) but significant (p < or = 0.001) portions of the variation existed between the populations and that there was no significant difference between the Scottish and the mainland European populations. Evidence of population substructure was found in the Rannoch population, which exhibited two subgroups. Finally, one of the loci studied exhibited an allele distribution uncharacteristic of the stepwise mutation model of evolution of simple-sequence repeats, and sequencing of the PCR products revealed that this was due to a duplication rather than slippage in the repeat region. An examination of the distribution of this mutation suggests that it may have occurred fairly recently in the Wester Ross region or that it may be evidence of a refugial population.

Alleles↗

Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in Scots pine roots.

To investigate whether Cd induces common plant defense pathways or unspecific necrosis, the temporal sequence of physiological reactions, including hydrogen peroxide (H(2)O(2)) production, changes in ascorbate-glutathione-related antioxidant systems, secondary metabolism (peroxidases, phenolics, and lignification), and developmental changes, was characterized in roots of hydroponically grown Scots pine (Pinus sylvestris) seedlings. Cd (50 microM, 6 h) initially increased superoxide dismutase, inhibited the systems involved in H(2)O(2) removal (glutathione/glutathione reductase, catalase [CAT], and ascorbate peroxidase [APX]), and caused H(2)O(2) accumulation. Elongation of the roots was completely inhibited within 12 h. After 24 h, glutathione reductase activities recovered to control levels; APX and CAT were stimulated by factors of 5.5 and 1.5. Cell death was increased. After 48 h, nonspecific peroxidases and lignification were increased, and APX and CAT activities were decreased. Histochemical analysis showed that soluble phenolics accumulated in the cytosol of Cd-treated roots but lignification was confined to newly formed protoxylem elements, which were found in the region of the root tip that normally constitutes the elongation zone. Roots exposed to 5 microM Cd showed less pronounced responses and only a small decrease in the elongation rate. These results suggest that in cells challenged by Cd at concentrations exceeding the detoxification capacity, H(2)O(2) accumulated because of an imbalance of redox systems. This, in turn, may have triggered the developmental program leading to xylogenesis. In conclusion, Cd did not cause necrotic injury in root tips but appeared to expedite differentiation, thus leading to accelerated aging.

Antioxidants↗

Consistency of polyamine profiles and expression of arginine decarboxylase in mitosis during zygotic embryogenesis of Scots pine.

In this study, we show that both arginine decarboxylase (ADC) protein and mRNA transcript are present at different phases of mitosis in Scots pine (Pinus sylvestris) zygotic embryogenesis. We also examined the consistency of polyamine (PA) profiles with the effective temperature sum, the latter indicating the developmental stage of the embryos. PA metabolism was analyzed by fitting statistical regression models to the data of free and soluble conjugated PAs, to the enzyme activities of ADC and ornithine decarboxylase (ODC), as well as to the gene expression of ADC. According to the fitted models, PAs typically had the tendency to increase at the early stages but decrease at the late stages of embryogenesis. Only the free putrescine fraction remained stable during embryo development. The PA biosynthesis strongly preferred the ADC pathway. Both ADC gene expression and ADC enzyme activity were substantially higher than putative ODC gene expression or ODC enzyme activity, respectively. ADC gene expression and enzyme activity increased during embryogenesis, which suggests the involvement of transcriptional regulation in the expression of ADC. Both ADC mRNA and ADC protein localized in dividing cells of embryo meristems and more specifically within the mitotic spindle apparatus and close to the chromosomes, respectively. The results suggest the essential role of ADC in the mitosis of plant cells.

Carboxy-Lyases↗

Indole-3-acetic acid controls cambial growth in scots pine by positional signaling

The vascular cambium produces secondary xylem and phloem in plants and is responsible for wood formation in forest trees. In this study we used a microscale mass-spectrometry technique coupled with cryosectioning to visualize the radial concentration gradient of endogenous indole-3-acetic acid (IAA) across the cambial meristem and the differentiating derivatives in Scots pine (Pinus sylvestris L.) trees that had different rates of cambial growth. This approach allowed us to investigate the relationship between growth rate and the concentration of endogenous IAA in the dividing cells. We also tested the hypothesis that IAA is a positional signal in xylem development (C. Uggla, T. Moritz, G. Sandberg, B. Sundberg [1996] Proc Natl Acad Sci USA 93: 9282-9286). This idea postulates that the width of the radial concentration gradient of IAA regulates the radial number of dividing cells in the cambial meristem, which is an important component for determining cambial growth rate. The relationship between IAA concentration in the dividing cells and growth rate was poor, although the highest IAA concentration was observed in the fastest-growing cambia. The radial width of the IAA concentration gradient showed a strong correlation with cambial growth rate. The results indicate that IAA gives positional information in plants.

Journal Article↗

Developmental regulation of indole-3-acetic acid turnover in Scots pine seedlings.

Indole-3-acetic acid (IAA) homeostasis was investigated during seed germination and early seedling growth in Scots pine (Pinus sylvestris). IAA-ester conjugates were initially hydrolyzed in the seed to yield a peak of free IAA prior to initiation of root elongation. Developmental regulation of IAA synthesis was observed, with tryptophan-dependent synthesis being initiated around 4 d and tryptophan-independent synthesis occurring around 7 d after imbibition. Induction of catabolism to yield 2-oxindole-3-acetic acid and irreversible conjugation to indole-3-acetyl-N-aspartic acid was noticed at the same time as de novo synthesis was first detected. As a part of the homeostatic regulation IAA was further metabolized to two new conjugates: glucopyranosyl-1-N-indole-3-acetyl-N-aspartic acid and glucopyranosyl-1-N-indole-3-acetic acid. The initial supply of IAA thus originates from stored pools of IAA-ester conjugates, mainly localized in the embryo itself rather than in the general nutrient storage tissue, the megagametophyte. We have found that de novo synthesis is first induced when the stored pool of conjugated IAA is used up and additional hormone is needed for elongation growth. It is interesting that when de novo synthesis is induced, a distinct induction of catabolic events occurs, indicating that the seedling needs mechanisms to balance synthesis rates for the homeostatic regulation of the IAA pool.

Biotransformation↗