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Pycnogenol does not impact the antioxidant or vitamin C status of healthy young adults.

OBJECTIVE: The objectives of this study were to determine if Pycnogenol (PYC), a water-processed extract made from the bark of Pinus maritima, interacts with vitamin C to increase its concentration and to increase total antioxidant capacity of serum and urine. DESIGN: The study design was a nonrandom intervention. SUBJECTS: Subjects (N=27; 15 women, 12 men) were aged 19 to 42 years. INTERVENTION: Subjects consumed a placebo twice daily with meals for the first 2 weeks (baseline) and PYC (200 mg/day) for the second 2 weeks. Main outcome measures On days 15 and 29, subjects had a fasting blood sample collected and then consumed a daily dose of placebo or PYC with a 310-calorie beverage. One hour later a second blood sample was collected. Blood samples were analyzed for vitamin C and total antioxidant capacity using the ORAC (oxygen radical absorbance capacity) assay. Twenty-four-hour urine samples were collected on days 14-15 and 28-29 and analyzed for total phenolics, FRAP (ferric reducing antioxidant potential), and ORAC. Statistical analyses Paired t tests were used to test the impact of PYC on the outcome variables. A univariate ANOVA was used to determine the influence of gender. Pearson's correlation analysis was used to explore the relationships between dietary factors and outcome measures. RESULTS: There was no apparent increase in fasting vitamin C concentration (P=.18) 2 weeks after supplementing the diet with PYC. Fasting ORAC values actually declined (P=.005). One hour after the ingestion of a daily dose of placebo or PYC, the total antioxidant capacity of serum increased by 15% to 19%, but the increase after ingesting PYC was not significantly (P=.80) more than after placebo. Antioxidant results from 24-hour urine samples were similar. APPLICATIONS/CONCLUSIONS: The present findings fail to support the vitamin C or antioxidant claims made for PYC. PYC does not impact the antioxidant or vitamin C status of healthy young adults.

Adult↗

Gene expression during formation of earlywood and latewood in loblolly pine: expression profiles of 350 genes.

The natural variability of wood formation in trees affords opportunities to correlate transcript profiles with the resulting wood properties. We have used cDNA microarrays to study transcript abundance in developing secondary xylem of loblolly pine (Pinus taeda) over a growing season. The cDNAs were selected from a collection of 75 000 ESTs that have been sequenced and annotated (http://web.ahc.umn.edu/biodata/nsfpine/). Cell wall thickness and climatic data were related to earlywood and latewood formation at different time points during the growing season. Seventy-one ESTs showed preferential expression in earlywood or latewood, including 23 genes with no significant similarity to genes in GenBank. Seven genes involved in lignin synthesis were preferentially expressed in latewood. The studies have provided initial insights into the variation of expression patterns of some of the genes related to the wood formation process.

Expressed Sequence Tags↗

Involvement of nuclear factor-kappaB in the inhibition of pro-inflammatory mediators by pinosylvin.

Nuclear factor-kappaB (NF-kappaB) is a critical transcription factor for maximal expression of many of the cytokines that are involved in the pathogenesis of inflammatory diseases. In this study, we found that pinosylvin, a natural stilbenoid that is a component of the pine leaf (Pinus densiflora), significantly inhibited LPS-induced NF-kappaB activation in a concentration-dependent manner. Additionally, pinosylvin was found to inhibit the LPS-induced phosphorylation and degradation of IkappaB alpha in THP-1 cells. Therefore, we have attempted to determine whether pinosylvin can inhibit the expression of cytokines possessing NF-kappaB binding sites in their promoter regions. In a consistent result, pinosylvin inhibited LPS-induced production of TNF alpha and interleukin-8 (IL-8). Taken together, these results show that pinosylvin suppresses the production of pro-inflammatory mediators through the inhibition of the NF-kappaB signaling pathway.

Anti-Inflammatory Agents↗

Loblolly pine abietadienol/abietadienal oxidase PtAO (CYP720B1) is a multifunctional, multisubstrate cytochrome P450 monooxygenase.

Cytochrome P450 monooxygenases (P450s) are important enzymes for generating some of the enormous structural diversity of plant terpenoid secondary metabolites. In conifers, P450s are involved in the formation of a suite of diterpene resin acids (DRAs). Despite their important role in constitutive and induced oleoresin defense, a P450 gene of DRA formation has not yet been identified. By using phylogenetic cluster analysis of P450-like ESTs from loblolly pine (Pinus taeda), functional cDNA screening in yeast (Saccharomyces cerevisiae), and in vitro enzyme characterization, we cloned and identified a multifunctional and multisubstrate cytochrome P450 enzyme, CYP720B1 [abietadienol/abietadienal oxidase (PtAO)]. PtAO catalyzes an array of consecutive oxidation steps with several different diterpenol and diterpenal intermediates in loblolly pine DRA biosynthesis. Recombinant PtAO oxidized the respective carbon 18 of abietadienol, abietadienal, levopimaradienol, isopimara-7,15-dienol, isopimara-7,15-dienal, dehydroabietadienol, and dehydroabietadienal with apparent Michaelis-Menten (K(m)) values of 0.5-5.3 muM. PtAO expressed in yeast also catalyzed in vivo oxidation of abietadiene to abietic acid, but with activity much lower than with abietadienol or abietadienal. Consistent with a role of DRAs in conifer defense, PtAO transcript levels increased upon simulated insect attack using methyl jasmonate treatment of loblolly pine. The multisubstrate, multifunctional P450 diterpene oxidase PtAO, in concert with expression of a family of single-product and multiproduct diterpene synthases, allows for formation of a diverse suite of DRA defense metabolites in long-lived conifers.

Abietanes↗

Aboveground sink strength in forests controls the allocation of carbon below ground and its [CO2]-induced enhancement.

The partitioning among carbon (C) pools of the extra C captured under elevated atmospheric CO2 concentration ([CO2]) determines the enhancement in C sequestration, yet no clear partitioning rules exist. Here, we used first principles and published data from four free-air CO2 enrichment (FACE) experiments on forest tree species to conceptualize the total allocation of C to below ground (TBCA) under current [CO2] and to predict the likely effect of elevated [CO2]. We show that at a FACE site where leaf area index (L) of Pinus taeda L. was altered through nitrogen fertilization, ice-storm damage, and droughts, changes in L, reflecting the aboveground sink for net primary productivity, were accompanied by opposite changes in TBCA. A similar pattern emerged when data were combined from the four FACE experiments, using leaf area duration (LD) to account for differences in growing-season length. Moreover, elevated [CO2]-induced enhancement of TBCA in the combined data decreased from approximately 50% (700 g C m(-2) y(-1)) at the lowest LD to approximately 30% (200 g C m(-2) y(-1)) at the highest LD. The consistency of the trend in TBCA with L and its response to [CO2] across the sites provides a norm for predictions of ecosystem C cycling, and is particularly useful for models that use L to estimate components of the terrestrial C balance.

Atmosphere↗

A novel multifunctional O-methyltransferase implicated in a dual methylation pathway associated with lignin biosynthesis in loblolly pine.

S-adenosyl-L-methionine (SAM)-dependent O-methyltransferases (OMTs) catalyze the methylation of hydroxycinnamic acid derivatives for the synthesis of methylated plant polyphenolics, including lignin. The distinction in the extent of methylation of lignins in angiosperms and gymnosperms, mediated by substrate-specific OMTs, represents one of the fundamental differences in lignin biosynthesis between these two classes of plants. In angiosperms, two types of structurally and functionally distinct lignin pathway OMTs, caffeic acid 3-O-methyltransferases (CAOMTs) and caffeoyl CoA 3-O-methyltransferases (CCoAOMTs), have been reported and extensively studied. However, little is known about lignin pathway OMTs in gymnosperms. We report here the first cloning of a loblolly pine (Pinus taeda) xylem cDNA encoding a multifunctional enzyme, SAM:hydroxycinnamic Acids/hydroxycinnamoyl CoA Esters OMT (AEOMT). The deduced protein sequence of AEOMT is partially similar to, but clearly distinguishable from, that of CAOMTs and does not exhibit any significant similarity with CCoAOMT protein sequences. However, functionally, yeast-expressed AEOMT enzyme catalyzed the methylation of CAOMT substrates, caffeic and 5-hydroxyferulic acids, as well as CCoAOMT substrates, caffeoyl CoA and 5-hydroxyferuloyl CoA esters, with similar specific activities and was completely inactive with substrates associated with flavonoid synthesis. The lignin-related substrates were also efficiently methylated in crude extracts of loblolly pine secondary xylem. Our results support the notion that, in the context of amino acid sequence and biochemical function, AEOMT represents a novel SAM-dependent OMT, with both CAOMT and CCoAOMT activities and thus the potential to mediate a dual methylation pathway in lignin biosynthesis in loblolly pine xylem.

Amino Acid Sequence↗

Utility of the polymerase chain reaction-restriction fragment length polymorphisms of the intergenic spacer region of the rDNA for characterizing Gibberella fujikuroi isolates.

In the present report, a total of thirty-one isolates of Gibberella fujikuroi (Sawada) Wollenw. species complex of Fusarium (section Liseola) morphologically classified as F. moniliforme according to the taxonomy of Nelson, Toussoun and Marasas (1983) were analyzed for their ability to produce fumonisin B1 and fumonisin B2 by an optimized liquid chromatographic method. They were isolated from three hosts (Zea mays, Musa sapientum and Pinus pinea). The results indicate that M. sapientum is a preferential host for G. fujikuroi isolates with low or null capacity for producing fumonisins, while isolates from Z. mays and P. pinea are generally high fumonisin producers. The molecular characterization of isolates was carried out in parallel using an optimized, simple and low-cost method for isolating DNA from filamentous fungi and polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) of the rDNA intergenic spacer (IGS) region. The haplotypes obtained with Hha I enzyme and combinations of Hha I, EcoR I, Alu I, Pst I and Xho I enzymes provided very characteristic groupings of G. fujikuroi isolates as a function of host type and fumonisin B1 and B2 producing capacity. IGS region restriction patterns showed no relationship to isolate geographical origin. This is the first report on this method's capacity to detect polymorphism permitting discrimination between G. fujikuroi isolates from different hosts and with different toxigenic profiles.

Chromatography↗

Food-plant effects on larval performance do not translate into differences in fitness between populations of Panolis flammea (Lepidoptera: Noctuidae).

In the UK, Panolis flammea (Denis & Schiffermüller) is a pest of monocultures of non-native lodgepole pine Pinus contorta Douglas, but not of the indigenous host Scots pine P. sylvestrisL. This difference in population dynamics may be due to the adaptation of P. flammea populations to the phenology, chemical composition and natural enemy complement of lodgepole pine. To ascertain if there was local adaptation of P. flammea populations to lodgepole pine, this study tested for improved performance of both larvae and adults on the host plant species from which they were sourced, compared with their performance on the alternative host plant species. No difference was found in the relative mean performance of populations sourced from Scots pine or lodgepole pine plantations, when fed on Scots or lodgepole pine foliage. Larvae grew faster on Scots pine but this difference did not translate into differences in pupal weight, female body weight or fecundity. Indeed, those insects that had fed on lodgepole pine had a longer lifespan than those that had fed on Scots pine, which, if translated into greater probability of mating or higher fecundity, could contribute to the observed outbreak dynamics in the field. The prediction that the observed outbreak dynamics of P. flammea can be explained by the existence of populations locally adapted to lodgepole pine was not supported. These results cast doubt on the use of larval growth parameters as surrogates of fitness in Lepidoptera.

Adaptation, Physiological↗

A common mode of attraction of larvae and adults of insect predators to the sex pheromone of their prey (Hemiptera: Matsucoccidae).

The attraction of several adult predators, genera Elatophilus, Hemerobius and Sympherobius, to the sex pheromones of pine bast scales, Matsucoccus Cockerell, has already been demonstrated. Here, the hypothesis that the larvae of these predators are similarly attracted to the host prey sex pheromone is tested. The response of predators was tested in field trials using pine tree arenas baited with the sex pheromones of M. josephi Bodenheimer & Harpaz, M. feytaudi Ducasse and M. matsumurae Kuwana. Experiments were conducted in Israel in stands of Pinus halepensis infested by M. josephi and in Portugal in stands of P. pinaster infested by M. feytaudi, respectively. The selectivity of larvae for the three sex pheromones was tested in Petri dish arenas in the laboratory. In the field, the larval stages exhibited similar modes of attraction to those of the conspecific adults: Elatophilus hebraicus Pericart in Aleppo pine forest, E. crassicornis Reuter and Hemerobius stigma Stephens in the maritime pine forests. Laboratory choice tests confirmed the kairomonal selectivity of larvae. Both forest and laboratory tests demonstrated the response of a coccinellid of the genus Rhyzobius to the sex pheromones of M. feytaudi and M. matsumurae. A unique chemical communication system among several taxa of predators of Matsucoccus spp. was highlighted that may be attributed to their coevolution on a geological time scale.

Animal Communication↗

Nickel adsorption on the modified pine tree materials.

Removal of nickel ions from aqueous solutions containing 1-100 mg l(-1), using pine tree (Pinus nigra) materials modified with HCl, was investigated on a laboratory scale. For this purpose, two natural adsorbents such as the modified pine bark (MPB) and the modified pine cone (MPC) materials with HCl solution were studied. At first, the required concentration level of the HCl solution for the modification was observed, and then this was followed by the determinations of optimum levels of adsorbent amount, stirring rate, contact time and pH values. Various adsorption isotherms were also obtained by using different concentrations of the heavy metal cations tested in the experiment. As a result, the maximum removal efficiency levels obtained were as follows; 97% for the modified pine bark at pH 8 and 80% for the modified pine cone at pH 8.

Adsorption↗

Potential for assessing long-term dynamics in soil nitrogen availability from variations in delta15N of tree rings.

Numerous researchers have used the isotopic signatures of C, H, and O in tree rings to provide a long-term record of changes in the physiological status, climate, or water-source use of trees. The frequently limiting element N is also found in tree rings, and variation in its isotopic signature may provide insight into long-term changes in soil N availability of a site. However, research has suggested that N is readily translocated among tree ring of different years; such infidelity between the isotopic compositions of the N taken up from the soil and the N contained in the ring of that growth year would obscure the long-term N isotopic record. We used a 15-year 15N-tracer study to assess the degree of N translocation among tree rings in ponderosa pine (Pinus ponderosa) trees growing in a young, mixed-conifer plantation. We also measured delta13C and delta15N values in unlabeled trees to assess the degree of their covariance in wood tissue, and to explore the potential for a biological linkage between them. We found that the maximum delta15N values in rings from the labeled trees occurred in the ring formed one-year after the 15N was applied to the roots. The delta15N value of rings from labeled trees declined exponentially and bidirectionally from this maximum peak, toward younger and older rings. The unlabeled trees showed considerable interannual variation in the delta15N values of their rings (up to 3 and 5 per thousand), but these values correlated poorly between trees over time and differed by as much as 6 per thousand. Removal of extractives from the wood reduced their delta15N value, but the change was fairly small and consistent among unlabeled trees. The delta13C and delta15N values of tree rings were correlated over time in only one of the unlabeled trees. Across all trees, both delta13C values of tree rings and annual stem wood production were well correlated with annual precipitation, suggesting that soil water balance is an important environmental factor controlling both net C gain and transpirational water loss at this site. Our results suggest that interannual translocation of N among tree rings is substantial, but may be predictable enough to remove this source of variation from the tree-ring record, potentially allowing the assessment of long-term changes in soil N availability of a site.

Environmental Monitoring↗

Effects of clear-cutting and soil preparation on natural 15N abundance in the soil and needles of two boreal conifer tree species.

This study presents the impacts of clear-cutting and site preparation on soil and needle 15N-fractionation of Scots pine (Pinus sylvestris, L.) and Norway spruce (Picea abies (L.), Karst). Three microsites on different methods of site preparation were used: (i) mound (broken O/E/B horizons piled upside down over undisturbed humus), (ii) deep (exposed C-horizon) and (iii) shallow (exposed E/B horizon). We found significant differences between species, between closed forest and clear-cuts as well as between different site preparations. For instance, in the context of interspecific variations, the mean needle nitrogen concentrations of both seedlings (1.15,+/-0.10 %) and mature (1.09,+/-0.07 %) pine trees were significantly higher compared to corresponding needle concentrations of seedlings (0.88,+/-0.06 %) and mature trees (0.79,+/-0.02 %) of spruce. Similarly, we observed significantly more 15N-enriched needles of mature spruces (-4.0,+/-0.20 per thousand) as well as of seedlings (-5.0,+/-0.11 per thousand) relative to that of mature pine needles (-5.6,+/-0.10 per thousand) and seedlings (-6.0,+/-0.31 per thousand). These variations were assumed to be caused by the variation in mycorrhizal associations between the species. We assume that the proportion of mycorrhizal N-uptake of pines might have been larger than that of spruce. Regarding the clear-cut effects on N and 15N of both tree species, we observed that, in the mature natural stand, needle N concentrations of both pine (1.09,+/-0.07 %) and spruce (0.79,+/-0.02 %) tree species did not change significantly after clear-cutting (pine: 1.01,+/-0.06 %; spruce: 0.74,+/-0.04 % ). However, clear-cutting resulted in the significant increase in needle 15N natural abundance of both pine (-2.70,+/-0.06 per thousand) and spruce (-2.09,+/-0.05 per thousand) in comparison to that of natural stand (pine:-5.60,+/-0.10 per thousand; spruce:-4.00,+/-0.20 per thousand), which is assumed to be due to the increased level of nitrification and leaching of nitrate after clear-cutting. In the context of site preparation methods, soil and needle N were observed to be more 15N-enriched in deep and shallow treatment sites compared to that of closed forest site and untreated clear-cut site, which indicated that the main source of N uptake seems to be mainly directly from the soil of the corresponding horizons of mineral soil with higher delta15N.

Forestry↗

Comparative effects of enzogenol and vitamin C supplementation versus vitamin C alone on endothelial function and biochemical markers of oxidative stress and inflammation in chronic smokers.

Chronic smoking is associated with endothelial dysfunction and inflammation, with oxidative stress contributing to both these processes. In this study, we investigated the effect of combined antioxidant treatment with Enzogenol, a flavonoid extract from the bark of Pinus radiata and vitamin C, over and above vitamin C alone, on endothelial function, plasma markers of inflammation and oxidative stress, blood pressure (BP) and anthropometrics. Forty-four chronic smokers without established cardiovascular disease were assigned randomly to receive either 480 mg Enzogenol and 60 mg vitamin C, or 60 mg vitamin C alone daily for 12 weeks. Endothelial function in the brachial artery was assessed by flow-mediated vasodilation (FMD). FMD improved in both treatment groups (p < 0.001), with no significant difference between the two groups (p = 0.84). In the group receiving Enzogenol and vitamin C, protein carbonyl levels were significantly reduced compared to the group taking vitamin C alone (p = 0.03). Enzogenol and vitamin C resulted in a significant reduction in fibrinogen levels in heavy smokers compared with vitamin C alone (p < 0.009). These findings demonstrated that co-supplementation with Enzogenol and vitamin C in smokers conferred no additional beneficial effect on macrovascular endothelial function over and above that seen in the vitamin C alone group. However, Enzogenol did demonstrate additional favourable effects on protein oxidative damage and fibrinogen levels.

Adult↗

Use of sewage sludge compost as the restoration agent on the degraded soil of Tatarstan.

One of the characteristics of soils in Tatarstan is their low organic matter content. The decrease in soil organic matter is paralleled by declines in soil fertility. One method to reverse this degradation in soil quality is the addition of organic matter. The use of sewage sludge on soils intended for growing of plant seedlings provides an alternative for sewage sludge disposal. Therefore, the evaluation of the feasibility of using compost from the municipal sewage sludge produced in Kazan for the soil restoration and growth of Pinus silvestris seedlings was carried out. The grey forest soil (Haplic Greyzem) was amended with compost at application rate 30, 60 and 90 Mg ha(-1) on a dry matter basis. Organic matter content increased with the increase in sludge amendment. The concentrations of individual heavy metal were below the current limits established for Russia and European countries. Sludge amendments enhanced the germination and the number of the seedlings and the increase were more obvious for the soil with highest sludge treatment. The application of composted sludge to soil was followed by the increase in microbial biomass and basal respiration.

Biomass↗

Reciprocal selection causes a coevolutionary arms race between crossbills and lodgepole pine.

Few studies have shown both reciprocal selection and reciprocal adaptations for a coevolving system in the wild. The goal of our study was to determine whether the patterns of selection on Rocky Mountain lodgepole pine (Pinus contorta spp. latifolia) and red crossbills (Loxia curvirostra complex) were concordant with earlier published evidence of reciprocal adaptations in lodgepole pine and crossbills on isolated mountain ranges in the absence of red squirrels (Tamiasciurus hudsonicus). We found that selection (directional) by crossbills on lodgepole pine where Tamiasciurus are absent was divergent from the selection (directional) exerted by Tamiasciurus on lodgepole pine. This resulted in divergent selection between areas with and without Tamiasciurus that was congruent with the geographic patterns of cone variation. In the South Hills, Idaho, where Tamiasciurus are absent and red crossbills are thought to be coevolving with lodgepole pine, crossbills experienced stabilizing selection on bill size, with cone structure as the agent of selection. These results show that crossbills and lodgepole pine exhibit reciprocal adaptations in response to reciprocal selection, and they provide insight into the traits mediating and responding to selection in a coevolutionary arms race.

Adaptation, Physiological↗

Measuring and simulating crown respiration of Scots pine with increased temperature and carbon dioxide enrichment.

Acclimation to elevated atmospheric carbon dioxide concentration and temperature of respiration by the foliage in the crown of Scots pine (Pinus sylvestris) trees is measured and modelled. Starting in 1996, individual 20-year-old trees were enclosed in chambers and exposed to either normal ambient conditions (CON), elevated CO2 concentration (EC), elevated temperature (ET) or a combination of EC and ET (ECT). Respiration of individual leaves within the crown was measured in 2000. To extrapolate the response of respiration of individual leaves to the whole crown, a multi-layer model was developed and used to predict daily and annual crown respiration, in which the crown structure and corresponding microclimate data were used as input. Respiration measurements showed that EC led to higher Q10 values (4.6%) relative to CON, but lower basal respiration rates at 20 degrees C [R1.d(20)] (-7.1%) during the main growth season (days 120-240), whereas ET and ECT both reduced Q10 (-12.0 and -9.8%, respectively) throughout the year but increased R1.d(20) (27.2 and 21.6%, respectively) during the period of no-growth, and slightly reduced R1.d(20) (-1.7 and -2.8%, respectively) during the main growth season. Model computations showed that annual crown respiration increased: (1) by 16% in EC, with 92% of this increase attributable to the increase in foliage area; (2) by 35% in ET, with 66% related to the increase in foliage area and 17% to the rise in ambient temperature; and (3) by 27% in the case of ECT, with 43% attributable to the increase in foliage area and 29% to the rise in ambient temperature. Changed respiration parameters for individual leaves, induced by treatments, made only a small contribution to the annual crown respiration compared with the increased foliage area. The effects of changes in crown architecture and nitrogen distribution, caused by treatments, on the daily and annual course of crown respiration are discussed.

Carbon Dioxide↗

Light and water-use efficiencies of pine shoots exposed to elevated carbon dioxide and temperature.

An automatic gas exchange system was used to continuously measure water and carbon fluxes of attached shoots of Scots pine trees (Pinus sylvestris L.) grown in environment-controlled chambers for a 3-year period (1998-2000) and exposed to either normal ambient conditions (CON), elevated CO2 (+350 micro mol mol-1; EC), elevated temperature (+2-6 degrees C; ET) or a combination of EC and ET (ECT). EC treatment enhanced the mean daily total carbon flux per unit projected needle area (Fc.d) by 17-21 %, depending on the year. This corresponds to a 16-24 % increase in light-use efficiency (LUE) based on incident photosynthetically active radiation. The EC treatment reduced the mean daily total water flux (Fw.d) by 1-12 %, corresponding to a 13-35 % increase in water-use efficiency (WUE). The ET treatment increased Fc.d by 10-18 %, resulting in an 8-19 % increase in LUE, and Fw.d by 48-74 %, resulting in a reduction of WUE by 19-34 %. There was no interaction between CO2 and temperature elevation in connection with either carbon or water fluxes, as the carbon flux responded similarly in both ECT and EC, while the water flux in the ECT treatment was similar to that in ET. Regressions indicated that the increase in maximum LUE was greater with increasing air temperature, whereas changes in WUE were related only to high vapour pressure deficit. Furthermore, changes in LUE and WUE caused by ECT treatment displayed strong diurnal and seasonal variation.

Carbon Dioxide↗

Seasonal variation in respiration of 1-year-old shoots of scots pine exposed to elevated carbon dioxide and temperature for 4 years.

Sixteen 20-year-old Scots pine (Pinus sylvestris L.) trees growing in the field were enclosed for 4 years in environment-controlled chambers that maintained: (1) ambient conditions (CON); (2) elevated atmospheric CO2 concentration (ambient + 350 micro mol mol-1; EC); (3) elevated temperature (ambient +2-6 degrees C; ET); or (4) elevated CO2 and elevated temperature (ECT). The dark respiration rates of 1-year-old shoots, from which needles had been partly removed, were measured over the growing season in the fourth year. In all treatments, the temperature coefficient of respiration, Q10, changed with season, being smaller during the growing season than at other times. Respiration rate varied diurnally and seasonally with temperature, being highest around mid-summer and declining gradually thereafter. When measurements were made at the temperature of the chamber, respiration rates were reduced by the EC treatment relative to CON, but were increased by ET and ECT treatments. However, respiration rates at a reference temperature of 15 degrees C were reduced by ET and ECT treatments, reflecting a decreased capacity for respiration at warmer temperatures (negative acclimation). The interaction between season and treatment was not significant. Growth respiration did not differ between treatments, but maintenance respiration did, and the differences in mean daily respiration rate between the treatments were attributable to the maintenance component. We conclude that maintenance respiration should be considered when modelling respiratory responses to elevated CO2 and elevated temperature, and that increased atmospheric temperature is more important than increasing CO2 when assessing the carbon budget of pine forests under conditions of climate change.

Carbon↗