Ethylenediurea (EDU) effects on hybrid larch saplings exposed to ambient or elevated ozone over three growing seasons

Ground-level ozone (O3) pollution is a persis-tent environmental issue that can lead to adverse effects on trees and wood production,thus indicating a need for forestry interventions to mediate O3 effects.We treated hybrid larch (Larix gmelinii var.japonica × L.kaempferi)saplings grown in nutrient-poor soils with 0 or 400 mg L-1 water solutions of the antiozonant ethylenediurea(EDU0,EDU400) and exposed them to ambient O3 (AOZ;08:00-18:00 ≈ 30 nmol mol-1) or elevated O3 (EOZ;08:00-18:00≈ 60 nmol mo1-1) over three growing sea-sons.We found that EDU400 protected saplings against most effects of EOZ,which included extensive visible foliar injury,premature senescence,decreased photosyn-thetic pigment contents and altered balance between pig-ments,suppressed gas exchange and biomass production,and impaired leaf litter decay.While EOZ had limited effects on plant growth (suppressed stem diameter),it decreased the total number of buds per plant,an effect that was not observed in the first growing season.These results indicate that responses to EOZ might have implications to plant competitiveness,in the long term,as a result of decreased potential for vegetative growth.However,when buds were standardized per unit of branches biomass,EOZ significantly increased the number of buds per unit of biomass,suggest-ing a potentially increased investment to bud development,in an effort to enhance growth potential and competitiveness in the next growing season.EDU400 minimized most of these effects of EOZ,significantly enhancing plant health under O3-induced stress.The effect of EDU was attributed mainly to a biochemical mode of action.Therefore,hybrid larch,which is superior to its parents,can be significantly improved by EDU under long-term elevated O3 exposure,providing a perspective for enhancing afforestation practices.     

Effects of mist acidity and ambient ozone removal on montane red spruce

Recent forest studies have established that high-elevation (> 900 m) populations of red spruce (Picea rubens Sarg.) in the northeastern USA are declining. Because it has been suggested that changes in air quality are responsible for the decline, we examined the effects of acidic mists and ozone on several biochemical and growth parameters in mature montane red spruce. We used branch-sized environmental chambers to introduce mists of controlled composition and exclude ambient clouds and ozone from individual branches within a tree. Mists consisting of distilled water increased the end-of-season pigment concentration and shoot length of enclosed branches relative to ambient or artificial mists. Needle and twig weights and starch concentrations were not significantly altered by the acidic mist treatments. Removal of ambient ozone had no apparent effect on the variables measured.     

Detection and quantification of changes in membrane-associated calcium in red spruce saplings exposed to acid fog

Five-year-old red spruce saplings (Picea rubens Sarg.) were exposed to either (1) acid fog consisting of a mixture of H(2)SO(4) and HNO(3) adjusted to pH 2.5, (2) distilled-water fog at pH 5.6, or (3) no fog (dry control) for 3.5 hours per day, five times a week during the 1996 and 1997 growing seasons. The effect of fog on cell membrane-associated calcium (mCa) of leaf mesophyll cells was investigated with the fluorescence probe chlortetracycline (CTC). In both years, mean mCa concentrations were significantly less in needles exposed to acid fog than in needles exposed to distilled-water fog or in untreated needles. In 1997, acid-fog treatment resulted in 25 and 12% reductions in mCa in current-year needles, and 18 and 15% reductions in 1-year-old needles, compared with untreated needles and needles exposed to distilled-water fog, respectively, indicating that acid deposition induced calcium leaching from the membranes of photosynthetic mesophyll cells. Exposure to distilled-water fog also led to reductions in mCa in young needles, suggesting that water films on needle surfaces can induce losses by diffusion between the needle interior and surface. Consistent with the chamber studies, field data obtained from red spruce trees at two sites in Maine showed that low mCa concentrations in needles were associated with exposure to acid fog.     

Needle metabolome, freezing tolerance and gas exchange in Norway spruce seedlings exposed to elevated temperature and ozone concentration

Northern forests are currently experiencing increasing mean temperatures, especially during autumn and spring. Consequently, alterations in carbon sequestration, leaf biochemical quality and freezing tolerance (FT) are likely to occur. The interactive effects of elevated temperature and ozone (O(3)), the most harmful phytotoxic air pollutant, on Norway spruce (Picea abies (L.) Karst.) seedlings were studied by analysing phenology, metabolite concentrations in the needles, FT and gas exchange. Sampling was performed in September and May. The seedlings were exposed to a year-round elevated temperature (+1.3 °C), and to 1.4× ambient O(3) concentration during the growing season in the field. Elevated temperature increased the concentrations of amino acids, organic acids of the citric acid cycle and some carbohydrates, and reduced the concentrations of phenolic compounds, some organic acids of the shikimic acid pathway, sucrose, cyclitols and steroids, depending on the timing of the sampling. Although growth onset occurred earlier at elevated temperature, the temperature of 50% lethality (LT(50)) was similar in the treatments. Photosynthesis and the ratio of photosynthesis to dark respiration were reduced by elevated temperature. Elevated concentrations of O(3) reduced the total concentration of soluble sugars, and tended to reduce LT(50) of the needles in September. These results show that alterations in needle chemical quality can be expected at elevated temperatures, but the seedlings' sensitivity to autumn and spring frosts is not altered. Elevated O(3) has the potential to disturb cold hardening of Norway spruce seedlings in autumn, and to alter the water balance of the seedling through changes in stomatal conductance (g(s)), while elevated temperature is likely to reduce g(s) and consequently reduce the O(3)-flux inside the leaves.     

Seasonal air and soil temperature effects on photosynthesis in red spruce (Picea rubens) saplings

Net photosynthesis and stomatal conductance were measured in ten red spruce (Picea rubens Sarg.) saplings, growing near Ithaca, New York, throughout the early spring and late-fall growing periods. Gas exchange and daily minimum and maximum soil and air temperatures were also measured. Linear regression analysis showed that rates of net photosynthesis were positively correlated with both minimum daily soil and air temperatures but that minimum soil temperature was a better predictor of net photosynthesis. Moreover, net photosynthesis was more sensitive to changes in soil temperature than to changes in air temperature, and photosynthesis was approximately twice as sensitive to temperature changes during the fall than during the spring.     

Influence of nitrogen and phosphorous availability and ozone stress on Norway spruce seedlings

Four-year-old Norway spruce (Picea abies L. (Karst.)) seedlings were exposed to ambient and elevated (1.5 x ambient in 1997 and 1.6 x ambient in 1998) ozone concentrations [O3] and three nitrogen (N) and two phosphorus (P) availabilities: "optimal" values (control); 70% of the control N and P values (LN and LP); and 150% of the control N value (HN). Treatments were applied in an open-field ozone fumigation facility during the 1997 and 1998 growing seasons. Effects on growth, mineral and pigment concentrations, stomatal conductance and ultrastructure of needles were studied. The HN treatment increased growth significantly, whereas elevated [O3] had a slight or variable impact on growth and biomass allocation in all N treatments. Although there were no significant effects of the LP treatment on plant growth during the second year, there was a reduction in 1-year-old shoot dry mass in the elevated O3 + LP treatment at the end of the experiment. There were no significant treatment effects on mineral concentrations of current-year and 1-year-old needles at the final harvest. In response to the HN treatment, chlorophyll a and b and carotenoid concentrations increased significantly in current-year needles. Chlorophyll a/b ratio decreased in response to elevated [O3] alone, but increased in seedlings in the O(3) + LP treatment. Stomatal conductance of current-year needles decreased with increasing N availability, but increased in response to elevated [O3]. However, the O3-induced increase in stomatal conductance was less in the LN and LP treatments than in the control treatment. In chloroplasts of current-year needles, increased N availability decreased mean starch grain area, but increased the number of plastoglobuli. We conclude that Norway spruce seedlings are relatively tolerant to slightly elevated [O3], and that nitrogen and phosphorus imbalances do not greatly affect the influence of O3 on this species when the exposure lasts for two growing seasons or less.     

Seasonal variations in soluble sugars and starch within woody stems of Cornus sericea L

Carbohydrate composition changed seasonally in red osier dogwood (Cornus sericea L.) stem tissues. Starch concentration was highest in fall and decreased to a minimum in midwinter. Coincident with the breakdown of starch in fall, there was an increase in the concentrations of soluble sugars. Soluble sugars were present in highest concentrations in midwinter. Glucose, fructose, sucrose, and raffinose were the predominant soluble sugars present in both bark and wood tissues. In early spring, the soluble sugar concentration decreased and the concentration of starch increased. The seasonal interchange between sugars and starch did not simply reflect a general quantitative shift in the balance between sugars and starch because qualitative changes in soluble sugars were also noted. The most striking changes involved the trisaccharide raffinose. Raffinose was barely detectable in summer and early fall, but increased to one fifth and one third of the total soluble sugars in January samples of bark and wood tissues, respectively. The potential physiological role of raffinose in overwintering red osier dogwood tissue is discussed.     

Acclimation of shade-developed leaves on saplings exposed to late-season canopy gaps

We hypothesized that photoinhibition of shade-developed leaves of deciduous hardwood saplings would limit their ability to acclimate photosynthetically to increased irradiance, and we predicted that shade-tolerant sugar maple (Acer saccharum Marsh.) would be more susceptible to photoinhibition than intermediately shade-tolerant red oak (Quercus rubra L.). After four weeks in a canopy gap, photosynthetic rates of shade-developed leaves of both species had increased in response to the increase in irradiance, although final acclimation was more complete in red oak. However, photoinhibition occurred in both species, as indicated by short-term reductions in maximum rates of net photosynthesis and the quantum yield of oxygen evolution, and longer-term reductions in the efficiency of excitation energy capture by open photosystem II (PSII) reaction centers (dark-adapted F(v)/F(m)) and the quantum yield of PSII in the light (phi(PSII)). The magnitude and duration of this decrease were greater in sugar maple than in red oak, suggesting greater susceptibility to photoinhibition in sugar maple. Photoinhibition may have resulted from photodamage, but it may also have involved sustained rates of photoprotective energy dissipation (especially in red oak). Photosynthetic acclimation also appeared to be linked to an ability to increase leaf nitrogen content. Limited photosynthetic acclimation in shade-developed sugar maple leaves may reflect a trade-off between shade-tolerance and rapid acclimation to a canopy gap.     

Influence of water stress on the physiology and growth of red spruce seedlings

Two-year-old, container-grown red spruce (Picea rubens Sarg.) seedlings from a New Hampshire seed source were exposed to 10 or 11 drying cycles in which the seedlings were not watered until their midday (1400 h) xylem water potentials averaged -1.57 MPa. Control seedlings were kept well watered to maintain midday water potentials of about -0.73 MPa. After the final drying cycle, the water-stressed seedlings were rehydrated and osmotic potentials were determined by pressure-volume analysis. Gas exchange at ambient CO(2) concentration (338 ppm) and at an elevated CO(2) concentration (838 ppm) was measured on both groups of plants as they slowly dried down. No osmotic adjustment or photosynthetic acclimation occurred as a result of the water-stress treatment and both groups of seedlings maintained photosynthesis to water potentials as low as -3.0 MPa. Twenty-four hours after rehydration, the water-stressed seedlings had photosynthetic rates as high as the control seedlings. Estimated stomatal limitation to photosynthesis was approximately 30% down to water potentials of -1.4 MPa, but increased steadily as water potentials decreased further. At ambient CO(2) concentrations (338 ppm) and water potentials averaging -2.45 MPa, photosynthetic rates of water-stressed seedlings were 15% those of well-watered seedlings, whereas when the same water-stressed seedlings were measured in the presence of an elevated concentration of CO(2) (838 ppm) their photosynthetic rates were 73% those of well-watered seedlings measured at an ambient CO(2) concentration (338 ppm).     

Sap flux in pure aspen and mixed aspen-birch forests exposed to elevated concentrations of carbon dioxide and ozone

Elevated concentrations of atmospheric carbon dioxide ([CO2]) and tropospheric ozone ([O3]) have the potential to affect tree physiology and structure and hence forest water use, which has implications for climate feedbacks. We investigated how a 40% increase above ambient values in [CO2] and [O3], alone and in combination, affect tree water use of pure aspen and mixed aspen-birch forests in the free air CO2-O3 enrichment experiment near Rhinelander, Wisconsin (Aspen FACE). Measurements of sap flux and canopy leaf area index (L) were made during two growing seasons, when steady-state L had been reached after more than 6 years of exposure to elevated [CO2] and [O3]. Maximum stand-level sap flux was not significantly affected by elevated [O3], but was increased by 18% by elevated [CO2] averaged across years, communities and O(3) regimes. Treatment effects were similar in pure aspen and mixed aspen-birch communities. Increased tree water use in response to elevated [CO2] was related to positive CO2 treatment effects on tree size and L (+40%). Tree water use was not reduced by elevated [O3] despite strong negative O3 treatment effects on tree size and L (-22%). Elevated [O3] predisposed pure aspen stands to drought-induced sap flux reductions, whereas increased tree water use in response to elevated [CO2] did not result in lower soil water content in the upper soil or decreasing sap flux relative to control values during dry periods. Maintenance of soil water content in the upper soil in the elevated [CO2] treatment was at least partly a function of enhanced soil water-holding capacity, probably a result of increased organic matter content from increased litter inputs. Our findings that larger trees growing in elevated [CO2] used more water and that tree size, but not maximal water use, was negatively affected by elevated [O3] suggest that the long-term cumulative effects on stand structure may be more important than the expected primary stomatal closure responses to elevated [CO2] and [O3] in determining stand-level water use under possible future atmospheric conditions.     

Effects of elevated nitrate and aluminum on the growth and nutrition of red spruce (Picea rubens) seedlings

Acidic deposition in high-elevation forests in the Appalachian Mountains of the eastern United States has been implicated in the decline of red spruce (Picea rubens Sarg.). Elevated soil acidity may increase soil Al availability and toxicity to roots. Enhanced soil solution NO(3) (-) concentrations, resulting from precipitation inputs and enhanced soil organic matter mineralization, may exacerbate Al toxicity by increasing root Al uptake. We exposed red spruce seedlings to 350, 500, 800 or 1400 micro M NO(3) (-) and 0 or 200 micro M Al in a factorial design in sand-nutrient solution culture to test if increased NO(3) (-) concentrations enhance Al uptake and toxicity. In addition to significant reductions in seedling growth parameters resulting from Al exposure, we found significant interactions between NO(3) (-) and Al for seedling height growth rate, needle weight, shoot weight and root weight. Differences in these parameters between Al treatments became more pronounced as solution NO(3) (-) concentration increased and reflected an Al-mediated inhibition of seedling response to increasing NO(3) (-) concentration. Solution NO(3) (-) concentrations above 500 micro M induced root nitrate reductase (NR) activity, whereas shoot NR activity increased in response to NO(3) (-) up to 500 micro M and declined above that concentration. In contrast, exposure to Al depressed NR activity of roots but tended to stimulate needle NR activity. Foliar N concentrations increased in seedlings grown in cultures containing between 350 and 500 micro M NO(3) (-), with no change above 500 micro M. Increasing concentrations of NO(3) (-) depressed foliar P concentrations, with reductions being greatest in seedlings exposed to 1400 micro M NO(3) (-). Exposure to Al increased foliar Ca, K and Al concentrations, decreased foliar P concentrations, and inhibited increases in foliar Mg concentration in response to increasing NO(3) (-). The consistent interactions between NO(3) (-) and Al for growth, root NR activity and foliar Mg concentration were the result of an inhibition of seedling response to NO(3) (-) mediated by Al in solution, rather than enhanced Al toxicity resulting from growth in the presence of elevated NO(3) (-) concentrations.     

Growth responses and related biochemical and ultrastructural changes of the photosynthetic apparatus in birch (Betula pendula) saplings exposed to low concentrations of ozone

Saplings of ozone-sensitive and ozone-tolerant birch (Betula pendula Roth.), clones B and C, respectively, were exposed to ozone concentrations that were 1.7-fold higher than ambient for one growing season under open-field conditions. Ambient air was used as the control treatment. In the ozone-sensitive clone B, there was an initial stimulation of leaf area growth in response to the ozone treatment, but further ozone exposure caused reductions in leaf and stem biomass growth, Rubisco and chlorophyll a contents, net photosynthesis, water use efficiency and chloroplast size. It also caused an alteration in chloroplast shape and injury to thylakoid membranes. In the ozone-tolerant clone C, ozone fumigation did not affect growth rate, and there were no consistent changes in chlorophyll content, photosynthesis or water use efficiency. There were also fewer ultrastructural abnormalities in the chloroplasts of clone C than of clone B. Based on the observed biochemical, physiological and structural changes in chloroplasts of clone B in response to low concentrations of ozone, we conclude that the increasing concentration of tropospheric ozone represents a risk to natural birch populations.     

Age-related changes in foliar morphology and physiology in red spruce and their influence on declining photosynthetic rates and productivity with tree age

The contribution of changes in meristem behavior to age-related decline in forest productivity is poorly understood. We studied age-related trends in needle morphology and gas exchange in a population of red spruce (Picea rubens Sarg.) growing in a multi-cohort stand where trees ranged from first-year germinants to trees over 150 years old, as well as in grafted scions from these trees. In the field study, age-related trends in foliar morphology were determined in six cohorts ranging in age from 2 to 120 years, and differences in gas exchange characteristics were compared between 60- and 120-year age classes. In a common-rootstock study, scions from trees representing 20-, 60-, and 120-year cohorts were grafted onto juvenile rootstock and maintained for three growing seasons, after which morphological and physiological foliar attributes were evaluated. The field study revealed significant age-related trends in foliar morphology, including decreasing specific leaf area, and increasing needle width, projected area, and width/length ratio. Similar trends were apparent in foliage from the grafted scions. Both in situ foliage and shoots of grafted scions from the oldest cohort showed significantly lower photosynthetic rates than their counterparts from younger trees; however, differences in stomatal conductance and internal CO(2) concentrations were not significant. These results suggest that: (1) foliage of red spruce exhibits age-related trends in both morphology and physiology; (2) age-related decreases in photosynthetic rates contribute to declining productivity in old red spruce; (3) declines in photosynthetic rates result from nonstomatal limitations; and (4) age-related changes in morphology and physiology are inherent in meristems and persist for at least 3 years in scions grafted to juvenile rootstock.     

Effects of acidic fog and ozone on the growth and physiological functions of Fagus crenata saplings

The effects of simulated acid fog (SAF) and ozone (O₃) stress on the growth and physiology of beech (Fagus crenata) saplings were investigated. Three-year-old beech saplings were exposed to SAFs of pH 3 and pH 5 (control) during May 2007 to July 2008. In each SAF treatment group, half of the saplings were exposed to 60 ppb of O₃ during September 2007 to July 2008. In comparison to the control saplings, those from the pH 3 treatment had lower total plant biomasses, epicuticular wax amounts, Ca²⁺ concentrations in their leaves, and lower starch concentrations in their leaves and roots. The effect of O₃ was significant only for the starch concentration in the roots, but the O₃ exposure also negatively affected the growth and physiology of beech saplings. Results show that acid fog exerts various severe effects, and that both chronic acid fog and O₃ exposure suppressed the physiological functions of beech saplings.     

Chemical and structural characteristics of conifer needles exposed to ambient air pollution

Conifer needles exposed to ambient air pollutants were observed by a scanning electron microscope (SEM). Predominant changes on needles were the breakdown and the aggregation of wax structures on stomatal chambers. However, the mode of destruction in stomatal wax structures was dependent on the pollutants. Ginkgo biloba seemed to be resistant to air pollutants despite higher amounts of sulfur in the foliage.     

Photosynthetic and transpirational responses of red spruce understory trees to light and temperature

Understory red spruce (Picea rubens Sarg.) trees, between 20 and 50 cm in height and 12 years or more in age, were collected from mid- and high-elevation stands in north-central Vermont and placed in a closed-cuvette system to measure photosynthetic and transpirational responses to photosynthetic photon flux density (PPFD) and temperature. Photosynthesis, dark respiration, transpiration and water-use efficiency of trees from both stands responded to changes in PPFD and temperature in similar ways. Trees from both stands exhibited maximum rates of net photosynthesis at temperatures between 15 and 20 degrees C, and exposure to higher temperatures resulted in reduced rates of photosynthesis and increased rates of respiration. Net photosynthetic rates generally increased with increasing light intensity but began to level off at 250 micro mol m(-2) s(-1). Water-use efficiency was maximal when temperature and PPFD were at 15 degrees C and above 400 micro mol m(-2) s(-1), respectively.     

秋枫和木棉对大气臭氧浓度升高的生理响应

以秋枫Bischofi a javanica Bl.和木棉Bombax malabaricum DC.苗木为试验材料,采用自行设计的开顶式熏气室装置,研究在4种体积分数的臭氧(O3)[E200,Φ(O3)=200×10-9;E100,Φ(O3)=100×10-9;E50,Φ(O3)=50×10-9;环境大气NF,Φ(O3)=10×10-9~20×10-9]下2种植物叶片可溶性糖含量、可溶性蛋白质含量、保护酶活性和膜脂过氧化程度的变化情况,旨在探究在地表臭氧浓度升高的条件下2种植物的生理生化指标的变化情况及其响应机理。结果显示:随臭氧浓度的增加,2种植物膜透性和丙二醛含量均逐渐上升,且与臭氧浓度呈显著正相关关系,说明2种植物膜脂过氧化程度加剧;秋枫可溶性蛋白含量均逐渐下降,木棉先降低后大幅上升;二者可溶性糖含量均是先上升后小幅度下降;POD活性均是先降低后升高;秋枫CAT活性呈先升高再降低的趋势,木棉则是逐渐降低。对2种植物各生理指标进行主成分分析表明,2种植物耐臭氧能力顺序为秋枫木棉。     

Pilot study on the effects of elevated air temperature and CO_2 on artificially defoliated silver birch saplings

The impacts of elevated temperature and CO_2 on young silver birch(Betula pendula Roth) saplings after 0,25, 50 or 75% artificial defoliation were assessed by measuring plant height and dry mass of aboveground compartments and roots and various morphological and physiological variables. Defoliation either increased or decreased plant growth depending on the severity of damage and the climatic treatment. At 21 °C and400 mg L~(-1) CO_2, defoliated plants were not able to compensate for the lost foliage, but growth compensation and adaptation to the changed conditions were greater; growth of young defoliated silver birch saplings increased, which led to increased height and a tendency to enhance final aboveground and root biomass and leaf nitrogen and carbon content compared to the nondefoliated controls. Nevertheless, the short-term effect of the different climatic conditions did not result in a significant overgrowth of defoliated plants. A slight increase in temperature and CO_2 were the most acceptable conditions for defoliated plants;however, a 4 °C increase with correspondingly higher CO_2 was more stressful as shown by less growth in height and biomass allocation to leaves, stems and roots. The findings from the pilot experiment are more applicable to young birch trees, but stress on young trees may be reflected in future tree growth.