Toini Holopainen

Effects of nitrogen fertilization on secondary chemistry and ectomycorrhizal state of scots pine seedlings and on growth of grey pine aphid

Effects of nitrogen availability on secondary compounds, mycorrhizal infection, and aphid growth of 1-year-old Scots pine (Pinus sylvestris L.) seedlings were studied during one growing season. Seedlings were fertilized with nutrient solutions containing low, optimum, and two elevated (2 × and 4 × optimum) levels of NH4NO3. At the end of growing season foliar nitrogen concentration, needle biomass, needle length, water contents of needles, root collar diameter, and number of buds increased with enhanced nitrogen availability. Addition of nitrogen did not have effect on concentrations of monoterpenes in growing needles, but in mature needles significantly decreased concentrations of some individual and total monoterpenes were detected. In growing needles the concentrations of some individual resin acids decreased, and in mature needles concentrations of some individual and total resin acids increased with increased nitrogen fertilization. Higher numbers of resin ducts were found in mature needles with nitrogen fertilization. Nitrogen fertilization decreased total phenolic concentrations in growing and mature needles of the current year, but in needles of the previous year no significant differences occurred. Mycorrhizal infection was highest at medium (optimum and 2 × optimum) nitrogen fertilization levels. The relative growth rate (RGR) of grey pine aphid [Schizolachnus pineti (F.)] responded positively to the increase in foliar nitrogen content. However, the increase in aphid performance between optimum and the highest fertilization level was slight. This may indicate a deterring effect of resin acids on aphids. The results indicate that carbon/nutrient balance hypothesis fails to predict directly the effects of nitrogen availability on concentrations of carbon-based defensive compounds in mature foliage. Altered nitrogen supply affects allocation to secondary metabolites differently, depending on the developmental state of the plant and the biosynthesis pathway, cost of synthesis, and storage of compounds.

Variation in ozone sensitivity among clones of Betula pendula and Betula pubescens

Forty clones of Betula pendula and 6 clones of Betula pubescens, originating from southern and central Finland, were ranked in order of ozone sensitivity according to visible injuries, growth and leaf senescense under low ozone exposure. The plants were fumigated in natural climatic conditions using an open-air exposure system during two growing seasons. Control plants were grown under ambient air, and the elevated-ozone exposures were 1.6x the ambient in 1994 and 1.7x the ambient in 1995. The differences in ozone sensitivity among clones were large. Ozone tolerance was related to thicker leaves and higher stomatal density as compared to sensitive clones. Ultrastructural ozone-induced symptoms were found in chloroplasts of sensitive clones. Increased number of visibly injured leaves on fumigated plants was correlated with reduced leaf formation, foliage area, shoot dry wt and number of stomata, and increased yellowing of leaves. The results suggest that a considerable proportion of birch trees, showing high sensitivity to ozone, are at risk if ambient ozone exposures increase.

Membrane permeability response of lichen Bryoria fuscescens to wet deposited heavy metals and acid rain

Abstract Lichen Bryoria fuscescens were treated in the field experiment with heavy metal solutions, containing a mixture of Cu 2+ and Ni 2+ ions alone or in combination with acidity (H 2 SO 4 , pH 3), to study the effects of simulated acid rain and heavy metal deposition on the membrane integrity of lichen. Membrane responses to pollutant treatments were assessed by measurements of the leachate conductivity, the K + leakage, and the ergosterol concentration. Both Cu and Ni concentrations of lichen were significantly increased by metal treatments. In addition a significant reduction of Ni with acid solution was found. The conductivity of leachate was significantly increased by the acidity but not affected by the metal treatments alone. Increased thallus metal concentrations (>400 μg g −1 Cu, >100 μg g −1 Ni) significantly increased K + leakage and reduced ergosterol concentration. The results suggest that the differences in the membrane responses are associated with the different sensitivity of algal and fungal partners to the acidity of heavy metal solutions.

The influence of elevated CO2 and O3 concentrations on Scots pine needles: changes in starch and secondary metabolites over three exposure years

Abstract Scots pine (Pinus sylvestris L.) trees, aged about 20 years old, growing on a natural pine heath were exposed to two concentrations of CO2 (ambient CO2 and double-ambient CO2) and two O3 regimes (ambient O3 and double-ambient O3) and their combination in open-top chambers during growing seasons 1994, 1995 and 1996. Concentrations of foliar starch and secondary compounds are reported in this paper. Starch concentrations remained unaffected by elevated CO2 and/or O3 concentrations during the first 2 study years. But in the autumn of the last study year, a significantly higher concentration of starch was found in current-year needles of trees exposed to elevated CO2 compared with ambient air. There were large differences in concentrations of starch and secondary compounds between individual trees. Elevated concentrations of CO2 and/or O3 did not have any significant effects on the concentrations of foliar total monoterpenes, total resin acids or total phenolics. Significantly higher concentrations of monoterpenes and resin acids and mostly lower concentrations of starch were found in trees growing without chambers than in those growing in open-top chambers, while there were no differences in concentrations of total phenolics between trees growing without or in chambers. The results suggest that elevated concentrations of CO2 might increase foliar starch concentrations in Scots pine, while secondary metabolites remain unaffected. Realistically elevated O3 concentrations do not have clear effects on carbon allocation to starch and secondary compounds even after 3 exposure years.

Differences in growth, leaf senescence and injury, and stomatal density in birch (Betula pendula Roth.) in relation to ambient levels of ozone in Finland.

The differences in growth, leaf senescence, visible ozone injuries and stomatal density between one coastal site (natural ozone) and two inland sites (natural and elevated ozone) in Finland were determined for saplings of Betula pendula clones grown under open-field conditions during two growing seasons. Responses in growth, leaf senescence, visible injuries, and stomatal density were determined in relation to cumulative ozone exposure accumulated over the thresholds of 30, 40 and 50 ppb (10(9)) during the exposure period. In addition, the effects of the different ozone exposures on ultrastructure of chloroplasts were studied. Increasing ozone exposure resulted in reduced shoot dry weight, stimulated (first year) or reduced (second year) height growth, accelerated autumn yellowing of leaves, increased stomatal density, visible symptoms and chloroplast injuries, and increased number and size of plastoglobuli. Newly expanded mature leaves in midsummer were more sensitive to ozone episodes than younger developing leaves in the early growing season. In most parameters, the best correlation was achieved with the exposure index AOT30. Ozone risk for birch is highest in the southern coastal area of Finland, where background ozone concentrations are higher than in inland sites.

Emissions of volatile organic compounds and leaf structural characteristics of European aspen (Populus tremula) grown under elevated ozone and temperature

Northern forest trees are challenged to adapt to changing climate, including global warming and increasing tropospheric ozone (O(3)) concentrations. Both elevated O(3) and temperature can cause significant changes in volatile organic compound (VOC) emissions as well as in leaf anatomy that can be related to adaptation or increased stress tolerance, or are signs of damage. Impacts of moderately elevated O(3) (1.3x ambient) and temperature (ambient + 1 degrees C), alone and in combination, on VOC emissions and leaf structure of two genotypes (2.2 and 5.2) of European aspen (Populus tremula L.) were studied in an open-field experiment in summer 2007. The impact of O(3) on measured variables was minor, but elevated temperature significantly increased emissions of total monoterpenes and green leaf volatiles. Genotypic differences in the responses to warming treatment were also observed. alpha-Pinene emission, which has been suggested to protect plants from elevated temperature, increased from genotype 5.2 only. Isoprene emission from genotype 2.2 decreased, whereas genotype 5.2 was able to retain high isoprene emission level also under elevated temperature. Elevated temperature also caused formation of thinner leaves, which was related to thinning of epidermis, palisade and spongy layers as well as reduced area of palisade cells. We consider aspen genotype 5.2 to have better potential for adaptation to increasing temperature because of thicker photosynthetic active palisade layer and higher isoprene and alpha-pinene emission levels compared to genotype 2.2. Our results show that even a moderate elevation in temperature is efficient enough to cause notable changes in VOC emissions and leaf structure of these aspen genotypes, possibly indicating the effort of the saplings to adapt to changing climate.

Climatic warming increases isoprene emission from a subarctic heath

Emissions of isoprene, a reactive hydrocarbon, from Subarctic vegetation are not well documented. However, the Arctic is likely to experience the most pronounced effects of climatic warming, which may increase temperature-dependent isoprene emission. Here, we assessed isoprene emission from a Subarctic heath subjected to a 3-4 degrees C increase in air temperature and mountain birch (Betula pubescens ssp. czerepanovii) litter addition for 7-8 yr, simulating climatic warming and the subsequent expansion of deciduous shrub species and migration of the treeline. The measurements were performed using the dynamic chamber method on a wet heath with a mixture of shrubs, herbs and graminoids. Isoprene emissions averaged across the treatments were 36 +/- 5 microg m(-2) h(-1) in 2006 and 58 +/- 7 microg m(-2) h(-1) in 2007. The experimental warming increased the emissions by 83% in 2007 (P = 0.021) and by 56% in 2006 (P = 0.056), while litter addition had no significant effects. The net ecosystem CO(2) exchange was significantly decreased by warming in 2007. These results show that isoprene emissions from Subarctic heaths are comparable to those from Subarctic peatlands. Climatic warming will increase the emissions, and the amount of carbon lost as isoprene, from Subarctic heath ecosystems.

Responses of spruce seedlings (Picea abies) to exhaust gas under laboratory conditions--II. Ultrastructural changes and stomatal behaviour.

This study examines the effects of exhaust gas exposure on the epistomatal wax structure and mesophyll ultrastructure in needles of Norway spruce (Picea abies (L.) Karst.) seedlings. Stomatal diffusive resistance was also measured. Two independent exhaust gas fumigations were performed: 100 and 200 ppb measured as NO(x), for 10 days and 50, 100 and 200 ppb NO(x) for 19 days. The obstructive effect of exhaust gas exposure on epistomatal wax tubules was apparent. The stomata became covered by flat and solid wax resulting from the structural degradation of the wax crystalloids. Increasing the exhaust gas concentration in the chamber atmosphere exacerbated the degradation of the wax structure. Exhaust gas exposure induced aggregation and electron translucence of plastoglobuli, swelling of thylakoids, increase of cytoplasmic lipids and slight increase of vesiculation of cytoplasm in mesophyll cells of current and previous year needles. These changes were exemplified in current year needles. Damage to the epicuticular waxes and mesophyll ultrastructure of spruce needles most likely reflects the NO(x) and volatile hydrocarbon fraction. The alterations in epicuticular waxes and mesophyll ultrastructure can be related to accelerated senescence of the youngest, photosynthetically active, needle generation. The exhaust gas also resulted in decreased diffusive stomatal resistance at night which indicates that the exhaust gas exposure disturbed the gas exchange of spruce seedlings. The results show that even relatively short-term exposure to realistic concentrations of exhaust gas in the atmosphere can induce rather severe injuries to the needle surface structure as well as ultrastructure at the cellular level.

Differences in leaf characteristics between ozone-sensitive and ozone-tolerant hybrid aspen (Populus tremula × Populus tremuloides) clones

The authors analyzed a suite of leaf characteristics that might help to explain the difference between ozone-sensitive and ozone-tolerant hybrid aspen (Populus tremula L. x Populus tremuloides Michx.) clones. An open-field experiment comprising ambient ozone and 1.5x ambient ozone concentration (about 35 ppb) and two soil nitrogen regimes (60 and 140 kg N ha(-1) year(-1)) was conducted over two growing seasons on potted plants of eight hybrid aspen clones. Four of the clones had previously been determined to be ozone sensitive based on impaired growth in response to elevated ozone concentration. Photosynthetic rate, chlorophyll fluorescence, and concentrations of chlorophyll, protein and carbohydrates were analyzed three times during the second growing season, and foliar phenolic concentrations were measured at the end of the second growing season. Nitrogen amendment counteracted the effects of ozone, but had no effect on growth-related ozone sensitivity of the clones. Ozone-sensitive clones had higher photosynthetic capacity and higher concentrations of Rubisco and phenolics than ozone-tolerant clones, but the effects of ozone were similar in the sensitive and tolerant groups. Nitrogen addition had no effect on phenolic concentration, but elevated ozone concentration increased the concentrations of chlorogenic acid and (+)-catechin. This study suggests that condensed tannins and catechin, but not salicylates or flavonol glycosides, play a role in the ozone tolerance of hybrid aspen.

Ozone effects on Sphagnum mosses, carbon dioxide exchange and methane emission in boreal peatland microcosms

Microcosms of a boreal peatland originating from an oligotrophic fen in Eastern Finland were fumigated under four ozone concentrations (0, 50, 100 and 150 ppb O3) in laboratory growth chambers during two separate experiments (autumn and summer) for 4 and 6 weeks, respectively. Ozone effects on Sphagnum mosses and the fluxes of carbon dioxide and methane were evaluated. In both experiments, the three Sphagnum species studied showed only a few significant responses to ozone. In the autumn experiment, membrane permeability of S. angustifolium, measured as conductivity and magnesium leakage, was significantly higher under ozone fumigation (P = 0.005 and < 0.001, respectively), and there was a distinct dose-dependence. S. magellanicum showed no clear responses, either for membrane leakage or pigment content. There were no substantial ozone responses in the gross photosynthesis or net CO2 exchange during the 6-week-long summer experiment, but dark ecosystem respiration was transiently increased by ozone concentration of 100 ppb after 14 days of exposure (P < 0.05). Fumigation with 100 ppb of ozone, however, more than doubled (P < 0.05) methane emission from the peatland monoliths. Our results suggest that increasing tropospheric ozone concentration may cause substantial changes in the carbon gas cycling of boreal peatlands, even though these changes are not closely associated with the changes in Sphagnum vegetation.