Ozone exposure results in various carry-over effects and prolonged reduction in biomass in birch (Betula pendula Roth)

In the first experiment, saplings of ozone-sensitive and a more tolerant clone of Betula pendula Roth were exposed to ambient ozone (control treatment, accumulated exposure over a threshold 40 nmol mol ^{? 1} (AOT40) exposure of 1·0 μmol mol ^{? 1} h) and 1·5 × ambient ozone (elevated-ozone treatment, AOT40 of 17·3 μmol mol ^{? 1} h) over one growing season, 1996. After over-wintering, the dormant elevated-ozone saplings were transferred to the control blocks and assessed for short-term carry-over effects during the following growing season. In the second experiment, three sensitive, four intermediate and three tolerant clones were grown under ambient ozone (control treatment, AOT40 of 0·5–0·8 μmol mol ^{? 1} h per growing season) and 1·6–1·7 × ambient ozone (elevated-ozone treatment, AOT40 of 18·3–18·6 μmol mol ^{? 1} h per growing season) from May 1994 until May 1996, and were assessed for long-term carry-over effects during growing season 1997, after a 12–16 months recovery period. Deleterious short-term carry-over effects of ozone exposure included reduced contents of Rubisco, chlorophyll, carotenoids, starch and nutrients in leaves, lower stomatal conductance, and decreased new shoot growth and net assimilation rate, followed by a 7·5% (shoot dry weight (DW)), 15·2% (root DW) and 23·2% (foliage area) decreased biomass accumulation and yield over the long term, including a reduced root : shoot ratio. However, a slow recovery of relative growth rates during the following two seasons without elevated ozone was apparent. Several long-lasting structural, biochemical and stomatal acclimation, stress-defence and compensation reactions were observed in the ozone-tolerant clone, whereas in the sensitive clone allocation shifted from growth towards defensive phenolics such as chlorogenic acid. The results provide evidence of persistent deleterious effects of ozone which remain long after the ozone episode.     

Physiological, stomatal and ultrastructural ozone responses in birch (Betula pendula Roth.) are modified by water stress

The physiological, stomatal and ultrastructural responses to ozone and drought of ozone-sensitive and more ozone-tolerant birch ( Betula pendula Roth.) clones were studied singly and in combination, in a high-stress chamber experiment and in a low-stress open-field experiment. In the chamber experiment, well watered (WW), moderately watered (MW) or drought-stressed (DS) saplings were exposed for 36 d to 0 or 130 nmol mol^∠1 ozone. In the open-field experiment, well watered or drought-stressed saplings were grown for one growing season in ambient air or exposed to 1·8 × ambient ozone. Drought stress reduced growth rate, stomatal conductance, stomatal density and the proportion of starch and thylakoids in chloroplasts, but stimulated net photosynthesis, Rubisco and chlorophyll quantity at the end of the growing season, and increased the size and density of plastoglobuli. Ozone fumigations caused more variable, clone- and exposure-dependent responses in growth, decreased stomatal conductance and net photosynthesis, an increased number of stomata, visible and ultrastructural chloroplast injuries, and enhanced autumn yellowing of the leaves. Ozone-induced changes in plastoglobuli, starch and thylakoids resembled drought responses. The two experiments revealed that, depending on the experimental conditions and the variable, the response to drought and ozone stress can be independent, additive or interactive. Drought protected the plants from ozone injuries under high-stress conditions in the chamber experiment. In the low-stress, open-field experiment, however, enhanced ozone damage was observed in birch saplings grown under restricted water supply.     

Growth response to step-decrease in nutrient availability in small birch (Betula pendula Roth)

Abstract Changes in the uptake and allocation of carbon and nitrogen, after a step-decrease in nutrient availability, were investigated in small birch (Betula pendula Roth). By demonstrating stable nutrition, before and after the decrease in nutrient supply, it was possible to eliminate the effects of plant size and age. Immediately following the step-decrease in nutrient availability, net nitrogen uptake to leaves and the relative rate of increase in shoot area tended to zero. Although photosynthetic rate per shoot area decreased, carbon uptake remained in excess of that used in structural growth and respiration. More of the excess carbon was accumulated as starch in leaves than in roots. After a lag phase, the relative rates of increase in plant dry matter, starch amount, net nitrogen uptake to leaves and shoot area development equalled that of the reduced rate of nutrient supply. It is concluded that the reduction in plant relative growth rate was much more attributable to the reduced allocation of photosynthate to leaf area growth than to the reduction in photosynthesis per shoot area.     

Responses of selected birch (Betula pendula Roth) clones to ozone change over time

A long-term free air ozone fumigation experiment was conducted to study changes in physiological ozone responses during tree ontogeny and exposure time in ozone sensitive and tolerant clones of European white birch (Betula pendula Roth), originated from south and central Finland. The trees were grown in soil in natural microclimatic conditions under ambient ozone (control) and 1.4-1.7 x ambient (elevated) ozone from May 1996 to October 2001, and were measured for stem and foliage growth, net photosynthesis, stomatal conductance, stomatal density, visible injuries, foliar starch content and bud formation. After 6 years of exposure, the magnitude of ozone-induced growth reductions in the sensitive clone was 12-48% (significant difference), levels similar or greater than those reported earlier for 2- and 3-year-old saplings undergoing shorter exposures. In the tolerant clone, growth of these larger trees was reduced by 1-38% (significant difference in stem volume), although the saplings had previously been unaffected. In both clones, ozone stress led to significantly reduced leaf-level net photosynthesis but significantly increased stomatal conductance rates during the late summer, resulting in a lower carbon gain for bud formation and the onset of visible foliar injuries. Increasing ozone sensitivity with duration of exposure was explained by a change in growth form (relatively reduced foliage mass), a lower photosynthesis to stomatal conductance ratio during the late summer, and deleterious carry-over effects arising from the reduced number of over-wintering buds.     

Somatic embryogenesis in cell cultures of birch (Betula pendula Roth.)

Somatic embryogenesis was induced in cell cultures of birch (Betula pendula Roth.) derived from juvenile tissue of seed embryos and from mature leaf tissue. Embryos were formed in liquid and on solidified medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-furfurylaminopurine (kinetin). Sometimes somatic embryos formed only after transfer to medium devoid of growth regulators. The embryos germinated on hormone-free medium and were potted in soil and grown in the greenhouse.Finnish Forest Research InstituteUniversity of Helsinki, School of Pharmacy;     

Dependence of starch storage on nutrient availability and photon flux density in small birch Betula pendula Roth)

Abstract Small birch plants (Betula pendula Roth) were grown in a climate chamber at different levels of nutrient availability and at two photon flux densities. The extent to which starch storage was dependent upon nutrient availability and photon flux density was investigated. Acclimated values of starch concentration in leaves were highest at low nutrient availability and high photon flux density. Starch storage in roots was only found at the lowest nutrient availability. However, the relative rate of starch storage (starch stored per unit plant dry weight and time) was higher in plants with good nutrition. The data suggest that, at sub-optimal nutrient availability, the momentary rate of net shoot photosynthesis is unlikely to limit the structural (as opposed to carbon storage) growth of the plant. Although photosynthetic rate per unit leaf area (as measured at the growth climate) was slightly lower in plants with poor nutrient availability, photosynthetic rate per unit leaf nitrogen was higher. These data suggest a priority of leaf nitrogen usage in photosynthesis, with limiting amounts of leaf nitrogen (and possibly other nutrients) for subsequent growth processes. This argument is consistent with the higher concentrations of starch found in plants with poor nutrient availability.     

Characterization of a birch (Betula pendula Roth.) embryogenic gene,BP8

We have isolated the birch homologue (BP8) for the carrot embryogenic gene DC8 by heterologous hybridization. The birch BP8 gene encodes a putative protein of 53 kDa, showing 52% sequence identity with the DC8 gene at the amino acid level. The putative BP8 protein contains 20 repeats of 11 amino acids and thus belongs to the group of LEA proteins isolated from such plants as carrot, cotton and wheat. Northern hybridization of mRNA isolated from birch cells representing different stages of somatic embryogenesis and non-embryogenetic material with a PB8 probe gave no signals, suggesting a low expression level of the BP8 gene.     

Dependence of amino acid composition upon nitrogen availability in birch (Betula pendula)

Small birch plants ( Betula pendula Roth .) were grown at different rates of exponentially increasing nitrogen supply. This resulted in plants with different relative growth rates and different internal nitrogen concentrations. Within a nitrogen treatment, both of these variables remained constant with time.
Free amino acids were measured in leaves and roots of the seedlings at two different harvests. At greater nitrogen supply, higher concentrations of total amino acid nitrogen were found in roots and leaves. The ratio of amino acid nitrogen to total nitrogen was low albeit greater at higher nitrogen supply. Higher concentrations of amino acid nitrogen were mainly due to high concentrations of citrulline, glutamine, γ-aminobuitric acid and arginine.
Greater leaf concentrations of amino acid nitrogen at higher nitrogen supply may be related lo increased concentrations in the xylem sap and/or may be indicative of small excesses of nitrogen with respect to current nitrogen usage in protein synthesis.     

Structural characteristics and chemical composition of birch (Betula pendula) leaves are modified by increasing CO2 and ozone

Impacts of ozone and CO₂ enrichment, alone and in combination, on leaf anatomical and ultrastructural characteristics, nutrient status and cell wall chemistry in two European silver birch (Betula pendula Roth) clones were studied. The young soil‐growing trees were exposed in open‐top chambers over three growing seasons to 2 × ambient CO₂ and/or ozone concentrations in central Finland. The trees were measured for changes in altogether 35 variables of leaf structure, nutrients and cell wall chemistry of green leaves, and 20 of the measured variables were affected by CO₂ and/or O₃. Elevated CO₂ increased the size of chloroplasts and starch grains, number of mitochondria, P : N ratio, and contents of cell wall hemicellulose. Elevated CO₂ decreased the total leaf thickness, specific leaf area, concentrations of N, K, Cu, S and Fe, and contents of cell wall α‐cellulose, uronic acids, acid‐soluble lignin and acetone‐soluble extractives. Elevated ozone led to thinner leaves, higher palisade to spongy ratio, increased number of peroxisomes and mitochondria, reduced content of Mn, Zn, Cu, hemicellulose and uronic acids, and lower Mn : N and Zn : N ratios. In the combined exposure, interactions were antagonistic. Ultrastructural changes became more evident towards the end of the exposure. Young leaves were tolerant against ozone‐caused oxidative stress, whereas oxidative H₂O₂ accumulation was found in older leaves. CO₂ enrichment improved ozone tolerance not only through increased photosynthesis rates, but also through changes in cell wall chemistry (hemicellulose, in particular). However, nutrient imbalances due to ozone and/or CO₂ may predispose the trees to other biotic and abiotic stresses. Down‐regulation and up‐regulation of photosynthesis under elevated CO₂ through anatomical changes is discussed.     

Response of small birch plants (Betula pendula Roth.) to elevated CO2 and nitrogen supply

Small birch plants were grown for up to 80 d in a climate chamber at varied relative addition rates of nitrogen in culture solution, and at ambient (350 μmol mol^-1) or elevated (700 μmol mol^-1) concentrations of CO₂. The relative addition rate of nitrogen controlled relative growth rate accurately and independently of CO₂ concentration at sub-optimum levels. During free access to nutrients, relative growth rate was higher at elevated CO₂. Higher values of relative growth rate and net assimilation rate were associated with higher values of plant N-concentration. At all N-supply rates, elevated CO₂ resulted in higher values of net assimilation rate, whereas leaf weight ratio was independent of CO₂. Specific leaf area (and leaf area ratio) was less at higher CO₂ and at lower rates of N-supply. Lower values of specific leaf area were partly because of starch accumulation. Nitrogen productivity (growth rate per unit plant nitrogen) was higher at elevated CO₂. At sub-optimal N-supply, the higher net assimilation rate at elevated CO₂ was offset by a lower leaf area ratio. Carbon dioxide did not affect root/shoot ratio, but a higher fraction of plant dry weight was found in roots at lower N-supply. In the treatment with lowest N-supply, five times as much root length was produced per amount of plant nitrogen in comparison with optimum plants. The specific fine root length at all N-supplies was greater at elevated CO₂. These responses of the root system to lower N-supply and elevated CO₂ may have a considerable bearing on the acquisition of nutrients in depleted soils at elevated CO₂. The advantage of maintaining steady-state nutrition in small plants while investigating the effects of elevated CO₂ on growth is emphasized.     

Variation of non-structural carbohydrates in silver birch (Betula pendula Roth) wood

Non-structural carbohydrates in silver birch (Betula pendula Roth) wood were analysed in a 7-year-old clone and in five mature stems. The analysis was conducted to obtain more detailed information on seasonal fluctuation of these components and of the tree-to-tree variation and within stem variation. The sugars were analysed by GLC-MS. The smallest total soluble sugar amounts (consisting of sucrose, fructose, glucose, raffinose and myo-inositol) in young trees were measured during mid-summer (ca. 0.3%) and the largest while in dormancy (ca. 1.6% on wood dry weight basis). Raffinose was detected in autumn as a minor component. The proportion of monosaccharides and the amount of myo-inositol were largest during growth. Compared to other studies silver birch showed more evident seasonal fluctuation in soluble sugars than evergreen tree species. The sugar amount in mature stems was approximately at the same level as in young trees that had the same felling time. Tree-to-tree variation in the non-structural carbohydrates in the mature wood was fairly large. However, the amount of total soluble sugars, sucrose and glucose showed significant variation within the stem. The amount of these sugars was largest in samples that were taken close to the cambium. Starch was also detected close to pith. According to the heartwood definition and starch measurement results in this paper, it could be stated that silver birch does not form heartwood.     

Source-sink relations affect growth but not the allocation pattern of birch (Betula pendula Roth) seedlings under elevated [CO2]

ABSTRACT

Birch (Betula pendula Roth.) seedlings were kept for two growing seasons under ambient (～350 µmol mol^-1) and elevated (～700 µmol mol^-1) [CO₂]. The present study was designed to examine the effects of [CO₂] and pot size on growth and carbon allocation under conditions of non-limiting water and nutrient supply, in order to separate the effects of source-sink interaction from the effects of nutrient deficiency. The manipulation of the source-sink relations had a strong influence on the growth response to elevated [CO₂]. When the rooting volume was inadequate, it resulted in a source-sink imbalance which constrained growth under elevated [CO₂]. When root exploration was unconstrained, total dry mass was significantly increased (by about 24%) under elevated [CO₂]. However, the allometric relationships in allocation pattern and in morphogenetic development were not affected by either [CO₂] or pot treatments when the saplings were of the same size. Thus, by constraining dry mass production, small sinks affected the magnitude of the growth responses to elevated [CO₂], but did not affect the plant allocation pattern and allometric relationships when nutrient supply was non-limiting. However, by slowing down growth, sink restrictions counteract the speed-up of ontogeny which is the main effect of elevated [CO₂] on tree growth.     

The consequences of leaf damage for subsequent insect grazing on birch (Betula spp.)

Summary Low levels of artificial damage were imposed on the leaves of Betla pubescens and B. pendula in early summer 1985. Sixteen tress were used and the damage was distributed throughout the canopy in two ways. It was either randomly dispersed on the tree, or restricted to a localised region of the canopy. Up to 250 leaves/tree were damaged and the position of control leaves was marked as appropriate. At the end of the summer the experimental and control leaves were collected for analysis. Subsequent patterns of grazing by naturally-occurring herbivores were affected by the previous damage to the foliage. Fewer artificially-damaged leaves received subsequent insect grazing damage. The frequecy distribution of insect damage to previously-damaged leaves was significantly different from that of the control leaves, and less leaf tissue was removed from those experimental leaves which did receive subsequent insect attack. The evidence presented suggests that wound-induced changes in leaf palatability do occur in B. pubescens and B. pendula in the field and that a major role is to disperse the damage throughout the canopy.     

Effects of ultraviolet-B radiation on growth, mycorrhizas and mineral nutrition of silver birch (Betula pendula Roth) seedlings grown in low-nutrient conditions

The effects of increased ultraviolet‐B (UV‐B) radiation on the growth, mycorrhizas and mineral nutrition of silver birch (Betula pendula Roth) seedlings were studied in greenhouse conditions. Seedlings—planted in a birch‐forest top soil and sand substrate—were grown without additional nutrient supply. Ultraviolet treatment started immediately after the seedlings emerged and the daily integrated biologically effective UV‐B irradiance on the UV‐B‐treated plants was equivalent to a 25% depletion of stratospheric ozone under clear sky conditions. Visible symptoms of UV‐B damage or nutrient deficiency were not observed throughout the experiment. Seedling height and dry weight (DW) (measured after 58 days and 76 days of treatment) were not affected by increased UV‐B. However, a significant shift in DW allocation toward roots resulted in a lower shoot/root ratio and leaf area ratio in UV‐B‐treated plants compared to control plants. At the first harvest (after 58 days of treatment), the percentage of various mycorrhizal morphotypes and the number of short roots per unit of root length or weight were not affected by increased UV‐B despite significantly increased DW allocation toward roots. Subtle reduction in the allocation of nitrogen (N) to leaves and increased allocation of phosphorus (P) to roots may suggest cumulative effects that could affect the plant performance over the long‐term.     

Effects of elevated carbon dioxide concentration on photosynthesis and growth of small birch plants (Betula pendula Roth.) at optimal nutrition

Small birch plants (Betula pendula Roth.) were grown from seed for periods of up to 70d in a climate chamber at optimal nutrition and at present (350 μmol mol^?1) or elevated (700 μmol mol^?1) concentrations of atmospheric CO₂. Nutrients were sprayed over the roots in Ingestad-type units. Relative growth rate and net assimilation rate were slightly higher at elevated CO₂, whereas leaf area ratio was slightly lower. Smaller leaf area ratio was associated with lower values of specific leaf area. Leaves grown at elevated CO₂ had higher starch concentrations (dry weight basis) than leaves grown at present levels of CO₂. Biomass allocation showed no change with CO₂, and no large effects on stem height, number of side shoots and number of leaves were found. However, the specific root length of fine roots was higher at elevated CO₂. No large difference in the response of carbon assimilation to intercellular CO₂ concentration (A/C_i curves) were found between CO₂ treatments. When measured at the growth environments, the rates of photosynthesis were higher in plants grown at elevated CO₂ than in plants grown at present CO₂. Water use efficiency of single leaves was higher in the elevated treatment. This was mainly attributable to higher carbon assimilation rate at elevated CO₂. The difference in water use efficiency diminished with leaf age. The small treatment difference in relative growth rate was maintained throughout the experiment, which meant that the difference in plant size became progressively greater. Thus, where plant nutrition is sufficient to maintain maximum growth, small birch plants may potentially increase in size more rapidly at elevated CO₂.