The effect of elevated carbon dioxide and fertilization on primary and secondary metabolites in birch,Betula pendula (Roth)

Seedlings of European white birch (Betula pendula Roth) were grown in growth chambers for one growth season under four carbon dioxide regimes (350, 700, 1050 and 1400 ppm) and at three fertilization levels (0, 100 and 500 kg ha^–1 monthly). The soluble carbohydrates and secondary phenolics in the leaves and stems were analysed. It was found that fertilizer addition reduced the amounts of glucose and fructose while sucrose remained almost unaffected. The sugar content of leaves increased at 700 ppm and 1050 ppm of CO₂ and decreased at the highest CO₂ concentration (1400 ppm). The amounts of proanthocyanidins and flavonoids in leaves decreased with fertilization addition and increased with CO₂ enrichment. The production of simple phenolic glucosides varied according to the fertilization and CO₂ treatments. The triterpenoid content of stems seemed to increase with fertilization and CO₂-addition. Our results indicate that the production of phytochemicals in the birch seedlings is very sensitive to both fertilization and CO₂ addition, which is in agreement with earlier studies, and thus provide some support for the hypothesis of carbon allocation to plant defence when there is an excess of carbon and nutrient. The considerable variation in the production of secondary components may indicate that the synthesis of these defensive metabolites can be regulated by a plant to certain extent, depending on the ability of the plant to acclimate to changes in the physical environment.     

The effects of long-term elevated UV-B on the growth and phenolics of field-grown silver birch (Betula pendula)

The effects of long‐term elevated UV‐B radiation on silver birch (Betula pendula Roth) seedlings were studied over three growing seasons in an outdoor experiment in Finland started 64 days after germination. One group of seedlings was exposed to a constant 50% increase in UV‐B_CIE radiation, which corresponds to 20–25% of ozone depletion; another group received a small increase in UV‐A radiation and a third (the control group) received ambient solar radiation. Changes in growth appeared during the third growing season; the stems of the UV‐B treated seedlings were thinner and their height tended to be shorter compared with that of the control seedlings. In contrast, there were no UV‐B effects on biomass, bud burst, bud dry weights, leaf area, rust frequency index or chlorophyll concentrations in any of the summers. During the three‐year study, the flavonols were significantly increased by the elevated UV‐B only in the first growing season. The responses varied greatly among individual compounds; the most induced were the quercetin glycosides, while the main flavonols, myricetins, were reduced by the UV‐A control treatment. In the second summer phenolic acids, such as 3,4′‐dihydroxypropiophenone‐3‐glucoside, neochlorogenic acid and 5‐coumarylquinic acid, were increased by the UV‐B treatment. In the third year, the constitutive concentrations of phenolics were not affected by the UV‐B treatment.     

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.     

Characterization of wound responses of stems of paper birch (Betula papyrifera) and European white birch (Betula pendula)

Plants respond to feeding injury by chewing insects by inducing both a general response to mechanical wounding and a specific response to herbivore-associated elicitors. In both cases, plant response involves complex biochemical and physiological changes. We compared chemical and physical responses of paper birch (B. papyrifera) and European white birch (B. pendula) stems to mechanical injury to determine if aspects of their wound response correspond with the much higher resistance of paper birch to bronze birch borer (Agrilus anxius). We also characterized stem responses to mechanical wounding plus bronze birch borer larval homogenate to determine if larval cues elicited a more specific response than mechanical wounding alone. In both species, wounding decreased concentrations of individual phenolics, total phenolics, and condensed tannins, perhaps because they were diverted to lignin biosynthesis, the concentration of which increased. Nitrogen concentration increased in both species while free amino acid concentrations declined, perhaps because they were utilized to synthesize proteins. Application of larval homogenate did not elicit a response different from that induced by mechanical injury. When comparing wound responses of the two birch species, phenolic profiles differed most conspicuously. However, multivariate analyses revealed no differences between constitutive and wound-induced phenolic profiles within each species, and the rate of wound periderm growth was equivalent between species. These results suggest that components of the wound response we measured may not contribute to interspecific variation in bronze birch borer resistance of paper birch and European white birch.     

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.     

Nutrition and the ozone sensitivity of birch (Betula pendula)

 Cuttings of a single birch clone (Betula pendula) were grown in field fumigation chambers throughout the growing season in either filtered air (control) or 90/40 nl O₃ l^–1 (day/night). Both regimes were split into plants under high and low nutrient supply (macro- and micronutrients). The stomatal density of leaves was increased by ozone but was lowered at high nutrition, while the inner air space was hardly affected by the treatments. Ozone induced macroscopic leaf injury regardless of nutrition, but leaf shedding was delayed in the low-fertilized plants, despite O₃ uptake being similar to that in high-fertilized plants. The leaf turn-over was enhanced in the O₃-exposed high-fertilized plants, but length growth and leaf formation of stems were not affected by ozone in either nutrient regime. Leaves of high-fertilized plants showed O₃-caused decline in photosynthetic capacity, water-use efficiency, apparent carbon uptake efficiency and quantum yield earlier as compared with low-fertilized plants, whereas chlorophyll fluorescence (F_V/F_M) and leaf nitrogen concentration were rather stable. CO₂ uptake rate and rubisco activity of young leaves compensated for the O₃ injury in the ageing leaves of the low-fertilized plants. In 8-week-old leaves, however, the O₃-induced decline in CO₂ uptake did not differ between the nutrient regimes and was associated with increased dark respiration rather than changed photorespiration. The balance between CO₂ supply and demand was lost, as was stomatal limitation on CO₂ uptake. High nutrition did not help leaves to maintain a high photosynthetic capacity and life span under O₃ stress. Received: 6 July 1996 / Accepted: 4 June 1997     

Control of potassium influx in roots of birch (Betula pendula) seedlings exposed to cadmium

The nutrient status of the root may be a factor of vital importancefor plant tolerance to changes in the environment. In this studythe effect of Cd on the K+ uptake of birch plants, starved andnon-starved of potassium, was examined. Changes in potassiuminflux of birch [Betula pendula) roots, with time, were examinedin birch plants introduced to a nutrient solution containing125/jM K+ after a K+-starvation period. Cadmium was introducedduring the K+-recovery period and the effects of the heavy metalon the development of the uptake pattern was studied. When noCd was present K+ influx, in previously K+-starved plants, increasedwith time and reached a maximum after 6 h. When 2 or 5//M Cdwas supplied simultaneously with potassium to the root, K+ influxwas unchanged during an 8 h K+-recovery period, and the K concentrationin the roots did not increase as was the case when the plantswere given K+ without Cd. In another experiment Cd supply (2//M)to K+-fed plants gave an immediate 50% decrease in K+ influx.However, after prolonged exposure to Cd (up to 20 h), K+ influxrecovered to the control value. Key words: Birch, cadmium, influx, potassium, regulation     

The effects of nitrogen fertilisation and elevated CO2 on the lipid biosynthesis and carbon isotopic discrimination in birch seedlings (Betula pendula)

The effects of nitrogen (N) fertilisation and elevated [CO₂] on lipid biosynthesis and carbon isotope discrimination in birch (Betula pendula Roth.) transplants were evaluated using seedlings grown with and without N fertiliser, and under two concentrations of atmospheric CO₂ (ambient and ambient+250 μmol mol^-1) in solar dome systems. N fertilisation decreased n-fatty acid chain length (18:0/16:0) and the ratios of α-linolenate (18:2)/linoleate (18:1), whereas elevated [CO₂] showed little effect on n-fatty acid chain length, but decreased the unsaturation (18:2+18:1)/18:0. Both N fertilisation and elevated [CO₂] increased the quantity of leaf wax n-alkanes, whilst reducing that of n-alkanols by 20–50%, but had no simple response in fatty acid concentrations. ¹³C enrichment by 1–2.5‰ under N fertilisation was observed, and can be attributed to both reduced leaf conductance and increased photosynthetic consumption of CO₂. Individual n-alkyl lipids of different chain length show consistent pattern of δ¹³C values within each homologue, but are in general 5–8‰ more depleted in ¹³C than the bulk tissues. Niether nitrogen fertilisation and elevated CO₂ influenced the relationship between carbon isotope discrimination of the bulk tissue and the individual lipids. This revised version was published online in June 2006 with corrections to the Cover Date.     

The effect of elevated CO2 and temperature on the secondary chemistry of Betula pendula seedlings

The effect of elevated atmospheric CO₂ and temperature on resource allocation and secondary chemistry of white birch (Betula pendula Roth) under a non-limiting nutrient and water supply was investigated. Birch seedlings were grown in closed-top chambers exposed to ambient CO₂ and temperature, elevated atmospheric CO₂ (700 ppm), elevated temperature (2°C above ambient) and a combination of elevated CO₂ and temperature for one growing season. Elevated CO₂ significantly increased the total biomass of the seedlings. The combined effect of the elevated CO₂ and temperature treatments further increased the total biomass, but not significantly. The content of nitrogen and water decreased, while some secondary compounds (such as condensed tannins and flavonol glycosides) increased in leaves subjected to CO₂ enrichment. Elevated temperature increased the concentration of total flavone aglycones and decreased that of total HPLC-phenolics in the leaves, due to the decrease in individual flavonol glycosides, cinnamoylquinic acids and (+)-catechin. There were no significant interactive effects between CO₂ and temperature in the phenolic concentrations of the leaves and in the stems, while the number of resin droplets in the top part of the stems showed significant interaction. This clearly implies that carbon allocation into secondary metabolites in the leaves and stems differ under enhanced CO₂ and temperature, and the combined effect of CO₂ and temperature on the herbivore resistance of birches, is lower than that of CO₂ and temperature alone.     

Allocation of carbon to growth and secondary metabolites in birch seedlings under UV-B radiation and CO2 exposure

In plants, the allocation of carbon to secondary metabolites has been shown to be determined by both the availability of resources (e.g., CO₂ concentration) and by specific stress factors (e.g., ultraviolet [UV]‐radiation). It has been suggested that, in combination, CO₂ and UV‐B radiation may differentially affect plant growth and morphogenic parameters, and elevated CO₂ may ameliorate the effects of UV‐B radiation. In the present work, the effects of increased atmospheric CO₂ concentration and UV‐B radiation on growth and the accumulation of different types of secondary metabolites were studied in silver birch (Betula pendula Roth). Seedlings were exposed to 350 and 700 μmol mol^?1 of CO₂ in a greenhouse. At both CO₂ levels, additional UV‐B was either present (8.16 kJ m^?2 day^?1 of biologically effective UV‐B irradiance) or absent. The time course of accumulation of individual secondary compounds and the shifts in allocation of carbon between biomass and the secondary metabolites (phenolic acids, flavonoids, condensed tannins) were studied during a 1‐month‐long exposure. Additionally, the activities of enzymes ( l ‐phenylalanine ammonia‐lyase [PAL], EC 4.3.1.5; peroxidase, EC 1.11.1.7; polyphenol oxidase, EC 1.10.3.1) were determined for leaves. UV‐B radiation significantly increased biomass, PAL activity, and the accumulation of phenolic acids and flavonoids in seedlings. Elevated CO₂ concentration increased the activities of all the enzymes studied and the accumulation of condensed tannins in leaves, especially with UV‐B radiation. Because the observed UV‐B induction of flavonoids was smaller under a high CO₂ concentration, it was suggested that the excess of carbon in the atmosphere may moderate the effect of UV‐B by increasing the metabolic activity of leaves (high enzyme activities) and by changing the allocation of internal carbon between different primary and secondary metabolites in the plant. Our results demonstrate the significant increase in the allocation of carbon to secondary metabolites without any large change in growth due to the elevation of CO₂ concentration and UV‐B radiation. There also was a stronger impact of CO₂ than UV‐B on the phenolic metabolism of birch seedlings.     

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.     

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.     

Combination treatment of elevated UVB radiation,CO2 and temperature has little effect on silver birch (Betula pendula) growth and phytochemistry

Elevations of carbon dioxide, temperature and ultraviolet‐B (UBV) radiation in the growth environment may have a high impact on the accumulation of carbon in plants, and the different factors may work in opposite directions or induce additive effects. To detect the changes in the growth and phytochemistry of silver birch (Betula pendula) seedlings, six genotypes were exposed to combinations of ambient or elevated levels of CO₂, temperature and UVB radiation in top‐closed chambers for 7 weeks. The genotypes were relatively similar in their responses, and no significant interactive effects of three‐level climate factors on the measured parameters were observed. Elevated UVB had no effect on growth, nor did it alter plant responses to CO₂ and/or temperature in combined treatments. Growth in all plant parts increased under elevated CO₂, and height and stem biomass increased under elevated temperature. Increased carbon distribution to biomass did not reduce its allocation to phytochemicals: condensed tannins, most flavonols and phenolic acids accumulated under elevated CO₂ and elevated UVB, but this effect disappeared under elevated temperature. Leaf nitrogen content decreased under elevated CO₂. We conclude that, as a result of high genetic variability in phytochemicals, B. pendula seedlings have potential to adapt to the tested environmental changes. The induction in protective flavonoids under UVB radiation together with the positive impact of elevated CO₂ and temperature mitigates possible UVB stress effects, and thus atmospheric CO₂ concentration and temperature are the climate change factors that will dictate the establishment and success of birch at higher altitudes in the future.     

Within-stand variation in silver birch (Betula pendula Roth) phenology

Key message

Within a local population genotypes differ in the timing of bud burst, but genotypes with early bud burst unfold their leaves slower, resulting in an equal period of carbon gain.

Abstract

The ability of local populations to cope with disturbances like adverse weather events or a changing climate depends on the genotypic richness of such populations, emphasising the importance of differences between genotypes in traits related to growth and survival at this scale. Due to their longevity, these differences are of special importance in trees, yet for trees, differences between genotypes within local populations remain unexplored. The phenological cycle is important in this respect, since a correct timing of phenological events is critical for growth and survival of trees, especially in environments with strong seasonality and changes in the timing of phenological events has consequences for, among others, net ecosystem productivity and the climate system as a whole. In this light accounting for differences in the timing of phenological events within species is currently identified as a research challenge. This study contributes to the knowledge of differences between genotypes on the small spatial scale of a local population. We examined the timing of phenological events of 15 micropropagated silver birch (Betula pendula Roth) genotypes representing a natural population. Measurements covered bud burst (7 years) and leaf unfolding in spring and chlorophyll degradation in autumn (2 years for both). These data were used to estimate the period of carbon gain. Differences between genotypes in the temperature sum required for bud burst were present, with genotypes showing ‘early’ (i.e. a low temperature sum requirement for bud burst) and ‘late’ bud burst across the 7-year study period. Differences were small in most years (i.e. 3 days), but differences of 16 days were recorded within the 7-year study period as well. Genotypes with ‘early’ bud burst were less sensitive to variations in environmental conditions in spring compared to genotypes with ‘late’ bud burst. Differences in bud burst were not carried over to the estimated period of carbon gain. Due to faster leaf expansion in genotypes with ‘late’ bud burst and the lack of differences between genotypes in autumn senescence the estimated period of carbon gain was similar among genotypes.     

Expression of photosynthesis- and senescence-related genes during leaf development and senescence in silver birch (Betula pendula) seedlings

Leaf development and senescence were studied in greenhouse-grown silver birch ( Betula pendula Roth) seedlings over a period of 7 weeks. Prior to the experiment, leaves from 100 seedlings were marked for five sampling dates. Timing of the developmental phases was studied with biochemical analyses of total soluble protein, Rubisco protein, chlorophyll concentration and at the level of gene expression related to photosynthesis, energy metabolism, ethylene synthesis and protein degradation. During the sampling period, an initial increase in photosynthetic capacity could be seen, when expression of the Rubisco small subunit gene ( RbcS ) and Rubisco protein (EC 4.1.1.39) were examined. Down-regulation of photosynthesis, visible as a decrease in Rubisco protein and RbcS mRNA, started soon after full expansion of the leaves and processes related to senescence followed. mRNA accumulation for the ethylene-forming enzyme 1-aminocyclopropane-1-carboxylic acid oxidase increased first during the onset of senescence. Protein degradation was observed as a loss of soluble proteins, with a simultaneous increase in the leucine aminopeptidase (EC 3.4.11.1) mRNA levels. The mRNA levels of ribonuclease-like pathogenesis-related protein 10 also increased clearly during senescence, but the mitochondrial phosphate translocator mRNA showed only a slight increase. Chlorophyll concentration of the leaves decreased after the mRNA levels of these senescence-related genes had become more abundant.     

Report of Quambalaria cyanescens in association with the birch (Betula pendula)

Long-term microbiological investigation of the pollen of silver birch (Betula pendula) in the Moscow and Moscow oblast areas revealed that almost one-third of the analyzed samples contained the fungus identified by morphological, cultural, and molecular genetic techniques as Quambalaria cyanescens (de Hoog & G.A. de Vries) Z.W. de Beer, Begerow & R. Bauer. This species was previously known mostly as a symbiont of tropical plants of the genera Eucalyptus and Corymbia and have not been isolated in Russia. We revealed a close association between Quambalaria cyanescens and silver birch. The micromycete was regularly detected in pollen samples, as well as on the inside and outside of the aments, and on the surface of leaves and branches. It was never isolated from other plant species in the investigated area. The data on the morphological and cultural characteristics of the fungus, its cell ultrastructure, and occurrence are presented, as well as the phylogenetic analysis of the isolated strains.     

Cadmium and copper interactions on the accumulation and distribution of Cd and Cu in birch (Betula pendula Roth) seedlings

The effect of different external cadmium (Cd) and copper (Cu) regimes on the concentration of Cd and Cu in roots and shoots of birch (Betula pendula Roth.) seedlings was investigated. The seedlings were grown for 12 days in a weak nutrient solution (containing all essential nutrient elements including 0.025 µM Cu) at pH 4.2 with combinations of additional 0–2 µM CdCl₂ and 0–2 µM CuCl₂. Root and shoot concentrations of Cu were decreased by Cd in all treatments which included 0.1–2 µM of additional Cu in the treatment solution. When no extra Cu was added, only the shoot concentration of Cu was decreased by Cd whereas the root concentration was not affected. The shoot concentration of Cd was decreased by 0.5 and 2 µM of additional Cu in the treatment solution. The root concentration of Cd was decreased by Cu only when the concentration of additional Cu in the treatment solution was equal to or exceeded the concentration of Cd.     

Comparisons of the efficiency of some promoters in silver birch (Betula pendula)

The efficiency of several promoters (pin2 from potato, ubiquitin from sunflower, rolC from Agrobacterium rhizogenes, act1 from rice and CaMV 35S from cauliflower mosaic virus) fused to the uidA reporter gene was measured after biolistic bombardment of birch leaves (Betula pendula L.). The highest level of β-glucuronidase (GUS) activity was achieved with the pin2 promoter and the lowest activity with the CaMV 35S promoter. The activity of the potato wound-inducible promoter (pin2) was also tested in stably transformed birch. The promoter showed induced activity after mechanical wounding and feeding by leaf weevils. The systemic effect was confirmed by enhanced GUS activity in non-wounded leaves. The results of this study indicated that the potato wound-inducible promoter maintains its function in birch and would be a suitable promoter in studies of insect-birch interaction at the molecular level. Received: 17 October 1996 / Revision received: 7 February 1997 / Accepted: 1 March 1997