Birch Leaf Research Articles

Ecological role of reindeer summer browsing in the mountain birch (Betula pubescens ssp. czerepanovii) forests: effects on plant defense, litter decomposition, and soil nutrient cycling

Mammalian herbivores commonly alter the concentrations of secondary compounds in plants and, by this mechanism, have indirect effects on litter decomposition and soil carbon and nutrient cycling. In northernmost Fennoscandia, the subarctic mountain birch (Betula pubescens ssp. czerepanovii) forests are important pasture for the semidomestic reindeer (Rangifer tarandus). In the summer ranges, mountain birches are intensively browsed, whereas in the winter ranges, reindeer feed on ground lichens, and the mountain birches remain intact. We analyzed the effect of summer browsing on the concentrations of secondary substances, litter decomposition, and soil nutrient pools in areas that had been separated as summer or winter ranges for at least 20 years, and we predicted that summer browsing may reduce levels of secondary compounds in the mountain birch and, by this mechanism, have an indirect effect on the decomposition of mountain birch leaf litter and soil nutrient cycling. The effect of browsing on the concentration of secondary substances in the mountain birch leaves varied between different years and management districts, but in some cases, the concentration of condensed tannins was lower in the summer than in the winter ranges. In a reciprocal litter decomposition trial, both litter origin and emplacement significantly affected the litter decomposition rate. Decomposition rates were faster for the litter originating from and placed into the summer range. Soil inorganic nitrogen (N) concentrations were higher in the summer than in the winter ranges, which indicates that reindeer summer browsing may enhance the soil nutrient cycling. There was a tight inverse relationship between soil N and foliar tannin concentrations in the winter range but not in the summer range. This suggests that in these strongly nutrient-limited ecosystems, soil N availability regulates the patterns of resource allocation to condensed tannins in the absence but not in the presence of browsing.

Chemical Composition and Decomposition of Silver Birch Leaf Litter Produced under Elevated CO2 and O3

Two field-growing silver birch (Betula pendula Roth) clones (clone 4 and 80) were exposed to elevated CO2 and O3 over three growing seasons (1999–2001). In each year, the nutrients and cell wall chemistry of naturally abscised leaf litter were analyzed in order to determine the possible CO2- and O3-induced changes in the litter quality. Also CO2 and O3 effects on the early leaf litter decomposition dynamics (i.e. decomposition before the lignin decay has started) were studied with litter-bag experiments (Incubation 1 with 1999 leaf litter, Incubation 2 with 2000 leaf litter, and Incubation 3 with 2001 leaf litter) in a nearby silver birch forest. Elevated CO2 decreased N, S, C:P and α-cellulose concentrations, but increased P, hemicellulose and lignin+polyphenolic concentrations, C:N and lignin+polyphenolic:N in both clones. CO2 enrichment decreased the subsequent decomposition of leaves of clone 4 transiently (in Incubations 1 and 2), whereas elevated CO2 effects on the subsequent leaf decomposition of clone 80 were inconsistent. In contrast to CO2, O3 decreased P concentrations and increased C:P, but both of these trends were visible in elevated O3 treatment only. O3-induced decreases in Mn, Zn and B concentrations were observed also, but O3 effects on the cell wall chemistry of leaf litter were minor. Some O3-induced changes either became more consistent in leaf litter collected during 2001 (decrease in B concentrations) or appeared only in this litter lot (decrease in N concentrations, decrease in decomposition at the end of Incubation 3). In conclusion, in northern birch forests elevated CO2 and O3 levels have the potential to affect leaf litter quality, but consistent CO2 and O3 effects on the decomposition process remain to be validated.

Wound-induced Oxidative Responses in Mountain Birch Leaves

The aim of the study was to examine oxidative responses in subarctic mountain birch, Betula pubescens subsp. czerepanovii, induced by herbivory and manual wounding. Herbivory-induced changes in polyphenoloxidase, peroxidase and catalase activities in birch leaves were determined. A cytochemical dye, 3,3-diaminobenzidine, was used for the in situ and in vivo detection of H2O2 accumulation as a response to herbivory and wounding. To localize peroxidase activity in leaves, 10 mm H2O2 was applied to the dye reagent. Feeding by autumnal moth, Epirrita autumnata, larvae caused an induction in polyphenoloxidase and peroxidase activities within 24 h, and a concomitant decrease in the activity of antioxidative catalases in wounded leaves. Wounding also induced H2O2 accumulation, which may have both direct and indirect defensive properties against herbivores. Wound sites and guard cells showed a high level of peroxidase activity, which may efficiently restrict invasion by micro-organisms. Birch oxidases together with their substrates may form an important front line in defence against herbivores and pathogens.

Delayed induced responses of birch glandular trichomes and leaf surface lipophilic compounds to mechanical defoliation and simulated winter browsing

Changes in morphology and chemistry of leaf surface in response to herbivore damage may increase plant resistance to subsequent herbivore attack; however, there is lack of studies on induced responses of glandular trichomes and their exudates in woody plants and on effects of these changes on herbivores. We studied delayed induced responses in leaf surface traits of five clones of silver birch (Betula pendula Roth) subjected to various types of mechanical defoliation and simulated winter browsing. Glandular trichome density and concentrations of the majority of surface lipophilic compounds increased in trees defoliated during the previous summer. This induced response was systemic, since control branches in branch defoliated trees responded to the treatments similarly to defoliated branches, but differently from control trees. In contrast to defoliation treatments, simulated winter browsing reduced glandular trichome density on the following summer and had fewer effects on individual surface lipophilic compounds. Moreover, constitutive density of glandular trichomes was negatively correlated with induced total amount of lipophilic compounds per trichome, indicating a trade-off between constitutive and induced resistance in silver birch. Induced changes in leaf surface traits had no significant effect on leaf damage by chewers, miners and gall mites, but increased susceptibility of birch trees to aphids. However, leaf damage by chewers, miners and gall mites in defoliated (but not in control) trees was correlated with concentrations of some fatty acids and triterpenoids, although the direction of relationships varied among herbivore species. This indicates that induction of surface lipophilic compounds may influence birch resistance to herbivores. Our study thus demonstrated both specificity of elicitation of induced responses of birch leaf surface traits by different types of damage and specificity of the effects of these responses on different types of herbivores.

Biochemical transformation of birch leaf phenolics in larvae of six species of sawflies

We investigated the biochemical transformation of individual phenolic compounds of mountain birch leaves in larvae of six birch-feeding sawfly species: Amauronematus amplus, Pristiphora alpestris, Nematus brevivalvis, Priophorus pallipes, Arge sp. and Nematus viridis by comparing the phenolic residues in larval faeces to those of their leaf diet. Partial hydrolysis of hydrolysable tannins, isomerisation of chlorogenic acid and glycosylation of flavonoid aglycones were observed in all studied species. Moreover, we found considerable among-species variation in the composition of phenolic compounds in larval faeces. In addition to foliar phenolics, seventeen non-foliar phenolic metabolites, including eight phenolic acids and nine flavonoid glycosides were detected from the faeces. Of the non-foliar phenolic acids, four were egested species-specifically and only two by all six sawfly species. We also detected differences in the ratios of chlorogenic acid isomers in the faeces of different species, which can indicate different physiological conditions in the guts of studied larvae. In addition to the qualitative differences, quantitative differences were detected in the egestion of chlorogenic acids, possible o-quinone precursors in the larvae. Detected differences, either qualitative or quantitative, could not be explained by seasonal changes in the content of compounds in the leaf diet.

Temperature dependence of leaf‐level CO2 fixation: revising biochemical coefficients through analysis of leaf three‐dimensional structure

CO2 fixation in a leaf is determined by biochemical and physical processes within the boundaries set by leaf structure. Traditionally determined temperature dependencies of biochemical processes include physical processes related to CO2 exchange that result in inaccurate estimates of parameter values. A realistic three-dimensional model of a birch (Betula pendula) leaf was used to distinguish between the physical and biochemical processes affecting the temperature dependence of CO2 exchange, to determine new chloroplastic temperature dependencies for V c(max) and Jmax based on experiments, and to analyse mesophyll diffusion in detail. The constraint created by dissolution of CO2 at cell surfaces substantially decreased the CO2 flux and its concentration inside chloroplasts, especially at high temperatures. Consequently, newly determined chloroplastic V c(max) and Jmax were more temperature dependent than originally. The role of carbonic anhydrase in mesophyll diffusion appeared to be minor under representative mid-day nonwater-limited conditions. Leaf structure and physical processes significantly affect the apparent temperature dependence of CO2 exchange, especially at optimal high temperatures when the photosynthetic sink is strong. The influence of three-dimensional leaf structure on the light environment inside a leaf is marked and affects the local choice between Jmax and V c(max)-limited assimilation rates.

Defensive Effect of Surface Flavonoid Aglycones of Betula pubescens Leaves Against First Instar Epirrita autumnata Larvae

The surface of birch leaves contains glandular trichomes that secrete exudates containing flavonoid aglycones. We investigated the biological activities of white birch (Betula pubescens) leaf surface exudates against larvae of the autumnal moth, Epirrita autumnata, a common insect pest of birch. We found that tree-specific mortality (up to 100%) of first instar larvae correlated strongly with the tree-specific contents of surface flavonoid aglycones (r(s) = 0.905) in emerging leaves. We also found that first instars clearly preferred birch buds from which surface exudates had been removed. In addition, the duration of the first instar was shortened by 29%, and the weights and relative growth rates of first instars improved by 8% and 52%, respectively, as a result of removal of the exudates from their leaf diet. The correlation of tree-specific foliar contents of flavonoid aglycones, especially 5-hydroxy-4',7-dimethoxyflavanone, with changes in larval performance, suggests that flavonoid aglycones are responsible for the changes observed in first instar larval performance. The results show that chemical characteristics of birch leaves are effective against neonate E. autumnata larvae. However, the removal of leaf surface exudates from fully expanded leaves did not affect the leaf acceptance for the voracious fifth instars. This is probably a result of reduction in contents of flavonoid aglycones compared to those of emerging leaves.

Activated carbon amendments to soil alters nitrification rates in Scots pine forests

The influence of charcoal on biotic processes in soils remains poorly understood. Charcoal is a natural product of wildfires that burned on a historic return interval of ∼100 years in Scots pine ( Pinus sylvestris L.) forests of northern Sweden. Fire suppression and changes in forest stand management have resulted in a lack of charcoal production in these ecosystems. It is thought that charcoal may alter N mineralization and nitrification rates, however, previous studies have not been conclusive. Replicated field studies were conducted at three late-succession field sites in northern Sweden and supporting laboratory incubations were conducted using soil humus collected from these sites. We used activated carbon (AC), as a surrogate for natural-occurring fire-produced charcoal. Two rates of AC (0 and 2000 kg ha −1), and glycine (0 and 100 kg N as glycine ha −1) were applied in factorial combination to field microplots in a randomized complete block pattern. Net nitrification, N mineralization, and free phenol concentrations were measured using ionic and non-ionic resin capsules, respectively. These same treatments and also two rates of birch leaf litter (0 and 1000 kg ha −1) were applied in a laboratory incubation and soils from this incubation were extracted with KCl and analyzed for NH 4 + and NO 3 −. Nitrification rates increased with AC amendments in laboratory incubations, but this was not supported by field studies. Ammonification rates, as measured by NH 4 + accumulation on ionic resins, were increased considerably by glycine applications, but some NH 4 + was apparently lost to surface sorption to the AC. Phenolic accumulation on non-ionic resin capsules was significantly reduced by AC amendments. We conclude that charcoal exhibits important characteristics that affect regulating steps in the transformation and cycling of N.

Progressive Soil Drought Promotes the Export Function in the Birch (Betula platyphylla) Leaf

A greenhouse experiment, which imitated a short (4-day-long) and progressive (3-week-long) soil drought, was employed to assess, with an IR gas analyzer, leaf CO2 exchange rate (CER) in intact one-year-old seedlings of Betula platyphylla as related to the flux of photosynthetically active radiation ranging from 0 to 1400 μE/(m2 s). The registered indices comprised leaf temperature, leaf transpiration conductivity, and the average daily increment of the leaf area. Within a week period following the transition from the short severe soil drought (20% H2O per soil weight) to the conditions of sufficient water content (35–40%), the plants completely regained the initial leaf CER. Under the progressive soil drought, leaf CER was reduced by 30–35%, as compared to the conditions of sufficient water content, evidently due to a 3.7-fold drop in the transpiration conductivity as compared to the control plants. The apparent constant of Rubisco carboxylation and leaf respiration in the light were not affected by the drought period. The rate of leaf growth under the progressive drought was reduced by 64% as compared to the sufficient moisture conditions. Thus, under the progressive drought, the diminished stomatal conductivity reduced CO2 concentration inside the leaf and lowered carbon photosynthetic assimilation. Meanwhile, the leaf source activity considerably increased in spite of diminished photosynthesis.

Processing of pine (Pinus sylvestris) and birch (Betula pubescens) leaf material in a small river system in the northern Cairngorms, Scotland

Abstract. Processing rates, and macroinvertebrate colonisation, of pine needles and birch leaves were studied at eight sites on the river Nethy, a small river system in the Cairngorm region of north-eastern Scotland. Throughout this river system, processing rates were slow for pine (k values 0.0015-0.0034 day-1) and medium to fast for birch (k values 0.0085-0.0331 day-1). Plecopteran shredders dominated both pine and birch leaf packs during the early part of the experiment while chironomids were more important in the latter stages. It is suggested that the slow processing rate of pine needles could adversely affect the productivity of streams, particularly where needles provide the major allochthonous energy source and retentive features are limited. Forest managers should consider this when creating new pinewoods in treeless areas as it will take many years for the trees to reach a size at which they can effectively contribute retentive features, in the form of woody debris, to streams. Keywords: leaf processing, pine needles, shredders, Pinus sylvestris, Betula pubescens, Scotland.

Metabolic Modifications of Birch Leaf Phenolics by an Herbivorous Insect: Detoxification of Flavonoid Aglycones via Glycosylation

The metabolic modifications of birch (Betula pubescens Ehrh.) leaf phenolics in the digestive tract of its major defoliator, larvae of the autumnal moth Epirrita autumnata, were studied. The main phenolic acids of birch, i.e. chlorogenic and p-coumaroylquinic acids, were isomerised in the alkaline digestive tract. Moreover, only 16 to 92% of the ingested amounts of chlorogenic acid were found in the faeces of individual larvae; the missing portion is possibly being used in the formation of reactive o-quinones. Water-soluble flavonoid glycosides were mostly excreted unaltered. In contrast, lipophilic flavonoid aglycones were not excreted as such, but as glycosides after being detoxified by E. autumnata via glycosylation. When the larvae were fed with leaf-painted acacetin and kaempferide, i.e. two naturally occurring birch leaf flavonoid aglycones, acacetin-7-O-glucoside and kaempferide-3-O-glucoside appeared in larval faeces as major metabolites. However, the efficiency of aglycone glycosylation varied-, ranging from 17 to 33%, depending on the aglycone and its dietary level. There was also large variation in the efficiency of glycosylation--from 2 to 57%--among individual larvae. These results demonstrate a compound-specific metabolism of phenolic compounds, leading to different phenolic profiles in the insect gut compared to its leaf diet.

Decomposition of mountain birch leaf litter at the forest-tundra ecotone in the Fennoscandian mountains in relation to climate and soil conditions

Litter decomposition is a key process in terrestrial ecosystems, releasing nutrients, returning CO2 to the atmosphere, and contributing to the formation of humus. Litter decomposition is strongly controlled both by climate and by litter quality: global warming scenarios involving shifts in vegetation communities are therefore of particular interest in this context. The objective of the present study was to quantify the role of climatic environment and underlying substrate chemistry for the decomposition of standard mountain birch (Betula pubescens Ehrh. spp. czerepanovii) leaf litter at four sites, spanning the forest-tundra ecotone, in the Fennoscandian mountain range. Litter quality effects were thus held constant, but the study incorporated systematic changes in (i) latitude/altitude, (ii) `continentality', and (iii) vegetation community at each site, together with (iv) experimental manipulation of temperature using passive warming systems. The study was undertaken during a 3 year period, and forms part of a broader investigation of forest-tundra ecotone dynamics in the Fennoscandian mountains. Our results showed (1) higher decomposition rates in forest sites compared to tundra, (2) that the difference between the two vegetation communities was most pronounced at the more maritime sites, and (3) that chemistry of litter remaining after the three years experiment varied according to site and vegetation community (e.g. at the most southerly site, more lignin had decomposed at tundra communities compared with the forest). (4) Surface temperature explained 58% of the variation in mass loss at forest sites; at tundra sites, however, we hypothesise that litter moisture content was the more important factor. (5) Experimental warming lent weight to this hypothesis by reducing rates of mass loss: this reduction was likely the result of surface soil drying, an artefact of the warming treatment. We conclude that a replacement of tundra by forest would likely accelerate litter decomposition both via changes in surface and near-surface temperature and moisture regimes, although the strength of this response will vary between maritime and continental parts of the mountain range.

Effect of natural and artificial defoliation on concentration and composition of extractive substances in leaves of birch (Betula pendula Roth.): methods of analysis and results.

Effect of natural and artificial defoliation on concentration and composition of extractive substances in leaves of birch (Betula pendula Roth.): methods of analysis and results.

Preparation of chromosomes from plant leaf meristems for karyotype analysis and in situ hybridization

A reliable method for preparing metaphase chromosomes from plant leaf tissues is described. The chromosomes are suitable for karyotype analysis and gene mapping by fluorescence in situ hybridisation (FISH). The method is based on enzymatic digestion of young leaf tissues (shoot-tips) after which the resulting protoplasts are treated hypotonically before being dropped onto microscopic slides. Compared to root-tip chromosomes, leaf chromosomes tend to be longer, or less condensed, and hence more karyotypically differentiated. Metaphase index in young leaf tissues is also very high. Metaphase spread consists of evenly and well-distributed chromosomes and this allows accurate counting. The plant used to demonstrate this method is birch (Betula L.), a group of tree species that has extremely small chromosomes. Root-tip chromosomes of these plants are difficult to obtain, as cutting does not produce roots readily. Seedling chromosomes do not represent the same genomic constitution as their mother trees due to introgressive hybridisation. Furthermore, sample collection in the field is convenient and actively growing leaf buds are available throughout the growing season. FISH experiments with these leaf chromosomes also give good results comparable to those obtained with root-tip chromosomes or even better as mapping on long or extended chromosomes has high resolution in general. Mapping of the 16S-28S ribosomal genes on birch leaf chromosomes has been shown to differentiate between birch species and therefore can accurately confirm their interspecific hybrids.

How browsing by red deer impacts on litter decomposition in a native regenerating woodland in the Highlands of Scotland

Herbivores can indirectly affect ecosystem productivity and processes such as nutrient cycling and decomposition by altering the quantity and quality of resource inputs into the decomposer subsystem. Here, we tested how browsing by red deer impacts on the decomposition of, and nutrient loss from, birch leaf litter (Betula pubescens), and tested whether effects of browsing on these measures were direct, via alteration of the quality of leaf litter, or indirect through long term impacts of deer browsing on soil biological properties. This was tested in a microcosm experiment using soil and litter taken from inside and outside three individual fenced exclosures located at Creag Meagaidh National Nature Reserve, Scotland. We found that litter of un-browsed trees decomposed faster than that from browsed trees, irrespective of whether soil was sourced from inside or outside exclosures. These findings suggest that effects of browsing on litter quality, rather than on soil biological properties, are the key determinant of enhanced decomposition in un-browsed areas of this ecosystem. Despite this, we found no consistent impact of browsing on litter C:N, a key indicator of litter quality; however, the rate of litter decomposition was linearly and negatively related to litter C:N when analysed across all the sites, indicating that this measure, in part, contributed to variation in rates of decomposition in this ecosystem. Our findings indicate that herbivores impact negatively on rates of decomposition in this ecosystem, ultimately retarding nutrient cycling rates, and that these effects are, in part, related to changes in litter quality.

Putting the insect into the birch-insect interaction.

Leaf maturation in mountain birch (Betula pubescens ssp. czerepanovii) is characterized by rapid shifts in the types of dominant phenolics: from carbon-economic flavonoids aglycons in flushing leaves, via hydrolysable tannins and flavonoid glycosides, to carbon-rich proanthocyanidins (condensed tannins) in mature foliage. This shift accords with the suggested trade-offs between carbon allocation to plant defense and growth, but may also relate to the simultaneous decline in nutritive leaf traits, such as water, proteins and sugars, which potentially limit insect growth. To elucidate how birch leaf quality translates into insect growth, I introduce a simple model that takes into account defensive compounds but also acknowledges insect demand for nutritive compounds. The effects of defensive compounds on insect growth depend strongly on background variation in nutritive leaf traits: compensatory feeding on low nutritive diets increases the intake of defensive compounds, and the availability of growth-limiting nutritive compounds may modify the effects of defenses. The ratio of consumption to larval growth (both in dry mass) increases very rapidly with leaf maturation: from 2.9 to 9.8 over 2 weeks in June-July, and to 15 by August. High concentrations in mature birch leaves of "quantitative" defenses, such as proanthocyanidins (15-20% of dry mass), presumably prevent further consumption. If the same compounds had also protected half-grown leaves (which supported the same larval growth with only one third of the dry matter consumption of older leaves), the same intake of proanthocyanidins would have demanded improbably high concentrations (close to 50%) in young leaves. The model thus suggests an adaptive explanation for the high levels of "quantitative" defenses, such as proanthocyanidins, in low-nutritive but not in high-nutritive leaves because of the behavioral responses of insect feeding to leaf nutritive levels.

Effects of sample drying and storage, and choice of extraction solvent and analysis method on the yield of birch leaf hydrolyzable tannins.

In this study, I investigated the effects of different methods of sample drying and storage, and the choice of extraction solvent and analysis method on the concentrations of 14 individual hydrolyzable tannins (HTs), and insoluble ellagitannins in birch (Betula pubescens) leaves. Freeze- and vacuum-drying of birch leaves were found to provide more reliable results than air- or oven-drying. Storage of leaves at -20 degrees C for 3 months before freeze-drying did not cause major changes in tannin content, although levels of 1,2,3,4,6-penta-O-galloylglucose and isostrictinin were altered. Storage of dried leaf material at -20 degrees C is preferred because 1 year storage of freeze-dried leaves at 4 degrees C and at room temperature decreased the concentration of the pedunculagin derivative, one of the main ellagitannins of birch. Furthermore, storage at room temperature increased the levels of isostrictinin and 2,3-(S)-HHDP-glucose, indicating possible HT catabolism. Of the extraction solvents tested, aqueous acetone was superior to pure acetone, or aqueous or pure methanol. The addition of 0.1% ascorbic acid into 70% acetone significantly increased the yield of ellagitannins. presumably by preventing their oxidation. By comparing the conventional rhodanine assay and the HPLC-ESI-MS assay for quantification of leaf galloylglucoses, the former tends to underestimate total concentrations of galloylglucoses in birch leaf extract. On the basis of the outcomes of all the method and solvent comparisons, their suitability for qualitative and quantitative analysis of plant HTs is discussed, emphasizing that each plant species, with its presumably unique HT composition, is likely to have a unique combination of ideal conditions for tissue preservation and extraction.

A three‐dimensional model of CO2transport in airspaces and mesophyll cells of a silver birch leaf

AbstractA realistic numerical three‐dimensional (3D) model was constructed to study CO2transport inside a birch leaf. The model included chloroplasts, palisade and spongy mesophyll cells, airspaces, stomatal opening and the leaf boundary layer. Diffusion equations for CO2were solved for liquid(mesophyll) and gaseous(air) phases. Simulations were made in typical ambient field conditions varying stomatal opening, photosynthetic capacity and temperature. Doubled ambient CO2concentration was also considered. Changes in variables caused non‐linear effects in the total flux, especially when compared with the results of double CO2concentration. The reduction in stomatal opening size had a smaller effect on the total flux in doubled concentrations than ambient CO2. The reduced photosynthetic capacity had a similar effect on the flux in both cases. The palisade and spongy mesophyll cells had unequal roles mainly due to the light absorption profile. Results from the 3D simulation were also compared to the classical one‐dimensional resistance approach. Liquid and gas phase resistances were estimated and found strongly variable according to changes in temperature and degree of stomatal opening. For the birch leaves modelled, intercellular airspace resistance was small (2% of the total resistance in saturating irradiance conditions at 25 °C at stomatal opening diameter of 4 µm) whereas the liquid phase resistance was significant (23% for mesophyll and chloroplasts in the same ‘base case’). The absorption of CO2into water at cell surfaces caused additional (strongly temperature dependent) resistance which accounted for 36% of the total resistance in the base case.

Distribution of hydrolysable tannins in the foliage of Finnish birch species.

On the basis of qualitative and quantitative analysis with liquid chromatography--electrospray ionisation mass spectrometry, the foliage of dwarf birch (Betula nana L.), silver birch (B. pendula Roth) and mountain birch (B. pubescens ssp. czerepanovii (Orlova) Hämet-Ahti) were found, for the first time, to contain the same individual HTs that were described earlier for white birch (B. pubescens Ehrh.). In addition, one previously unidentified ellagitannin was preliminarily identified from the leaves of white and mountain birches, being totally absent from the foliage of the other two species. There were large variations in the contents of HTs between species. Seasonal variation affected significantly the contents of some individual HTs within species, and these changes were mainly in accordance with the biosynthetic pathway of HTs. All species converted galloylglucoses (GGs) into ellagitannins (ETs), dwarf birch being the only one that's efficient ET synthesis resulted in seasonally increased contents of ETs and thereof total HTs as well. The presence of insoluble ETs as well as the absence of insoluble GGs was confirmed in all four birch species for the first time. Furthermore, the amounts of insoluble ETs per one birch leaf were found to accumulate during the growing season. These findings complemented our knowledge of the biosynthetic pathway of birch leaf HTs: from soluble GGs via soluble ETs into insoluble ETs. The possible role of HTs in the herbivore defence of these species is discussed.

Seasonal changes in birch leaf chemistry: are there trade-offs between leaf growth and accumulation of phenolics?

Several plant-herbivore hypotheses are based on the assumption that plants cannot simultaneously allocate resources to growth and defence. We studied seasonal patterns in allocation to growth and putatively defensive compounds by monitoring several chemical and physical traits in the leaves of mountain birch from early June (budburst) to late September (leaf senescence). We found significant seasonal changes in all measured characteristics, both in terms of concentrations (mg g-1) and amounts (mg leaf-1). Changes were very rapid in the spring, slow in the middle of the season, and there was another period of fast changes in the senescing leaves. Co-occurring changes in physical leaf traits and concentrations of several compounds indicated a seasonal decline in foliage suitability for herbivores. Concentrations of protein and free amino acids declined through the growing season whereas individual sugars showed variable seasonal patterns. The seasonal trends of phenolic groups differed drastically: concentrations of soluble proanthocyanidins increased through the season, whereas cell wall-bound proanthocyanidins, gallotannins and flavonoid glycosides declined after an initial increase in young leaves. We failed to find proof that the seasonal accumulation of phenolics would have been seriously compromised by leaf or shoot growth, as assumed by the growth/differentiation balance hypothesis and the protein competition model hypothesis. On the contrary, there was a steady increase in the total amount of phenolics per leaf even during the most active leaf growth.