Kyösti Lempa

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.

Biosynthetic origin of carbon-based secondary compounds: cause of variable responses of woody plants to fertilization?

Summary. We propose that variation in the responses of carbon-based secondary compounds to fertilization in woody plants has a biosynthetic cause. The synthesis of phenylpropanoids and derived compounds (e.g., condensed tannins) competes directly with the synthesis of proteins, and therefore with plant growth, because of a common precursor, phenylalanine. In contrast, the biosynthesis of terpenoids and of hydrolyzable tannins proceeds presumably without direct competition with protein synthesis. Therefore, accelerated plant growth induced by fertilization may cause a reduction in concentrations of phenylpropanoids but may affect less or not at all the levels of other classes of secondary compounds. A meta-analysis based on fertilization experiments with 35 woody plant species supported the predicted differences fertilizing significantly decreased concentrations of phenylpropanoids but not of terpenoids or hydrolyzable tannins.

Multiplicity of biochemical factors determining quality of growing birch leaves

Abstract Due to rapidly changing physical and biochemical characteristics of growing leaves, correlations between traits of foliage biochemistry and the performance indices of flush feeding herbivores may vary considerably following relatively minor changes in experimental conditions. We examined the effects of the seasonal and inter-tree variation of a comprehensive array of biochemical compounds on the success of an early season geometrid, Epirrita autumnata, feeding on maturing foliage of mountain birch, Betula pubescens ssp. czerepanovii. We monitored the concentrations of individual phenolics, sugars, total nitrogen, nitrogen of proteins, and nitrogen of soluble compounds, water and acetone-insoluble residue. Simultaneously we recorded larval consumption, physiological performance, growth, and pupal mass of E. autumnata. We found significant phenological changes in almost all leaf traits measured. In bioassays with half-grown leaves, leaf gallotannin concentrations showed a nonlinear effect: in trees with high foliar gallotannin concentrations (over 10 mg g−1), physiological performance was strongly reduced by high gallotannin concentrations. In trees with lower gallotannin concentrations, on the other hand, larval growth was reduced by soluble proanthocyanidins, not gallotannins. Differences between high and low gallotannin trees largely depended on phenology, i.e., on the age of leaves. However, not all the differences in leaf traits between late (with high gallotannin concentrations at the time of the bioassay) and early flushing trees disappeared with leaf maturation, indicating that there is also phenology-independent variance in the tree population. In the full-grown leaves of all the study trees, low concentrations of water and of nitrogen of proteins (but not nitrogen of soluble compounds) were the main factors reducing pupal masses of E. autumnata, while neither gallotannin nor proanthocyanidins now played a significant role. The observed change in the factors underlying leaf quality (from gallotannins and proanthocyanidins to nitrogen and water) relate to the activity of the shikimate pathway and the formation of cell walls: gallotannins and proanthocyanidins are both produced in the pathway, and these tannins are assumed to contribute – via binding into cell walls – to tough and durable cell walls. Interestingly, low quality of leaves did not automatically translate into low foliar consumption (i.e., benefits to the tree). On the trees with young, high gallotannin leaves, larvae actually increased consumption on low quality foliage. In the group of trees with slightly more developed, low gallotannin leaves, the quality of leaves did not clearly modify amounts consumed. In full-grown leaves, low leaf quality strongly reduced leaf consumption. These results emphasize the strong influence of tree phenology on the relationships between biochemical compounds and the herbivore.

Covariation of fluctuating asymmetry, herbivory and chemistry during birch leaf expansion

Abstract Fluctuating asymmetry (FA) is used to describe developmental instability in bilateral structures. In trees, high FA of leaves has been assumed to indicate the level of environmental or genetic stress, and for herbivores leaves from such trees have been shown to be in some cases (though not invariably) of higher quality compared to trees with symmetrical leaves. We demonstrated that FA of birch leaves correlated positively with growth rate of leaves, and with the amount of leaf biomass consumed by larvae of the geometrid Epirrita autumnata. Since asymmetry per se cannot define leaf quality for a herbivore, we determined the biochemical compounds which covary with the degree of foliage FA, in order to elucidate relationships between leaf FA, chemistry and herbivory. High foliar FA was characteristic of birches with high initial concentrations, and rapid seasonal decline in the concentrations of gallic acid and hydrolysable tannins, and with rapid seasonal changes in the concentrations of flavonoid-glycosides and sugars. In contrast, leaf FA was not related to concentrations of proanthocyanidins, protein-bound amino acids or soluble phenylalanine, the precursor of proanthocyanidins and proteins with aromatic amino acids. The positive correlation between leaf FA and consumption by E. autumnata was presumably related to the previously demonstrated compensatory consumption of E. autumnata to high concentrations of foliar gallotannins. Furthermore, sugars are well-known feeding stimulants. We propose that the variable results in studies correlating leaf FA and herbivory may stem from variable chemical associations of FA in different plants and of species-specific effects of compounds on insects.

Interactive effects of leaf maturation and phenolics on consumption and growth of a geometrid moth

Phenolic compounds are commonly regarded as the main chemical defenses of deciduous woody plants against insects. To examine how indices of leaf maturation (water content, toughness, and sugar/protein ratio) modified larval consumption and growth relative to phenolics and phenolic‐related leaf traits, we measured consumption and growth of fourth‐instar Epirrita autumnata (Bkh.) (Lepidoptera: Geometridae) larvae on three different days on young, normal, and mature leaves, respectively, from the same mountain birch (Betula pubescens ssp. czerepanovii (Orlova) Hämet‐Ahti) trees. The larvae achieved the same growth rates on young and normal leaves, but had to consume 40% more on the latter. On more mature leaves, larval growth was poorer and was positively correlated with sugar/protein ratios (although the ratio peaked at that time). Indices of leaf maturation correlated with several phenolics in data pooled over the three study days, but poorly in any individual day. Similarly, in the pooled data, larval consumption and growth correlated with several leaf traits, but correlations between leaf and insect traits were few on any of the three days, and no trait was significant on each of the three days.

Effects of stress and rapid growth on fluctuating asymmetry and insect damage in birch leaves

We studied how fluctuating asymmetry (FA), which is commonly used as an non-specific stress indicator, is influenced by abiotic stress along a moisture gradient and by rapid growth, i.e. the production of larger than normal leaves after simulated browsing, in white birch (Betula pubescens) growing in a mire. Variation of abiotic stress (plots near vs far from open, wet boundary of tree stand) was obvious as interpreted from smaller leaf size and lower stature of trees growing closest to the wettest. open center of the mire, However, we did not find increased leaf FA near the wettest area, possibly because of the high background level of stress in the mire habitat or the presence of flood-tolerant genotypes. Simulated winter browsing increased leaf size in the following growing season, and the strength of the response increased with the severity of the browsing treatment Likewise, larger leaves of browsed ramets had higher developmental instability, as measured by size-corrected FA. Moreover, the effects of simulated winter browsing on FA were proportionally greater than the size effects in expanding leaves, Simulated winter browsing also increased insect damage incurred by leaves Our results demonstrate that, in addition to previously shown effects of stress, excess leaf growth can also increase developmental instability and FA. We also demonstrated a higher rate of developmental errors in the later than early stages of leaf expansion.

Ranking of individual mountain birch trees in terms of leaf chemistry: seasonal and annual variation

Summary.The quality of tree leaves as food for herbivores changes rapidly especially during the spring and early summer. However, whether the quality of an individual tree in relation to other trees in the population changes during the growing season and between years is less clear. We studied the seasonal and annual stability of chemical and physical traits affecting leaf quality for herbivores. Rankings of trees in terms of the contents of two major groups of phenolics in their leaves, hydrolyzable tannins and proanthocyanidins (condensed tannins), were very stable from the early spring to the end of the growing season. There were also strong positive within-season correlations in the levels of some other groups of phenolics in the leaves (kaempferol glycosides, myricetin glycosides and p-coumaroylquinic acid derivatives). The contents of individual sugars and the sum content of protein-bound amino acids showed patterns of seasonal consistency in mature leaves, but not in young developing leaves. The seasonal correlations in leaf water content and toughness were also strongest in mature leaves. The correlations between two years at corresponding times of the growing season were strongly positive for the major groups of phenolics throughout the season, but were more variable for the contents of proteins and some sugars. Leaf toughness and water content showed strong positive correlations in mature leaves. Despite the consistency of tree ranking in terms of leaf phenolics, the relative resistance status of trees may, however, change during a growing season because there was a negative correlation between the content of hydrolyzable tannins (early-season resistance compounds) in leaves early in the season and the content of proanthocyanidins (late-season resistance compounds) late in the season, and vice versa. Thus, assuming that phenolics affect herbivore preference and performance, different plants may suffer damage at different times of the growing season, and the overall variation between trees in the fitness consequences may be low. In addition, the adaptation of herbivorous insects to mountain birch foliage in general, as well as to specific tree individuals, may be constrained by variation in the relative resistance status of the trees.

Effects of hydrolysable tannins on a herbivorous insect: fate of individual tannins in insect digestive tract

Summary. We investigated the effects of four chemically characterised galloylglucoses (GGs, a subgroup of hydrolysable tannins) and their hydrolysis product, gallic acid (GA), on consumption and performance of larvae of the autumnal moth Epirrita autumnata. Larvae were fed with birch (Betula pubescens) leaves that had been painted individually with each of the compounds at two levels, 5 and 20 mg/g. In addition, we investigated the fates of the leaf-painted GGs and GA in the E. autumnata digestive tract by comparing phenolics in leaves consumed and in faeces. In general, GGs reduced leaf consumption by E. autumnata during the second and fourth instars, although there was high compound- and instar-specific variation. However, GGs did not affect the leaf consumption rates by the most voracious fifth instar larvae. This resulted in approximately the same loss of total biomass by the experimental tree, regardless of the nature and level of GGs enriched to its foliage. The characteristic fate of hydrolysable tannins, i.e. hydrolysis, was evidenced in the larval digestive tract for three of the four leaf-painted GGs. In addition to hydrolysis, the almost total absence of GGs in larval faeces was presumably related to the oxidation of GGs. The dose-dependent excretion percentage of ingested GA showed that its faecal content should not be used, although it commonly is, to calculate the level of GG hydrolysis. Moreover, by comparing the non-uniform appearance of faecal tetragalloylglucoses, whether ingested as such or hydrolysed from pentagalloylglucose, we concluded that a major part of oxidation of GGs occurs before their hydrolysis in the digestive tract of E. autumnata. Criticism against the common use of tannic acid, a heterogeneous mixture of GA and GGs, in ecological studies is presented.

Patterns in content of phenolic compounds in leaves of mountain birches along a strong pollution gradient.

The contents of individual low-molecular weight phenolic compounds (LMWPs) in mountain birch, Betula pubescens ssp. czerepanovii, leaves collected during 1996-1998 in six plots 7-65 km south of the nickel-copper smelter at Monchegorsk, Kola Peninsula, NW Russia, were reported. A high-performance liquid chromatography-electrospray ionisation-mass spectrometry (HPLC-ESI-MS) was used for the rapid identification of low-molecular weight phenolics. Quantification was performed by the analytical high-performance liquid chromatography with UV-detection. Contents of (+)-catechin and some gallic acid derivatives decreased significantly, and contents of flavonol glycosides slightly increased with the distance from the smelter. Hydroxycinnamic acid derivatives remained unaffected. These changes in birch leaf phenolics are probably related to the effect of environmental contamination on the biosynthetic reactions both in the shikimate and phenylpropanoid pathways.

Concentrations and among-compound correlations of individual phenolics in white birch leaves under air pollution stress

Abstract To detect early symptoms of heavy metal pollution in birch ( Betula pubescens Ehrh.) trees, we studied foliar concentrations of phenolic compounds in polluted and control areas around the Harjavalta copper-nickel smelter, SW Finland. Phenolic compounds were quantified by analytical high performance liquid chromatography. The total content of individual phenolics was 21% higher in samples from the heavily polluted (400–500 m from the smelter) area than on the control area. Differences in foliar concentrations of individual compounds between the study areas were significant for (+)-catechin, trans -3- p -coumaroylquinic acid, quercetin-3- O -β-D-galactopyranoside and one gallotannin. Variation in contents of phenolics was quite large. Among tree variances of individual phenolics between control and polluted area differed significantly in the case of seven compounds: 1- O -galloyl-β-D-(2- O -acetyl)-glucopyranose, four gallotannins, neochlorogenic acid and quercetin-3-O-α-L-(4″- O -acetyl)-rhamnopyranoside. Differences in pairwise correlations between phenolic compounds in polluted and control areas indicated competition between some gallotannins and p -coumaroylquinic acids on the polluted but not on control site. Concluding, in strongly polluted areas atmospheric stress factors appear to affect accumulation of some plant phenolics in birch leaves, their variability as well as between-compound correlations.