Fagus sylvatica L. provenances maintain different leaf metabolic profiles and functional response

Most temperate forest tree species will suffer important environmental changes as result of the climate change. Adaptiveness to local conditions could change at different sites in the future. In this context, the study of intra-specific variability is important to clarify the singularity of different local populations. Phenotypic differentiation between three beech provenances covering a wide latitudinal range (Spain/ES, Germany/DE and Sweden/SE), was studied in a greenhouse experiment. Non-target leaf metabolite profiles and ecophysiological response was analyzed in well-watered and water stressed seedlings. There was a provenance-specific pattern in the relative concentrations of some leaf metabolites regardless watering treatment. The DE and SE from the center and north of the distribution area of the species showed a clear differentiation from the ES provenance in the relative concentration of some metabolites. Thus the ES provenance from the south maintained larger relative concentration of some organic and amino acids (e.g. fumaric and succinic acids or valine and isoleucine), and in some secondary metabolites (e.g. kaempferol, caffeic and ferulic acids). The ecophysiological response to mild water stress was similar among the three provenances as a consequence of the moderate water stress applied to seedlings, although leaf N isotope composition (δ¹⁵N) and leaf C:N ratio were higher and lower respectively in DE than in the other two provenances. This would suggest potential differences in the capacity to uptake and post-process nitrogen according to provenance. An important focus of the study was to address for the first time inter-provenance leaf metabolic diversity in beech from a non-targeted metabolic profiling approach that allowed differentiation of the three studied provenances.     

Mechanism of freeze-induced embolism in Fagus sylvatica L.

 The mechanism of freeze stress-induced embolism in Fagus sylvatica L. branches was analyzed under controlled conditions. Excised branches were exposed to successive freeze-thaw cycles in temperature controlled chambers. Thermocouples were placed on the bark to detect sap freezing exotherms. The degree of xylem embolism was estimated after each cycle by the loss of hydraulic conductivity. After one freeze-thaw cycle the degree of embolism was found to decrease with xylem specific hydraulic conductivity, small apical shoots being more susceptible to embolism. Exotherms revealed that distal shoots were freezing first and exuded sap as a result of water expansion. The lower water content in apical shoots upon freezing probably induced higher sap tensions which promoted air bubble expansion and vessel cavitation preferentially near the apices. When the decrease in water content was experimentally prevented, embolism developed to a lesser extent. The higher vulnerability of shoot apices may protect the rest of the branch from winter damage. Received: 29 May 1998 / Accepted: 15 August 1998     

Characterization of microsatellite markers in Fagus sylvatica L. and Fagus orientalis Lipsky

Using an enrichment procedure, we cloned microsatellite repeats from European beech (Fagus sylvatica L.) and developed primers for the amplification of microsatellite markers. Six polymorphic loci were characterized which produced 3–21 alleles in 70 individuals from one Italian population, with an observed heterozygosity between 0.58 and 0.85. All six loci amplified fragments which were polymorphic in the closely related species, Fagus orientalis, also. Owing to their very high degree of variation, these markers should be very useful in gene flow studies of these species.     

Callus formation from mesophyll protoplasts of Fagus sylvatica L.

Viable protoplasts of European beech (Fagus sylvatica L.) were isolated from sterilized young leaves of juvenile (3–5 years) and mature (40 years) trees. Isolation in a saline solution containing 0.5% (w/v) Pectinol and 2% (w/v) Cellulase R-10 yielded 3×10⁷ protoplasts per gram fresh weight. Protoplast culture in modified Kao and Michayluk (1975) medium resulted in cell wall regeneration and sustained cell divisions with the formation of colonies and microcalli.Abbreviations KM Kao and Michayluk (1975) - LS Linsmaier and Skoog (1965) - MES 2(N-morpholino)-ethane-sulfonic acid - NAA 1-naphthalene acetic acid - PVP Polyvinylpyrrolidone - Tween 80 Polyoxy-ethylene-sorbitan-monooleate Dedicated to Landesforstmeister Prof. Dr. Hans-Joachim Fröhlich on the occasion of his 65th birthday.     

Blue radiation stimulates photosynthetic induction in Fagus sylvatica L.

This study was designed to test the hypothesis that the spectral composition of incident radiation, as defined by the relative proportions of blue (B; λ_max = 455 nm) and red (R; λ_max = 625 nm) photons, can affect photosynthetic induction, since B photons stimulate stomatal opening and are more effectively absorbed by leaves than R photons. Different stages of photosynthetic induction, primarily determined by the photo-modulation of Rubisco activity and stomata opening, were investigated in dark-adapted leaves of Fagus sylvatica transferred to saturating irradiance [800µmol(photon) m^?2 s^?1] at B/R ratios of 1/3, 1/1, or 3/1. In agreement with our hypothesis, photosynthesis was induced faster by irradiance with a high B/R ratio (3/1); as demontrated by a higher IS₆₀ (induction state 60 s after leaf illumination) and lower T ₉₀ (the time period required to reach 90 % of maximum steady-state photosynthesis). However, there were no differences in induction between leaves receiving equal (1/1) and low (1/3) B/R ratios. Electron transport was highly sensitive to radiation quality, exhibiting faster induction kinetics with increasing B/R ratio. Such stimulation of carbon-assimilatory processes corresponds with faster activation of Rubisco and lower non-photochemical quenching (NPQ) as the proportion of B photons is increased. In contrast, the kinetics of stomatal opening was independent of the spectral composition of incoming radiation. Since slightly higher absorption efficiency of high B/R radiation does not fully explain the changes in induction kinetics, the other possible mechanisms contributing to the stimulation of electron transport and Rubisco activity are discussed.     

On the ultrastructure of resting cambium of Fagus sylvatica L.

Summary Cells of the resting cambium contain strands of smooth endoplasmic reticulum which is often arranged as a series of parallel membranes. Single membrane bound bodies of different types including lipid droplets, protein bodies, many different intermediate forms, and structures that are empty and vacuolelike are present. The possibility that vacuoles are formed by the progressive dissolution of stored lipids and protein, during the onset of cambial activity, is suggested.     

Towards an understanding of the Holocene distribution of Fagus sylvatica L.

Aim Understanding the driving forces and mechanisms of changes in past plant distribution and abundance will help assess the biological consequences of future climate change scenarios. The aim of this paper is to investigate whether modelled patterns of climate parameters 6000 years ago can account for the European distribution of Fagus sylvatica at that time. Consideration is also given to the role of non‐climatic parameters as driving forces of the Holocene spread and population expansion of F. sylvatica. Location Europe. Methods European distributions were simulated using a physiologically‐based bioclimatic model (STASH) driven by three different atmospheric general circulation model (AGCM) outputs for 6000 years ago. Results The three simulations generally showed F. sylvatica to have potentially been as widespread 6000 years ago as it is today, which gives a profound mismatch with pollen‐based reconstructions of the F. sylvatica distribution at that time. The results indicate that drier conditions during the growing season 6000 years ago could have caused a restriction of the range in the south. Poorer growth conditions with consequently reduced competitive ability were modelled for large parts of France. Main conclusions Consideration of the entire European range of F. sylvatica showed that no single driving force could account for the observed distributional limits 6000 years ago, or the pattern of spread during the Holocene. Climatic factors, particularly drought during the growing season, are the likely major determinants of the potential range. Climatic factors are regionally moderated by competition, disturbance effects and the intrinsically slow rate of population increase of F. sylvatica. Dynamic vegetation modelling is needed to account for potentially important competitive interactions and their relationship with changing climate. We identify uncertainties in the climate and pollen data, as well as the bioclimatic model, which suggest that the current study does not identify whether or not climate determined the distribution of F. sylvatica 6000 years ago. Pollen data are better suited for comparison with relative abundance gradients rather than absolute distributional limits. These uncertainties from a study of the past, where we have information about plant distribution and abundance, argue for extreme caution in making forecasts for the future using equilibrium models.     

Gene Flow in European Beech (Fagus sylvatica L.)

Three relatively isolated stands were used to study gene flow in European beech (Fagus sylvatica L.) in Northern Germany. Nine allozyme loci (Got-B, Idh-A, Lap-A, Mdh-B, Mdh-C, Mnr-A, 6-pgdh-A, Pgi-B and Pgm-A) were utilized for multilocus-genotyping adult trees and seeds. Expected heterozygosity (He) ranged from 0.325 to 0.351 for the three stands. F(ST) revealed that there was small differentiation among stands (mean F(ST) = 0.013). The indirect estimates of gene flow (Nm) based on the mean F(ST) were high and the average Nm was 19.14. External gene flow by pollen ranged from 0.7 to 1.2% inferred from new alleles in seed samples. Moreover, paternity analysis was used to assess effective pollen dispersal by inferring paternity of offspring. The weighted mean distances of pollen dispersal for these three stands were 36.8 and 37.1 m based on simple exclusion procedure and most-likely method, respectively. Two of the trees in one stand had rare allozyme alleles (Lap-A1 and Idh-A4, respectively), which were used to directly measure pollen movement away from those trees. The frequency of the rare Lap and Idh alleles in seeds declines as the distance from the source tree increases. The weighted mean distance of pollen dispersal with rare allele Lap-A1 or Idh-A4 was 26.3 m.     

欧洲水青冈(Fagus sylvatical L.)构筑型与形态多样性研究

通过对植物构件和构筑型的分析,可以了解植物的整体结构与系统演化关系。高等植物构筑型的研究以树木最为深入,植物学家对全球的树木进行了构筑模式的分类,并确定了２３个基本的构筑类型。植物的构筑型和其形态的多样性是密不可分的。构筑奕型确定了植物所属的整体形态类型,而对每种植物具体的形态多样性分析可以深入了解每种构筑型的数量特征及其形成过程。在概述了树木树筑型研究的基础上,系统说明了欧洲水青冈构筑才形态多样性分析的步骤和步骤和分析的意义。水青冈属（Fagus)为Ｔroll型构筑模式,处于不同生态条件下的同一种的不同种群之间,在形态、解剖、生理和遗传多样性方面都存在着差异。树木的形态多样性虽然主要受遗传因素的控制,但生态条件对其形态和遗传多样性也有非常显著的影响。在对树木进行构筑型分析时,道德要根据构筑要素（主要是形态与生长特性）确定它所属的构筑类型,其次是分析它的形态特征与环境条件的关系,进行进一步分析其遗传多样性。     

Photosynthetic UV-B Response of Beech (Fagus sylvatica L.) Saplings

Cloned saplings of beech (7-y-old) were exposed to enhanced UV-B irradiation (+25 %) continuously over three growing seasons (1999–2001). Analysis of CO₂ assimilation, variable chlorophyll (Chl) a fluorescence, and pigment composition was performed in late summer of the third growing season to evaluate the influence of long-term elevated UV-B irradiation. This influence was responsible for the stimulation of the net assimilation rate (P _N) over a range of irradiances. The increase in P _N was partially connected to increase of the area leaf mass, and thus to the increased leaf thickness. Even a higher degree of UV-B induced stimulation was observed at the level of photosystem 2 (PS2) photochemistry as judged from the irradiance response of electron transport rate and photochemical quenching of Chl a. The remarkably low irradiance-induced non-photochemical quenching of maximum Chl a fluorescence (NPQ) in the UV-B plants over the entire range of applied irradiances was attributed both to the reduced demand on non-radiative dissipation processes and to the considerably reduced contribution of the quenching localised in the inactivated PS2 reaction centres. Neither the content of Chls and total carotenoids expressed per leaf area nor the contents of lutein, neoxanthin, and the pool of xanthophyll cycle pigments (VAZ) were affected under the elevated UV-B. However, the contributions of antheraxanthin (A) and zeaxanthin (Z) to the entire VAZ pool in the dark-adapted UV-B treated plants were 1.61 and 2.14 times higher than in control leaves. Surprisingly, the retained A+Z in UV-B treated plants was not accompanied with long-term down-regulation of the PS2 photochemical efficiency, but it facilitated the non-radiative dissipation of excitation energy within light-harvesting complexes (LHC) of PS2. Thus, in the beech leaves the accumulation of A+Z, induced by other factors than excess irradiance itself, supports the resistance of PS2 against combined effects of high irradiance and elevated UV-B.     

Factors influencing the storability of Fagus sylvatica L. seeds after release from dormancy

Seeds of beech (Fagus sylvatica L.) that have been subjected to dormancy breaking consisting of 10 weeks of prechilling at 3 °C and 34 % water content (WC) and then desiccation to 10 % WC, are non-dormant (ND). ND seeds are characterised by greater sensitivity to storage conditions, than no prechilled, dormant (D) seeds. The aim of the present work was to investigate factors affecting the loss of seed viability during storage of D and ND beech seeds at different temperatures (4 and 20 °C) and humidity levels (45 and 75 % RH) for 3 weeks. In general, both D and ND seeds maintained a high germination capacity after storage at 4 °C. At 20 °C and 45 and 75 % RH the germination capacity of D seeds diminished to 80 and 28 %, respectively. Under the same conditions, ND seeds lost germination capacity to a greater degree, with only 62 and 7 % germinated seeds, respectively. At 20 °C, an increase in production of reactive oxygen species was observed, and the increase was significantly higher in ND seeds. The loss of germination capacity was coincident with an increase in electrolyte leakage and accumulation of free fatty acids, which suggests that membrane deterioration was the cause of the decline in germinability. ND seeds stored at 20 °C and 45 and 75 % RH showed a greater decrease than D seeds in contents of the primary phospholipids phosphatidylcholine (PC) and phosphatidylethanolamine (PE) as well as in polyunsaturated fatty acids (18:2 and 18:3). ND seeds possessed more unsaturated fatty acids, especially 18:3, than D seeds in the phospholipid fraction before storage. D seeds were characterised by a significantly higher level of α-tocopherol and UV-absorbing phenols. The level of ascorbate was similar in D and ND seeds. D seeds contained glutathione in both reduced (GSH) and oxidised (GSSG) forms, and GSSG dominated GSH. ND seeds contained more GSSG form than D seeds. We concluded that the membranes of ND seeds are exposed to greater oxidative stress during storage due to higher levels of unsaturation and lower levels of α-tocopherol, the main antioxidant that protects membranes against free radical attack.     

Assessment of genetic relationships and population discrimination among Fagus sylvatica L. by RAPD

 We assessed the genetic relationships between members of the Fagaceae family by RAPDs in order to better ascertain the taxonomic status of a very particular population of Fagus sylvatica, the ‘tortuosa’ variety. Intra- and inter-population Nei and Li’s mean genetic distances were compared, and the genetic relationships between individuals were clarified on dendrograms by the Neighbor joining method. RAPD analysis was first conducted on three species from three genera, Quercus petraea, Castanea sativa, and Fagus sylvatica, in order to develop an efficient RAPD protocol. The variety level was then studied, and a general tendency of the individuals to cluster by variety was observed. Individuals also clustered by geographic locations, but the genetic distances between populations were not correlated to the distances between sites. Finally, we compared the common beech and ‘tortuosa’ varieties from two different locations, Verzy and Süntel. Both populations from one location were closer than the same variety from two sites. This last result is in agreement with those previously obtained with isozymes. Hypotheses concerning the origin of the ‘tortuosa’ variety are discussed. Received: 9 January 1998 / Accepted: 25 February 1998     

The effect of age on height growth in even-sized saplings of Fagus sylvatica L.

Key message

After controlling for the effects of size and light, partial regression revealed that height growth of common beech saplings was negatively affected by sapling age.

Abstract

Common beech (Fagus sylvatica L.) saplings were studied along gradients of light availability (4–82 % of full sunlight), initial size (9–290 cm), and age (2–25 years) to examine the interactive effect of these variables on saplings’ annual height growth. Although age was non-significant as a main effect in a linear model, sapling age had a significant interaction with the other variables. After controlling for the effects of size and light, partial regression revealed that height growth was negatively affected by sapling age. Observed growth decline in older common-sized saplings may be explained not as effect of age per se, but as indirect age-related effect probably induced through plastic response of saplings to past growth conditions.     

Humidity response of cuticular conductance of beech (Fagus sylvatica L.) leaf discs maintained at high relative water content

Cuticular conductance of adaxial (astomatous) and abaxial (stomatous)surfaces ofFagus sylvatica L. leaves was measured under varyingvapour pressure deficits (D). Conductance was determined fromgravimetric measurements of water flux made using a leaf discenvelope specially designed to maintain leaf relative watercontent and minimize reduction in cuticular hydration. The adaxialsurface provided a determination of ‘true’ cuticularconductance (g_c) and transpiration (E_c). The abaxial surfacewas used to estimate minimum leaf surface conductance (g^MIN_sur)and transpiration (E^MIN_sur). In experiment I, leaf discs wereplaced under one of a range of water vapour pressure deficits(0.4-2.0 kPa). Both g_c and g^MIN_sur decreased approximately 2-foldwith an increase in D between 0.4-2.0 kPa. The decrease in g_cwas linear, but g^MIN_sur declined more steeply at D between 0.4-0.95kPa than at D between 0.95-2.0 kPa. In experiment II, leaf discswere exposed to a stepwise change in D. After a period of acclimationto D of 0.95 kPa, responses of g_c and g^MIN_sur to an increaseor decrease in D were recorded. The response time of g_c to increasingor decreasing D were similar (<60 min). By contrast, g^MIN_surresponded more slowly to increasing than to decreasing D. Thesesignificant responses of g_c and g^MIN_sur to increasing and decreasingD are discussed in relation to hydration state of the cuticleand current knowledge of cuticle structure. Key words: Cuticle, cuticular conductance, cuticular membrane, Fagus sylvatica, humidity, vapour pressure deficit     

Vapour Pressure Deficit Response of Cuticular Conductance in Intact Leaves of Fagus sylvatica L.

The cuticular conductance (g_c) of the astomatous adaxial surfaceof Fagus sylvatica L. leaves was determined under varying vapourpressure deficits at a constant temperature of 20 °C. Cuticularconductance was determined from the weight loss of detachedleaves after both the stomatous abaxial surface and the cutend of the petiole had been sealed using low melting-point paraffinwax. Cuticular conductance was found to decrease as the watervapour pressure was increased in steps. No response was observedwhen vapour pressure deficit was decreased from an initiallyhigh value. It is concluded that these results are consistentwith the hypothesis that cuticular conductance is influencedby the water content of the cuticle and that the apparent hysteresisis likely to be a result of the long time-constant for the processof cuticle rehydration in intact leaves. Key words: Cuticle, cuticular conductance, Fagus sylvatica L., Fagaceae, vapour pressure deficit, VPD     

Light-flecks cause non-uniform stomatal opening – studies with special emphasis on Fagus sylvatica L.

The appearance of stomatal patchiness in response to rapid (seconds) changes in light has been studied in European beech, Fagus sylvatica L., and, by comparison, in a further 17 different woody species from the understorey of a European beech forest, using a simple water infiltration method. Water infiltrated areoles indicate open stomata. Since infiltration changes optical characteristics of a leaf section it can be analysed by photography, computer-aided image analysis and by weighing. For F. sylvatica clear differences were found between infiltration of cotyledons (no patchy pattern) and any other leaf type. Despite identical cultivation, leaves of the same type and age from different individual plants responded differently to application of 30 s of light after darkness. In contrast, the patchiness patterns were very similar for leaves of the same type originating from the same plant. Infiltration patterns after a light-fleck, observed on different leaves as a series of momentary clusters, probably indicate waves of opening stomata moving across the leaf blade. During and after a 30 s light-fleck infiltration increased and it continued to increase in the dark up to 10 min, indicating increasing stomatal opening over that period. In general, shade leaves became more infiltrated (by weight) than half-shade or sun leaves, due to larger intercellular air spaces. All species, without exception, showed patchy infiltration and, thus, non-uniform stomatal opening. Measuring leaf gas exchange (as ”quasi-steady states” using a fast responding system) during photosynthetic induction resulted in very similar CO₂ responses of net photosynthesis (A/c _i) as in the true steady state, proving that, in shade and half-shade leaves, the presence of stomatal patchiness does not necessarily affect the calculation of intercellular CO₂ concentrations. Causes and consequences of stomatal patchiness are discussed. Received: 18 November 1998 / Accepted: 1 July 1999