The effect of ozone on the emission of carbonyls from leaves of adult Fagus sylvatica

Under the site conditions of a temperate forest, the exchange of short-chained oxygenated carbonyls (aldehydes, ketones) was assessed from leaves of adult European beech trees. The crowns of the trees were either exposed to an elevated O₃ regime as released by a free-air fumigation system (2 × O₃) or to the unchanged O₃ regime at the site (1 × O₃, ‘control’). Daily fluctuations of oxygenated carbonyls were quantified in relation to environmental and physiological factors. In particular, the effect of O₃ on carbonyl exchange was studied. Measurements of leaf gas exchange were performed with a dynamic cuvette system, and carbonyl fluxes were determined using 2,4-dinitrophenylhydrazine (DNPH)-coated silica gel cartridges. Leaves mainly emitted acetaldehyde, formaldehyde and acetone. Acetaldehyde dominated the emissions, amounting up to 100 nmol m⁻² min⁻¹, followed by formaldehyde (approximately 80 nmol m⁻² min⁻¹) and acetone (approximately 60 nmol m⁻² min⁻¹). Carbonyl emissions were highest during midday and significantly lowered at night, irrespective of the O₃ exposure regime. Trees exposed to 2 × O₃ emitted acetaldehyde and acetone at enhanced rates. The findings are of particular significance for future climate change scenarios that assume increased O₃ levels.     

Seasonal Changes of the Antioxidative Systems in Foliar Buds and Leaves of Field-grown Beech Trees (Fagus sylvatica,L.) in a Stressful Climate

The activities of superoxide dismutase, ascorbate peroxidase, monodehydroascorbate radical reductase, and dehydroascorbate reductase and the contents of ascorbate, chlorophyll and soluble protein were determined in beech (Fagus sylvatica, L.) foliage over two or three seasons. Four important stages of leaf development were distinguished: resting buds, emerging, mature and senescent leaves. Foliar buds in spring, prior to the emergence of new leaves, contained a lower chlorophyll content but a higher protein content and higher activities of ascorbate peroxidase and monodehydroascorbate radical reductase than mature leaves in summer. By contrast, superoxide dismutase and glutathione reductase activities and ascorbate contents were higher in mature leaves than in swollen foliar buds. Dehydroascorbate reductase activity was low in all developmental stages. Resting buds in winter contained activities of superoxide dismutase, ascorbate peroxidase and monodehydroascorbate radical reductase that were similar to those found in mature leaves in summer, whereas the contents of total and reduced ascorbate were 6- and 20-times lower, respectively, in buds than in mature leaves. The low foliar concentration of reduced ascorbate in resting buds, despite high monodehydroascorbate radical reductase activity, suggests that the regeneration of ascorbate might be limited by the availability of reductant. High antioxidative capacity was conferred by mature beech leaves and may be an important protection measure for coping with the large fluctuations in temperature and exposure to elevated ozone concentrations in summer.     

Genetic variation of European beech (Fagus sylvatica L.) along an altitudinal transect at Mount Vogelsberg in Hesse, Germany

Allelic and genotypic variation at 13 different enzyme loci of autochthonous European beech (Fagus sylvatica L.) was investigated in six 110-160-year-old stands growing at elevations between 150 and 660 m above sea level on the western slope of mount Vogelsberg in central Germany. The highest elevated population showed the highest number of effective alleles (Ne), the highest total heterozygosity (He) and the highest population differentiation deltaT. Also, the genotype SKD-A2A3 of shikimate dehydrogenase was significantly more frequent at the two highest elevated stands (P = 11%) than at the three lowest elevated stands (P = 1%). Further differences in genotype frequencies between 11 of 15 stand pairs were elevation independent.     

Interpretation of seasonal changes of xylem embolism and plant hydraulic resistance in Fagus sylvatica

The annual course of xylem embolism in twigs of adult beech trees was monitored, and compared to concurrent changes of tree water status and hydraulic resistances. Xylem embolism was quantified in 1-year-old apical twigs by the hydraulic conductivity as a percentage of the maximum measured after removal of air emboli. Tree and root hydraulic resistances were estimated from water potential differences and sap flux measurements. The considerable degree of twig embolism found in winter (up to 90% loss of hydraulic conductivity) may be attributed to the effect of freeze-thaw cycles in the xylem. A partial recovery from winter embolism occurred in spring, probably because of the production of new functional xylem. Xylem embolism fluctuated around 50% throughout the summer, without significant changes. Almost complete refilling of apical twigs was observed early in autumn. A significant negative correlation was found between xylem embolism and precipitation; thus, an active role of rainfall in embolism reversion is hypothesized. Tree and root hydraulic resistances were found to change throughout the growing period. A marked decrease of hydraulic resistance preceded the refilling of apical twigs in the autumn. Most of the decrease in total tree resistance was estimated to be located in the root compartment.     

Acclimation to changing light conditions of long-term shade-grown beech (Fagus sylvatica L.) seedlings of different geographic origins

 Effects of changing light conditions on the ecophysiological condition behind survival were examined on beech from two different populations. Plants were grown in a greenhouse under simulated understorey and canopy gap light conditions. Upon exposure to high light maximum photosynthesis of shade-acclimated leaves increased followed by a reduction over several days to between high- and low-light control rates. In the reciprocal transfer, the decrease in maximum photosynthesis was rapid during the first 2–3 days and then levelled off to values comparable to low-light controls. Seedlings from Sicily (Madonie) showed generally higher maximum photosynthetic rates than those from Abetone. Leaf conductance varied in the same direction as photosynthesis in high- to low-light seedlings but to a lesser degree. Leaves grown under low light and exposed to high light experienced photoinhibition. The Abetone population was more susceptible to photoinhibitory damage than the seedlings from Sicily. Exposure to high light of shade-acclimated seedlings resulted in intermediate chlorophyll concentrations between levels of the high-light and low-light seedlings. Carotenoid concentration was unaffected by treatments. Seedlings grew more in high light, but had a lower leaf area ratio. Light-limited seedlings showed a shift in carbon allocation to foliage. Leaves formed in the new light regime maintained the same anatomy that had been developed before transfer. Seedlings from Sicily had thicker leaves than those of seedlings from Abetone. Seedlings from Abetone were found to be more susceptible to changing light conditions than seedlings from Sicily. We conclude that small forest gaps may represent a favorable environment for photosynthesis and growth of beech regeneration as a result of the limited ability of seedlings to acclimate to sudden increases in high irradiance and because of the moderate levels of light stress in small gaps. Received: 11 April 1997 / Accepted: 11 December 1997     

The effect of shoot architecture on hydraulic conductance in beech (Fagus sylvatica L.)

 Shoot hydraulic conductance was measured in beech (Fagus sylvatica L.) that had previously been exposed to high levels of nitrogen input. Whole-shoot hydraulic conductance, conductance per unit pressure gradient and leaf specific conductance were negatively correlated with the number of bud scars per unit length, a morphological parameter of tree decline. We propose a negative feedback mechanism by which stress induced alterations in shoot morphology can cause a lasting reduction of tree vigour. Received: 2 July 1997 / Accepted: 25 June 1998     

Significance of ozone exposure for inter-annual differences in primary metabolites of old-growth beech (Fagus sylvatica L.) and Norway spruce (Picea abies L.) trees in a mixed forest stand

The influence of long-term free-air ozone fumigation and canopy position on leaf contents of total glutathione, its redox state, non-structural proteins (NSP), soluble amino compounds, and total soluble sugars in old-growth beech (FAGUS SYLVATICA) and spruce (PICEA ABIES) trees were determined over a period of five years. Ozone fumigation had weak effects on the analysed metabolites of both tree species and significant changes in the contents of total glutathione, NSP, and soluble sugars were observed only selectively. Beech leaves were affected by crown position to a higher extent than spruce needles and exhibited lower contents of total glutathione and NSP and total soluble sugars, but enhanced contents of oxidised glutathione and amino compounds in the shade compared to the sun crown. Contents of total soluble sugars generally were decreased in shade compared to sun needles of spruce trees. Interspecific differences between beech and spruce were more distinct in the sun compared to the shade crown. Contents of total glutathione were increased whilst contents of amino compounds and total soluble sugars were lower in spruce needles compared to beech leaves. The metabolites determined showed individual patterns in the course of the five measurement years. Contents of total glutathione and its redox state correlated with air temperature and global radiation, indicating an important role for the antioxidant at low temperatures. Correlations of glutathione with instantaneous ozone concentrations seem to be a secondary effect. Differences in proteins and/or amino compounds in the inter-annual course are assumed to be a consequence of alterations in specific N uptake rates.     

Storage and remobilisation of sulphur in beech trees (Fagus sylvatica)

Three-year-old beech trees were fed ³⁵S-sulphate in August 1993 via a flap in a mature leaf of an upper branch. Harvest of beech trees was performed 24 h after feeding ³⁵S-sulphate, before leaf senescence, after leaf abscission, in early winter (January 1994). in late winter (March 1994). before bud break and after bud break. Twenty-four h after feeding ³⁵S-sulphate, 0.7 ± 0.5% of the ³⁵S-radioactivity taken up was exported out of the fed leaf. When trees were analysed 2 months later, i.e., before leaf senescence, this value had increased to 22 ± 7%. The exported ³⁵S-radioactivity was located in the branch containing the fed leaf (2.8 ± 13%). in basipetal parts of the trunk (41 ± 77%) and in the main rool (21 ± 6%). Leaves and apical parts of the trunk were no sink organs for the exported sulphur. Along the tree axis the main proportion of the radiolabel was located in the wood, predominantly in the acid soluble fraction. In the bark the greater portion of the radiolabel was found in the acid insoluble fraction. In both tissues the bulk of the ³⁵S of the soluble fraction was sulphate together with small amounts of glutathione. This pattern did not change until bud break. After bud break, basipetal parts of the trunk lost part of its ³⁵S-radioactivity. Of the ³⁵S-radioactivity which had been exported out of the fed leaf during the previous autumn, 16 ± 2% remained in the trunk, whereas 47 ± 7% of the ³⁵S was found in branches, mainly in the newly developed leaves. The present results show that sulphur, mainly in the form of sulphate, is stored along the tree axis in both bark and wood of beech trees and is re-mobilised during leaf development in spring.     

Carbon and nitrogen dynamics in acid detergent fibre lignins of beech (Fagus sylvatica L.) during the growth phase

To study the incorporation of carbon and nitrogen in different plant fractions, 3‐year‐old‐beech (Fagus sylvatica L.) seedlings were exposed in microcosms to a dual‐labelling experiment employing ¹³C and ¹⁵N throughout one season. Leaves, stems, coarse and fine roots were harvested 6, 12 and 18 weeks after bud break (June to September) and used to isolate acid‐detergent fibre lignins (ADF lignin) for the determination of carbon and nitrogen and their isotope ratios. Lignin concentrations were also determined with the thioglycolic acid method. The highest lignin concentrations were found in fine roots. ADF lignins of all tissues analysed, especially those of leaves, also contained significant concentrations of nitrogen. This suggests that lignin‐bound proteins constitute an important cell wall fraction and shows that the ADF method is not suitable to determine genuine lignin. ADF lignin should be re‐named as ligno‐protein fraction. Whole‐leaf biomass was composed of 50 to 70% newly assimilated carbon and about 7% newly assimilated nitrogen; net changes in the isotope ratios were not observed during the experimental period. In the other tissues analysed, the fraction of new carbon and nitrogen was initially low and increased significantly during the time‐course of the experiment, whereas the total tissue concentrations of carbon remained almost unaffected and nitrogen declined. At the end of the experiment, the whole‐tissue biomass and ADF lignins of fine roots contained about 65 and 50% new carbon and about 50 and 40% new nitrogen, respectively. These results indicate that significant metabolic activity was related to the formation of structural biopolymers after leaf growth, especially below‐ground and that this activity also led to a substantial binding of nitrogen to structural compounds.     

Factors toxic to beech (Fagus sylvatica L.) seedlings in acid soils

The effects of highly and moderately acid soils on total biomass, biomass partitioning, fine root characteristics and nutritional status of beech seedlings (Fagus sylvatica L.) were studied in a growth chamber experiment. In Haplic Arenosols seedlings grew slowly but equally well without damage symptoms in a highly acid and a moderately acid soil horizon. The moderately acid A_h+B_w-horizon of a Eutric Cambisol was favourable to seedling growth. The fine root development was reduced in the highly acid A+B_w-horizon of a Dystric Cambisol and in the A_h+E-horizon of a Haplic Podzol, the latter of which also caused increased mortality. Seedling growth in the B₂-horizon of the Haplic Podzol was vigorous, in spite of a higher level of extractable Al and lower base saturation as compared with the A_h+E-horizon. These results are interpreted in relation to soil acidity, soil Al and nutritional status of the seedlings. We conclude that neither Al-toxicity nor nutrient deficiency cause the damage symptoms observed in the A_h+E-horizon of a Haplic Podzol and the fine root reduction in the A+B_w-horizon of a Dystric Cambisol. The damage symptoms of the PZh_A treatment seems to be more the result of H-toxicity or H-related factors other than nutrient shortage or Al-toxicity. Other pH-related toxic factors are discussed.     

The variability of vessel size in beech (Fagus sylvatica L.) and its ecophysiological interpretation

The vessel areas of ten beech trees growing on a dry site were measured separately for all tree rings using automatic image analysis. These data were correlated with the monthly amount of precipitation from the July prior to the growing season until the August of the current growing season. It is evident that vessel formation at the beginning of cambial activity is mainly controlled by internal factors. The rainfall in the previous summer and autumn and in the contemporary May had only a slight influence. Vessel formation towards the end of the cambial activity is strongly influenced by the July rainfall and is thus determined to a greater degree by external factors. These results are discussed on the basis of hypotheses of tree physiology.     

Phosphate and calcium uptake in beech (Fagus sylvatica) in the presence of aluminium and natural fulvic acids

In the present study we examine the effects of Al on the uptake of Ca²⁺ and H₂PO^-₄ in beech (Fagus sylvatica L.) grown in inorganic nutrient solutions and nutrient solutions supplied with natural fulvic acids (FA). All the solutions used were chemically well characterized. The uptake of Al by roots of intact plants exposed to solutions containing 0, 0.15 or 0.3 mM AlCl₃ for 24 h, was significantly less if FA (300 mg l⁻¹) were also present in the solutions. The Ca²⁺(⁴⁵Ca²⁺) uptake was less affected by Al in solutions supplied with FA than in solutions without FA. There was a strong negative correlation between the Al and Ca²⁺ uptake (r²=0.98). When the Al and Ca²⁺ (⁴⁵Ca²⁺) uptake were plotted as a function of the Al³⁺ activity (or concentration of inorganic mononuclear Al), almost the same response curves were obtained for the -FA and +FA treatments. We conclude that FA-complexed Al was not available for root uptake and therefore could not affect the Ca²⁺ uptake. The competitive effect of Al on the Ca²⁺ uptake was also shown in a 5-week cultivation experiment, where the Ca concentration in shoots decreased at an AlCl₃ concentration of 0.3 mM. The effect of Al on H₂PO⁻₄ uptake was more complex. The P content in roots and shoots was not significantly affected, compared with the control, by cultivation for 5 weeks in a solution supplied with 0.3 mM AlCl₃, despite a reduction of the H₂PO⁻₄ concentration in the nutrient solution to about one-tenth. At this concentration Al obviously had a positive effect on H₂PO⁻₄ uptake. The presence of FA decreased ³²P-phosphate uptake by more than 60% during 24 h, and the addition of 0.15 or 0.3 mM AlCl₃ to these solutions did not alter the uptake of ³²P-phosphate.     

Identifying the pathways for foliar water uptake in beech (Fagus sylvatica L.): a major role for trichomes

Foliar water uptake (FWU), the direct uptake of water into leaves, is a global phenomenon, having been observed in an increasing number of plant species. Despite the growing recognition of its functional relevance, our understanding of how FWU occurs and which foliar surface structures are implicated, is limited. In the present study, fluorescent and ionic tracers, as well as microcomputed tomography, were used to assess potential pathways for water entry in leaves of beech, a widely distributed tree species from European temperate regions. Although none of the tracers entered the leaf through the stomatal pores, small amounts of silver precipitation were observed in some epidermal cells, indicating moderate cuticular uptake. Trichomes, however, were shown to absorb and redistribute considerable amounts of ionic and fluorescent tracers. Moreover, microcomputed tomography indicated that 72% of empty trichomes refilled during leaf surface wetting and microscopic investigations revealed that trichomes do not have a cuticle but are covered with a pectin‐rich cell wall layer. Taken together, our findings demonstrate that foliar trichomes, which exhibit strong hygroscopic properties as a result of their structural and chemical design, constitute a major FWU pathway in beech.