Effects of elevated CO2 on leaf gas exchange in beech and oak at two levels of nutrient supply: consequences for sensitivity to drought in beech

Beech (Fagus sylvatica L.) and pedunculate oak (Quercus robur L.) were grown from seed for two whole seasons at two CO₂ concentrations (ambient and ambient + 250 μmol mol^?1) with two levels of soil nutrient supply. Measurements of net leaf photosynthetic rate (A) and stomatal conductance (g_s) of well-watered plants were taken over both seasons; a drought treatment was applied in the middle of the second growing season to a separate sample of beech drawn from the same population. The net leaf photosynthetic rate of well-watered plants was stimulated in elevated CO₂ by an average of 75% in beech and 33% in oak; the effect continued through both growing seasons at both nutrient levels. There were no interactive effects of CO₂ concentration and nutrient level on A or g_s in beech or oak. Stomatal conductance was reduced in elevated CO₂ by an average of 34% in oak, but in beech there were no significant reductions in g_s except under cloudy conditions (–22% in elevated CO₂). During drought, there was no effect of CO₂ concentration on g_s in beech grown with high nutrients, but for beech grown with low nutrients, g_s was significantly higher in elevated CO₂, causing more rapid soil drying. With high nutrient supply, soil drying was more rapid at elevated CO₂ due to increased leaf area. It appears that beech may substantially increase whole-plant water consumption in elevated CO₂, especially under conditions of high temperature and irradiance when damage due to high evaporative demand is most likely to occur, thereby putting itself at risk during periods of drought.     

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.     

Irradiance and temperature affect the competitive interference of blackberry on the physiology of European beech seedlings

The potential negative influence of competition from early successional species like blackberry (Rubus fruticosus) may be decisive for the natural regeneration success of drought-sensitive beech (Fagus sylvatica), especially in the light of climate change. With a split plot glasshouse experiment, we investigated the influence of two air temperature and irradiance levels on the competitive interference of blackberry on the water, nitrogen (N) and carbon (C) balance of beech seedlings under moderate drought. When increased temperature was accompanied by low irradiance the biomass, root-to-shoot ratio, N uptake and assimilation rates of blackberry were lower compared with beech, either grown alone or with blackberry. By contrast, when elevated temperature and high irradiance were combined, the root-to-shoot ratio and specific N uptake rate of blackberry were substantially increased, while the N acquisition of beech was impaired. Under lower temperature, with either full light or shade, the presence of blackberry had no significant effects on beech, for almost all tested parameters. Under elevated air temperature beech was impaired by the presence of blackberry at high irradiance. These findings emphasize the interacting effects between environmental factors and competition on the establishment of beech regeneration, which should be considered for future forest management in the frame of climate change.     

Antioxidants in the apoplast and symplast of beech (Fagus sylvatica L.) leaves: seasonal variations and responses to changing ozone concentrations in air

Concentrations of the antioxidants ascorbate and glutathione were measured in the apoplast of beech (Fagus sylvatica L.) leaves and in leaf tissue. During early leaf development, reduced ascorbate (ASC) was almost absent from the apoplast, whereas levels of oxidized ascorbate (DHA) were high. Less than 20% of the apoplastic ascorbate was reduced. ASC increased towards midsummer, reaching top levels of about 4molm^?3 apoplast volume in July and August. Reduction increased to 60–75% in summer. Neither DHA reductase nor glutathione was detected in the apoplast of beech leaves. Levels of apoplastic ascorbate were compared with ambient concentrations of ozone in air. Statistical analysis indicated a significant interrelation between atmospheric ozone and apoplastic ascorbate. In midsummer of 1993, contents of DHA were increased in the apoplast when ozone concentrations were high. Apoplastic ASC was also positively correlated with ambient ozone concentrations, but with a delay of 3 to 7d. In leaf tissue, levels of ascorbate were between 17 and 21 μmolg^?1 FW in summer. Except for late April and November, more than 95% of the intracellular ascorbate was reduced. Glutathione contents were lowest during the summer. Oxidation was increased in spring and autumn, when apoplastic ascorbate was also largely oxidized. Usually, 80 to 90% of the glutathione was reduced. During the summer, intracellular concentrations of oxidized glutathione (GSSG) were increased, with a delay of about 1d following periods of high ambient ozone concentrations. The transitory accumulation of GSSG may be explained by slow enzymatic regeneration of glutathione.     

Above-ground space sequestration determines competitive success in juvenile beech and spruce trees

A 2-yr phytotron study was conducted to investigate the intra- and inter-specific competitive behaviour of juvenile beech (Fagus sylvatica) and spruce (Picea abies). Competitiveness was analysed by quantifying the resource budgets that occur along structures and within occupied space of relevance for competitive interaction. Ambient and elevated CO(2) and ozone (O(3)) regimes were applied throughout two growing seasons as stressors for provoking changes in resource budgets, growth and allocation to facilitate the competition analysis. The hypothesis tested was that the ability to sequester space at low structural cost will determine the competitive success. Spruce was a stronger competitor than beech, as displayed by its higher above-ground biomass increments in mixed culture compared with monoculture. A crucial factor in the competitive success of spruce was its ability to enlarge crown volume at low structural costs, supporting the hypothesis. Interspecific competition with spruce resulted in a size-independent readjustment of above-ground allocation in beech (reduced leaf : shoot biomass ratio). The efficient use of resources for above-ground space sequestration proved to be a parameter that quantitatively reflects competitiveness.     

A new scenario for the quaternary history of European beech populations: palaeobotanical evidence and genetic consequences

Here, palaeobotanical and genetic data for common beech (Fagus sylvatica) in Europe are used to evaluate the genetic consequences of long-term survival in refuge areas and postglacial spread. Four large datasets are presented, including over 400 fossil-pollen sites, 80 plant-macrofossil sites, and 450 and 600 modern beech populations for chloroplast and nuclear markers, respectively. The largely complementary palaeobotanical and genetic data indicate that: (i) beech survived the last glacial period in multiple refuge areas; (ii) the central European refugia were separated from the Mediterranean refugia; (iii) the Mediterranean refuges did not contribute to the colonization of central and northern Europe; (iv) some populations expanded considerably during the postglacial period, while others experienced only a limited expansion; (v) the mountain chains were not geographical barriers for beech but rather facilitated its diffusion; and (vi) the modern genetic diversity was shaped over multiple glacial-interglacial cycles. This scenario differs from many recent treatments of tree phylogeography in Europe that largely focus on the last ice age and the postglacial period to interpret genetic structure and argue that the southern peninsulas (Iberian, Italian and Balkan) were the main source areas for trees in central and northern Europe.     

Responses to ozone in 12 provenances of European beech (Fagus sylvatica): genotypic variation and chamber effects on photosynthesis and dry-matter partitioning

Seedlings of 12 provenances of European beech ( Fagus sylvatica ) were exposed to ambient, non-filtered air (NF) or NF+50 nl l⁻¹ ozone (NF50) for 8 h d⁻¹ in open-top chambers (OTCs), from 1 June to 4 October 1995. In 1996 exposure was continued from 31 May to 1 October at four levels: charcoal-filtered air (CF), NF, NF50 and NF+100 nl l⁻¹ ozone (NF100). Provenances were grown for both seasons in outside reference plots. All treatments were replicated. Ozone did not affect gas exchange in the provenances until late in the second season. NF100 reduced photosynthesis by 18% in August 1996 compared to CF. In September, photosynthesis was reduced by 22% in NF50 and by 29% in NF100. After two seasons, ozone reduced the root:shoot ratio by 24% when comparing CF and NF100; this was caused by reductions in the root biomass. Ozone did not affect height growth or stem diameter, and there were no ozone×provenance interactions for any growth parameter. There was, however, a significant ozone×provenance interaction for photosynthesis, showing northwest European provenances to be more sensitive to ozone than southeast European provenances when comparing dose–response estimates. This is interpreted in terms of genetic adaptation of the photosynthetic apparatus to regional growing conditions. Seedlings in the chambers grew 45% taller, and had 28% more shoot biomass and 29% smaller root biomass, resulting in a 44% reduction of root:shoot ratios compared to seedlings outside. Increased temperature and decreased PAR inside the chambers relative to the outside were probably the main causes for the differences. The magnitude of the chamber effects in OTCs raises doubts about conclusions drawn from ozone exposures in such chambers. This and previous ozone experiments with OTCs may have reached inaccurate conclusions concerning the size of ozone responses due to chamber effects.     

Tree stem phosphoenolpyruvate carboxylase (PEPC): lack of biochemical and localization evidence for a C4-like photosynthesis system

Here, the kinetic properties and immunolocalization of phosphoenolpyruvate carboxylase (PEPC) and ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in young stems of Fagus sylvatica were investigated. The aim of the study was to test the hypothesis that there is a C4-like photosynthesis system in the stems of this C3 tree species. The activity, optimal pH and L-malate sensitivity of PEPC, and the Michaelis-Menten constant (Km) for phosphoenolpyruvate (PEP), were measured in protein extracts from current-year stems and leaves. A gel blot experiment and immunolocalization studies were performed to examine the isozyme complexity of PEPC and the tissue distribution of PEPC and Rubisco in stems. Leaf and stem PEPCs exhibited similar, classical values characteristic of C3 PEPCs, with an optimal pH of c. 7.8, a Km for PEP of c. 0.3 mM and a IC50 for L-malate (the L-malate concentration that inhibits 50% of PEPC activity at the Km for PEP) of c. 0.1 mM. Western blot analysis showed the presence of two PEPC subunits (molecular mass c. 110 kDa) both in leaves and in stems. Immunogold labelling did not reveal any differential localization of PEPC and Rubisco, neither between nor inside cells. This study suggests that C4-type photosynthesis does not occur in stems of F. sylvatica and underlines the importance of PEPC in nonphotosynthetic carbon fixation by most stem tissues (fixation of respired CO2 and fixation via the anaplerotic pathway).     

Competition increasingly dominates the responsiveness of juvenile beech and spruce to elevated CO2 and/or O3 concentrations throughout two subsequent growing seasons

Saplings of Fagus sylvatica and Picea abies were grown in mono‐ and mixed cultures in a 2‐year phytotron study under all four combinations of ambient and elevated ozone (O₃) and carbon dioxide (CO₂) concentrations. The hypotheses tested were (1) that the competitiveness of beech rather than spruce is negatively affected by the exposure to enhanced O₃ concentrations, (2) spruce benefits from the increase of resource availability (elevated CO₂) in the mixed culture and (3) that the responsiveness of plants to CO₂ and O₃ depends on the type of competition (i.e. intra vs. interspecific). Beech displayed a competitive disadvantage when growing in mixture with spruce: after two growing seasons under interspecific competition, beech showed significant reductions in leaf gas exchange, biomass development and crown volume as compared with beech plants growing in monoculture. In competition with spruce, beech appeared to be nitrogen (N)‐limited, whereas spruce tended to benefit in terms of its plant N status. The responsiveness of the juvenile trees to the atmospheric treatments differed between species and was dominated by the type of competition: spruce growth benefited from elevated CO₂ concentrations, while beech growth suffered from the enhanced O₃ regime. In general, interspecific competition enhanced these atmospheric treatment effects, supporting our hypotheses. Significant differences in root : shoot biomass ratio between the type of competition under both elevated O₃ and CO₂ were not caused by readjustments of biomass partitioning, but were dependent on tree size. Our study stresses that competition is an important factor driving plant development, and suggests that the knowledge about responses of plants to elevated CO₂ and/or O₃, acquired from plants growing in monoculture, may not be transferred to plants grown under interspecific competition as typically found in the field.     

Functional characteristics of corticolous lichens in the understory of a tropical lowland rain forest

In tropical lowland forests, corticolous crustose green algal lichens are abundant and highly diverse. This may be related to adaptation to prevailing microenvironmental conditions including, for example, high precipitation and low light intensities. In the understory of a tropical lowland rain forest in French Guiana, we studied the morphology of crustose green algal lichens and measured gas exchange and chlorophyll a fluorescence. We found that (i) periods of thallus suprasaturation with water were reduced by the presence of water-repelling surface structures of mycobiont hyphae at the thallus surface and the medulla; (ii) photosynthesis was adapted to the low light intensities present in the understory; and (iii) photosynthesis was rapidly activated in fluctuating light. The combination of these three mechanisms enables corticolous lichens to implement specific morphological and physiological strategies, which may favour growth in the limiting understory habitat of tropical lowland rain forests.     

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.     

Contributions of diffusional limitation, photoinhibition and photorespiration to midday depression of photosynthesis in Arisaema heterophyllum in natural high light

Diurnal changes in photosynthetic gas exchange and chlorophyll fluorescence were measured under full sunlight to reveal diffusional and non‐diffusional limitations to diurnal assimilation in leaves of Arisaema heterophyllum Blume plants grown either in a riparian forest understorey (shade leaves) or in an adjacent deforested open site (sun leaves). Midday depressions of assimilation rate (A) and leaf conductance of water vapour were remarkably deeper in shade leaves than in sun leaves. To evaluate the diffusional (i.e. stomatal and leaf internal) limitation to assimilation, we used an index [1–A/A₃₅₀], in which A₃₅₀ is A at a chloroplast CO₂ concentration of 350 μ mol mol ^{? 1}. A₃₅₀ was estimated from the electron transport rate (J_T), determined fluorometrically, and the specificity factor of Rubisco (S), determined by gas exchange techniques. In sun leaves under saturating light, the index obtained after the ‘peak’ of diurnal assimilation was 70% greater than that obtained before the ‘peak’, but in shade leaves, it was only 20% greater. The photochemical efficiency of photosystem II ( Δ F/F_m ′ ) and thus J_T was considerably lower in shade leaves than in sun leaves, especially after the ‘peak’. In shade leaves but not in sun leaves, A at a photosynthetically active photon flux density (PPFD) > 500 μ mol m ^{? 2} s ^{? 1} depended positively on J_T throughout the day. Electron flows used by the carboxylation and oxygenation (J_O) of RuBP were estimated from A and J_T. In sun leaves, the J_O/J_T ratio was significantly higher after the ‘peak’, but little difference was found in shade leaves. Photorespiratory CO₂ efflux in the absence of atmospheric CO₂ was about three times higher in sun leaves than in shade leaves. We attribute the midday depression of assimilation in sun leaves to the increased rate of photorespiration caused by stomatal closure, and that in shade leaves to severe photoinhibition. Thus, for sun leaves, increased capacities for photorespiration and non‐photochemical quenching are essential to avoid photoinhibitory damage and to tolerate high leaf temperatures and water stress under excess light. The increased Rubisco content in sun leaves, which has been recognized as raising photosynthetic assimilation capacity, also contributes to increase in the capacity for photorespiration.