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Herbivores and mycorrhizal fungi interactively influence growth, resource utilization, and plant defense responses. We studied these interactions in a tritrophic system comprising Quercus robur, the herbivore Lymantria dispar, and the ectomycorrhizal fungus Piloderma croceum under controlled laboratory conditions at the levels of gene expression and carbon and nitrogen (C/N) allocation. Taking advantage of the endogenous rhythmic growth displayed by oak, we thereby compared gene transcript abundances and resource shifts during shoot growth with those during the alternating root growth flushes. During root flush, herbivore feeding on oak leaves led to an increased expression of genes related to plant growth and enriched gene ontology terms related to cell wall, DNA replication, and defense. C/N-allocation analyses indicated an increased export of resources from aboveground plant parts to belowground. Accordingly, the expression of genes related to the transport of carbohydrates increased upon herbivore attack in leaves during the root flush stage. Inoculation with an ectomycorrhizal fungus attenuated these effects but, instead, caused an increased expression of genes related to the production of volatile organic compounds. We conclude that oak defense response against herbivory is strong in root flush at the transcriptomic level but this response is strongly inhibited by inoculation with ectomycorrhizal fungi and it is extremely weak at shoot flush.

The ability of treeline conifers in the Central European Alps to cope with recent climate warming and increasing CO2 concentration is still poorly understood. We determined basal area increment (BAI) and tree ring stable carbon isotope ratios (δ13C) of Pinus cembra trees from 1925 through 2013. Stable isotope ratios and BAI were compared with leaf level gas exchange measurements carried out in situ between 1934 and 2012, and thus, provided new insights into long-term trends of tree-ring derived intrinsic water-use efficiency (iWUE). Mature P. cembra trees at treeline responded to increasing Ca and air temperature with a parallel increase in maximum net CO2 uptake rate at ambient CO2 (Amax) and tree-ring-derived intercellular CO2 partial pressure (Ci). Amax tripled and was positively correlated to BAI and Ci. The latter increased in parallel with ambient CO2 concentration and stomatal conductance. In contrast to the instantaneous gas exchange parameters, δ13C derived iWUE informs about the long-term changes in the carbon water relations. These data showed three changes in the iWUE chronosequences, which could be identified with different long term gas exchange patterns: (1) from stomatal controlled functioning from 1925 to 1981, to a situation where (2) both net CO2 fixation (A) and leaf conductance for water vapour (gw), responded to the environment from 1982 to 1997, and (3) back to a stomata controlled pattern over iWUE from 1998 onwards. This temporal pattern was also mirrored in leaf level gas exchange assessments, suggesting a parallel increase of A and gw of P. cembra at treeline during the last nine decades.

Tree roots and mycorrhizal fungi form an important resource for soil microinvertebrates; however, there is little knowledge on the role of root feeders and microbial grazers on belowground carbon flux of forest trees. Using oak (Quercus robur) microcuttings (Clone DF159) inoculated with an ectomycorrhizal symbiont partner (Piloderma croceum), the present study investigated the impact of multitrophic interactions in the rhizosphere on the carbon flow to the soil food web in a pulse-chase labelling experiment. Changes in plant biomass, microbial communities (soil phospholipid fatty acids - PLFAs) and Collembola lipid pattern (total lipid fatty acids - TLFAs) as well as the 13C incorporation into fatty acids were assessed at 2, 5 and 20 days post labelling of oak microcuttings. Microcosms were inoculated with Pratylenchus penetrans (root-feeding nematode), Protaphorura armata (fungal-feeding Collembola) or their combination. The interaction of P. penetrans and P. armata synergistically reduced oak biomass. Both soil animals independently impacted growth of microorganisms in the rhizosphere, P. penetrans inhibited whilst P. armata promoted bacterial biomass. Irrespective of treatment and time, Gram-positive bacteria derived most of the recent photoassimilated oak carbon, with amounts ranging between 80 and 92% of the total 13C allocated in microbial PLFAs. Presence of either P. armata or P. penetrans enhanced the incorporation of plant carbon in bacteria, whereas collective effects of these two functional groups was scarce. Predominantly the impact of the root-feeding nematode on belowground carbon flow diminished with time. In sum, root feeders and detritivores played independent roles in carbon allocation patterns and community structure of microorganisms in the rhizosphere of oaks.

Hydraulic redistribution (HR) of soil water through plant roots is a crucial phenomenon improving the water balance of plants and ecosystems. It is mostly described under severe drought, and not yet studied under moderate drought. We tested the potential of HR under moderate drought, hypothesizing that (H1) tree species redistribute soil water in their roots even under moderate drought and that (H2) neighboring plants are supported with water provided by redistributing plants. Trees were planted in split-root systems with one individual (i.e., split-root plant, SRP) having its roots divided between two pots with one additional tree each. Species were 2- to 4-year-old English oak (Quercus robur L.), European beech (Fagus sylvatica L.) and Norway spruce (Picea abies (L.) Karst). A gradient in soil water potential (ψsoil) was established between the two pots (-0.55 ± 0.02 MPa and -0.29 ± 0.03 MPa), and HR was observed by labeling with deuterium-enriched water. Irrespective of species identity, 93% of the SRPs redistributed deuterium enriched water from the moist to the drier side, supporting H1. Eighty-eight percent of the plants in the drier pots were deuterium enriched in their roots, with 61 ± 6% of the root water originating from SRP roots. Differences in HR among species were related to their root anatomy with diffuse-porous xylem structure in both beech and-opposing the stem structure-oak roots. In spruce, we found exclusively tracheids. We conclude that water can be redistributed within roots of different tree species along a moderate ψsoil gradient, accentuating HR as an important water source for drought-stressed plants, with potential implications for ecohydrological and plant physiological sciences. It remains to be shown to what extent HR occurs under field conditions in Central Europe.

Mixed stands of European beech (Fagus sylvatica L.) and Norway spruce (Picea abies (L.) Karst.) frequently over-yield, when compared to respective monospecific stands. Over-yielding is attributed to enhanced resource uptake efficiency through niche complementarity alleviating species competition, for example through enhanced root stratification in mixture. Under severe and frequent summer drought, however, water limitation may become crucial in modifying the prevailing competitive interaction in mixed beech-spruce forests. We hypothesize, therefore, that under drought (H I) inter-specific interaction with beech reduces water accessibility for spruce more than intra-specific conditions, thus (H II) exacerbating drought susceptibility of spruce in terms of reduced photosynthesis and stem growth. Reactions at the organ (leaf, fine root), tree and stand scale were analysed in a mature forest with beech-spruce group mixture. Under inter-specific conditions spruce’s fine-root production and depth of water uptake (assessed via δ18O of xylem water) shifted to shallow, drought-prone soil horizons, in agreement with H I. Overall, lowered fine root production and ramification along with a reduction in long-distance explorative ectomycorrhizal types resulted in decreased soil exploitation in spruce when growing together with beech. Spruce’s drought sensitivity was exemplified by a distinct decrease in stomatal conductance, net CO2 uptake rate and stem growth during periods of water limitation. Notwithstanding, species interaction effects were absent in leaf gas exchange and stem diameter growth, during a six-week summer drought period in 2013 as well as in the extremely dry year of 2003, hence rejecting H II. Based on results from soil moisture measurements and water uptake depth, we interpret the conflicting findings for H I and H II to result from: (i) seasonal shifts between positive (during spring drought) and negative (during summer drought) effects of beech neighbourhood on soil water availability for spruce, possibly overriding each other in their effect on annual stem diameter growth and (ii) the group-wise mixture pattern, where spruce is exposed to competition with beech only along group edges, i.e. laterally only, so that the putatively adverse beech effect on water accessibility stays limited. Our results suggest, compared to single tree mixture, group-wise mixture of beech and spruce to be a favourable silvicultural option in the face of climate change.

The (13/12) C ratio in plant roots is likely dynamic depending on root function (storage versus uptake), but to date, little is known about the effect of season and root order (an indicator of root function) on the isotopic composition of C-rich fractions in roots. To address this, we monitored the stable isotopic composition of one evergreen (Picea abies) and one deciduous (Fagus sylvatica), tree species' roots by measuring δ(13) C of bulk, respired and labile C, and starch from first/second and third/fourth order roots during spring and fall root production periods. In both species, root order differences in δ(13) C were observed in bulk organic matter, labile, and respired C fractions. Beech exhibited distinct seasonal trends in δ(13) C of respired C, while spruce did not. In fall, first/second order beech roots were significantly depleted in (13) C, whereas spruce roots were enriched compared to higher order roots. Species variation in δ (13) C of respired C may be partially explained by seasonal shifts from enriched to depleted C substrates in deciduous beech roots. Regardless of species identity, differences in stable C isotopic composition of at least two root order groupings (first/second, third/fourth) were apparent, and should hereafter be separated in belowground C-supply-chain inquiry.

Highlight Increased resource availability has no impact on endogenous rhythmic growth and related allocation shifts in oak trees; an internal clock regulates core genes during shoot and root growth cessation phases., Common oak trees display endogenous rhythmic growth with alternating shoot and root flushes. To explore the mechanisms involved, microcuttings of the Quercus robur L. clone DF159 were used for 13C/15N labelling in combination with RNA sequencing (RNASeq) transcript profiling of shoots and roots. The effect of plant internal resource availability on the rhythmic growth of the cuttings was tested through inoculation with the ectomycorrhizal fungus Piloderma croceum. Shoot and root flushes were related to parallel shifts in above- and below-ground C and, to a lesser extent, N allocation. Increased plant internal resource availability by P. croceum inoculation with enhanced plant growth affected neither the rhythmic growth nor the associated resource allocation patterns. Two shifts in transcript abundance were identified during root and shoot growth cessation, and most concerned genes were down-regulated. Inoculation with P. croceum suppressed these transcript shifts in roots, but not in shoots. To identify core processes governing the rhythmic growth, functions [Gene Ontology (GO) terms] of the genes differentially expressed during the growth cessation in both leaves and roots of non-inoculated plants and leaves of P. croceum-inoculated plants were examined. Besides genes related to resource acquisition and cell development, which might reflect rather than trigger rhythmic growth, genes involved in signalling and/or regulated by the circadian clock were identified. The results indicate that rhythmic growth involves dramatic oscillations in plant metabolism and gene regulation between below- and above-ground parts. Ectomycorrhizal symbiosis may play a previously unsuspected role in smoothing these oscillations without modifying the rhythmic growth pattern.

In the present study, we performed a large-scale protein analysis based on 2-DE DIGE to examine the effects of ozone on the leaves of juvenile European beech ( Fagus sylvatica L.), one of the most important deciduous tree species in Central Europe. To this end, beech trees were grown under field conditions and subjected to ambient and twice ambient ozone concentrations during the vegetation periods of four consecutive years. The twice ambient ozone concentration altered the abundance of 237 protein spots, which showed relative ratios higher than 30% compared to the ambient control trees. A total of 74 protein spots were subjected to mass spectrometry identification (LC–MS/MS), followed by homology-driven searches. The differentially expressed proteins participate in key biological processes including the Calvin cycle and photosynthesis, carbon metabolism, defense- and stress-related responses, detoxification mechanisms, protein folding and degradation, and mechanisms involved in senescence. The ozone-induced responses provide evidence of a changing carbon metabolism and counteraction against increased levels of reactive oxygen species. Biological significance This study provides useful information on how European beech, an economically and ecologically important tree species, reacts on the molecular level to increased ozone concentrations expected in the near future. The main emphasis in the present study was placed on identifying differentially abundant proteins after long-term ozone exposure under climatically realistic settings, rather than short-term responses or reactions under laboratory conditions. Additionally, using nursery-grown beech trees, we took into account the natural genotypic variation of this species. As such, the results presented here provide information on molecular responses to ozone in an experimental plant system at very close to natural conditions. Furthermore, this proteomic approach was supported by previous studies on the present experiment. Ultimately, the combination of this proteomic approach with several approaches including transcriptomics, analysis of non-structural carbohydrates, and morphological effects contributes to a more global picture of how beech trees react under increased ozone concentrations.

Summary The root-rot pathogen Phytophthora quercina is a key determinant of oak decline in Europe. The susceptibility of pedunculate oak (Quercus robur) to this pathogen has been hypothesized to depend on the carbon availability in roots as an essential resource for defense. Microcuttings of Q. robur undergo an alternating rhythm of root and shoot growth. Inoculation of mycorrhizal (Piloderma croceum) and nonmycorrhizal oak roots with P. quercina was performed during both growth phases, that is, root flush (RF) and shoot flush (SF). Photosynthetic and morphological responses as well as concentrations of nonstructural carbohydrates (NSC) were analyzed. Infection success was quantified by the presence of pathogen DNA in roots. Concentrations of NSC in roots depended on the alternating root/shoot growth rhythm, being high and low during RF and SF, respectively. Infection success was high during RF and low during SF, resulting in a significantly positive correlation between pathogen DNA and NSC concentration in roots, contrary to the hypothesis. The alternating growth of roots and shoots plays a crucial role for the susceptibility of lateral roots to the pathogen. NSC availability in oak roots has to be considered as a benchmark for susceptibility rather than resistance against P. quercina.

We hypothesize that reductions in stand density through thinning improve the recovery of radial stem growth in Norway spruce trees ( Picea abies ) from severe drought. However, thinning may not lead to higher relative radial growth during drought. Annual stem growth and stable carbon and oxygen isotopes in early- and latewood were assessed in trees from heavily thinned (HT), moderately thinned (MT) and un-thinned control stands at two sites in southern Germany. Physiological performance of trees as inferred from stable isotope analysis was used to interpret annual stem growth in response to the drought events in 1976 and 2003. Only in recently thinned stands, trees maintained growth probably through higher soil water availability during the drought year when compared to controls. In contrast, thinning improved the growth recovery in the years following the drought irrespective of the time span between thinning and drought. We conclude that thinning improves drought recovery response in the short and long term and should be considered as an effective management strategy to increase drought tolerance of Norway spruce stands.

Summary Oaks (Quercus spp.), which are major forest trees in the northern hemisphere, host many biotic interactions, but molecular investigation of these interactions is limited by fragmentary genome data. To date, only 75 oak expressed sequence tags (ESTs) have been characterized in ectomycorrhizal (EM) symbioses. We synthesized seven beneficial and detrimental biotic interactions between microorganisms and animals and a clone (DF159) of Quercus robur. Sixteen 454 and eight Illumina cDNA libraries from leaves and roots were prepared and merged to establish a reference for RNASeq transcriptomic analysis of oak EMs with Piloderma croceum. Using the Mimicking Intelligent Read Assembly (MIRA) and Trinity assembler, the OakContigDF159.1 hybrid assembly, containing 65 712 contigs with a mean length of 1003 bp, was constructed, giving broad coverage of metabolic pathways. This allowed us to identify 3018 oak contigs that were differentially expressed in EMs, with genes encoding proline-rich cell wall proteins and ethylene signalling-related transcription factors showing up-regulation while auxin and defence-related genes were down-regulated. In addition to the first report of remorin expression in EMs, the extensive coverage provided by the study permitted detection of differential regulation within large gene families (nitrogen, phosphorus and sugar transporters, aquaporins). This might indicate specific mechanisms of genome regulation in oak EMs compared with other trees.

Climate simulations anticipate an increase in mean summer temperature with synchronous decrease in summer precipitation during the course of the current century in Central Europe. As a consequence, transpiration of forest trees and stands may be altered along with soil water availability. In this study, the effect of reduced plant water availability to conifers was investigated into an open Pinus sylvestris forest (Erico-Pinetum typicum; P. sylvestris 60 %, Picea abies 20 %; and Larix decidua 20 %) within the inner alpine dry valley of the Inn River in Tyrol, Austria. For reducing plant water availability, we installed a transparent roof construction above the forest floor to prevent precipitation to reach the soil. The roofed area covered 240 m² and included 10 trees. A respective number of 11 trees served as controls in the absence of any manipulation. Roofing significantly reduced plant water availability as indicated in lower predawn needle water potentials. Sap flow density (Q s) was 63, 47, and 24 % lower in roofed P. sylvestris, P. abies, and L. decidua trees, respectively, as compared to control trees. Our findings suggest that P. sylvestris and P. abies behaves “isohydric” as they close their stomata relatively early under conditions of reduced plant water availability and thus stabilize their water relations, whereas L. decidua behaves “anisohydric” and maintains high transpiration rates.

• The CO(2) efflux of adult trees is supplied by recent photosynthates and carbon (C) stores. The extent to which these C pools contribute to growth and maintenance respiration (R(G) and R(M), respectively) remains obscure. • Recent photosynthates of adult beech (Fagus sylvatica) and spruce (Picea abies) trees were labeled by exposing whole-tree canopies to (13) C-depleted CO(2). Label was applied three times during the year (in spring, early summer and late summer) and changes in the stable C isotope composition (δ(13) C) of trunk and coarse-root CO(2) efflux were quantified. • Seasonal patterns in C translocation rate (CTR) and fractional contribution of label to CO(2) efflux (F(Label-Max)) were found. CTR was fastest during early summer. In beech, F(Label-Max) was lowest in spring and peaked in trunks during late summer (0.6 ± 0.1, mean ± SE), whereas no trend was observed in coarse roots. No seasonal dynamics in F(Label-Max) were found in spruce. • During spring, the R(G) of beech trunks was largely supplied by C stores. Recent photosynthates supplied growth in early summer and refilled C stores in late summer. In spruce, CO(2) efflux was constantly supplied by a mixture of stored (c. 75%) and recent (c. 25%) C. The hypothesis that R(G) is exclusively supplied by recent photosynthates was rejected for both species.

Respiration is a substantial driver of carbon (C) flux in forest ecosystems and stable C isotopes provide an excellent tool for its investigation. We studied seasonal dynamics in δ¹³C of CO₂ efflux (δ¹³C(E)) from non-leafy branches, upper and lower trunks and coarse roots of adult trees, comparing deciduous Fagus sylvatica (European beech) with evergreen Picea abies (Norway spruce). In both species, we observed strong and similar seasonal dynamics in the δ¹³C(E) of above-ground plant components, whereas δ¹³C(E) of coarse roots was rather stable. During summer, δ¹³C(E) of trunks was about -28.2‰ (Beech) and -26.8‰ (Spruce). During winter dormancy, δ¹³C(E) increased by 5.6-9.1‰. The observed dynamics are likely related to a switch from growth to starch accumulation during fall and remobilization of starch, low TCA cycle activity and accumulation of malate by PEPc during winter. The seasonal δ¹³C(E) pattern of branches of Beech and upper trunks of Spruce was less variable, probably because these organs were additionally supplied by winter photosynthesis. In view of our results and pervious studies, we conclude that the pronounced increases in δ¹³C(E) of trunks during the winter results from interrupted access to recent photosynthates.

Stable carbon (C) isotopes, in particular employed in labeling experiments, are an ideal tool to broaden our understanding of C dynamics in trees and forest ecosystems. Here, we present a free-air exposure system, named isoFACE, designed for long-term stable C isotope labeling in the canopy of 25 m tall forest trees. Labeling of canopy air was achieved by continuous release of CO2 with a δ13C of −46.9‰. To this end, micro-porous tubes were suspended at c. 1 m distance vertically through the canopy, minimizing CO2 gradients from the exterior to the interior and allowing for C labeling exposure during periods of low wind speed. Target for CO2 concentration ([CO2]) increase was ambient +100 μmol mol−1. Canopy [CO2] stayed within 10% of the target during more than 57% of the time and resulted in a drop of δ13C in canopy air by 7.8‰. After 19 labeling days about 50% of C in phloem sugars and stem CO2 efflux were turned over and 20–30% in coarse root CO2 efflux and soil CO2. The isoFACE system successfully altered δ13C of canopy air for studying turn-over of C pools in forest trees and soils, highlighting their slow turn-over rates.

The impact of chronic free air ozone (O3) exposure and belowground pathogen stress on growth and total biomass development of young beech trees (Fagus sylvatica L.) was investigated in a lysimeter study. Plants were growing during four years under ambient or elevated atmospheric O3 concentrations. Additionally, in the last vegetation period the root rot pathogen Phytophthora citricola was introduced to study the interaction of ozone exposure and pathogen stress in the soil-plant system. A complete harvest at the end of the experiment enabled for the first time the assessment of fine and coarse root biomass of individual trees with a high vertical resolution down to two meter depth. Plant growth was significantly reduced by elevated ozone but not affected by P. citricola. Biomass partitioning between fine and coarse roots as well as vertical root distribution were significantly affected by both factors, whereas changes in root/shoot biomass ratio were not observed.

Combining measurements of electric potential and pH with such of chlorophyll fluorescence and leaf gas exchange showed heat stimulation to evoke an electrical signal (propagation speed: 3-5 mm s(-1)) that travelled through the leaf while reducing the net CO(2) uptake rate and the photochemical quantum yield of both photosystems (PS). Two-dimensional imaging analysis of the chlorophyll fluorescence signal of PS II revealed that the yield reduction spread basipetally via the veins through the leaf at a speed of 1.6 +/- 0.3 mm s(-1) while the propagation speed in the intervein region was c. 50 times slower. Propagation of the signal through the veins was confirmed because PS I, which is present in the bundle sheath cells around the leaf vessels, was affected first. Hence, spreading of the signal along the veins represents a path with higher travelling speed than within the intervein region of the leaf lamina. Upon the electrical signal, cytoplasmic pH decreased transiently from 7.0 to 6.4, while apoplastic pH increased transiently from 4.5 to 5.2. Moreover, photochemical quantum yield of isolated chloroplasts was strongly affected by pH changes in the surrounding medium, indicating a putative direct influence of electrical signalling via changes of cytosolic pH on leaf photosynthesis.

Based on the growth-differentiation balance theory (GDB) and the influence of tropospheric ozone (O3) on plants, we hypothesized that pre-conditioning with elevated O3 reduces adverse effects of the root rot pathogen Phytophthora citricola Sawada. To this end a 2-year phytotron study with juvenile European beech (Fagus sylvatica L.) and (Picea abies [L.] Karst.) grown in mixture was performed. The hypothesis was tested on phenological, leaf and root morphological as well as physiological aspects of plant performance. Contrasting with spruce, elevated O3 limited leaf and root biomass development, photosynthetic performance and N uptake of beech. The growth limitation by O3 conveyed increased resistance in beech against the pathogen. Conversely, spruce displayed enhanced susceptibility in the combined O3/P. citricola treatment. The hypothesis was supported in the case of beech rather than spruce. Nevertheless, conclusions support GDB regarding the trade-off between growth and stress defense, although compliance appears to be species-specific.

The effects of elevated O3 on photosynthetic properties in adult beech trees (Fagus sylvatica) were investigated in relation to leaf mass per area as a measure of the gradually changing, within-canopy light availability. Leaves under elevated O3 showed decreased stomatal conductance at unchanged carboxylation capacity of Rubisco, which was consistent with enhanced delta 13C of leaf organic matter, regardless of the light environment during growth. In parallel, increased energy demand for O3 detoxification and repair was suggested under elevated O3 owing to enhanced dark respiration. Only in shade-grown leaves,light-limited photosynthesis was reduced under elevated O3, this effect being accompanied by lowered F(v)/F(m). These results suggest that chronic O3 exposure primarily caused stomatal closure to adult beech trees in the field regardless of the within-canopy light gradient. However, light limitation apparently raised the O3 sensitivity of photosynthesis and accelerated senescence in shade leaves.

Saplings of Fagus sylvatica and Picea abies were grown under conditions of intra and interspecific competition in a 2-year phytotron study under combinations of ambient and elevated ozone (+O3 which is 2 × O3, but

A lysimeter study was performed to monitor effects of elevated ozone on juvenile trees of Fagus sylvatica L. as well as on the plant–soil system. During a fumigation period over almost three growing seasons, parameters related to plant growth, phenological development and physiology as well as soil functions were studied. The data analyses identified elevated ozone to delay leaf phenology at early and to accelerate it at late developmental stages, to reduce growth, some leaf nutrients (Ca, K) as well as some soluble phenolics (hydroxycinnamic acid derivatives, total flavonol glycosides). No or very weak ozone effects were found in mobile carbon pools of leaves (starch, sucrose), and other phenolic compounds (flavans). Altered gene expression related to stress and carbon cycling corresponded well with findings from leaf phenology and chemical composition analyses indicating earlier senescence and oxidative stress in leaves under elevated ozone. Conversely in the soil system, no effects of ozone were detected on soil enzyme activities, rates of litter degradation and lysimeter water balances. Despite the fact that the three reported years 2003–2005 were climatically very contrasting including a hot and dry as well as an extremely wet summer, and also mild as well as cold winters, the influence of ozone on a number of plant parameters is remarkably consistent, further underlining the phytotoxic potential of elevated tropospheric ozone levels.

In Mimosa pudica L., heat stimulation triggers leaflet folding in local, neighbouring and distant leaves. Stomatal movements were observed microscopically during this folding reaction and electrical potentials, chlorophyll fluorescence, and leaf CO(2)/H(2)O-gas exchange were measured simultaneously. Upon heat stimulation of a neighbouring pinna, epidermal cells depolarized and the stomata began a rapid and pronounced transient opening response, leading to an approximately 2-fold increase of stomatal aperture within 60 s. At the same time, net CO(2) exchange showed a pronounced transient decrease, which was followed by a similar drop in photochemical quantum yield at photosystem (PS) II. Subsequently, CO(2)-gas exchange and photochemical quantum yield recovered and stomata closed partly or completely. The transient and fast stomatal opening response is interpreted as a hydropassive stomatal movement caused by a sudden loss of epidermal turgor. Thus, epidermal cells appear to respond in a similar manner to heat-induced signals as the pulvinar extensor cells. The subsequent closing of the stomata confirms earlier reports that stomatal movements can be induced by electrical signals. The substantial delay (several minutes) of guard cell turgor loss compared with the immediate response of the extensor and epidermal cells suggests a different, less direct mechanism for transmission of the propagating signal to the guard cells.

To gain an understanding of the role of electrical signaling in trees, poplar (Populus trichocarpa, Populus tremula × P. tremuloides) shoots were stimulated by chilling as well as flaming. Two kinds of signal propagation were detected by microelectrode measurements (aphid technique) in the phloem of leaf veins: (1) basipetal, short-distance signaling that led to rapid membrane hyperpolarization caused by K+-efflux within the leaf lamina; and (2) acropetal, long-distance signaling that triggered depolarization of the membrane potential in the leaf phloem. In the latter, the depolarizing signals travel across the stem from the manipulated leaves to adjacent leaves where the net CO2 uptake rate is temporarily depressed toward compensation. With regard to photosystem II, both heat-induced long-distance and short-distance signaling were investigated using two-dimensional “imaging” analysis of chlorophyll fluorescence. Both types of signaling significantly reduced the quantum yield of electron transport through photosystem II. Imaging analysis revealed that the signal that causes yield reduction spreads through the leaf lamina. Coldblocking of the stem proved that the electrical signal transmission via the phloem becomes disrupted, causing the leaf gas exchange to remain unaffected. Calcium-deficient trees showed a marked contrast inasmuch as the amplitude of the electrical signal was distinctly reduced, concomitant with the absence of a significant response in leaf gas exchange upon flame wounding. In summary, the above results led us to conclude that calcium as well as potassium is involved in the propagation of phloem-transmitted electrical signals that evoke specific responses in the photosynthesis of leaves.

To test the involvement of ethylene in mediating ozone- induced cell death and leaf loss in European beech ( Fagus sylvatica L.), tree seedlings were exposed to proportionally increased or decreased field ozone levels for up to 6 months. Ozone treatment caused cell death and acceler- ated leaf loss at higher than ambient levels, but had only minor effects at ambient and no effects at subambient ozone levels. The emission of ethylene, the levels of its precursor, 1-aminocyclopropane-1-carboxylate (ACC), and mRNA levels of specific ACC synthase ( FS-ACS2 ) and ACC oxidase ( FS-ACO1 ) isoforms showed a persistent increase and preceded cell death by approximately 2 weeks. Inhibition of ethylene biosynthesis led to reduced lesion formation whereas application of ACC accelerated ozone- induced cell death and leaf loss. Similar results were obtained when adult beech trees were exposed to 2 ¥ ¥ ¥ ozone by a whole tree free-air canopy exposure system. The results suggest a role of ethylene in amplifying ozone effects under field conditions in this major European broad-leaved tree species.

Summary • In the sensitive species Mimosa pudica electric signals arise when the leaves are stimulated by touching or wounding. Experiments reported here provide information about a photosynthetic response that results from heat-induced electrical signalling in leaves. • Electric potential measurements, combined with chlorophyll fluorescence, as well as gas exchange measurements showed that wounding evokes an electrical signal that travels rapidly into the neighbouring leaf pinna to eliminate the net-CO 2 uptake rate. At the same time the PSII quantum yield of electron transport is reduced from c . 0.6 to 0.2. Two-dimensional imaging analysis of the chlorophyll fluorescence signal revealed that the yield reduction spreads acropetally through the pinna and via the veins through the leaflets. •T o determine the speed of a chemical signal, a part of a pinna was exposed to 14 CO 2 . The remaining parts of the leaf were provided with label only when the translocation was extended to 12 h, indicating that a chemical signal is much too slow to account for the photosynthetic response after heat stimulation. • The results provide evidence for a role of the electrical signal in the regulation of photosynthesis because the high speed of the signal transduction rules out the involvement of a chemical signal, and the photosynthetic response occurs after the arrival of the electrical signal in the leaf pinna.

International audience; Microclimate and tree age have been suggested to be factors influencing the defense capacity against oxidative stress. Therefore, 5-year-old Fagus sylvatica seedlings were grown on a scaffolding in the sun and shade crown of 55-year-old trees throughout one growing season. Independent of tree age sun leaves had a lower specific leaf area, lower pigment contents and a more capacitive antioxidative system than shade leaves. In addition, in the sun crown leaves of seedlings displayed a higher specific leaf area than leaves of adult trees. Age dependent changes in leaf morphology were related to changes in the defense capacity against oxidative stress, with area based concentrations of antioxidants and pigments increasing with tree age. Thus our results suggest that differences in the response to oxidative stress may be attributed to age and crown position related differences in the specific leaf area, the latter influencing the biochemical and physiological performance of Fagus sylvatica leaves.; L'influence du microclimat et de l'âge de l'arbre sur la capacité défensive du hêtre européen (Fagus sylvatica L.) contre le stress oxidatif. Le microclimat et l'âge de l'arbre sont considerés comme des facteurs qui influencent la capacité défensive contre le stress oxidatif. Par conséquent, des plants de Fagus sylvatica de cinq ans ont grandi sur un échafaudage à la couronne des arbres âgés de 55 ans, une fois au soleil, une autre fois à l'ombre pendant une période de croissance. Indépendamment de l'âge de l'arbre, le feuillage exposé au soleil avait une surface spécifique de la feuille inférieure, un contenu de pigment inférieur et un système antioxidatif plus puissant que le feuillage à l'ombre. De plus, au niveau de la couronne la plus haute, le feuillage des plants a développé une surface spécifique plus élevée que celui des arbres adultes. Des transformations de morphologie du feuillage, qui dépendent de l'âge des feuilles, ont été comparées avec des transformations de la capacité contre le stress oxidatif. À cela la teneur en substances antioxidatives relative à la surface de la feuille a augmenté avec l'âge de l'arbre. Donc nos résultats suggèrent que les différences de réaction au stress oxidatif peuvent être attribuées aux différences de la surface spécifique de la feuille - dépendant de l'âge et de la position à la couronne - ce qui influence la performance biochimique et physiologique des feuilles du Fagus sylvatica.

summary Young trees of European beech (Fagus sylvatica) acclimated for one growing season to ambient (c. 367 l ll ’" )o r elevated CO # levels (c. 660 l ll ’") were exposed during the subsequent year to combinations of the same CO # regimes and ambient or twice-ambient ozone (O $ ) levels (generated from the database of a rural site). By the end of June, before the development of macroscopic leaf injury, the raised O $ levels had not aected the light and dark reactions of photosynthesis. However, acclimation to elevated CO # had resulted in lowered chlorophyll and nitrogen concentrations, whereas photosynthetic performance, examined over a wide range of parameters from light and dark reactions, remained unchanged or showed only slight reductions (e.g. apparent electron transport rate, ETR; apparent quantum yield of CO # gas exchange, U CO# ; apparent carboxylation eciency, CE; and photosynthetic capacity at light and CO # saturation, PC). In August, after the appearance of leaf necroses, plants grown under ambient CO # and twice-ambient O $ conditions declined in both the photosynthetic light reactions (optimum electron quantum yield, F v }F m , non-photochemical energy quenching, NPQ, reduction state of Q A , apparent electron quantum yield, U PSI I, maximum electron transport rates) and the dark reactions as reflected by CE, U CO# , as well as the maximum CO # uptake rate (i.e. PC). CE, U CO# and PC were reduced by c. 75, 40 and 75%, respectively, relative to plants exposed to ambient CO # and O $ levels. By contrast, plants exposed to twice-ambient O $ and elevated CO # levels maintained a photosynthetic performance similar to individuals grown either under ambient CO # and ambient O $ , or elevated CO # and ambient O $ conditions. The long-term exposure to elevated CO # therefore tended to counteract adverse chronic eects of enhanced O $ levels on photosynthesis. Possible reasons for this compensatory eect in F. sylvatica are discussed.

The large majority of the Ca. 150 species of the neotropical shrub and tree genus Clusia have the potential to perform Crassulacean acid metabolism (CAM). They are either obligate CAM plants or C3/CAM intermediate plants. Only a very small percentage of the plants studied so far are apparently obligate C3 species. Among these was C. criuva, until recent laboratory studies showed that it may also have a certain CAM capacity under artificial stress conditions. Measurements of stomatal conductance (porometry) and chlorophyll fluorescence variables of C. criuva occurring along a transect from deep shade inside a gallery forest across the semi-shaded ecotone towards a cerrado and into the exposed cerrado itself in central Brazil now show that it can adapt its photosynthetic apparatus to effective performance of C3 photosynthesis under highly different photosynthetic photon flux densities. In addition, however, it does have a certain potential for CAM and cannot be considered as a strictly obligate C3 plant. Should a basic capacity for performing CAM be a general property of the genus, the quest for CAM traits in other remaining putatively obligate C3 species of the genus ought to be pursued.

Allocation of recent photoassimilates of juvenile beech and spruce in response to twice-ambient ozone (2 × O(3)) and plant competition (i.e. intra vs. inter-specific) was examined in a phytotron study. To this end, we employed continuous (13)CO(2)/(12)CO(2) labeling during late summer and pursued tracer kinetics in CO(2) released from stems. In beech, allocation of recent photoassimilates to stems was significantly lowered under 2 × O(3) and increased in spruce when grown in mixed culture. As total tree biomass was not yet affected by the treatments, C allocation reflected incipient tree responses providing the mechanistic basis for biomass partitioning as observed in longer experiments. Compartmental modeling characterized functional properties of substrate pools supplying respiratory C demand. Respiration of spruce appeared to be exclusively supplied by recent photoassimilates. In beech, older C, putatively located in stem parenchyma cells, was a major source of respiratory substrate, reflecting the fundamental anatomical disparity between angiosperm beech and gymnosperm spruce.

Drought reduces the carbon (C) assimilation of trees and decouples aboveground from belowground carbon fluxes, but little is known about the response of drought-stressed trees to rewetting. This study aims to assess dynamics and patterns of C allocation in beech saplings under dry and rewetted soil conditions. In October 2010, 5-year-old beech saplings from a forest site were transplanted into 20 l pots. In 2011, the saplings were subjected to different levels of soil drought ranging from non-limiting water supply (control) to severe water limitation with soil water potentials of less than -1.5 MPa. As a physiologically relevant measure of drought, the cumulated soil water potential (i.e., drought stress dose (DSD)) was calculated for the growing season. In late August, the saplings were transferred into a climate chamber and pulse-labeled with (13)C-depleted CO2 (δ(13)C of -47‰). Isotopic signatures in leaf and soil respiration were repeatedly measured. Five days after soil rewetting, a second label was applied using 99 atom% (13)CO2. After another 12 days, the fate of assimilated C in each sapling was assessed by calculating the (13)C mass balance. Photosynthesis decreased by 60% in saplings under severe drought. The mean residence time (MRT) of recent assimilates in leaf respiration was more than three times longer than under non-limited conditions and was positively correlated to DSD. Also, the appearance of the label in soil respiration was delayed. Within 5 days after rewetting, photosynthesis, MRT of recent assimilates in leaf respiration and appearance of the label in soil respiration recovered fully. Despite the fast recovery, less label was recovered in the biomass of the previously drought-stressed plants, which also allocated less C to the root compartment (45 vs 64% in the control). We conclude that beech saplings quickly recover from extreme soil drought, although transitional after-effects prevail in C allocation, possibly due to repair-driven respiratory processes.

Stomata tend to narrow under ozone (O(3)) impact, leading to limitation of stomatal O(3) influx. Here, we review stomatal response under recently conducted free-air O(3) exposure experiments on two species of the same tree genus: Fagus sylvatica at Kranzberg Forest (Germany) and F. crenata at Sapporo Experimental Forest (Japan). Both beeches exhibited reduction in stomatal conductance (gs) by 10-20% under experimentally enhanced O(3) regimes throughout the summer relative to ambient-air controls. Stomatal narrowing occurred, in early summer, in the absence of reduced carboxylation capacity of Rubisco, although photosynthetic net CO(2) uptake rate temporarily reflected restriction to some minor extent. Observed stomatal narrowing was, however, diminished in autumn, suggesting gradual loss of stomatal regulation by O(3). Monotonic decline in gs with cumulative O(3) exposure or flux in current modeling concepts appear to be unrealistic in beech.

Diurnal measurements of chlorophyll fluorescence parameters and stomatal conductance to water vapour were carried out in five woody species and a bromeliad from two coastal vegetation types in Brazil, the rock outcrop of Pao de Acucar and the sand dunes of Macae. The environmental conditions of both study sites are characterized by high temperatures and light levels. The studied species comprised C3 plants with different degrees of stomatal closure during the day, overall daily stomatal conductance and a plant with a typical CAM pattern. Plants on Pao de Acucar exhibited only a small decline in potential quantum yield throughout the day. The non-photochemical quenching and the approximate photosynthetic electron transport rates were maximal during the peak of irradiance. In Macae, light response curves of fluorescence parameters in the CAM-tree Clusia hilariana showed a clear differentiation between phases III and IV of CAM. In phase III, decarboxylation of organic acids probably maintained high internal CO2 levels and there was only a small decrease in photochemical quenching with saturating light levels. In phase IV, the depletion of the organic acid pool and low stomatal conductances resulted in much lower levels of effective quantum yield and a substantial increase in both 1-qP and non-photochemical quenching with increasing light levels. This behaviour during phase IV in the CAM-tree C. hilariana was comparable to the performance of the C3-ClusiaC. aff parviflora at Pao de Acucar. It is seen that both the C3 species and the CAM tree occurring in the two coastal communities effectively adjust their photochemical efficiency to environmental conditions and especially to diurnal variations of stress.

Changes in chlorophyll a fluorescence during the day and diurnal-changes of net CO2-exchange and organic acid contents were determined in two species of the genus Clusia during the dry season in Venezuela. The investigations included plants of the C3/CAM intermediate species Clusia minor and the C3 species C. multiflora growing at exposed and shaded sites. Both species showed a C3 pattern of net CO2-exchange at the exposed site. In the shade under extreme drought stress C. minor showed a weak expression of CAM without CO2-uptake during the afternoon (phase IV of CAM). C. multiflora growing in the shade exhibited a C3-pattern of net CO2-exchange and a small but significant nocturnal accumulation of citrate. Shaded plants of C. minor were able to double their light utilisation for electron transport and to reduce non-photochemical quenching during phase III compared to phase II of CAM. Furthermore, increase of electron transport rate through photosystem II in phase III of CAM is correlated to decarboxylation of malate. At the exposed site C. multiflora was less negatively affected by high PPFD than C. minor. This was shown by a lower reduction of potential electron quantum yield (Fv/Fm) and higher light utilisation of electron transport of C. multiflora compared to C. minor. At the exposed site C. minor did not make use of the CAM option to increase light utilisation of electron transport and to reduce non-photochemical quenching as did the plants growing in the shade.

Four different Clusia species showing C 3 photosynthesis (C. multiflora) or crassulacean acid metabolism (CAM) (C. rosea, an unidentified Clusia sp. and C. alata) co-occur in the same habitat in the northwest of Venezuela. The aim of this field study was to correlate the adaptive changes in chlorophyll a fluorescence with gas exchange patterns and deacidification of nocturnally accumulated organic acids. The results indicate that in C. rosea and Clusia sp. a lowering of the potential quantum yield, measured as F v /F m after 10 min of dark adaptation, is restricted to the early morning hours. With the onset of deacidification there was no further decline in F v /F m , although the irradiance upon the leaves remained high. For the C 3 -plant, C. multiflora, changes in F v /F m were correlated with changes in irradiance incident on the leaves before predarkening for 10 min for F v /F m measurements. The fast relaxation of F v /F m under low light indicates that C. multiflora was not suffering photoinhibitory damage. Effective quantum yield as determined by ΔF/F' m , was low under high irradiances for C. rosea and Clusia sp. and did not change significantly for C. rosea during phase III of CAM. During the time of low incident irradiance C. alata and C. multiflora reached an effective quantum yield that was close to the optimal quantum yield of electron transport. It is noteworthy that apparent electron transport rates through photosystem II (PS II) ((ΔF/F' m ) x PPFD) were highest for C. rosea, Clusia sp. and C. alata during phase III of CAM. Electron transport rates in C. multiflora were lower under high irradiance and did not differ from those found in phases II and IV of CAM. The possible role of CO 2 evolution from acid decarboxylation with respect to photosynthetic efficiency is discussed.

Samples of the Clusiaceae generaClusia, Oedematopus andDystovomita were collected at various sites and different altitudes in northern and south-western Venezuela. Analyses of stable isotopes of carbon and hydrogen and of leaf-nitrogen levels were performed on the dried samples. Correlations among these variables, i.e. carbon isotope discrimination (Δ), hydrogen isotope ratio (δD) and N-levels, and with altitude were assessed. In the samples, where values of Δ above 15‰ indicate predominant performance of C3 photosynthesis, there were slight tendencies of increasing Δ, δD and N-levels with increasing altitude and of increasing Δ with increasing N. Although these correlations taken separately were not statistically significant, they support each other and indicate increasing transpiration and increased leaf-nutrient supply at increasing altitude. Performance of crassulacean acid metabolism (CAM) in species ofClusia appears to be restricted to altitudes below 1500 m a.s.l. There was a significant negative correlation of Δ with altitude in the samples, where values of Δ below 10‰ indicated predominant performance of CAM. This suggests that phases II and IV of CAM are progressively suppressed towards the upper altitudinal limit of CAM inClusia in northern Venezuela. It is concluded that among the large number of environmental factors and combinations thereof, which determine the expression of CAM inClusia and trigger C3-CAM transitions in C3/CAM intermediate species, low availability of water is the most important.

Sandy plains are characteristic of the coastal region of Brazil. We investigated the diel patterns of changes in organic acid levels, leaf conductance and chlorophylla fluorescence for sun-exposed and shaded plants ofClusia hilariana, one of the dominant woody species in the sandy coastal plains of northern Rio de Janeiro state. Both exposed and shaded plants showed a typical CAM pattern with considerable diel oscillations in organic acid levels. The degradation of both malic and citric acids during the midday stomatal closure period could lead to potential CO2 fixation rates of 28 μmol m-2 s-1 in exposed leaves. Moreover, exposed leaves exhibited large increases in total non-photochemical quenching (qN) accompanied by a substantial decrease in effective quantum yield during the course of the day. However, these potential high rates of CO2 fixation and the increases inqn of exposed plants were not enough to maintain the primary electron acceptor of photosystem II (qA) in a low reduction state, similar to that of shaded plants. As a result, there was a moderate increase in the reduction state of qA throughout the day. Most of the decline in photochemical efficiency of exposed leaves ofC. hilariana was reversible, as evidenced by the high levels of pre-dawn potential quantum yields (Fv/Fm) and their rapid recovery after sunset. However, the depletion of the organic acid pool in the afternoon resulted in an accentuated subsequent drop in Fv/Fm, suggesting that prolonged periods of water stress accompanied by high irradiance levels may expose plants ofC. hilariana in unprotected habitats to the danger of photoinhibition.

Canopies of adult European beech (Fagus sylvatica) and Norway spruce (Picea abies) were labeled with CO(2) depleted in (13)C to evaluate carbon allocation belowground. One-half the trees were exposed to elevated O(3) for 6 yrs prior to and during the experiment. Soil-gas sampling wells were placed at 8 and 15 cm and soil CO(2) was sampled during labeling in mid-late August, 2006. In beech, delta(13)CO(2) at both depths decreased approximately 50 h after labeling, reflecting rapid translocation of fixed C to roots and release through respiration. In spruce, label was detected in fine-root tissue, but there was no evidence of label in delta(13)CO(2). The results show that C fixed in the canopy rapidly reaches respiratory pools in beech roots, and suggest that spruce may allocate very little of recently-fixed carbon into root respiration during late summer. A change in carbon allocation belowground due to long-term O(3) exposure was not observed.

In this study, the effects of different light intensities either in direct sunlight or in the shade crown of adult beech (Fagus sylvatica L.) trees on delta13C and Delta18O were determined under ambient (1 x O3) and twice-ambient (2 x O3) atmospheric ozone concentrations during two consecutive years (2003 and 2004). We analysed the isotopic composition in leaf bulk, leaf cellulose, phloem and xylem material and related the results to (a) meteorological data (air temperature, T and relative humidity, RH), (b) leaf gas exchange measurements (stomatal conductance, g(s); transpiration rate, E; and maximum photosynthetic activity, A(max)) and (c) the outcome of a steady-state evaporative enrichment model. Delta13C was significantly lower in the shade than in the sun crown in all plant materials, whilst Delta18O was increased significantly in the shade than in the sun crown in bulk material and cellulose. Elevated ozone had no effect on delta13C, although Delta18O was influenced by ozone to varied degrees during single months. We observed significant seasonal changes for both parameters, especially in 2004, and also significant differences between the study years. Relating the findings to meteorological data and gas exchange parameters, we conclude that the differences in Delta18O between the sun and the shade crown were predominantly caused by the Péclet effect. This assumption was supported by the modelled Delta18O values for leaf cellulose. It was demonstrated that independent of RH, light-dependent reduction of stomatal conductance (and thus transpiration) and of A(max) can drive the pattern of Delta18O increase with the concomitant decrease of delta13C in the shade crown. The effect of doubling ozone levels on time-integrated stomatal conductance and transpiration as indicated by the combined analysis of Delta18O and delta13C was much lower than the influence caused by the light exposure.

Combined delta(13)C and delta(18)O analyses of leaf material were used to infer changes in photosynthetic capacity (A(max)) and stomatal conductance (g(l)) in Fagus sylvatica and Picea abies trees growing under natural and controlled conditions. Correlation between g(l) and delta(18)O in leaf cellulose (delta(18)O(cel)) allowed us to apply a semi-quantitative model to infer g(l) from delta(18)O(cel) and also interpret variation in delta(13)C as reflecting variation in A(max). Extraction of leaf cellulose was necessary, because delta(18)O from leaf organic matter (delta(18)O(LOM)) and delta(18)O(cel) was not reliably correlated. In juvenile trees, the model predicted elevated carbon dioxide (CO(2)) to reduce A(max) in both species, whereas ozone (O(3)) only affected beech by reducing CO(2) uptake via lowered g(l). In adult trees, A(max) declined with decreasing light level as g(l) was unchanged. O(3) did not significantly affect isotopic signatures in leaves of adult trees, reflecting the higher O(3) susceptibility of juvenile trees under controlled conditions. The isotopic analysis compared favourably to the performance of leaf gas exchange, underlining that the semi-quantitative model approach provides a robust way to gather time-integrated information on photosynthetic performance of trees under multi-faced ecological scenarios, in particular when information needed for quantitative modelling is only scarcely available.

Root respiration has been shown to increase with temperature, but less is known about how this relationship is affected by the fungal partner in mycorrhizal root systems. In order to test respiratory temperature dependence, in particular Q (10) of mycorrhizal and non-mycorrhizal root systems, seedlings of PICEA ABIES (L.) Karst. (Norway spruce) were inoculated with the ectomycorrhizal fungus PILODERMA CROCEUM (Eriksson and Hjortstam, SR430; synonym: PILODERMA FALLAX: [Libert] Stalpers) and planted in soil respiration cuvettes (mycocosms). Temperature dependence of hyphal respiration in sterile cultures was determined and compared with respiration of mycorrhizal roots. Respiration rates of mycorrhizal and non-mycorrhizal root systems as well as sterile cultures were sensitive to temperature. Q (10) of mycorrhizal root systems of 3.0 +/- 0.1 was significantly higher than that of non-mycorrhizal systems (2.5 +/- 0.2). Q (10) of P. CROCEUM in sterile cultures (older than 2 months) was similar to that of mycorrhizal root systems, suggesting that mycorrhizae may have a large influence on the temperature sensitivity of roots in spite of their small biomass. Our results stress the importance of considering mycorrhization when modeling the temperature sensitivity of spruce roots.

In a two-year phytotron study, juvenile trees of European beech (Fagus sylvatica) and Norway spruce (Picea abies) were grown in mixture under ambient and twice ambient ozone (O3) and infected with the root pathogen Phytophthora citricola. We investigated the influence of O3 on the trees' susceptibility to the root pathogen and assessed, through a 15N-labelling experiment, the impact of both treatments (O3 exposure and infection) on belowground competitiveness. The hypotheses tested were that: (1) both P. citricola and O3 reduce the belowground competitiveness (in view of N acquisition), and (2) that susceptibility to P. citricola infection is reduced through acclimation to enhanced O3 exposure. Belowground competitiveness was quantified via cost/benefit relationships, i.e., the ratio of structural investment in roots relative to their uptake of 15N. Beech had a lower biomass acquisition and captured less 15N under enhanced O3 and P. citricola infection alone than spruce, whereas the latter species appeared to profit from the lower resource acquisition of beech in these treatments. Nevertheless, in the combined treatment, susceptibility to P. citricola of spruce was increased, while beech growth and 15N uptake were not further reduced below the levels found under the single treatments. Potential trade-offs between stress defence, growth performance, and associated nitrogen status are discussed for trees affected through O3 and/or pathogen infection. With respect to growth performance, it is concluded that O3 enhances susceptibility to the pathogen in spruce, but apparently raises the defence capacity in beech.

The aim was to analyze functional changes in the mycorrhizosphere (MR) of juvenile spruce and beech grown in a mixture under ambient and twice ambient ozone and inoculated with the root pathogen Phytophthora citricola. The phytotron experiment was performed over two vegetation periods, adding the pathogen at the end of the first growing season. Root biomass data suggest that the combined treatment affected spruce more than beech and that the negative influence of ozone on stress tolerance against the root pathogen P. citricola was greater for spruce than for beech. In contrast, beech was more affected when the pathogen was the sole stressor. The functional soil parameter chosen for studies of MR soil samples was activity of extracellular enzymes. After the first year of ozone exposure, MR soil samples of both species showed increased activity of almost all measured enzymes (acid phosphatase, chitinase, beta-glucosidase, cellobiohydrolase) in the O3 treatment. Species-specific differences were observed, with a stronger effect of P. citricola on beech MR and a stronger ozone effect on spruce MR. In the second year, the effects of the combined treatment (ozone and P. citricola) were a significant increase in the activity of most enzymes (except cellobiohydrolase) for both tree species. The results indicated that responsiveness of MR soils towards ozone and P. citricola was related to the severity of infection of the plants and the reduction of belowground biomass, suggesting a strong, direct influence of plant stress on MR soil enzyme activity. Additional research is needed using different species and combined stresses to determine the broader ecological relevance of shifts in rhizosphere enzymes.

Three and four-year-old saplings of beech (Fagus sylvatica L.) and spruce (Picea abies (L.) Karst.) grown in monoculture and mixed culture were exposed in phytotrons to (1) ambient air, (2) elevated pO3, (3) elevated pCO2, or (4) elevated pCO2 plus elevated pO3. After 5 months, the contents of soluble sugars, starch, soluble amino compounds, non-structural proteins (NSP), as well as reduced (GSH) and oxidized (GSSG) glutathione were determined in the leaves of both species in order to assess the effects of the gaseous regimes on primary metabolism. Elevated pO3 did not affect sugar and starch levels in beech leaves in monoculture, but significantly increased sugar levels in beech leaves grown in mixed culture. In spruce needles, sugar levels tended to be enhanced in both culture types. Individual and combined exposure of elevated pCO2 led to an increase in non-structural carbohydrate (soluble sugars plus starch) levels in beech and spruce leaves of both culture types. Differences in the responses of non-structural carbohydrate levels to elevated pCO2 between beech and spruce were apparent from different contributions of sugars and starch to the increase in carbohydrate levels. Exposure to elevated pCO2 and/or elevated pO3 did not affect the levels of soluble amino compounds and NSP in beech leaves, but reduced amino compound levels in spruce needles of both culture types. Elevated pO3 increased GSH levels in the leaves of both tree species in both culture types, while GSSG levels in monoculture were reduced in beech leaves, but significantly enhanced in spruce needles. Elevated pCO2 reduced GSSG levels in beech and spruce leaves in monoculture, and GSH levels in spruce needles of both culture types. The combination of elevated pCO2 and pO3 increased GSSG levels in beech leaves of both culture types and in spruce needles in monoculture, but reduced GSH levels in spruce needles of both culture types. Apparently, under each gaseous regime, the culture type significantly altered primary metabolism of the leaves of beech and spruce. © 2005 Elsevier SAS. All rights reserved.

This chapter deals with the processes that affect the carbon and hydrogen isotope ratios of plant cellulose. Under natural conditions the influence of a single climate parameter on the isotope composition of tree-rings cannot be evaluated exactly, because of the strong interdependences of the climatic parameters. Results from growth chamber experiments with bean plants (Vicia faba) are presented, in which temperature, relative humidity and irrigation were varied independently from each other. All these parameters were found to be negatively correlated with the δ13C values of stem cellulose. The δD values of stem cellulose were influenced mainly by the humidity of the ambient air and by the isotope signal of the source water. The experimental results may help to explain δD and δ13C values in plant material for the purpose of climate reconstructions. As an example, stable isotope chronologies from tree-rings of spruces (Picea abies) from two different sites in Southern Germany are interpreted in the light of the experimental results.

Plants of the CAM species Kalanchoe daigremontiana grown at elevated temperatures (34/25°C day/night) did not show endogenous net CO 2 -exchange rhythm at 34°C whereas at 30-32°C the rhythm was present. In contrast, the endogenous rhythm never occurs above 30°C in plants grown at 25/15°C day/night.

Cumulative ozone (O3) uptake and O3 flux were related to physiological, morphological and biochemical characteristics of Norway spruce (Picea abies (L.) Karst.) trees of different ages. Under ambient CO2 conditions, photosynthetic capacity (Amax) declined in mature trees when cumulative O3 uptake into needles, which provides a measure of effective O3 dose, exceeded 21 mmol m-2 of total needle surface area. A comparable decline in Amax of seedlings occurred when cumulative O(3) uptake was only 4.5 mmol m-2. The threshold O3 flux causing a significant decline in Amax ranged between 2.14 and 2.45 nmol m-2 s-1 in mature trees and seedlings subjected to exposure periods of > or = 70 and > or = 23 days, respectively. The greater O3 sensitivity of young trees compared with mature trees was associated with needle morphology. Biomass of a 100-needle sample increased significantly with tree age, whereas a negative correlation was found for specific leaf area, these changes parallel those observed during differentiation from shade-type to sun-type needles with tree ontogeny. Age-dependent changes in leaf morphology were related to changes in detoxification capacity, with area-based concentrations of ascorbate increasing during tree ontogeny. These findings indicate that the extent of O3-induced injury is related to the ratio of potentially available antioxidants to O3 influx. Because this ratio, when calculated for ascorbate, increased with tree age, we conclude that the ratio may serve as an empirical basis for characterizing age-related differences in tree responses to O3.

Summary Most of the species in the genus Clusia are very flexible in their photosynthetic pathway, and are C 3 -photosynthesis/Crassulacean acid metabolism (CAM) intermediate species; therefore, the identification and clear assignment of Clusia -species to C 3 - or CAM-pathways is difficult. We made an attempt to characterize obligate C 3 -photosynthesis species within that genus. Therefore, we selected five species of the family Clusiacea (four of the genus Clusia and one of the genus Oedematopus ) on the basis of their low leaf succulence as potential obligate C 3 -photosynthesis species. All of them showed a C 3 -photosynthesis pattern of gas exchange under well-watered conditions and their δ 13 C-values were also found to be in the range of obligate C 3 -photosynthesis plants. Under drought stress two of them ( C. articulata and C. cruiva ) proved to be C 3 -photosynthesis/CAM intermediate species, while C. parviflora, C. multiflora and most likely O. mirandensis did not show any nocturnal CO 2 -fixation. However, in C. parviflora and O. mirandensis some nighttime accumulation of malate by internal CO 2 -recycling might occur. So far, C. multiflora seems to be the only known Clusia species with neither nocturnal net CO 2 -uptake nor malate accumulation.

Summary The facultative epiphytic bromeliads Nidularium procerum and N. innocentii grow terrestrially in distinct but neighbouring patches of a swamp forest understory in southeastern Brazil. N. innocentii (C 3 ) is restricted to a shaded, periodically flooded patch, and N. procerum (CAM) to a semi-exposed, permanently flooded patch. Semi-exposed and shaded leaves of both species were used to evaluate the efficiency of photochemical energy conversion in photosystem II and the extent of non-radiative energy dissipation of these two contrasting photosynthetic pathways. By late afternoon, both semi-exposed C 3 and CAM plants were still unable to fully recover from photoinhibition suffered earlier at the peak of irradiance, although this effect was more marked in the C 3 -species. Light response curves of fluorescence parameters obtained on a cloudy and a sunny day also suggested that semi-exposed conditions are more favourable to the CAM than to the C 3 species. Both the C 3 and the CAM Nidularium are shade-adapted, but CAM allows colonization by N. procerum of patches eventually exposed to higher irradiance. This partially explains the habitat segregation of these species in this swamp forest. It remains to be seen how rhizome flood-tolerance relates to this pattern.