Coordination between degree of isohydricity and depth of root water uptake in temperate tree species.

In an increasingly dry environment, it is crucial to understand how tree species use soil water and cope with drought. However, there is still a knowledge gap regarding the relationships between species-specific stomatal behaviour, spatial root distribution, and root water uptake (RWU) dynamics. Our study aimed to investigate above- and below-ground aspects of water use during soil drying periods in four temperate tree species that differ in stomatal behaviour: two isohydric tracheid-bearing conifers, Scots pine and Norway spruce, and two more anisohydric deciduous species, the diffuse-porous European beech, and the ring-porous Downy oak. From 2015 to 2020, soil-tree-atmosphere-continuum parameters were measured for each species in monospecific forests where trees had no access to groundwater. The hourly time series included data on air temperature, vapor pressure deficit, soil water potential, soil hydraulic conductivity, and RWU to a depth of 2 m. Analysis of drought responses included data on stem radius, leaf water potential, estimated osmotically active compounds, and drought damage. Our study reveals an inherent coordination between stomatal regulation, fine root distribution and water uptake. Compared to conifers, the more anisohydric water use of oak and beech was associated with less strict stomatal closure, greater investment in deep roots, four times higher maximum RWU, a shift of RWU to deeper soil layers as the topsoil dried, and a more pronounced soil drying below 1 m depth. Soil hydraulic conductivity started to limit RWU when values fell below 10-3 to 10-5 cm/d, depending on the soil. As drought progressed, oak and beech may also have benefited from their leaf osmoregulatory capacity, but at the cost of xylem embolism with around 50 % loss of hydraulic conductivity when soil water potential dropped below -1.25 MPa. Consideration of species-specific water use is crucial for forest management and vegetation modelling to improve forest resilience to drought.

Unravelling the impact of soil data quality on species distribution models of temperate forest woody plants.

Soil properties influence plant physiology and growth, playing a fundamental role in shaping species niches in temperate forest ecosystems. Here, we investigated the impact of soil data quality on the performance of species distribution models (SDMs) of 41 woody plant species in Swiss forests. We compared models based on measured soil properties with those based on digitally mapped soil properties on regional (Swiss Forest Soil Maps) and global scales (SoilGrids). We first calibrated topo-climatic SDMs with measured soil data and plant species presences and absences from mature temperate forest stand plots. We developed further models using the same soil predictors, but with values extracted from digital soil maps at the nearest neighbouring plots of the Swiss National Forestry Inventory. The predictive power of SDMs without soil information compared to those with soil information, as well as measured soil information vs digitally mapped, was evaluated with metrics of model performance and variable contribution. On average, models with measured and digitally mapped soil properties performed significantly better than those without soil information. SDMs based on measured and Swiss Forest Soil Maps showed higher performance, especially for species with an 'extreme' niche position (e.g., preference for high or low pH), compared to those using SoilGrids. Nevertheless, if no regional soil maps are available, SoilGrids should be tested for their potential to improve SDMs. Moreover, among the tested soil predictors, pH, and clay content of the topsoil layers most improved the predictive power of SDMs for forest woody plants. In conclusion, we demonstrate the value of regional soil maps for predicting the distribution of woody species across strong environmental gradients in temperate forests. The improved accuracy of SDMs and insights into drivers of distribution may support forest managers in strategies supporting e.g. biodiversity conservation, or climate adaptation planning.

Dendrometers challenge the 'moon wood concept' by elucidating the absence of lunar cycles in tree stem radius oscillation.

Wood is a sustainable natural resource and an important global commodity. According to the 'moon wood theory', the properties of wood, including its growth and water content, are believed to oscillate with the lunar cycle. Despite contradicting our current understanding of plant functioning, this theory is commonly exploited for marketing wooden products. To examine the moon wood theory, we applied a wavelet power transformation to series of 2,000,000 hourly stem radius records from dendrometers. We separated the influence of 74 consecutive lunar cycles and meteorological conditions on the stem variation of 62 trees and six species. We show that the dynamics of stem radius consist of overlapping oscillations with periods of 1 day, 6 months, and 1 year. These oscillations in stem dimensions were tightly coupled to oscillations in the series of air temperature and vapour pressure deficit. By contrast, we revealed no imprint of the lunar cycle on the stem radius variation of any species. We call for scepticism towards the moon wood theory, at least as far as the stem water content and radial growth are concerned. We foresee that similar studies employing robust scientific approaches will be increasingly needed in the future to cope with misleading concepts.

Elevation dependent response of soil organic carbon stocks to forest windthrow.

Storms represent a major disturbance factor in forest ecosystems, but the effects of windthrows on soil organic carbon (SOC) stocks are poorly quantified. Here, we assessed the SOC stocks of windthrown forests at 19 sites across Switzerland spanning an elevation gradient from 420 to 1550 m, encompassing a strong climatic gradient. Results show that the effect size of disturbance on SOC stocks increases with the size of the initial SOC stocks. The largest windthrow-induced SOC losses of up to 29 t C ha-1 occurred in high-elevation forests with a harsh climate developing thick organic layers. In contrast, SOC stocks of low-elevation forests with thin organic layers were hardly affected. A mineralization study further revealed high elevation forests to store higher amounts of easily mineralizable C in thick organic layers that got lost following windthrow. These findings are supported by a meta-analysis of available windthrow studies, showing an increase of storm-induced SOC losses with the size of the initial SOC stocks. Modelling simulations further indicate longer-lasting SOC losses and a slower recovery of SOC stocks after windthrow at high compared to low elevations, due to a slower regeneration of mountain forests and associated lower C inputs into soils in a harsh climate. Upscaling the experimental findings/observed patterns by linking them to a data base of Swiss forest soils shows a total SOC loss of ∼0.4 Mt. C for the whole forested area of Switzerland after two major storm events, counteracting the forest net carbon sink of decades. Our study provides strong evidence that the vulnerability of SOC stocks to windthrow is particularly high in forests featuring thick and slowly forming organic layers, such as mountain soils. Thus, the risk of losing SOC to more frequent windthrows in mountain forests strongly limits their potential to mitigate climate change.

Soil-plant interactions modulated water availability of Swiss forests during the 2015 and 2018 droughts.

Central Europe has been experiencing unprecedented droughts during the last decades, stressing the decrease in tree water availability. However, the assessment of physiological drought stress is challenging, and feedback between soil and vegetation is often omitted because of scarce belowground data. Here we aimed to model Swiss forests' water availability during the 2015 and 2018 droughts by implementing the mechanistic soil-vegetation-atmosphere-transport (SVAT) model LWF-Brook90 taking advantage of regionalized depth-resolved soil information. We calibrated the model against soil matric potential data measured from 2014 to 2018 at 44 sites along a Swiss climatic and edaphic drought gradient. Swiss forest soils' storage capacity of plant-available water ranged from 53 mm to 341 mm, with a median of 137 ± 42 mm down to the mean potential rooting depth of 1.2 m. Topsoil was the primary water source. However, trees switched to deeper soil water sources during drought. This effect was less pronounced for coniferous trees with a shallower rooting system than for deciduous trees, which resulted in a higher reduction of actual transpiration (transpiration deficit) in coniferous trees. Across Switzerland, forest trees reduced the transpiration by 23% (compared to potential transpiration) in 2015 and 2018, maintaining annual actual transpiration comparable to other years. Together with lower evaporative fluxes, the Swiss forests did not amplify the blue water deficit. The 2018 drought, characterized by a higher and more persistent transpiration deficit than in 2015, triggered widespread early wilting across Swiss forests that was better predicted by the SVAT-derived mean soil matric potential in the rooting zone than by climatic predictors. Such feedback-driven quantification of ecosystem water fluxes in the soil-plant-atmosphere continuum will be crucial to predicting physiological drought stress under future climate extremes.

Shotgun Metagenomics of Deep Forest Soil Layers Show Evidence of Altered Microbial Genetic Potential for Biogeochemical Cycling.

Soil microorganisms such as Bacteria and Archaea play important roles in the biogeochemical cycling of soil nutrients, because they act as decomposers or are mutualistic or antagonistic symbionts, thereby influencing plant growth and health. In the present study, we investigated the vertical distribution of soil metagenomes to a depth of 1.5 m in Swiss forests of European beech and oak species on calcareous bedrock. We explored the functional genetic potential of soil microorganisms with the aim to disentangle the effects of tree genus and soil depth on the genetic repertoire, and to gain insight into the microbial C and N cycling. The relative abundance of reads assigned to taxa at the domain level indicated a 5-10 times greater abundance of Archaea in the deep soil, while Bacteria showed no change with soil depth. In the deep soil there was an overrepresentation of genes for carbohydrate-active enzymes, which are involved in the catalyzation of the transfer of oligosaccharides, as well as in the binding of carbohydrates such as chitin or cellulose. In addition, N-cycling genes (NCyc) involved in the degradation and synthesis of N compounds, in nitrification and denitrification, and in nitrate reduction were overrepresented in the deep soil. Consequently, our results indicate that N-transformation in the deep soil is affected by soil depth and that N is used not only for assimilation but also for energy conservation, thus indicating conditions of low oxygen in the deep soil. Using shotgun metagenomics, our study provides initial findings on soil microorganisms and their functional genetic potential, and how this may change depending on soil properties, which shift with increasing soil depth. Thus, our data provide novel, deeper insight into the "dark matter" of the soil.

Number of growth days and not length of the growth period determines radial stem growth of temperate trees.

Radial stem growth dynamics at seasonal resolution are essential to understand how forests respond to climate change. We studied daily radial growth of 160 individuals of seven temperate tree species at 47 sites across Switzerland over 8 years. Growth of all species peaked in the early part of the growth season and commenced shortly before the summer solstice, but with species-specific seasonal patterns. Day length set a window of opportunity for radial growth. Within this window, the probability of daily growth was constrained particularly by air and soil moisture, resulting in intermittent growth to occur only on 29 to 77 days (30% to 80%) within the growth period. The number of days with growth largely determined annual growth, whereas the growth period length contributed less. We call for accounting for these non-linear intra-annual and species-specific growth dynamics in tree and forest models to reduce uncertainties in predictions under climate change.

Why trees grow at night.

The timing of diel stem growth of mature forest trees is still largely unknown, as empirical data with high temporal resolution have not been available so far. Consequently, the effects of day-night conditions on tree growth remained uncertain. Here we present the first comprehensive field study of hourly-resolved radial stem growth of seven temperate tree species, based on 57 million underlying data points over a period of up to 8 yr. We show that trees grow mainly at night, with a peak after midnight, when the vapour pressure deficit (VPD) is among the lowest. A high VPD strictly limits radial stem growth and allows little growth during daylight hours, except in the early morning. Surprisingly, trees also grow in moderately dry soil when the VPD is low. Species-specific differences in diel growth dynamics show that species able to grow earlier during the night are associated with the highest number of hours with growth per year and the largest annual growth increment. We conclude that species with the ability to overcome daily water deficits faster have greater growth potential. Furthermore, we conclude that growth is more sensitive than carbon uptake to dry air, as growth stops before stomata are known to close.

Deep Soil Layers of Drought-Exposed Forests Harbor Poorly Known Bacterial and Fungal Communities.

Soil microorganisms such as bacteria and fungi play important roles in the biogeochemical cycling of soil nutrients, because they act as decomposers or are mutualistic or antagonistic symbionts, thereby influencing plant growth and health. In the present study, we investigated the vertical distribution of the soil microbiome to a depth of 2 m in Swiss drought-exposed forests of European beech and oaks on calcareous bedrock. We aimed to disentangle the effects of soil depth, tree (beech, oak), and substrate (soil, roots) on microbial abundance, diversity, and community structure. With increasing soil depth, organic carbon, nitrogen, and clay content decreased significantly. Similarly, fine root biomass, microbial biomass (DNA content, fungal abundance), and microbial alpha-diversity decreased and were consequently significantly related to these physicochemical parameters. In contrast, bacterial abundance tended to increase with soil depth, and the bacteria to fungi ratio increased significantly with greater depth. Tree species was only significantly related to the fungal Shannon index but not to the bacterial Shannon index. Microbial community analyses revealed that bacterial and fungal communities varied significantly across the soil layers, more strongly for bacteria than for fungi. Both communities were also significantly affected by tree species and substrate. In deep soil layers, poorly known bacterial taxa from Nitrospirae, Chloroflexi, Rokubacteria, Gemmatimonadetes, Firmicutes and GAL 15 were overrepresented. Furthermore, archaeal phyla such as Thaumarchaeota and Euryarchaeota were more abundant in subsoils than topsoils. Fungal taxa that were predominantly found in deep soil layers belong to the ectomycorrhizal Boletus luridus and Hydnum vesterholtii. Both taxa are reported for the first time in such deep soil layers. Saprotrophic fungal taxa predominantly recorded in deep soil layers were unknown species of Xylaria. Finally, our results show that the microbial community structure found in fine roots was well represented in the bulk soil. Overall, we recorded poorly known bacterial and archaeal phyla, as well as ectomycorrhizal fungi that were not previously known to colonize deep soil layers. Our study contributes to an integrated perspective on the vertical distribution of the soil microbiome at a fine spatial scale in drought-exposed forests.

From the comfort zone to crown dieback: Sequence of physiological stress thresholds in mature European beech trees across progressive drought.

Drought responses of mature trees are still poorly understood making it difficult to predict species distributions under a warmer climate. Using mature European beech (Fagus sylvatica L.), a widespread and economically important tree species in Europe, we aimed at developing an empirical stress-level scheme to describe its physiological response to drought. We analysed effects of decreasing soil and leaf water potential on soil water uptake, stem radius, native embolism, early defoliation and crown dieback with comprehensive measurements from overall nine hydrologically distinct beech stands across Switzerland, including records from the exceptional 2018 drought and the 2019/2020 post-drought period. Based on the observed responses to decreasing water potential we derived the following five stress levels: I (predawn leaf water potential >-0.4 MPa): no detectable hydraulic limitations; II (-0.4 to -1.3): persistent stem shrinkage begins and growth ceases; III (-1.3 to -2.1): onset of native embolism and defoliation; IV (-2.1 to -2.8): onset of crown dieback; V (<-2.8): transpiration ceases and crown dieback is >20%. Our scheme provides, for the first time, quantitative thresholds regarding the physiological downregulation of mature European beech trees under drought and therefore synthesises relevant and fundamental information for process-based species distribution models. Moreover, our study revealed that European beech is drought vulnerable, because it still transpires considerably at high levels of embolism and because defoliation occurs rather as a result of embolism than preventing embolism. During the 2018 drought, an exposure to the stress levels III-V of only one month was long enough to trigger substantial crown dieback in beech trees on shallow soils. On deep soils with a high water holding capacity, in contrast, water reserves in deep soil layers prevented drought stress in beech trees. This emphasises the importance to include local data on soil water availability when predicting the future distribution of European beech.

Determinants of legacy effects in pine trees - implications from an irrigation-stop experiment.

Tree responses to altered water availability range from immediate (e.g. stomatal regulation) to delayed (e.g. crown size adjustment). The interplay of the different response times and processes, and their effects on long-term whole-tree performance, however, is hardly understood. Here we investigated legacy effects on structures and functions of mature Scots pine in a dry inner-Alpine Swiss valley after stopping an 11-yr lasting irrigation treatment. Measured ecophysiological time series were analysed and interpreted with a system-analytic tree model. We found that the irrigation stop led to a cascade of downregulations of physiological and morphological processes with different response times. Biophysical processes responded within days, whereas needle and shoot lengths, crown transparency, and radial stem growth reached control levels after up to 4 yr only. Modelling suggested that organ and carbon reserve turnover rates play a key role for a tree's responsiveness to environmental changes. Needle turnover rate was found to be most important to accurately model stem growth dynamics. We conclude that leaf area and its adjustment time to new conditions is the main determinant for radial stem growth of pine trees as the transpiring area needs to be supported by a proportional amount of sapwood, despite the growth-inhibiting environmental conditions.

Tree species distribution in temperate forests is more influenced by soil than by climate.

Knowledge of the ecological requirements determining tree species distributions is a precondition for sustainable forest management. At present, the abiotic requirements and the relative importance of the different abiotic factors are still unclear for many temperate tree species. We therefore investigated the relative importance of climatic and edaphic factors for the abundance of 12 temperate tree species along environmental gradients. Our investigations are based on data from 1,075 forest stands across Switzerland including the cold-induced tree line of all studied species and the drought-induced range boundaries of several species. Four climatic and four edaphic predictors represented the important growth factors temperature, water supply, nutrient availability, and soil aeration. The climatic predictors were derived from the meteorological network of MeteoSwiss, and the edaphic predictors were available from soil profiles. Species cover abundances were recorded in field surveys. The explanatory power of the predictors was assessed by variation partitioning analyses with generalized linear models. For six of the 12 species, edaphic predictors were more important than climatic predictors in shaping species distribution. Over all species, abundances depended mainly on nutrient availability, followed by temperature, water supply, and soil aeration. The often co-occurring species responded similar to these growth factors. Drought turned out to be a determinant of the lower range boundary for some species. We conclude that over all 12 studied tree species, soil properties were more important than climate variables in shaping tree species distribution. The inclusion of appropriate soil variables in species distribution models allowed to better explain species' ecological niches. Moreover, our study revealed that the ecological requirements of tree species assessed in local field studies and in experiments are valid at larger scales across Switzerland.

Distinct genecological patterns in seedlings of Norway spruce and silver fir from a mountainous landscape.

Understanding the genecology of forest trees is critical for gene conservation, for predicting the effects of climate change and climate change adaptation, and for successful reforestation. Although common genecological patterns have emerged, species-specific details are also important. Which species are most vulnerable to climate change? Which are the most important adaptive traits and environmental drivers of natural selection? Even though species have been classified as adaptive specialists vs. adaptive generalists, large-scale studies comparing different species in the same experiment are rare. We studied the genecology of Norway spruce (Picea abies) and silver fir (Abies alba), two co-occurring but ecologically distinct European conifers in Central Europe. For each species, we collected seed from more than 90 populations across Switzerland, established a seedling common-garden test, and developed genecological models that associate population variation in seedling growth and phenology to climate, soil properties, and site water balance. Population differentiation and associations between seedling traits and environmental variables were much stronger for Norway spruce than for silver fir, and stronger for seedling height growth than for bud phenology. In Norway spruce, height growth and second flushing were strongly associated with temperature and elevation, with seedlings from the lowlands being taller and more prone to second flush than seedlings from the Alps. In silver fir, height growth was more weakly associated with temperature and elevation, but also associated with water availability. Soil characteristics explained little population variation in both species. We conclude that Norway spruce has become an adaptive specialist because trade-offs between rapid juvenile growth and frost avoidance have subjected it to strong diversifying natural selection based on temperature. In contrast, because silver fir has a more conservative growth habit, it has evolved to become an adaptive generalist. This study demonstrates that co-occurring tree species can develop very different adaptive strategies under identical environmental conditions, and suggests that Norway spruce might be more vulnerable to future maladaptation due to rapid climate change than silver fir.

Spring water deficit and soil conditions matter more than seed origin and summer drought for the establishment of temperate conifers.

In anticipation of more severe summer droughts, forestry in temperate Europe is searching for drought-resistant ecotypes of native tree species that might maintain ecosystem services in the future. We investigated how spring precipitation and soil conditions interact with summer drought and affect the establishment of conifer seedlings from different climatic origin. Emergence, establishment and subsequent performance of seedlings originating from autochthonous, Central Alpine, continental Eastern European, and Mediterranean Pinus sylvestris and Picea abies populations were studied in the dry Alpine Rhine valley, Switzerland, at three sites with differing soil water holding capacities and in 3 years with contrasting weather conditions. In addition to this natural inter-annual variation, precipitation was manipulated within sites with throughfall reduction roofs. Seedling establishment and growth were principally affected by the spring weather in the year of emergence. In years with average to positive spring water balance, seedlings grown at the site with the highest water holding capacity had 2-5 times more aboveground biomass than seedlings grown at sites with less favourable soils. Effects of seed origin were marginal and only detectable at the drier sites: contrary to our expectations, seedlings from the Central Alpine Rhone valley, where the climatic spring water deficit is large, outperformed those from the Mediterranean. Consequently, plantation of non-native populations from dryer origin will mitigate the effects of increased summer drought at driest sites only, while the inter-annual variability of spring precipitation will continue to enable temperate conifers to regenerate on a wide range of forest soils independent of seed origin.

Signatures of local adaptation in candidate genes of oaks (Quercus spp.) with respect to present and future climatic conditions.

Testing how populations are locally adapted and predicting their response to their future environment is of key importance in view of climate change. Landscape genomics is a powerful approach to investigate genes and environmental factors involved in local adaptation. In a pooled amplicon sequencing approach of 94 genes in 71 populations, we tested whether >3500 single nucleotide polymorphisms (SNPs) in the three most common oak species in Switzerland (Quercus petraea, Q. pubescens, Q. robur) show an association with abiotic factors related to local topography, historical climate and soil characteristics. In the analysis including all species, the most frequently associated environmental factors were those best describing the habitats of the species. In the species-specific analyses, the most important environmental factors and associated SNPs greatly differed among species. However, we identified one SNP and seven genes that were associated with the same environmental factor across all species. We finally used regressions of allele frequencies of the most strongly associated SNPs along environmental gradients to predict the risk of nonadaptedness (RONA), which represents the average change in allele frequency at climate-associated loci theoretically required to match future climatic conditions. RONA is considerable for some populations and species (up to 48% in single populations) and strongly differs among species. Given the long generation time of oaks, some of the required allele frequency changes might not be realistic to achieve based on standing genetic variation. Hence, future adaptedness requires gene flow or planting of individuals carrying beneficial alleles from habitats currently matching future climatic conditions.

A decade of monitoring at Swiss Long-Term Forest Ecosystem Research (LWF) sites: can we observe trends in atmospheric acid deposition and in soil solution acidity?

Trends in atmospheric acid deposition and in soil solution acidity from 1995 or later until 2007 were investigated at several forest sites throughout Switzerland to assess the effects of air pollution abatements on deposition and the response of the soil solution chemistry. Deposition of the major elements was estimated from throughfall and bulk deposition measurements at nine sites of the Swiss Long-Term Forest Ecosystem Research network (LWF) since 1995 or later. Soil solution was measured at seven plots at four soil depths since 1998 or later. Trends in the molar ratio of base cations to aluminum (BC/Al) in soil solutions and in concentrations and fluxes of inorganic N (NO(3)-N + NH(4)-N), sulfate (SO(4)-S), and base cations (BC) were used to detect changes in soil solution chemistry. Acid deposition significantly decreased at three out of the nine study sites due to a decrease in total N deposition. Total SO(4)-S deposition decreased at the nine sites, but due to the relatively low amount of SO(4)-S load compared to N deposition, it did not contribute to decrease acid deposition significantly. No trend in total BC deposition was detected. In the soil solution, no trend in concentrations and fluxes of BC, SO(4)-S, and inorganic N were found at most soil depths at five out of the seven sites. This suggests that the soil solution reacted very little to the changes in atmospheric deposition. A stronger reduction in base cations compared to aluminum was detected at two sites, which might indicate that acidification of the soil solution was proceeding faster at these sites.

Atmospheric deposition and ozone levels in Swiss forests: are critical values exceeded?

Air pollution affects forest health through atmospheric deposition of acidic and nitrogen compounds and elevated levels of tropospheric ozone (O3). In 1985, a monitoring network was established across Europe and various research efforts have since been undertaken to define critical values. We measured atmospheric deposition of acidity and nitrogen as well as ambient levels of O3 on 12, 13, and 14 plots, respectively, in the framework of the Swiss Long-Term Forest Ecosystem Research (LWF) in the period from 1995 to 2002. We estimated the critical loads of acidity and of nitrogen, using the steady state mass balance approach, and calculated the critical O3 levels using the AOT40 approach. The deposition of acidity exceeded the critical loads on 2 plots and almost reached them on 4 plots. The median of the measured molar ratio of base nutrient cations to total dissolved aluminium (Bc/Al) in the soil solution was higher than the critical value of 1 for all depths, and also at the plots with an exceedance of the critical load of acidity. For nitrogen, critical loads were exceeded on 8 plots and deposition likely represents a long-term ecological risk on 3 to 10 plots. For O3, exceedance of critical levels was recorded on 12 plots, and led to the development of typical O3-induced visible injury on trees and shrubs, but not for all plots due to (1) the site specific composition of O3 sensitive and tolerant plant species, and (2) the influence of microclimatic site conditions on the stomatal behaviour, i.e., O3 uptake.

Callose in root apices of European chestnut seedlings: a physiological indicator of aluminum stress.

We determined if callose formation in 1-cm-long root apices of European chestnut (Castanea sativa Mill.) seedlings is affected by naturally occurring high concentrations of aluminum (Al) under laboratory conditions and by low base saturation (BS) of soils under forest field conditions. Under controlled conditions in the laboratory, seedlings were treated with simulated soil solutions in the presence or absence (control) of Al. One day after exposing seedlings to a simulated soil solution containing 168 microM Al, callose formation in the root apices had increased significantly. After 7 days, callose concentration in root apices was significantly correlated with the concentration of free Al3+ ions recovered in the simulated soil solution and with the concentration of Al in fine roots. At four field sites in southern Switzerland, seedlings were grown for five months in either A-horizon soil with a BS above 17% or in a B-horizon soil with a BS below 5%. Callose concentrations were significantly higher in root apices in the B horizon than in the A horizon. Callose concentrations in root apices were negatively correlated with Ca/Al molar ratio in fine roots. We conclude that callose in the root apices of European chestnut could be a useful physiological parameter for assessing Al toxicity under both laboratory and field conditions.