Author Correction: Environment and host as large-scale controls of ectomycorrhizal fungi.

Change history: In the HTML version of this Article, author 'Filipa Cox' had no affiliation in the author list, although she was correctly associated with affiliation 3 in the PDF. In addition, the blue circles for 'oak' were missing from Extended Data Fig. 1. These errors have been corrected online.

Environment and host as large-scale controls of ectomycorrhizal fungi.

Explaining the large-scale diversity of soil organisms that drive biogeochemical processes-and their responses to environmental change-is critical. However, identifying consistent drivers of belowground diversity and abundance for some soil organisms at large spatial scales remains problematic. Here we investigate a major guild, the ectomycorrhizal fungi, across European forests at a spatial scale and resolution that is-to our knowledge-unprecedented, to explore key biotic and abiotic predictors of ectomycorrhizal diversity and to identify dominant responses and thresholds for change across complex environmental gradients. We show the effect of 38 host, environment, climate and geographical variables on ectomycorrhizal diversity, and define thresholds of community change for key variables. We quantify host specificity and reveal plasticity in functional traits involved in soil foraging across gradients. We conclude that environmental and host factors explain most of the variation in ectomycorrhizal diversity, that the environmental thresholds used as major ecosystem assessment tools need adjustment and that the importance of belowground specificity and plasticity has previously been underappreciated.

Sidestream materials show potential as top-dressed soil improvers for peatland forests.

The effects of top-dressing of several industrial and farming sidestream materials on the growth of downy birch (Betula pubescens Ehrh.) and Scots pine (Pinus sylvestris L.) seedlings in natural sphagnum peat soil were evaluated. Wood fly ash, industrial filter cake waste, mine tailings sand (quartz feldspar from lithium orebody), and digestate and liquid reject of cow manure from a biogas plant were studied for their physical and chemical properties, as well as for their effects as soil ameliorants on seedling growth during one growing period in a greenhouse. Each material was top-dressed on unfertilised peat in pots in quantities that corresponded to the amounts of ash used in Finnish peatland forest fertilisation (2-6 t ha-1). During growing, the pH of percolate water from the growing pots was below 4, and in the treatments with filter cake even below 3. However, no clear impairment of seedling growth due to acidity was observed. In all treatments, birch and pine seedlings grew at least as well as in the unfertilised peat (control treatment). Growth was strongest in the peat top-dressed with additives originating from cow manure, in which the high N and P contents promoted growth so much that foliar N was found to be diluted with respect to a high P content in the birch seedlings. No harmful concentrations of heavy metal residues were observed from the materials used. Overall, the results suggest that all the used sidestream materials show potential as soil improvers on forested peatlands.

Tree mineral nutrition is deteriorating in Europe.

The response of forest ecosystems to increased atmospheric CO2 is constrained by nutrient availability. It is thus crucial to account for nutrient limitation when studying the forest response to climate change. The objectives of this study were to describe the nutritional status of the main European tree species, to identify growth-limiting nutrients and to assess changes in tree nutrition during the past two decades. We analysed the foliar nutrition data collected during 1992-2009 on the intensive forest monitoring plots of the ICP Forests programme. Of the 22 significant temporal trends that were observed in foliar nutrient concentrations, 20 were decreasing and two were increasing. Some of these trends were alarming, among which the foliar P concentration in F. sylvatica, Q. Petraea and P. sylvestris that significantly deteriorated during 1992-2009. In Q. Petraea and P. sylvestris, the decrease in foliar P concentration was more pronounced on plots with low foliar P status, meaning that trees with latent P deficiency could become deficient in the near future. Increased tree productivity, possibly resulting from high N deposition and from the global increase in atmospheric CO2, has led to higher nutrient demand by trees. As the soil nutrient supply was not always sufficient to meet the demands of faster growing trees, this could partly explain the deterioration of tree mineral nutrition. The results suggest that when evaluating forest carbon storage capacity and when planning to reduce CO2 emissions by increasing use of wood biomass for bioenergy, it is crucial that nutrient limitations for forest growth are considered.

Exploring continental-scale stand health - N : P ratio relationships for European forests.

Understanding the relationship between nitrogen (N) availability and stand health in forest ecosystems is crucial, because a large proportion of European forests is subjected to N-deposition levels beyond their retention capacity. We used data from a long-term forest monitoring programme (ICP Forests) to test the relationship between an index of N availability, foliar nitrogen : phosphorus (N : P) ratios, tree defoliation and discoloration. We hypothesized a segmented response of stand health to N : P ratios and an improved model-fit after correcting for climatic covariates. In accordance with the hypothesis, we found a segmented response with a breakpoint for conifer defoliation at N : P ratios as low as 7.3. Inclusion of climatic variables improved the fit of the models, but there was significant collinearity with N : P. Increases in N availability appear, at least for conifers, to have a negative effect on tree health even under N-limiting conditions. Regulation of N-deposition levels is consequently as timely as ever. We propose that increases in tree defoliation, other than resulting in serious plant fitness issues, may represent early diagnostic symptoms of N-addition related imbalances.

Fungal community composition predicts forest carbon storage at a continental scale.

Forest soils harbor hyper-diverse microbial communities which fundamentally regulate carbon and nutrient cycling across the globe. Directly testing hypotheses on how microbiome diversity is linked to forest carbon storage has been difficult, due to a lack of paired data on microbiome diversity and in situ observations of forest carbon accumulation and storage. Here, we investigated the relationship between soil microbiomes and forest carbon across 238 forest inventory plots spanning 15 European countries. We show that the composition and diversity of fungal, but not bacterial, species is tightly coupled to both forest biotic conditions and a seven-fold variation in tree growth rates and biomass carbon stocks when controlling for the effects of dominant tree type, climate, and other environmental factors. This linkage is particularly strong for symbiotic endophytic and ectomycorrhizal fungi known to directly facilitate tree growth. Since tree growth rates in this system are closely and positively correlated with belowground soil carbon stocks, we conclude that fungal composition is a strong predictor of overall forest carbon storage across the European continent.

Changing winter climate and snow conditions induce various transcriptional stress responses in Scots pine seedlings.

In northern boreal forests the warming winter climate leads to more frequent snowmelt, rain-on-snow events and freeze-thaw cycles. This may be harmful or even lethal for tree seedlings that spend even a half of the year under snow. We conducted a snow cover manipulation experiment in a natural forest to find out how changing snow conditions affect young Scots pine (Pinus sylvestris L.) seedlings. The ice encasement (IE), absence of snow (NoSNOW) and snow compaction (COMP) treatments affected ground level temperature, ground frost and subnivean gas concentrations compared to the ambient snow cover (AMB) and led to the increased physical damage and mortality of seedlings. The expression responses of 28 genes related to circadian clock, aerobic and anaerobic energy metabolism, carbohydrate metabolism and stress protection revealed that seedlings were exposed to different stresses in a complex way depending on the thickness and quality of the snow cover. The IE treatment caused hypoxic stress and probably affected roots which resulted in reduced water uptake in the beginning of the growing season. Without protective snowpack in NoSNOW seedlings suffered from cold and drought stresses. The combination of hypoxic and cold stresses in COMP evoked unique transcriptional responses including oxidative stress. Snow cover manipulation induced changes in the expression of several circadian clock related genes suggested that photoreceptors and the circadian clock system play an essential role in the adaptation of Scots pine seedlings to stresses under different snow conditions. Our findings show that warming winter climate alters snow conditions and consequently causes Scots pine seedlings various abiotic stresses, whose effects extend from overwintering to the following growing season.

Defoliation Change of European Beech (Fagus sylvatica L.) Depends on Previous Year Drought.

European beech (Fagus sylvatica L.) forests provide multiple essential ecosystem goods and services. The projected climatic conditions for the current century will significantly affect the vitality of European beech. The expected impact of climate change on forest ecosystems will be potentially stronger in southeast Europe than on the rest of the continent. Therefore, our aim was to use the long-term monitoring data of crown vitality indicators in Croatia to identify long-term trends, and to investigate the influence of current and previous year climate conditions and available site factors using defoliation (DEF) and defoliation change (ΔDEF) as response variables. The results reveal an increasing trend of DEF during the study period from 1996 to 2017. In contrast, no significant trend in annual ΔDEF was observed. The applied linear mixed effects models indicate a very strong influence of previous year drought on ΔDEF, while climate conditions have a weak or insignificant effect on DEF. The results suggest that site factors explain 25 to 30% DEF variance, while similar values of conditional and marginal R2 show a uniform influence of drought on ΔDEF. These results suggest that DEF represents the accumulated impact of location-specific stressful environmental conditions on tree vitality, while ΔDEF reflects intense stress and represents the current or recent status of tree vitality that could be more appropriate for analysing the effect of climate conditions on forest trees.

The Effect of Environmental Factors on the Nutrition of European Beech (Fagus sylvatica L.) Varies with Defoliation.

Despite being adapted to a wide range of environmental conditions, the vitality of European beech is expected to be significantly affected by the projected effects of climate change, which we attempted to assess with foliar nutrition and crown defoliation, as two different, yet interlinked vitality indicators. Based on 28 beech plots of the ICP Forests Level I network, we set out to investigate the nutritional status of beech in Croatia, the relation of its defoliation and nutrient status, and the effects of environmental factors on this relation. The results indicate a generally satisfactory nutrition of common beech in Croatia. Links between defoliation and nutrition of beech are not very direct or very prominent; differences were observed only in some years and on limited number of plots. However, the applied multinomial logistic regression models show that environmental factors affect the relationship between defoliation and nutrition, as climate and altitude influence the occurrence of differences in foliar nutrition between defoliation categories.

Forest tree growth is linked to mycorrhizal fungal composition and function across Europe.

Most trees form symbioses with ectomycorrhizal fungi (EMF) which influence access to growth-limiting soil resources. Mesocosm experiments repeatedly show that EMF species differentially affect plant development, yet whether these effects ripple up to influence the growth of entire forests remains unknown. Here we tested the effects of EMF composition and functional genes relative to variation in well-known drivers of tree growth by combining paired molecular EMF surveys with high-resolution forest inventory data across 15 European countries. We show that EMF composition was linked to a three-fold difference in tree growth rate even when controlling for the primary abiotic drivers of tree growth. Fast tree growth was associated with EMF communities harboring high inorganic but low organic nitrogen acquisition gene proportions and EMF which form contact versus medium-distance fringe exploration types. These findings suggest that EMF composition is a strong bio-indicator of underlying drivers of tree growth and/or that variation of forest EMF communities causes differences in tree growth. While it may be too early to assign causality or directionality, our study is one of the first to link fine-scale variation within a key component of the forest microbiome to ecosystem functioning at a continental scale.

Induced accumulation of phenolics and sawfly performance in Scots pine in response to previous defoliation.

Phenolic compounds often accumulate in foliar tissues of deciduous woody plants in response to previous insect defoliation, but similar responses have been observed infrequently in evergreen conifers. We studied the effects of defoliation on the foliar chemistry of Scots pine (Pinus sylvestris L.) and cocoon mass, and survival of the pine sawfly (Diprion pini L.). In two successive years, needles were excised early in the season leaving only the current-year shoot intact (defoliated trees); untreated entire shoots served as controls (control trees). A year after the second defoliation, pine sawfly larvae were transferred to the trees. Delayed induced resistance in Scots pine in response to defoliation was indicated by (1) reduced cocoon mass in defoliated trees and (2) increased concentrations of phenolics and soluble condensed tannins in the foliage of defoliated trees compared with controls. Myricetin-3-galactoside, which showed the strongest induced response (104% and 71% increase in current-year (C) and previous-year (C+1) needles) of the compounds analyzed, also entered the regression model explaining variation in sawfly performance. Other compounds that entered the model, e.g., (+)-catechin, showed weaker responses to defoliation than myricetin-3-galactoside. Hyperin, condensed tannins and quercitrin showed strong induced responses in C or C+1 needles, or both, but these compounds did not explain the variation in sawfly performance. Accumulation of phenolics is sometimes associated with the reduced foliage nitrogen (N) concentrations in deciduous trees, and our results suggest that this may also be the case in evergreen conifers. Based on the earlier findings that defoliation reduces needle N concentration and N deficiency results in the accumulation of the same phenolic compounds, i.e., myricetin and quercetin glycosides, and soluble condensed tannins, we suggest that the accumulation of phenolics in defoliated trees occurred in response to the reduced foliar N concentration.

Let it snow! Winter conditions affect growth of birch seedlings during the following growing season.

Air temperatures and precipitation are predicted to increase in the future, especially at high latitudes and particularly so during winter. In contrast to air temperatures, changes in soil temperatures are more difficult to predict, as the fate of the insulating snow cover is crucial in this respect. Soil conditions can also be affected by rain-on-snow events and warm spells during winter, resulting in freeze-thaw cycles, compacted snow, ice encasement and local flooding. These adverse conditions during winter could counteract the otherwise positive effects of climate change on forest growth and productivity. For studying the effects of different winter and snow conditions on young Downy birch (Betula pubescens Ehrh.) seedlings, we carried out a laboratory experiment with birch seedlings subjected to four different winter scenarios: snow covering the seedlings (SNOW), compressed snow and ice encasement (ICE), flooded and frozen soil (FLOOD) and no snow at all (NO SNOW). After the winter treatments we simulated a spring and early summer period of 9.5 weeks, and monitored the growth by measuring shoot and root biomass of the seedlings, and starch and soluble sugar concentrations. We also assessed the stress experienced by the seedlings by measuring leaf chlorophyll fluorescence and gas exchange. Although no difference in mortality was observed between the treatments, the seedlings in the SNOW and ICE treatments had significantly higher shoot and root biomass compared with those in the FLOOD and NO SNOW treatments. We found higher starch concentrations in roots of the seedlings in the SNOW and ICE treatments, compared with those in the FLOOD and NO SNOW treatments, although photosynthesis did not differ. Our results suggest a malfunction of carbohydrate distribution in the seedlings of the FLOOD and NO SNOW treatments, probably resulting from decreased sinks. The results underline the importance of an insulating and protecting snow cover for small tree seedlings, and that future winters with changed snow pattern might affect the growth of tree seedlings and thus possibly species composition and forest productivity.

Winter survival of Scots pine seedlings under different snow conditions.

Future climate scenarios predict increased air temperatures and precipitation, particularly at high latitudes, and especially so during winter. Soil temperatures, however, are more difficult to predict, since they depend strongly on the fate of the insulating snow cover. 'Rain-on-snow' events and warm spells during winter can lead to thaw-freeze cycles, compacted snow and ice encasement, as well as local flooding. These adverse conditions could counteract the otherwise positive effects of climatic changes on forest seedling growth. In order to study the effects of different winter and snow conditions on young Scots pine (Pinus sylvestris L.) seedlings, we conducted a laboratory experiment in which 80 1-year-old Scots pine seedlings were distributed between four winter treatments in dasotrons: ambient snow cover (SNOW), compressed snow and ice encasement (ICE), flooded and frozen soil (FLOOD) and no snow (NO SNOW). During the winter treatment period and a 1.5-month simulated spring/early summer phase, we monitored the needle, stem and root biomass of the seedlings, and determined their starch and soluble sugar concentrations. In addition, we assessed the stress experienced by the seedlings by measuring chlorophyll fluorescence, electric impedance and photosynthesis of the previous-year needles. Compared with the SNOW treatment, carbohydrate concentrations were lower in the FLOOD and NO SNOW treatments where the seedlings had almost died before the end of the experiment, presumably due to frost desiccation of aboveground parts during the winter treatments. The seedlings of the ICE treatment showed dead needles and stems only above the snow and ice cover. The results emphasize the importance of an insulating and protecting snow cover for small forest tree seedlings, and that future winters with changed snow patterns might affect the survival of tree seedlings and thus forest productivity.

Bacterial and fungal communities in boreal forest soil are insensitive to changes in snow cover conditions.

The northern regions are experiencing considerable changes in winter climate leading to more frequent warm periods, rain-on-snow events and reduced snow pack diminishing the insulation properties of snow cover and increasing soil frost and freeze-thaw cycles. In this study, we investigated how the lack of snow cover, formation of ice encasement and snow compaction affect the size, structure and activities of soil bacterial and fungal communities. Contrary to our hypotheses, snow manipulation treatments over one winter had limited influence on microbial community structure, bacterial or fungal copy numbers or enzyme activities. However, microbial community structure and activities shifted seasonally among soils sampled before snow melt, in early and late growing season and seemed driven by substrate availability. Bacterial and fungal communities were dominated by stress-resistant taxa such as the orders Acidobacteriales, Chaetothyriales and Helotiales that are likely adapted to adverse winter conditions. This study indicated that microbial communities in acidic northern boreal forest soil may be insensitive to direct effects of changing snow cover. However, in long term, the detrimental effects of increased ice and frost to plant roots may alter plant derived carbon and nutrient pools to the soil likely leading to stronger microbial responses.

The Snow Must Go On: Ground Ice Encasement, Snow Compaction and Absence of Snow Differently Cause Soil Hypoxia, CO2 Accumulation and Tree Seedling Damage in Boreal Forest.

At high latitudes, the climate has warmed at twice the rate of the global average with most changes observed in autumn, winter and spring. Increasing winter temperatures and wide temperature fluctuations are leading to more frequent rain-on-snow events and freeze-thaw cycles causing snow compaction and formation of ice layers in the snowpack, thus creating ice encasement (IE). By decreasing the snowpack insulation capacity and restricting soil-atmosphere gas exchange, modification of the snow properties may lead to colder soil but also to hypoxia and accumulation of trace gases in the subnivean environment. To test the effects of these overwintering conditions changes on plant winter survival and growth, we established a snow manipulation experiment in a coniferous forest in Northern Finland with Norway spruce and Scots pine seedlings. In addition to ambient conditions and prevention of IE, we applied three snow manipulation levels: IE created by artificial rain-on-snow events, snow compaction and complete snow removal. Snow removal led to deeper soil frost during winter, but no clear effect of IE or snow compaction done in early winter was observed on soil temperature. Hypoxia and accumulation of CO2 were highest in the IE plots but, more importantly, the duration of CO2 concentration above 5% was 17 days in IE plots compared to 0 days in ambient plots. IE was the most damaging winter condition for both species, decreasing the proportion of healthy seedlings by 47% for spruce and 76% for pine compared to ambient conditions. Seedlings in all three treatments tended to grow less than seedlings in ambient conditions but only IE had a significant effect on spruce growth. Our results demonstrate a negative impact of winter climate change on boreal forest regeneration and productivity. Changing snow conditions may thus partially mitigate the positive effect of increasing growing season temperatures on boreal forest productivity.

Seasonal and inter-annual variation in the chlorophyll content of three co-existing Sphagnum species exceeds the effect of solar UV reduction in a subarctic peatland.

We measured chlorophyll (chl) concentration and chl a/b ratio in Sphagnum balticum, S. jensenii, and S. lindbergii, sampled after 7 and 8 years of ultraviolet-B (UVB) and temperature manipulation in an open field experiment in Finnish Lapland (68°N). We used plastic filters with different transmittance of UVB radiation to manipulate the environmental conditions. The plants were exposed to (1) attenuated UVB and increased temperature, (2) ambient UVB and increased temperature and (3) ambient conditions. Chlorophyll was extracted from the capitula of the mosses and the content and a/b ratio were measured spectrophotometrically. Seasonal variation of chlorophyll concentration in the mosses was species specific. Temperature increase to 0.5-1 °C and/or attenuation of solar UVB radiation to ca. one fifth of the ambient (on average 12 vs. 59 uW/cm(2)) had little effect on the chlorophyll concentration or its seasonal variation. In the dominant S. lindbergii, UVB attenuation under increased temperature led to a transient decrease in chlorophyll concentration. Altogether, species-specific patterns of seasonal chlorophyll variation in the studied Sphagna were more pronounced than temperature and UVB treatment effects.

Total vs. internal element concentrations in Scots pine needles along a sulphur and metal pollution gradient.

Analysis of foliar elements is a commonly used method for studying tree nutrition and for monitoring the impacts of air pollutants on forest ecosystems. Interpretations based on the results of foliar element analysis may, however, be different in nutrition vs. monitoring studies. We studied the impacts of severe sulphur and metal (mainly Cu and Ni) pollution on the element concentrations (Al, Ca, Cu, Fe, K, Mg, Mn, Ni, P, Pb, S and Zn) in Scots pine (Pinus sylvestris L.) foliage along an airborne sulphur and metal pollution gradient. Emphasis was put on determining the contribution of air-borne particles that have accumulated on needle surfaces to the total foliage concentrations. A comparison of two soil extraction methods was carried out in order to obtain a reliable estimate of plant-available element concentrations in the soil. Element concentrations in the soil showed only a weak relationship with internal foliar concentrations. There were no clear differences between the total and internal needle S concentrations along the gradient, whereas at the plot closest to the metal smelter complex the total Cu concentrations in the youngest needles were 1.3-fold and Ni concentrations over 1.6-fold higher than the internal needle concentrations. Chloroform-extracted surface wax was found to have Ni and Cu concentrations of as high as 3000 and 600 microg/g of wax, respectively. Our results suggest that bioindicator studies (e.g. monitoring studies) may require different foliar analysis techniques from those used in studies on the nutritional status of trees.

Defoliation causes parallel temporal responses in a host tree and its fungal symbionts.

Growth of the host and its symbiont is often closely linked and so host damage may negatively affect the symbiont. While negative effects of aboveground herbivory on belowground fungal symbionts have been reported in several woody and herbaceous plants, here we report, for the first time, on differential effects of the timing of foliar damage on ectomycorrhizal (ECM) fungal symbionts. The phenologies of host trees and their ECM symbionts differ; the growth of the latter mainly occurs later in the season than that of the host. By removing Scots pine foliage on three occasions during the growing season (early, middle and late season defoliation) in one, two or three successive years, we demonstrate that, despite the differences in the seasonal growth dynamics of the tree and the symbionts, ECM fungi follow the host's response patterns to defoliation. Early season defoliation was most detrimental to the host and resulted in an increased proportion of low-biomass ectomycorrhizae which are presumed to require less carbon from the host tree. This may improve the recovery of the host, as most roots remained mycorrhizal in spite of the defoliation treatments repeated in successive years.

Associational effects of plant defences in relation to within- and between-patch food choice by a mammalian herbivore: neighbour contrast susceptibility and defence.

A basic idea of plant defences is that a plant should gain protection from its own defence. In addition, there is evidence that defence traits of the neighbouring plants can influence the degree of protection of an individual plant. These associational effects depend in part on the spatial scale of herbivore selectivity. A strong between-patch selectivity together with a weak within-patch selectivity leads to a situation where a palatable plant could avoid being grazed by growing in a patch with unpalatable plants, which is referred to as associational defence. Quite different associational effects will come about if the herbivore instead is unselective between patches and selective within a patch. We studied these effects in a manipulative experiment where we followed the food choice of fallow deer when they encountered two patches of overall different quality. One of the two patches consisted of pellets with low-tannin concentration in seven out of eight buckets and with high concentration in the remaining bucket. The other patch instead had seven high- and one low-tannin bucket. We performed the experiment both with individuals one at a time and with a group of 16-17 deer. We found that the deer were unselective between patches, but selective within a patch, and that the single low-tannin bucket among seven high-tannin buckets was used more than a low-tannin bucket among other low-tannin buckets. This corresponds to a situation where a palatable plant that grows among unpalatable plants is attacked more than if it was growing among its own kind, and for this effect we suggest the term neighbour contrast susceptibility, which is the opposite of associational defence. We also found that the high-tannin bucket in the less defended patch was less used than the high-tannin buckets in the other patch, which corresponds to neighbour contrast defence. The neighbour contrast susceptibility was present both for individual and group foraging, but the strength of the effect was somewhat weaker for groups due to weaker within-patch selectivity.

Defoliation increases carbon limitation in ectomycorrhizal symbiosis of Betula pubescens.

Boreal forest trees are highly dependent on root-colonizing mycorrhizal fungi. Since the maintenance of mycorrhizal symbiosis implies a significant carbon cost for the host plant, the loss of photosynthetic leaf area due to herbivory is expected to reduce the host investment in mycorrhizae. We tested this hypothesis in a common garden experiment by exposing ectomycorrhizal white birch (Betula pubescens Ehrh.) seedlings to simulated insect defoliation of 50 or 100% intensity during either the previous or the current summer or repeatedly during both seasons before harvest. The shoot and root growth of the seedlings were distinctly reduced by both 100% defoliation and repeated 50% defoliation, and they were more strongly affected by previous-year than current-year defoliation. The root to shoot ratio significantly decreased after 100% defoliation, indicating reduced proportional allocation to the roots. Ergosterol concentration (i.e. fungal biomass) in the fine roots decreased by 100% defoliation conducted either in the year of harvest or in both years. No such decrease occurred following the 100% defoliation conducted in the previous year, indicating the importance of current photosynthates for fungal symbionts. The trend was similar in the colonization percentage of thick-mantled mycorrhizae in the roots, the most marked decline occurring in the repeatedly defoliated seedlings. The present results thus support the prediction that the plant investment in ectomycorrhizae may decline as a response to foliage loss. Moreover, the colonization percentage of thick-mantled mycorrhizae correlated positively with the ratio of leaf to heterotrophic plant biomass in the defoliated birch seedlings, but not in the control ones. This tends to indicate a stronger carbon limitation of ectomycorrhizal colonization in defoliated seedlings.