Search

The supply of carbon (C) from tree photosynthesis to ectomycorrhizal (ECM) fungi is known to decrease with increasing plant nitrogen (N) supply, but how this affects fungal nutrition and growth remains to be clarified. We placed mesh-bags with quartz sand, with or without an organic N (N-15-, C-13-labeled) source, in the soil along a natural N supply gradient in boreal forest, to measure growth and use of N and C by ECM extramatrical mycelia. Mycelial C : N declined with increasing N supply. Addition of N increased mycelial growth at the low-N end of the gradient. We found an inverse relationship between uptake of added N and C; the use of added N was high when ambient N was low, whereas use of added C was high when C from photosynthesis was low. We propose that growth of ECM fungi is N-limited when soil N is scarce and tree belowground C allocation to ECM fungi is high, but is C-limited when N supply is high and tree belowground C allocation is low. This suggests that ECM fungi have a major role in soil N retention in nutrient-poor, but less so in nutrient-rich boreal forests.

Ectomycorrhizal symbiosis is omnipresent in boreal forests, where it is assumed to benefit plant growth. However, experiments show inconsistent benefits for plants and volatility of individual partnerships, which calls for a re-evaluation of the presumed role of this symbiosis. We reconcile these inconsistencies by developing a model that demonstrates how mycorrhizal networking and market mechanisms shape the strategies of individual plants and fungi to promote symbiotic stability at the ecosystem level. The model predicts that plants switch abruptly from a mixed strategy with both mycorrhizal and nonmycorrhizal roots to a purely mycorrhizal strategy as soil nitrogen availability declines, in agreement with the frequency distribution of ectomycorrhizal colonization intensity across a wide-ranging data set. In line with observations in field-scale isotope labeling experiments, the model explains why ectomycorrhizal symbiosis does not alleviate plant nitrogen limitation. Instead, market mechanisms may generate self-stabilization of the mycorrhizal strategy via nitrogen depletion feedback, even if plant growth is ultimately reduced. We suggest that this feedback mechanism maintains the strong nitrogen limitation ubiquitous in boreal forests. The mechanism may also have the capacity to eliminate or even reverse the expected positive effect of rising CO2 on tree growth in strongly nitrogen-limited boreal forests.

Interactions between Norway spruce trees and bacteria and fungi in nutrient limited boreal forests can be beneficial for tree growth and fitness. Tree-level effects of anthropogenic nutrient addition have been well studied, however understanding of the long-term effects on the associated microbiota is limited. Here, we report on the sensitivity of microbial community composition to the growing season and nutrient additions. Highthroughput sequencing of the bacterial 16S rRNA gene and fungal ITS1 region was used to characterise changes in the microbial community after application of a complete mineral nutrient mixture for five and 25 years. The experiment was conducted using the Flakaliden forest research site in northern boreal Sweden and included naturally low nutrient control plots. Needle and fine root samples of Norway spruce were sampled in addition to bulk soil during one growing season to provide comprehensive insight into phyllosphere and belowground microbiota community changes. The phyllosphere microbiota was compositionally distinct from the belowground communities and phyllosphere diversity increased significantly over the growing season but was not influenced by the improved nutrient status of the trees. In both root and soil samples, alpha diversity of fungal, in particular ectomycorrhizal fungi (EMF), and bacterial communities increased after long-term nutrient optimisation, and with increasing years of treatment the composition of the fungal and bacterial communities changed toward a community with a higher relative abundance of nitrophilic EMF and bacterial species but did not cause complete loss of nitrophobic species from the ecosystem. From this, we conclude that 25 years of continuous nutrient addition to a boreal spruce stand increased phylotype richness and diversity of the microbiota in the soil, and at the root-soil interface, suggesting that long-term anthropogenic nutrient inputs can have positive effects on belowground biodiversity that may enhance ecosystem robustness. Future studies are needed to assess the impact of these changes to the microbiota on ecosystem carbon storage and nitrogen cycling in boreal forests. Bio4Energy

Using a gradient of changing ground age, caused by glacial isostatic adjustment, we compared systems that spanned ages from 25 to 560-years-old. Illumina sequencing was applied to determine archaeal and bacterial composition, investigating how different phylogenetic groups change as ecosystems develop. Bacterial communities dramatically changed during early ecosystem development (p

The introduction of photosynthates through plant roots is a major source of carbon (C) for soil microbial biota and shapes the composition of fungal and bacterial communities in the rhizosphere. Although the importance of this process, especially to ectomycorrhizal fungi, has been known for some time, the extent to which plant belowground C allocation controls the composition of the wider soil community is not understood. A tree-girdling experiment enabled studies of the relationship between plant C allocation and microbial community composition. Girdling involves cutting the phloem of trees to prevent photosynthates from entering the soil. Four years after girdling, fungal and bacterial communities were characterized using DNA-based profiles and cloning and sequencing. Data showed that girdling significantly altered fungal and bacterial communities compared with the control. The ratio of ectomycorrhizal to saprobic fungal sequences significantly decreased in girdled treatments, and this decline was found to correlate with the fungal phospholipid fatty acid biomarker 18:2ω6,9. Bacterial communities also varied in the abundance of the two dominant phyla Acidobacteria and Alphaproteobacteria. Concomitant changes in fungal and bacterial communities suggest linkages between these two groups and point toward plant belowground C allocation as a key determinant of microbial community composition.

Ergosterol and the phospholipid fatty acid (PLFA) 18:2o6,9 are frequently used as fungal biomarkers in studies on soils, and in accordance with the ideal for biomarkers of microorganisms they are thought to turn over rapidly after cell death and lysis. These biomarkers should also show the same patterns and responses to perturbations of the studied system. Here, I report strong correlations, in natural boreal forests of contrasting fertility, between free ergosterol and PLFA 18:2o6,9 (r ¼ 0.821, P ¼ 0.007, n ¼ 9). Surprisingly, ergosterol, but not PLFA 18:2o6,9, appears non-responsive to both large-scale tree girdling, which interrupts tree belowground C allocation to ectomycorrhizal fungi, and to long-term N-loading, which may have negative effects on both mycorrhizal and saprotrophic fungi. These results, therefore, question the use of ergosterol to monitor effects of soil perturbations on fungi in the field, but do not put into question the use of the biomarker in natural forest ecosystems. r 2006 Elsevier Ltd. All rights reserved.

The low plant productivity of boreal forests in general has been attributed to low soil N supply and low temperatures. Exceptionally high productivity occurs in toe-slope positions, and has been ascribed to influx of N from surrounding areas and higher rates of soil N turnover in situ. Despite large apparent natural variations in forest productivity, rates of gross soil N mineralization and gross nitrification have never been compared in Fennoscandian boreal forests of contrasting productivity. We report contrasting patterns of soil N turnover in three model ecosystems, representing the range in soil C-to-N ratios (19-41) in Fennoscandian boreal forests and differences in forest productivity by a factor close to 3. Gross N mineralization was seven times higher when soil, microbial, and plant C-to-N ratios were the lowest compared to the highest. This process, nitrification and potential denitrification correlated with inorganic, total and microbial biomass N, but not microbial C. There was a constant ratio between soil and microbial C-to-N ratio of 3.7+/-0.2, across wide ratios of soil C-to-N and fungi-to-bacteria. Soil N-cycling should be controlled by the supplies of C and N to the microbes. In accordance with plant allocation theory, we discuss the possibility that the high fungal biomass at high soil C-to-N ratio reflects a particularly high supply of plant photosynthates, substrates of high-quality C, to mycorrhizal fungi. Methods to study soil N turnover and N retention should be developed to take into account the impact of mycorrhizal fungi on soil N-cycling.

• The natural abundance of 13C (δ13C) and 15N (δ15N) of saprotrophic and ectomycorrhizal (ECM) fungi has been investigated on a number of occasions, but the significance of observed differences within and between the two trophic groups remains unclear. • Here, we examine the influence of taxonomy, site, host and time upon isotopic data from 135 fungal species collected at two forest sites in Sweden. • Mean δ13C and δ15N values differed significantly between ECM and saprotrophic fungi, with only a small degree of overlap even at the species level. Among ECM fungi, intraspecific variation in δ15N was low compared with interspecific and intergeneric variation. Significant variation due to site, year and host association was found. • At broad scales a number of factors clearly influence δ13C and δ15N values making interpretation problematic. We suggest that values are essentially site-specific within the two trophic groups, but that species-level patterns exist potentially reflecting ecophysiological attributes of species. The species is therefore highlighted as the taxonomic level at which most information may be obtained from fungal δ13C and δ15N data.

In Fennoscandian boreal forests, in which productivity in general is N lim- ited, there are regular, topographically related variations in forest productivity and plant community composition. Regional surveys have demonstrated strong correlations among soil pH, N content, and base saturation on the one hand and plant productivity and com- munity composition on the other, but the nature of these relationships is poorly understood. We studied in detail the variation in and controls of soil acidity, availability of N and P, and changes in community composition and plant nutrition along a short (only 90 m long) but extreme forest productivity gradient in northern Sweden, which ranged from a ground- water recharge area with low productivity to a very productive discharge area. The pH in the soil solution of the mor layer ranged from 3.5 in the recharge area to 6.4 in the discharge area, and it was strongly correlated with the base saturation of the exchange complex. Neither the acid strength of organic matter, the ionic strength of the soil solution, nor the quantity of acids could explain more than a minor part of this variation in pH. There were strong correlations between total N in the mor layer and soil solution pH (r = 0.97) and base saturation of the exchange complex (r = 0.88). At the poor end of the transect the concentration of inorganic N was very low in the mor, and plants with either ectomycorrhizae (ECM) or ericoid mycorrhizae (EM) dominated. With increasing pH, there was an increase in NH4 concentrations, while plants that potentially have arbus- cular mycorrhizae (AM) became prominent along with ECM and EM species. In the dis- charge area, which comprised only the last 10 m of the transect, NO3 dominated over NH4 in the soil solution, the soil had a high capacity for net nitrification, and the vegetation was totally dominated by potentially AM or nonmycorrhizal herbs, some of which had high foliar nitrate reductase activity. Foliar and root N concentrations increased steeply towards the discharge area, but foliar P/N ratios declined below critical levels at the end of the transect. Root 32P uptake bioassays also indicated a P deficiency in the discharge area, where the soil total P content was high, while the concentration of P04 in the soil solution was very low. The high capacity of the mor in the discharge area to adsorb P04, due to the presence of organically complexed Fe and Fe-oxihydroxides, may explain the low P04 concentrations. Our data indicate that the underlying factors influencing both productivity and com- munity composition are pH and supply of base cations. Fundamental differences in exchange characteristics of soil and soil water underlie other related nutrient supply features, in particular the amount and availability of N. Our study of a single short topographic transect supported a previous suggestion based on a regional survey in Norway that variability in soil pH and the supply of base cations affects plant productivity and community composition via effects on N supply. Our data also encompass the interrelations between soil pH, soil N turnover, and the mycorrhizal type of dominant plant species, which, according to Read (1991), occur along long latitudinal or altitudinal climatic gradients. Through millennia discharge areas like the one observed by us have probably provided a relatively stable environment for plants demanding high soil pH and N supply, at the same time as sur- rounding recharge areas have been acidified naturally through podzolization.

Fine roots play a key role in the forest carbon balance, but their carbon dynamics remain largely unknown. We pulse labelled 50 m(2) patches of young boreal forest by exposure to (13)CO(2) in early and late summer. Labelled photosynthates were traced into carbon compounds of1 and 1-3 mm diameter roots (fine roots), and into bulk tissue of these and first-order roots (root tips). Root tips were the most strongly labelled size class. Carbon allocation to all size classes was higher in late than in early summer; mean residence times (MRTs) in starch increased from 4 to 11 months. In structural compounds, MRTs were 0.8 yr in tips and 1.8 yr in fine roots. The MRT of carbon in sugars was in the range of days. Functional differences within the fine root population were indicated by carbon allocation patterns and residence times. Pronounced allocation of recent carbon and higher turnover rates in tips are associated with their role in nutrient and water acquisition. In fine roots, longer MRTs but high allocation to sugars and starch reflect their role in structural support and storage. Accounting for heterogeneity in carbon residence times will improve and most probably reduce the estimates of fine root production.

Analysis of the isotopic composition of the CO 2 respired from soils may reveal information about the important component of the ecosystem C balance. This is crucial, since a large terrestrial sink for atmospheric CO 2 has been located in the northern hemisphere, and the vast boreal forests may be largely responsible. At the same time, boreal and arctic ecosystems have large amounts of C stored in the soil, and could potentially become a source of CO 2 in a warmer climate promoting more rapid decomposition of soil organic matter. Furthermore, the northern hemisphere has complex dynamics in terms of annual fluctuations in both the concentration of CO 2 in the atmosphere and its δ l3 C. It is of utmost importance to understand the causes of this variability, since it interferes with the partitioning between the ocean and the terrestrial contributions in global models. This chapter aims to provide an update on the reviews by Flanagan and Ehleringer and Ehleringer et al. on the causation of the δ 13 C of the soil CO 2 efflux and, in doing this, focuses on the boreal forests.

Soil-surface CO2 efflux ('soil respiration') accounts for roughly two-thirds of forest ecosystem respiration, and can be divided into heterotrophic and autotrophic components. Conventionally, the latter is defined as respiration by plant roots. In Boreal forests, however, fine roots of trees are invariably covered by ectomycorrhizal fungi, which by definition are heterotrophs, but like the roots, receive sugars derived from photosynthesis. There is also a significant leaching of labile carbon compounds from the ectomycorrhizal roots. It is, therefore, more meaningful in the context of carbon balance studies to include mycorrhizal fungi and other mycorrhizosphere organisms, dependent on the direct flux of labile carbon from photosynthesis, in the autotrophic component. Hence, heterotrophic activity becomes reserved for the decomposition of more complex organic molecules in litter and other forms of soil organic matter. In reality, the complex situation is perhaps best described as a continuum from strict autotrophy to strict heterotrophy. As a result of this, and associated methodological problems, estimates of the contribution of autotrophic respiration to total soil respiration have been highly variable. Based on recent stand-scale tree girdling experiments we have estimated that autotrophic respiration in boreal forest accounts for up to 50-65% of soil respiration during the snow-free part of the year. Girdling experiments and studies of the delta(13)C of the soil CO2 efflux show that there is a lag of a few days between the carbon uptake by photosynthesis and the release by autotrophic soil respiration of the assimilated carbon. In contrast, estimates of 'bomb 14C' and other approaches have suggested that it takes years to decades between carbon uptake via photosynthesis and the bulk of soil heterotrophic activity. Temperature is normally used as a driver in models of soil processes and it is often assumed that autotrophic soil activity is more sensitive to temperature than is heterotrophic activity, but this is questionable. It is inherently difficult to make a precise separation of autotrophic and heterotrophic respiration from soils. The partitioning between these two components is highly variable in space and time, and taxonomic autotrophs and heterotrophs may perform the function of the other group to some degree. Care should be taken to disturb as little as possible the delicate plant-microbe-soil system, and this speaks for non-intrusive isotopic methods. There are, however, problems in modelling the flux of isotopes through this complex system. Girdling of tree stands is a very robust alternative approach to make the distinction between autotrophic and heterotrophic activities, but ultimately kills the trees and cannot, therefore, always be used. A further development would be to block the phloem sugar transport reversibly. We propose that thus assumption needs further critical testing.

Soil microorganisms are considered C-limited, while plant productivity is frequently N-limited. Large stores of organic C in boreal forest soils are attributed to negative effects of low temperature, soil acidity and plant residue recalcitrance upon microbial activity. We examined microbial activity, biomass and community composition along a natural 90-m-long soil N supply gradient, where plant species composition varies profoundly, forest productivity three-fold and soil pH by three units. There was, however, no significant variation in soil respiration in the field across the gradient. Neither did microbial biomass C determined by fumigation-extraction vary, while other estimates of activity and biomass showed a weak increase with increasing N supply and soil pH. Simultaneously, a phospholipid fatty acid attributed mainly to mycorrhizal fungi declined drastically, while bacterial biomass increased. We hypothesize that low N supply and plant productivity, and hence low litter C supply to saprotrophs is associated with a high plant C supply to mycorrhizal fungi, while the reverse occurs under high N supply. This should mean that effects of N availability on C supply to these functional groups of microbes acts in opposing directions.

Preliminary attempts to make retrospective studies of N balances and water stress in forest fertilization experiments by analyzing changes in the abundances of 15N and 13C, respectively, are discussed. Most evidence is from the Swedish Forest Optimum Nutrition Experiments, which have been running for two decades. Annual additions of N have been given either alone or in combination with other elements, notably P and K, every third year. Processes leading to loss of N, e.g. volatilization of ammonia, nitrification followed by leaching or denitrification, and denitrification alone, discriminate against the heavy isotope 15N. A correlation was found between fractional losses of added N and the change in δ15N (‰) during 19 years in current needles in a Scots pine forest, irrespective of source of N. Isotope effects were larger on urea than on ammonium nitrate plots (2 as compared to 9 δ15N (‰)) because of ammonia volatilization and higher rates of nitrification. They developed gradually over time, which opens possibilities to analyse the development of N saturation. However, the analysis may be confounded by shifts in 15N abundance of fertilizer N. In another trial, N isotope effects could be seen in both plants and soils 10 years after the last fertilization; they were smaller in soils because of a large pretreatment memory effect, but we expect them to persist there for decades.