Ectomycorrhizal fungi integrate nitrogen mobilisation and mineral weathering in boreal forest soil.

Tree growth in boreal forests is driven by ectomycorrhizal fungal mobilisation of organic nitrogen and mineral nutrients in soils with discrete organic and mineral horizons. However, there are no studies of how ectomycorrhizal mineral weathering and organic nitrogen mobilisation processes are integrated across the soil profile. We studied effects of organic matter (OM) availability on ectomycorrhizal functioning by altering the proportions of natural organic and mineral soil in reconstructed podzol profiles containing Pinus sylvestris plants, using 13 CO2 pulse labelling, patterns of naturally occurring stable isotopes (26 Mg and 15 N) and high-throughput DNA sequencing of fungal amplicons. Reduction in OM resulted in nitrogen limitation of plant growth and decreased allocation of photosynthetically derived carbon and mycelial growth in mineral horizons. Fractionation patterns of 26 Mg indicated that magnesium mobilisation and uptake occurred primarily in the deeper mineral horizon and was driven by carbon allocation to ectomycorrhizal mycelium. In this horizon, relative abundance of ectomycorrhizal fungi, carbon allocation and base cation mobilisation all increased with increased OM availability. Allocation of carbon through ectomycorrhizal fungi integrates organic nitrogen mobilisation and mineral weathering across soil horizons, improving the efficiency of plant nutrient acquisition. Our findings have fundamental implications for sustainable forest management and belowground carbon sequestration.

Quantification of Biodriven Transfer of Per- and Polyfluoroalkyl Substances from the Aquatic to the Terrestrial Environment via Emergent Insects.

Emergent aquatic insects are important food subsidies to riparian food webs but can also transfer waterborne contaminants to the terrestrial environment. This study aimed to quantitatively assess this biodriven transfer for per- and polyfluoroalkyl substances (PFAS). Aquatic insect larvae, emergent aquatic insects, terrestrial consumers, sediment, and water were collected from a contaminated lake and stream and an uncontaminated pond, and analyzed for PFAS and stable isotopes of carbon and nitrogen. Top predators in this study were spiders, which showed the highest average ∑24PFAS concentration of 1400 ± 80 ng g-1 dry weight (dw) at the lake and 630 ng g-1 dw at the stream. The transfer of PFAS from the lake to the riparian zone, via deposition of emergent aquatic insects, was 280 ng ∑24PFAS m-2 d-1 in 2017 and only 23 ng ∑24PFAS m-2 d-1 in 2018. Because of higher production of emergent aquatic insects, the lake had higher PFAS transfer and higher concentrations in terrestrial consumers compared to the stream, despite the stream having higher PFAS concentration in water and aquatic insect larvae. Our results indicate that biodriven transfer of PFAS from the aquatic systems and subsequent uptake in terrestrial food webs depend more on emergence amounts, i.e., aquatic prey availability, rather than on PFAS concentrations in water and aquatic prey.

Per- and Polyfluoroalkyl-Contaminated Freshwater Impacts Adjacent Riparian Food Webs.

The occurrence of per- and polyfluoroalkyl substances (PFASs) in aquatic ecosystems is a global concern because of their persistence, potential bioaccumulation, and toxicity. In this study, we investigated a PFAS-contaminated pond in Sweden to assess the cross-boundary transfer of PFASs from the aquatic environment to the riparian zone via emergent aquatic insects. Aquatic and terrestrial invertebrates, surface water, sediments, soils, and plants were analyzed for 24 PFASs including branched isomers. Stable isotope analysis of carbon and nitrogen was performed to elucidate the importance of diet and trophic position for PFAS uptake. We present the first evidence that PFASs can propagate to the riparian food web via aquatic emergent insects. Elevated Σ24PFAS concentrations were found in aquatic insect larvae, such as dragon- and damselflies, ranging from 1100 to 4600 ng g-1 dry weight (dw), and remained high in emerged adults (120-3500 ng g-1 dw), indicating exposure risks for top predators that prey in riparian zones. In terrestrial invertebrate consumers, PFAS concentrations increased with the degree of aquatic-based diet and at higher trophic levels. Furthermore, stable isotope data together with calculated bioaccumulation factors indicated that bioconcentration of PFASs was the major pathway of exposure in the aquatic food web and bioaccumulation in the riparian food web.

Soil fertility in boreal forest relates to root-driven nitrogen retention and carbon sequestration in the mor layer.

Boreal forest soils retain significant amounts of carbon (C) and nitrogen (N) in purely organic layers, but the regulation of organic matter turnover and the relative importance of leaf litter and root-derived inputs are not well understood. We combined bomb 14 C dating of organic matter with stable isotope profiling for Bayesian parameterization of an organic matter sequestration model. C and N dynamics were assessed across annual depth layers (cohorts), together representing 256 yr of organic matter accumulation. Results were related to ecosystem fertility (soil inorganic N, pH and litter C : N). Root-derived C was estimated to decompose two to 10 times more slowly than leaf litter, but more rapidly in fertile plots. The amounts of C and N per cohort declined during the initial 20 yr of decomposition, but, in older material, the amount of N per cohort increased, indicating N retention driven by root-derived C. The dynamics of root-derived inputs were more important than leaf litter dynamics in regulating the variation in organic matter accumulation along a forest fertility gradient. N retention in the rooting zone combined with impeded mining for N in less fertile ecosystems provides evidence for a positive feedback between ecosystem fertility and organic matter turnover.

Identifying the Active Microbiome Associated with Roots and Rhizosphere Soil of Oilseed Rape.

RNA stable isotope probing and high-throughput sequencing were used to characterize the active microbiomes of bacteria and fungi colonizing the roots and rhizosphere soil of oilseed rape to identify taxa assimilating plant-derived carbon following 13CO2 labeling. Root- and rhizosphere soil-associated communities of both bacteria and fungi differed from each other, and there were highly significant differences between their DNA- and RNA-based community profiles. Verrucomicrobia, Proteobacteria, Planctomycetes, Acidobacteria, Gemmatimonadetes, Actinobacteria, and Chloroflexi were the most active bacterial phyla in the rhizosphere soil. Bacteroidetes were more active in roots. The most abundant bacterial genera were well represented in both the 13C- and 12C-RNA fractions, while the fungal taxa were more differentiated. Streptomyces, Rhizobium, and Flavobacterium were dominant in roots, whereas Rhodoplanes and Sphingomonas (Kaistobacter) were dominant in rhizosphere soil. "Candidatus Nitrososphaera" was enriched in 13C in rhizosphere soil. Olpidium and Dendryphion were abundant in the 12C-RNA fraction of roots; Clonostachys was abundant in both roots and rhizosphere soil and heavily 13C enriched. Cryptococcus was dominant in rhizosphere soil and less abundant, but was 13C enriched in roots. The patterns of colonization and C acquisition revealed in this study assist in identifying microbial taxa that may be superior competitors for plant-derived carbon in the rhizosphere of Brassica napusIMPORTANCE This microbiome study characterizes the active bacteria and fungi colonizing the roots and rhizosphere soil of Brassica napus using high-throughput sequencing and RNA-stable isotope probing. It identifies taxa assimilating plant-derived carbon following 13CO2 labeling and compares these with other less active groups not incorporating a plant assimilate. Brassica napus is an economically and globally important oilseed crop, cultivated for edible oil, biofuel production, and phytoextraction of heavy metals; however, it is susceptible to several diseases. The identification of the fungal and bacterial species successfully competing for plant-derived carbon, enabling them to colonize the roots and rhizosphere soil of this plant, should enable the identification of microorganisms that can be evaluated in more detailed functional studies and ultimately be used to improve plant health and productivity in sustainable agriculture.

Changes in turnover rather than production regulate biomass of ectomycorrhizal fungal mycelium across a Pinus sylvestris chronosequence.

In boreal forest soils, ectomycorrhizal fungi are fundamentally important for carbon (C) dynamics and nutrient cycling. Although their extraradical mycelium (ERM) is pivotal for processes such as soil organic matter build-up and nitrogen cycling, very little is known about its dynamics and regulation. In this study, we quantified ERM production and turnover, and examined how these two processes together regulated standing ERM biomass in seven sites forming a chronosequence of 12- to 100-yr-old managed Pinus sylvestris forests. This was done by determining ERM biomass, using ergosterol as a proxy, in sequentially harvested in-growth mesh bags and by applying mathematical models. Although ERM production declined with increasing forest age from 1.2 to 0.5 kg ha-1  d-1 , the standing biomass increased from 50 to 112 kg ha-1 . This was explained by a drastic decline in mycelial turnover from seven times to one time per year with increasing forest age, corresponding to mean residence times from 25 d up to 1 yr. Our results demonstrate that ERM turnover is the main factor regulating biomass across differently aged forest stands. Explicit inclusion of ERM parameters in forest ecosystem C models may significantly improve their capacity to predict responses of mycorrhiza-mediated processes to management and environmental changes.

Production and turnover of ectomycorrhizal extramatrical mycelial biomass and necromass under elevated CO2 and nitrogen fertilization.

Extramatrical mycelia (EMM) of ectomycorrhizal fungi are important in carbon (C) and nitrogen (N) cycling in forests, but poor knowledge about EMM biomass and necromass turnovers makes the quantification of their role problematic. We studied the impacts of elevated CO2 and N fertilization on EMM production and turnover in a Pinus taeda forest. EMM C was determined by the analysis of ergosterol (biomass), chitin (total bio- and necromass) and total organic C (TOC) of sand-filled mycelium in-growth bags. The production and turnover of EMM bio- and necromass and total C were estimated by modelling. N fertilization reduced the standing EMM biomass C to 57% and its production to 51% of the control (from 238 to 122 kg C ha(-1)  yr(-1) ), whereas elevated CO2 had no detectable effects. Biomass turnover was high (˜13 yr(-1) ) and unchanged by the treatments. Necromass turnover was slow and was reduced from 1.5 yr(-1) in the control to 0.65 yr(-1) in the N-fertilized treatment. However, TOC data did not support an N effect on necromass turnover. An estimated EMM production ranging from 2.5 to 6% of net primary production stresses the importance of its inclusion in C models. A slow EMM necromass turnover indicates an importance in building up forest humus.

Comparison of PFASs contamination in the freshwater and terrestrial environments by analysis of eggs from osprey (Pandion haliaetus), tawny owl (Strix aluco), and common kestrel (Falco tinnunculus).

The level of PFAS (per- and polyfluorinated alkyl substances) contamination in freshwater and terrestrial Swedish environments in 2013/2014 was assessed by analyzing a range of perfluorinated alkyl acids, fluorotelomer acids, sulfonamides, sulfonamidoethanols and polyfluoralkyl phosphate diesters (diPAPs) in predator bird eggs. Stable isotopes ((13)C and (15)N) were analyzed to elucidate the dietary source. The tawny owl (Strix aluco, n=10) and common kestrel (Falco tinnunculus, n=40), two terrestrial species, and the osprey (Pandion haliaetus, n=30), a freshwater specie were included. In addition, a temporal trend (1997-2001, 2008-2009, 2013) in osprey was studied as well. The PFAS profile was dominated by perfluorooctane sulfonic acid (PFOS) in eggs from osprey and tawny owl, while for common kestrel perfluorinated carboxylic acids (∑PFCA) exceeded the level of PFOS. PFOS concentration in osprey eggs remained at the same level between 1997 and 2001 and 2013. For the long-chained PFCAs, there were a significant increase in concentrations in osprey eggs between 1997 and 2001 and 2008-2009. The levels of PFOS and PFCAs were about 10 and five times higher, respectively, in osprey compared to tawny owl and common kestrel. Evidence of direct exposure from PFCA precursor compounds to birds in both freshwater and terrestrial environment was observed. Low levels of diPAPs were detected in a few samples of osprey (<0.02-2.4ng/g) and common kestrel (<0.02-0.16ng/g) eggs, and 6:2 FTSA was detected in a majority of the osprey eggs (<6.3-52ng/g). One saturated telomer acid (7:3 FTCA), which is a transformation marker from precursor exposure, was detected in all species (<0.24-2.7ng/g). The (15)N data showed higher levels in osprey eggs compared to tawny owl and common kestrel, indicating that they feed on a 2-3 times higher trophic level. We conclude that ospreys are continuously exposed to PFAS at levels where adverse toxic effects have been observed in birds.

Partial mycoheterotrophy in Pyroleae: nitrogen and carbon stable isotope signatures during development from seedling to adult.

Mycoheterotrophic plants (MHP) are divided into non-photosynthesizing full MHP and green-leaved partial or initial MHP. We investigated (13)C and (15)N isotope enrichment in five putatively partial MHP species in the tribe Pyroleae (Ericaceae): Chimaphila umbellata, Moneses uniflora, Orthilia secunda, Pyrola chlorantha and Pyrola minor, sampled from forest sites on Öland, Sweden. For M. uniflora and P. chlorantha, we investigated isotope signatures of subterranean seedlings (which are mycoheterotrophic), to examine how the use of seedlings instead of full MHP species (Hypopitys monotropa) as reference species affects the assessment of partial mycoheterotrophy. Our main findings were as follows: (1) All investigated Pyroleae species were enriched in (15)N compared to autotrophic reference plants. (2) significant fungal-derived C among the Pyroleae species was found for O. secunda and P. chlorantha. For the remaining species of C. umbellata, M. uniflora and P. minor, isotope signatures suggested adult autotrophy. (3) C and N gains, calculated using seedlings as a full MHP reference, yielded qualitatively similar results as when using H. monotropa as a reference. However, the estimated differences in C and N gains became larger when using seedlings as an MHP reference. (4) A previously unknown interspecific variation in isotope signature occurs during early ontogeny, from seed production to developing seedlings. Our findings suggest that there is a variation among Pyroleae species concerning partial mycoheterotrophy in adults. Adult autotrophy may be most common in Pyroleae species, and these species may not be as dependent on fungal-derived nutrients as some green orchids.

Increase in soil stable carbon isotope ratio relates to loss of organic carbon: results from five long-term bare fallow experiments.

Changes in the (12)C/(13)C ratio (expressed as δ(13)C) of soil organic C (SOC) has been observed over long time scales and with depth in soil profiles. The changes are ascribed to the different reaction kinetics of (12)C and (13)C isotopes and the different isotopic composition of various SOC pool components. However, experimental verification of the subtle isotopic shifts associated with SOC turnover under field conditions is scarce. We determined δ(13)C and SOC in soil sampled during 1929-2009 in the Ap-horizon of five European long-term bare fallow experiments kept without C inputs for 27-80 years and covering a latitudinal range of 11°. The bare fallow soils lost 33-65% of their initial SOC content and showed a mean annual δ(13)C increase of 0.008-0.024‰. The (13)C enrichment could be related empirically to SOC losses by a Rayleigh distillation equation. A more complex mechanistic relationship was also examined. The overall estimate of the fractionation coefficient (ε) was -1.2 ± 0.3‰. This coefficient represents an important input to studies of long-term SOC dynamics in agricultural soils that are based on variations in (13)C natural abundance. The variance of ε may be ascribed to site characteristics not disclosed in our study, but the very similar kinetics measured across our five experimental sites suggest that overall site-specific factors (including climate) had a marginal influence and that it may be possible to isolate a general mechanism causing the enrichment, although pre-fallow land use may have some impact on isotope abundance and fractionation.

Biochar and peat amendments affect nitrogen retention, microbial capacity and nitrogen cycling microbial communities in a metal and polycyclic aromatic hydrocarbon contaminated urban soil.

Soil contaminants may restrict soil functions. A promising soil remediation method is amendment with biochar, which has the potential to both adsorb contaminants and improve soil health. However, effects of biochar amendment on soil-plant nitrogen (N) dynamics and N cycling microbial guilds in contaminated soils are still poorly understood. Here, a metal- and polycyclic aromatic hydrocarbon (PAH) contaminated soil was amended with either biochar (0, 3, 6 % w/w) and/or peat (0, 1.5, 3 % w/w) in a full-factorial design and sown with perennial ryegrass in an outdoor field trial. After three months, N and the stable isotopic ratio δ15N was measured in soil, roots and leaves, along with microbial responses. Aboveground grass biomass decreased by 30 % and leaf N content by 20 % with biochar, while peat alone had no effect. Peat in particular, but also biochar, stimulated the abundance of microorganisms (measured as 16S rRNA gene copy number) and basal respiration. Microbial substrate utilization (MicroResp™) was altered differentially, as peat increased respiration of all carbon sources, while for biochar, respiration of carboxylic acids increased, sugars decreased, and was unaffected for amino acids. Biochar increased the abundance of ammonia oxidizing archaea, while peat stimulated ammonia oxidizing bacteria, Nitrobacter-type nitrite oxidizers and comB-type complete ammonia oxidizers. Biochar and peat also increased nitrous oxide reducing communities (nosZI and nosZII), while peat alone or combined with biochar also increased abundance of nirK-type denitrifiers. However, biochar and peat lowered leaf δ15N by 2-4 ‰, indicating that processes causing gaseous N losses, like denitrification and ammonia volatilization, were reduced compared to the untreated contaminated soil, probably an effect of biotic N immobilization. Overall, this study shows that in addition to contaminant stabilization, amendment with biochar and peat can increase N retention while improving microbial capacity to perform important soil functions.

An alpine treeline in a carbon dioxide-rich world: synthesis of a nine-year free-air carbon dioxide enrichment study.

We evaluated the impacts of elevated CO2 in a treeline ecosystem in the Swiss Alps in a 9-year free-air CO2 enrichment (FACE) study. We present new data and synthesize plant and soil results from the entire experimental period. Light-saturated photosynthesis (A max) of ca. 35-year-old Larix decidua and Pinus uncinata was stimulated by elevated CO2 throughout the experiment. Slight down-regulation of photosynthesis in Pinus was consistent with starch accumulation in needle tissue. Above-ground growth responses differed between tree species, with a 33 % mean annual stimulation in Larix but no response in Pinus. Species-specific CO2 responses also occurred for abundant dwarf shrub species in the understorey, where Vaccinium myrtillus showed a sustained shoot growth enhancement (+11 %) that was not apparent for Vaccinium gaultherioides or Empetrum hermaphroditum. Below ground, CO2 enrichment did not stimulate fine root or mycorrhizal mycelium growth, but increased CO2 effluxes from the soil (+24 %) indicated that enhanced C assimilation was partially offset by greater respiratory losses. The dissolved organic C (DOC) concentration in soil solutions was consistently higher under elevated CO2 (+14 %), suggesting accelerated soil organic matter turnover. CO2 enrichment hardly affected the C-N balance in plants and soil, with unaltered soil total or mineral N concentrations and little impact on plant leaf N concentration or the stable N isotope ratio. Sustained differences in plant species growth responses suggest future shifts in species composition with atmospheric change. Consistently increased C fixation, soil respiration and DOC production over 9 years of CO2 enrichment provide clear evidence for accelerated C cycling with no apparent consequences on the N cycle in this treeline ecosystem.

Nitrogen dynamics of decomposing Scots pine needle litter depends on colonizing fungal species.

In boreal ecosystems plant production is often limited by low availability of nitrogen. Nitrogen retention in below-ground organic pools plays an important role in restricting recirculation to plants and thereby hampers forest production. Saprotrophic fungi are commonly assigned to different decomposer strategies, but how these relate to nitrogen cycling remains to be understood. Decomposition of Scots pine needle litter was studied in axenic microcosms with the ligninolytic litter decomposing basidiomycete Gymnopus androsaceus or the stress tolerant ascomycete Chalara longipes. Changes in chemical composition were followed by 13C CP/MAS NMR spectroscopy and nitrogen dynamics was assessed by the addition of a 15N tracer. Decomposition by C. longipes resulted in nitrogen retention in non-hydrolysable organic matter, enriched in aromatic and alkylic compounds, whereas the ligninolytic G. androsaceus was able to access this pool, counteracting nitrogen retention. Our observations suggest that differences in decomposing strategies between fungal species play an important role in regulating nitrogen retention and release during litter decomposition, implying that fungal community composition may impact nitrogen cycling at the ecosystem level.

Contrasting effects of ectomycorrhizal fungi on early and late stage decomposition in a boreal forest.

Symbiotic ectomycorrhizal fungi have received increasing attention as regulators of below-ground organic matter storage. They are proposed to promote organic matter accumulation by suppressing saprotrophs, but have also been suggested to play an active role in decomposition themselves. Here we show that exclusion of tree roots and associated ectomycorrhizal fungi in a boreal forest increased decomposition of surface litter by 11% by alleviating nitrogen limitation of saprotrophs-a "Gadgil effect". At the same time, root exclusion decreased Mn-peroxidase activity in the deeper mor layer by 91%. Our results show that ectomycorrhizal fungi may hamper short-term litter decomposition, but also support a crucial role of ectomycorrhizal fungi in driving long-term organic matter oxidation. These observations stress the importance of ectomycorrhizal fungi in regulation of below-ground organic matter accumulation. By different mechanisms they may either hamper or stimulate decomposition, depending upon stage of decomposition and location in the soil profile.

Complex effects of mammalian grazing on extramatrical mycelial biomass in the Scandes forest-tundra ecotone.

Mycorrhizal associations are widespread in high-latitude ecosystems and are potentially of great importance for global carbon dynamics. Although large herbivores play a key part in shaping subarctic plant communities, their impact on mycorrhizal dynamics is largely unknown. We measured extramatrical mycelial (EMM) biomass during one growing season in 16-year-old herbivore exclosures and unenclosed control plots (ambient), at three mountain birch forests and two shrub heath sites, in the Scandes forest-tundra ecotone. We also used high-throughput amplicon sequencing for taxonomic identification to investigate differences in fungal species composition. At the birch forest sites, EMM biomass was significantly higher in exclosures (1.36 ± 0.43 g C/m2) than in ambient conditions (0.66 ± 0.17 g C/m2) and was positively influenced by soil thawing degree-days. At the shrub heath sites, there was no significant effect on EMM biomass (exclosures: 0.72 ± 0.09 g C/m2; ambient plots: 1.43 ± 0.94). However, EMM biomass was negatively related to Betula nana abundance, which was greater in exclosures, suggesting that grazing affected EMM biomass positively. We found no significant treatment effects on fungal diversity but the most abundant ectomycorrhizal lineage/cortinarius, showed a near-significant positive effect of herbivore exclusion (p = .08), indicating that herbivory also affects fungal community composition. These results suggest that herbivory can influence fungal biomass in highly context-dependent ways in subarctic ecosystems. Considering the importance of root-associated fungi for ecosystem carbon balance, these findings could have far-reaching implications.

Nitrogen fixation by Alnus incana and nitrogen transfer from A. incana to Pinus sylvestris influenced by macronutrients and ectomycorrhiza.

The aims of this study were to evaluate the effect of macronutrients on nitrogen fixation in mycorrhizal and non-mycorrhizal grey alder (Alnus incana (L.) Moench), and to evaluate the effect of ectomycorrhizal mycelium on the transfer of symbolically fixed nitrogen from grey alder to Scots pine (Pinus sylvestris L). One alder and one pine were grown together in pots with root systems separated by a 20µm mesh nylon filter which allowed hyphae but not roots to penetrate. Half the plants of both species were inoculated with Paxillus involutus (Ft.) Ft. and all alders were inoculated with Frankia. Nutrient solutions were added with macronutrient (N, K, P, Ca, Mg and S) concentrations varied according to a two-level fractional factorial design. The plants were harvested after two growing periods in a growth chamber. Nitrogen fixation by alder and transfer of symbiotically fixed N from alder to pine was measured by 15 N-dilution. Fixed N (mg) correlated with nodule biomass in both mycorrhizal and non-mycorrhizal alders. On average, specific nodule activity over the two periods was 510 mg N fixed R' nodule d. wt. This was not affected by mycorrhizal infection or by the different nutrient treatments. By contrast there was a strong nutrient effect on the proportion of N derived from fixation. These results indicate that the regulation of nitrogen fixation was via nodule growth rather than via nodule specific activity. Nitrogen had a strong negative effect and P a positive effect on the percentage of N derived from fixation (%Ndfa). However, the effect of N depended on the level of P. This N × P interaction resulted in a %Ndfa when N was high, of 5-10%, at low P and 45-48%, at high P. The highest value of 90% Ndfa was found at the combination of low N and high P. Potassium had a small but statistically significant effect on the %Ndfa but Ca, Mg and S had no significant effects. No mycorrhizal effect was found on the %Ndfa in alder. By contrast, the %Ndfa and biomass were lower in mycorrhizal than in non-mycorrhizal alders. The proportion of fixed N in pine, transferred from alder, was greatest (9%) when the pine was nitrogen starved and mycorrhizal and the alder was fixing maximally (low N and high P). However, the amount of fixed N transferred to pine was not statistically different from zero.

Fungal biomass in roots and extramatrical mycelium in relation to macronutrients and plant biomass of ectomycorrhizal Pinus sylvestris and Alnus incana.

We studied the effects of macronutrients on the production and distribution of fungal biomass and plant biomass in ectomycorrhizal (Paxillus involutus (Fr.) Fr.) or non-mycorrhiza] Pinus sylvestris L, and Alnus incana (L.) Moench. Fungal biomass was measured as ergosterol content in roots and extramatrical mycelium, Alnus infants was nodulated with Frankia. All six macronutrients were varied according to a two-level fractional factorial design, The plants were grown in pots during two growing periods in a growth chamber. Levels of N, P and sometimes K and interactions between them, had highly significant effects, whereas Ca. Mg and S had no significant effects. The production of extramatrical mycelial biomass peaked when P was low and other nutrients were high. This investment in extramatrical mycelium resulted in a 660%, higher biomass in mycorrhizal compared with non-mycorrhizal P. sylvestris at this nutrient regime. The proportion of fungal biomass in roots was stable in P. sylvestris hut more variable in A. incana. Alnus incana grew less when mycorrhizal then when non-mycorrhizal. The growth responses to mycorrhiza and to the different nutrient treatments were evident at the end of the first growing period. Non-mycorrhizal P. sylvestris did not respond to P limitation by a production of proportionally more roots. This might be a reflection of an obligate dependency on mycorrhiza for effective P uptake. By contrast, the root/shoot ratio in both mycorrhizal and non-mycorrhiza] P. sylvestris decreased strongly in response to increased N. The opposite root/shoot response was found in Alnus incana, and the ratio decreased strongly in response to increased P and increased in response to increased N.

13C-discrimination during microbial respiration of added C3-, C4- and 13C-labelled sugars to a C3-forest soil.

We tested whether 13C-discrimination during microbial respiration, or during CO2 sampling in the field, can explain changes observed in the δ13C of emitted CO2 that follow the addition of C4-sucrose, as a microbial substrate, to the soil of a C3-ecosystem. We approached this problem by adding C3-glucose (δ13C=-23.4‰), C4-sucrose (-10.8‰) or 13C-labelled glucose (103.7‰) to the intact mor layer, the upper organic soil (-26.5‰, bulk soil organic matter), of a boreal Pinus sylvestris L. forest. If 13C-discrimination is significant, it should generate illusory differences in the calculated contributions from the added C and endogenous C3-C to total soil respiration, when C4-sucrose or 13C-labelled glucose is added. Further, if discrimination occurs, we should also be able to detect a shift in the δ13C of respired CO2 after the addition of C3-glucose. The addition of the three sugar solutions gave similar increases in soil respiration (up to a doubling 1 h after the additions), while the addition of water gave no increase in respiration. There was no change in δ13C of the emitted CO2 after additions of H2O or C3-glucose. In contrast, the addition of C4-sucrose and 13C-labelled glucose gave δ13C values of evolved CO2 that were 4.5‰ and 30.3‰ higher than the pre-sugar values, respectively. The calculated respiration rates of the added carbon sources, C4-C or 13C-labelled C, were very similar. Also, we found very similar sugar-induced increases in respiration of endogenous C3-C in the plots supplied with C4-sucrose and 13C-labelled glucose, accounting for about 50% of the total increase in respiration 1 h after addition. Our results confirm that any microbial 13C-discrimination during respiration is minor.

Nitrogen and carbon reallocation in fungal mycelia during decomposition of boreal forest litter.

Boreal forests are characterized by spatially heterogeneous soils with low N availability. The decomposition of coniferous litter in these systems is primarily performed by basidiomycete fungi, which often form large mycelia with a well-developed capacity to reallocate resources spatially- an advantageous trait in heterogeneous environments. In axenic microcosm systems we tested whether fungi increase their biomass production by reallocating N between Pinus sylvestris (Scots pine) needles at different stages of decomposition. We estimated fungal biomass production by analysing the accumulation of the fungal cell wall compound chitin. Monospecific systems were compared with systems with interspecific interactions. We found that the fungi reallocated assimilated N and mycelial growth away from well-degraded litter towards fresh litter components. This redistribution was accompanied by reduced decomposition of older litter. Interconnection of substrates increased over-all fungal C use efficiency (i.e. the allocation of assimilated C to biomass rather than respiration), presumably by enabling fungal translocation of growth-limiting N to litter with higher C quality. Fungal connection between different substrates also restricted N-mineralization and production of dissolved organic N, suggesting that litter saprotrophs in boreal forest ecosystems primarily act to redistribute rather than release N. This spatial integration of different resource qualities was hindered by interspecific interactions, in which litters of contrasting quality were colonised by two different basidiomycete species. The experiments provide a detailed picture of how resource reallocation in two decomposer fungi leads to a more efficient utilisation of spatially separated resources under N-limitation. From an ecosystem point of view, such economic fungal behaviour could potentially contribute to organic matter accumulation in the litter layers of boreal forests.

Isotope fractionation and 13C enrichment in soil profiles during the decomposition of soil organic matter.

The mechanisms behind the (13)C enrichment of organic matter with increasing soil depth in forests are unclear. To determine if (13)C discrimination during respiration could contribute to this pattern, we compared delta(13)C signatures of respired CO(2) from sieved mineral soil, litter layer and litterfall with measurements of delta(13)C and delta(15)N of mineral soil, litter layer, litterfall, roots and fungal mycelia sampled from a 68-year-old Norway spruce forest stand planted on previously cultivated land. Because the land was subjected to ploughing before establishment of the forest stand, shifts in delta(13)C in the top 20 cm reflect processes that have been active since the beginning of the reforestation process. As (13)C-depleted organic matter accumulated in the upper soil, a 1.0 per thousand delta(13)C gradient from -28.5 per thousand in the litter layer to -27.6 per thousand at a depth of 2-6 cm was formed. This can be explained by the 1 per thousand drop in delta(13)C of atmospheric CO(2) since the beginning of reforestation together with the mixing of new C (forest) and old C (farmland). However, the isotopic change of the atmospheric CO(2) explains only a portion of the additional 1.0 per thousand increase in delta(13)C below a depth of 20 cm. The delta(13)C of the respired CO(2) was similar to that of the organic matter in the upper soil layers but became increasingly (13)C enriched with depth, up to 2.5 per thousand relative to the organic matter. We hypothesise that this (13)C enrichment of the CO(2) as well as the residual increase in delta(13)C of the organic matter below a soil depth of 20 cm results from the increased contribution of (13)C-enriched microbially derived C with depth. Our results suggest that (13)C discrimination during microbial respiration does not contribute to the (13)C enrichment of organic matter in soils. We therefore recommend that these results should be taken into consideration when natural variations in delta(13)C of respired CO(2) are used to separate different components of soil respiration or ecosystem respiration.