Erik A. Lilleskov

Effects of nitrogen deposition and empirical nitrogen critical loads for ecoregions of the United States

Human activity in the last century has led to a significant increase in nitrogen (N) emissions and atmospheric deposition. This N deposition has reached a level that has caused or is likely to cause alterations to the structure and function of many ecosystems across the United States. One approach for quantifying the deposition of pollution that would be harmful to ecosystems is the determination of critical loads. A critical load is defined as the input of a pollutant below which no detrimental ecological effects occur over the long-term according to present knowledge. The objectives of this project were to synthesize current research relating atmospheric N deposition to effects on terrestrial and freshwater ecosystems in the United States, and to estimate associated empirical N critical loads. The receptors considered included freshwater diatoms, mycorrhizal fungi, lichens, bryophytes, herbaceous plants, shrubs, and trees. Ecosystem impacts included: (1) biogeochemical responses and (2) individual species, population, and community responses. Biogeochemical responses included increased N mineralization and nitrification (and N availability for plant and microbial uptake), increased gaseous N losses (ammonia volatilization, nitric and nitrous oxide from nitrification and denitrification), and increased N leaching. Individual species, population, and community responses included increased tissue N, physiological and nutrient imbalances, increased growth, altered root : shoot ratios, increased susceptibility to secondary stresses, altered fire regime, shifts in competitive interactions and community composition, changes in species richness and other measures of biodiversity, and increases in invasive species.

Detection of forest stand-level spatial structure in ectomycorrhizal fungal communities

Ectomycorrhizal fungal (EMF) communities are highly diverse at the stand level. To begin to understand what might lead to such diversity, and to improve sampling designs, we investigated the spatial structure of these communities. We used EMF community data from a number of studies carried out in seven mature and one recently fire-initiated forest stand. We applied various measures of spatial pattern to characterize distributions at EMF community and species levels: Mantel tests, Mantel correlograms, variance/mean and standardized variograms. Mantel tests indicated that in four of eight sites community similarity decreased with distance, whereas Mantel correlograms also found spatial autocorrelation in those four plus two additional sites. In all but one of these sites elevated similarity was evident only at relatively small spatial scales (< 2.6 m), whereas one exhibited a larger scale pattern ( approximately 25 m). Evenness of biomass distribution among cores varied widely among taxa. Standardized variograms indicated that most of the dominant taxa showed patchiness at a scale of less than 3 m, with a range from 0 to < or =17 m. These results have implications for both sampling scale and intensity to achieve maximum efficiency of community sampling. In the systems we examined, cores should be at least 3 m apart to achieve the greatest sampling efficiency for stand-level community analysis. In some cases even this spacing may result in reduced sampling efficiency arising from patterns of spatial autocorrelation. Interpretation of the causes and significance of these patterns requires information on the genetic identity of individuals in the communities.

Nitrogen availability is a primary determinant of conifer mycorrhizas across complex environmental gradients

Global environmental change has serious implications for functional biodiversity in temperate and boreal forests. Trees depend on mycorrhizal fungi for nutrient uptake, but predicted increases in nitrogen availability may alter fungal communities. To address a knowledge gap regarding the effects of nitrogen availability on mycorrhizal communities at large scales, we examine the relationship between nitrogen and ectomycorrhizas in part of a European biomonitoring network of pine forest plots. Our analyses show that increased nitrogen reduces fungal diversity and causes shifts in mycorrhizal community composition across plots, but we do not find strong evidence that within-plot differences in nitrogen availability affect ectomycorrhizal communities. We also carry out exploratory analyses to determine the relative importance of other environmental variables in structuring mycorrhizal communities, and discuss the potential use of indicator species to predict nitrogen-induced shifts in fungal communities.

Fungal community composition and metabolism under elevated CO2 and O3

Atmospheric CO2 and O3 concentrations are increasing due to human activity and both trace gases have the potential to alter C cycling in forest ecosystems. Because soil microorganisms depend on plant litter as a source of energy for metabolism, changes in the amount or the biochemistry of plant litter produced under elevated CO2 and O3 could alter microbial community function and composition. Previously, we have observed that elevated CO2 increased the microbial metabolism of cellulose and chitin, whereas elevated O3 dampened this response. We hypothesized that this change in metabolism under CO2 and O3 enrichment would be accompanied by a concomitant change in fungal community composition. We tested our hypothesis at the free-air CO2 and O3 enrichment (FACE) experiment at Rhinelander, Wisconsin, in which Populus tremuloides, Betula papyrifera, and Acer saccharum were grown under factorial CO2 and O3 treatments. We employed extracellular enzyme analysis to assay microbial metabolism, phospholipid fatty acid (PLFA) analysis to determine changes in microbial community composition, and polymerase chain reaction–denaturing gradient gel electrophoresis (PCR–DGGE) to analyze the fungal community composition. The activities of 1,4-β-glucosidase (+37%) and 1,4,-β-N-acetylglucosaminidase (+84%) were significantly increased under elevated CO2, whereas 1,4-β-glucosidase activity (−25%) was significantly suppressed by elevated O3. There was no significant main effect of elevated CO2 or O3 on fungal relative abundance, as measured by PLFA. We identified 39 fungal taxonomic units from soil using DGGE, and found that O3 enrichment significantly altered fungal community composition. We conclude that fungal metabolism is altered under elevated CO2 and O3, and that there was a concomitant change in fungal community composition under elevated O3. Thus, changes in plant inputs to soil under elevated CO2 and O3 can propagate through the microbial food web to alter the cycling of C in soil.

Simulated nitrogen deposition affects community structure of arbuscular mycorrhizal fungi in northern hardwood forests

Our previous investigation found elevated nitrogen deposition caused declines in abundance of arbuscular mycorrhizal fungi (AMF) associated with forest trees, but little is known about how nitrogen affects the AMF community composition and structure within forest ecosystems. We hypothesized that N deposition would lead to significant changes in the AMF community structure. We studied the diversity and community structure of AMF in northern hardwood forests after more than 12 years of simulated nitrogen deposition. We performed molecular analyses on maple (Acer spp.) roots targeting the 18S rDNA region using the fungal‐specific primers AM1 and NS31. PCR products were cloned and identified using restriction fragment length polymorphism (RFLP) and sequencing. N addition significantly altered the AMF community structure, and Glomus group A dominated the AMF community. Some Glomus operational taxonomic units (OTUs) responded negatively to N inputs, whereas other Glomus OTUs and an Acaulospora OTU responded positively to N inputs. The observed effect on community structure implies that AMF species associated with maples differ in their response to elevated nitrogen. Given that functional diversity exists among AMF species and that N deposition has been shown to select less beneficial fungi in some ecosystems, this change in community structure could have implications for the functioning of this type of ecosystem.

Productivity and community structure of ectomycorrhizal fungal sporocarps under increased atmospheric CO2 and O3

Sporocarp production is essential for ectomycorrhizal fungal recombination and dispersal, which influences fungal community dynamics. Increasing atmospheric carbon dioxide (CO2) and ozone (O3) affect host plant carbon gain and allocation, which may in turn influence ectomycorrhizal sporocarp production if the carbon available to the ectomycorrhizal fungus is dependant upon the quantity of carbon assimilated by the host. We measured sporocarp production of ectomycorrhizal fungi over 4 years at the Aspen FACE (free air CO2 enrichment) site, which corresponded to stand ages seven to 10 years. Total mean sporocarp biomass was greatest under elevated CO2, regardless of O3 concentration, while it was generally lowest under elevated O3 with ambient CO2. Community composition differed significantly among the treatments, with less difference in the final year of the study. Whether this convergence was due to succession or environmental factors is uncertain. CO2 and O3 affect ectomycorrhizal sporocarp productivity and community composition, with likely effects on dispersal, colonization and sporocarp-dependent food webs.

Water sources and controls on water-loss rates of epigeous ectomycorrhizal fungal sporocarps during summer drought.

Access to deeper soil water and water-conserving traits should reduce water stress for ectomycorrhizal fungi, permitting function during drought. Here, we explored whether epigeous fruiting of ectomycorrhizal fungi during drought was facilitated by access to deep soil water, how much water was lost from sporocarps, and how sporocarp surface to volume ratios affected water-loss rates. We used oxygen stable isotope analysis of water combined with modeling of water sources used by ectomycorrhizal fungi; measured sporocarp water loss using a transient porometer, and related water loss to vapor pressure deficit (VPD) and sporocarp morphology. In drier soils sporocarps likely derived a significant portion (25-80%) of their water from deep (> 30 cm) or hydraulically lifted water. Amanita muscaria had water-loss rates over twice those of Suillus sp., Boletus edulis, Tricholoma spp. and Russula albonigra. Vapor pressure deficit was an excellent predictor of water-loss rates for individual mushrooms. Sporocarp surface to volume ratios explained much of the variation among mushrooms in the slope of VPD-water loss relationships. Access to deeper soil water might be a significant driver of ectomycorrhizal symbiotic function, sporocarp distribution, fruiting habit and morphology. Sporocarp morphology can affect water-loss rates and hence influences fungal ability to fruit during summer drought.

Advancing the use of minirhizotrons in wetlands

BackgroundWetlands store a substantial amount of carbon (C) in deep soil organic matter deposits, and play an important role in global fluxes of carbon dioxide and methane. Fine roots (i.e., ephemeral roots that are active in water and nutrient uptake) are recognized as important components of biogeochemical cycles in nutrient-limited wetland ecosystems. However, quantification of fine-root dynamics in wetlands has generally been limited to destructive approaches, possibly because of methodological difficulties associated with the unique environmental, soil, and plant community characteristics of these systems. Non-destructive minirhizotron technology has rarely been used in wetland ecosystems.ScopeOur goal was to develop a consensus on, and a methodological framework for, the appropriate installation and use of minirhizotron technology in wetland ecosystems. Here, we discuss a number of potential solutions for the challenges associated with the deployment of minirhizotron technology in wetlands, including minirhizotron installation and anchorage, capture and analysis of minirhizotron images, and upscaling of minirhizotron data for analysis of biogeochemical pools and parameterization of land surface models.ConclusionsThe appropriate use of minirhizotron technology to examine relatively understudied fine-root dynamics in wetlands will advance our knowledge of ecosystem C and nutrient cycling in these globally important ecosystems.

Fungal functioning in a pine forest: evidence from a 15N-labeled global change experiment

• We used natural and tracer nitrogen (N) isotopes in a Pinus taeda free air CO₂ enrichment (FACE) experiment to investigate functioning of ectomycorrhizal and saprotrophic fungi in N cycling. • Fungal sporocarps were sampled in 2004 (natural abundance and (15) N tracer) and 2010 (tracer) and δ(15)N patterns were compared against litter and soil pools. • Ectomycorrhizal fungi with hydrophobic ectomycorrhizas (e.g. Cortinarius and Tricholoma) acquired N from the Oea horizon or deeper. Taxa with hydrophilic ectomycorrhizas acquired N from the Oi horizon (Russula and Lactarius) or deeper (Laccaria, Inocybe, and Amanita). (15)N enrichment patterns for Cortinarius and Amanita in 2010 did not correspond to any measured bulk pool, suggesting that a persistent pool of active organic N supplied these two taxa. Saprotrophic fungi could be separated into those colonizing pine cones (Baeospora), wood, litter (Oi), and soil (Ramariopsis), with δ(15)N of taxa reflecting substrate differences. (15)N enrichment between sources and sporocarps varied across taxa and contributed to δ(15)N patterns. • Natural abundance and (15)N tracers proved useful for tracking N from different depths into fungal taxa, generally corresponded to literature estimates of fungal activity within soil profiles, and provided new insights into interpreting natural abundance δ(15)N patterns.

The Sphagnum microbiome: new insights from an ancient plant lineage

57 I. 57 II. 58 III. 59 IV. 59 V. 61 VI. 62 63 References 63 SUMMARY: Peat mosses of the genus Sphagnum play a major role in global carbon storage and dominate many northern peatland ecosystems, which are currently being subjected to some of the most rapid climate changes on Earth. A rapidly expanding database indicates that a diverse community of microorganisms is intimately associated with Sphagnum, inhabiting the tissues and surface of the plant. Here we summarize the current state of knowledge regarding the Sphagnum microbiome and provide a perspective for future research directions. Although the majority of the microbiome remains uncultivated and its metabolic capabilities uncharacterized, prokaryotes and fungi have the potential to act as mutualists, symbionts, or antagonists of Sphagnum. For example, methanotrophic and nitrogen-fixing bacteria may benefit the plant host by providing up to 20-30% of Sphagnum carbon and nitrogen, respectively. Next-generation sequencing approaches have enabled the detailed characterization of microbiome community composition in peat mosses. However, as with other ecologically or economically important plants, our knowledge of Sphagnum-microbiome associations is in its infancy. In order to attain a predictive understanding of the role of the microbiome in Sphagnum productivity and ecosystem function, the mechanisms of plant-microbiome interactions and the metabolic potential of constituent microbial populations must be revealed.