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Intensively managed agriculture land is a significant contributor to nitrous oxide (N2O) emissions, which adds to global warming and the depletion of the ozone layer. Recent studies have suggested that fungal dominant N2O production may be promoted by pathogenic fungi under high nitrogen fertilization and continuous cropping. Here, we measured the contribution of fungal communities to N2O production under intensively managed strawberry fields of three continuous cropping years (1, 5, and 10 years) and compared this adjacent bare soil. Higher N2O emission was observed from the 10-year field, of which fungi and prokaryotes accounted for 79.7% and 21.3%, respectively. Fungal population density in the 10-year field soil (4.25 × 105 colony forming units per g (CFU/g) of air-dried soil) was greater than the other cropping years. Illumina MiSeq sequencing of the nirK gene showed that long-term continuous cropping decreased the diversity of the fungal denitrifier community, but increased the abundance of Fusarium oxysporum. Additionally, F. oxysporum produced large amounts of N2O in culture and in sterile 10-year field soil. A systemic infection displayed by bioassay strawberry plants after inoculation demonstrated that F. oxysporum was a pathogenic fungus. Together, results suggest that long-term intensively managed monocropping significantly influenced the denitrifying fungal community and increased their biomass, which increased fungal contribution to N2O emissions and specifically by pathogenic fungi. KEY POINTS: • Distinguishing the role of fungi in long-term continuous cropping field. • Identifying the abundant fungal species with denitrifying ability.

Viruses shape microbial community structures, impacting metabolic pathways and influencing biogeochemical cycles. Despite their importance, the influence of biotic and abiotic factors on viral community structures across environmental gradients in soil is relatively unknown compared to their prokaryotic hosts. While soil pH strongly influences microbial community structure, it is unclear whether there is a similar influence on soil virus communities. In this study, prokaryotic and viral communities were characterized in soils sampled from the extremes of a long-term pH-manipulated soil gradient (pH 4.5 and 7.5), and viral populations were compared to those in a variety of soil ecosystems ranging in pH (4.0 – 7.5). Prokaryotic and viral community structure were significantly influenced by soil pH at the local scale. Of 1,910 viral operational taxonomic units (vOTUs), 99% were restricted to pH 4.5 or 7.5 soil only. These were compared in gene sharing networks of populations from six other European and North American soil systems. A selection of viral clusters from acidic and neutral pH soils were more associated with those from the local gradient pH 4.5 or 7.5 soils, respectively. Results indicate that as with prokaryotes, soil pH is a factor structuring viral communities at the local and global scale.

International audience; The concentration of atmospheric methane (CH 4 ) continues to increase with microbial communities controlling soil–atmosphere fluxes. While there is substantial knowledge of the diversity and function of prokaryotes regulating CH 4 production and consumption, their active interactions with viruses in soil have not been identified. Metagenomic sequencing of soil microbial communities enables identification of linkages between viruses and hosts. However, this does not determine if these represent current or historical interactions nor whether a virus or host are active. In this study, we identified active interactions between individual host and virus populations in situ by following the transfer of assimilated carbon. Using DNA stable-isotope probing combined with metagenomic analyses, we characterized CH 4 -fueled microbial networks in acidic and neutral pH soils, specifically primary and secondary utilizers, together with the recent transfer of CH 4 -derived carbon to viruses. A total of 63% of viral contigs from replicated soil incubations contained homologs of genes present in known methylotrophic bacteria. Genomic sequences of 13 C-enriched viruses were represented in over one-third of spacers in CRISPR arrays of multiple closely related Methylocystis populations and revealed differences in their history of viral interaction. Viruses infecting nonmethanotrophic methylotrophs and heterotrophic predatory bacteria were also identified through the analysis of shared homologous genes, demonstrating that carbon is transferred to a diverse range of viruses associated with CH 4 -fueled microbial food networks.

Crop harvest followed by a fallow period can act as a disturbance on soil microbial communities. Cropping systems intended to improve alpha-diversity of communities may also confer increased compositional stability during succeeding growing seasons. Over a single growing season in a long-term (18 year) agricultural field experiment incorporating conventional (CON), conservation (CA), organic (ORG) and integrated (INT) cropping systems, temporal changes in prokaryote, fungal and arbuscular mycorrhizal fungi (AMF) communities were investigated overwinter, during crop growth and at harvest. While certain prokaryote phyla were influenced by cropping system (e.g. Acidobacteria), the community as a whole was primarily driven by temporal changes over the growing season as distinct overwinter and crop-associated communities, with the same trend observed regardless of cropping system. Species- rich prokaryote communities were most stable over the growing season. Cropping system exerted a greater effect on fungal communities, with alpha-diversity highest and temporal changes most stable under CA. CON was particularly detrimental for alpha-diversity in AMF communities, with AMF alpha-diversity and stability improved under all other cropping systems. Practices that promoted alpha-diversity tended to also increase the similarity and temporal stability of soil fungal (and AMF) communities during a growing season, while prokaryote communities were largely insensitive to management.

Functional, physiological traits are the underlying drivers of niche differentiation. A common framework related to niches occupied by terrestrial prokaryotes is based on copiotrophy or oligotrophy, where resource investment is primarily in either rapid growth or stress tolerance, respectively. A quantitative trait-based approach sought relationships between taxa, traits and niche in terrestrial prokaryotes. With 175 taxa from 11 Phyla and 35 Families (n = 5 per Family), traits were considered as discrete counts of shared genome-encoded proteins. Trait composition strongly supported non-random functional distributions as preferential clustering of related taxa via unweighted pair-group method with arithmetic mean. Trait similarity between taxa increased as taxonomic rank decreased. A suite of Random Forest models identified traits significantly enriched or depleted in taxonomic groups. These traits conveyed functions related to rapid growth, nutrient acquisition and stress tolerance consistent with their presence in copiotroph-oligotroph niches. Hierarchical clustering of traits identified a clade of competitive, copiotrophic Families resilient to oxidative stress versus glycosyltransferase-enriched oligotrophic Families resistant to antimicrobials and environmental stress. However, the formation of five clades suggested a more nuanced view to describe niche differentiation in terrestrial systems is necessary. We suggest considering traits involved in both resource investment and acquisition when predicting niche.

Potential rate assays are used in soil microbial ecology to determine the rates of a functional process in environmental samples under a defined set of conditions. While they can be used appropriately to provide mechanistic insights, potential rates are also often used to estimate the abundance of specific taxonomic groups and their in situ activity. These estimates incorrectly assume that all contributing organisms in a community are active at a maximum rate under one set of ‘optimal’ incubation conditions and that potential rates reflect activity in the soil. While investigators now recognise that populations within communities are physiologically diverse, they often ignore the consequent suboptimal activity, or even inactivity, of the majority of community members performing that function. In this short perspective article, we discuss when potential assays can be informative and highlight the underlying conceptual problems under circumstances where potential assays are misused, using potential nitrification rate (PNR) as an example. PNR was originally developed to estimate the size of active ammonia oxidising communities in environmental samples. It is routinely determined in short-term shaken slurry incubations by measuring assumed maximum rates of nitrate or nitrite production under optimal, non-substrate-limiting conditions. As with other functional processes, it is now recognised that a broad diversity of organisms contribute to aerobic ammonia oxidation in terrestrial and other habitats, and this diversity represents a substantial range of physiologies, including variation in substrate affinity, ammonia tolerance, cell specific activity and substrate preference. Despite this, PNR, and other potential rate assays, are often inappropriately used in an attempt to determine an ecologically relevant measurement of activity in soil. As with any potential assay, PNR has inherent biases towards particular functional groups and its use in investigating the ecology of ammonia oxidisers in natural systems should be carefully considered.

Contents Summary 1122 I. Introduction 1122 II. Are there consistent patterns in diversity of mycorrhizal fungal genotypes and species across space? 1125 III. What is the variation in functional traits and genes of mycorrhizal fungi at different taxonomic scales? 1125 IV. How will environmental change impact the relationships between genotypic and species diversity of mycorrhizal fungi and ecosystem function? 1126 V. Conclusions: considerations for future MEF research 1127 Acknowledgements 1127 References 1127 SUMMARY: Both genotypes and species of mycorrhizal fungi exhibit considerable variation in traits, and this variation can result in their diversity regulating ecosystem function. Yet, the nature of mycorrhizal fungal diversity-ecosystem function (MEF) relationships for both genotypes and species is currently poorly defined. New experiments should reflect the richness of genotypes and species in nature, but we still lack information about the extent to which fungal populations in particular are structured. Sampling designs should quantify the diversity of mycorrhizal fungal genotypes and species at three key broad spatial scales (root fragment, root system and interacting root systems) in order to inform manipulation experiments and to test how mycorrhizal fungal diversity both responds, and confers resilience to, environmental drivers.

A major gap in our understanding of biodiversity–ecosystem function relationships concerns the role of intra- and interspecific diversity of mycorrhizal fungi, which are critical for plant fitness, biogeochemical cycling and other processes. Here, we test the hypothesis that the identity and richness of ectomycorrhizal (ECM) fungi at the intra- and interspecific levels affect ecosystem multifunctionality by regulating plant and fungal productivity, soil CO2 efflux and nutrient retention. Microcosms containing Scots pine (Pinus sylvestris) seedlings colonized by different ECM fungal isolates, in monocultures and mixtures, enabled us to test for both intra- and interspecific identity and richness effects, and transgressive overyielding. Intra- and interspecific identity had modest but significant effects on plant and fungal productivity and nutrient retention, but no effect on CO2 efflux. Intraspecific richness increased plant root productivity and ECM root tips but decreased hyphal length, whereas interspecific richness had no effects. Interspecific mixtures outperformed the most productive monocultures in only 10% of the cases, compared with 42% for the intraspecific mixtures. Both intra- and interspecific identity and richness of ECM fungi regulate ecosystem multifunctionality, but their effects on the direction and magnitude of individual variables differ. Transgressive overyielding suggests that positive niche complementarity effects are driving some of the responses to intraspecific richness.

Landspreading of biosolids (treated sewage sludge) in agroecosystems is a common waste management practice worldwide. Evidence suggests biosolids may be detrimental to arbuscular mycorrhizal fungi (AMF); however, previous studies focused on arable systems and often unrealistically high biosolids application levels. We investigated the effects of biosolids on AMF communities in grassland and arable agroecosystems, in the context of the natural seasonal dynamics of AMF community composition and diversity. A pasture and arable system under commercial farming management were amended annually with two different types of biosolids, applied at levels meeting current European Union regulations, in a factorial, replicated field-scale plot experiment. AMF root colonisation and community composition were measured in Lolium perenne roots from the pasture and Trifolium repens roots growing in arable soil across the seasons of two years. AMF community compositions were assessed by terminal-restriction fragment length polymorphism analyses. Biosolids had no significant effect on AMF root colonisation or community composition in either agroecosystem. Soil chemical analyses indicated several changes in the top 0–5 cm layer of the pasture soil, including small increases in heavy metal concentrations in biosolids relative to control plots. Temporal AMF dynamics were detected in soils from both agroecosystem indicating that the effect of seasonality outweighed that of biosolids application.

The first species of Rhizopogon subgenus Amylopogon identified from eastern North America is described here as Rhizopogon kretzerae Grubisha, Dowie, & Mill. sp. nov. In nature, R. kretzerae has been identified only from DNA sequences of the nuclear ribosomal internal transcribed spacer region (ITS) from mycorrhizal roots of Pinus strobus L. and Pterospora andromedea Nutt. Phylogenetic analyses and nucleotide divergence estimates of Rhizopogon ITS sequences from Pterospora roots from Quebec, Michigan, New York, and New Hampshire combined with ITS sequences of Rhizopogon subgenus Amylopogon species type collections suggest R. kretzerae is a sister-group to Rhizopogon salebrosus A.H. Sm. An absence of shared mutations, two informative sequence gaps, and 12 fixed nucleotide differences differentiate R. kretzerae and R. salebrosus. Furthermore, ITS sequences of 40 Pterospora root samples from five populations in the province of Quebec provided the first record of the distribution of R. kretzerae, a potentially rare to endangered species, in eastern Canada.

Important steps in developing reliable bioindicators for soil quality are characterising soil biodiversity and determining the response of its components to environmental factors across a range of land uses and soil types. Baseline data from a national survey in Ireland were used to explore relationships between diversity and composition of micro-organisms (bacteria, fungi, mycorrhiza), and micro-, meso- and macro-fauna (nematodes; mites; earthworms, ants) across a general gradient representing dominant land uses (arable, pasture, rough-grazing, forest and bogland). These diversity data were also linked to soil physico-chemical properties. Differences in diversity and composition of meso- and macro-fauna, but not microbes, were clear between agriculturally-managed (arable and pasture) and extensively-managed (rough-grazing and bogland) soils corresponding to a broad division between ‘mineral’ and ‘organic’ soils. The abundance, richness and composition of nematode and earthworm taxa were significantly congruent with a number of the other groups. Further analysis, using significant indicator species from each group, identified potential target taxa and linked them to soil environmental gradients. This study suggests that there is potential surrogacy between the diversity of key soil taxa groups and that different sets of bioindicators may be most effective under agricultural and extensive land use.

Culture-independent molecular studies have provided new insights into the diversity of fungi associating with ericaceous plant roots. However, there is little understanding of the distribution of these fungi across landscapes, or the effects of environmental heterogeneity on ericoid mycorrhizal (ERM) fungal diversity and distribution. Terminal-restriction fragment length polymorphism and selective sequence analyses of the internal transcribed spacer regions of rDNA were used to infer fungal diversity of bait Vaccinium macrocarpon grown in soils from nine peatland sites in Ireland, representing three different land uses (bog, rough grazing and forest plantation) and the fungal communities of field-collected Calluna vulgaris for five of these nine sites. A diverse range of potential ERM fungi were found, and the sampling approach significantly affected the diversity of the fungal community. Despite significant site groupings of the fungal communities associated with V. macrocarpon and C. vulgaris, fungal communities were significantly dissimilar between sites with different land uses. Soil nitrogen content significantly explained 52% of the variation in the V. macrocarpon fungal communities. Evidence suggests that environmental heterogeneity has a role in shaping ERM fungal community composition at the landscape scale.

Arbuscular fungi have a major role in directing the functioning of terrestrial ecosystems yet little is known about their biogeographical distribution. The Baas-Becking hypothesis (‘everything is everywhere, but, the environment selects’) was tested by investigating the distribution of arbuscular mycorrhizal fungi (AMF) at the landscape scale and the influence of environmental factors and geographical distance in determining community composition. AMF communities in Trifolium repens and Lolium perenne roots were assessed in 40 geographically dispersed sites in Ireland representing different land uses and soil types. Field sampling and laboratory bioassays were used, with AMF communities characterised using 18S rRNA terminal-restriction fragment length polymorphism. Landscape-scale distribution of AMF was driven by the local environment. AMF community composition was influenced by abiotic variables (pH, rainfall and soil type), but not land use or geographical distance. Trifolium repens and L. perenne supported contrasting communities of AMF, and the communities colonising each plant species were consistent across pasture habitats and over distance. Furthermore, L. perenne AMF communities grouped by soil type within pasture habitats. This is the largest and most comprehensive study that has investigated the landscape-scale distribution of AMF. Our findings support the Baas-Becking hypothesis at the landscape scale and demonstrate the strong influence the local environment has on determining AMF community composition.

Like other myco-heterotrophic plants, Pterospora andromedea (pinedrops) is dependent upon its specific fungal symbionts for survival. The rarity of pinedrops fungal symbiont was investigated in the eastern United States where pinedrops are rare. Wild populations of eastern pinedrops were sampled, and the plant haplotypes and fungal symbionts were characterized with molecular techniques; these data were compared to those from the West with phylogenetic analyses. The frequency of the fungal symbiont in eastern white pine forests was assessed using a laboratory soil bioassay and in situ pinedrops seed baiting. Only one plant haplotype and fungal symbiont was detected. The plant haplotype was not unique to the East. The fungal symbiont appears to be a new species within the genus Rhizopogon, closely related to the western symbionts. This fungal species was not frequent in soils with or without pinedrops, but was less frequent in the latter and in comparison to the fungal symbionts in western forests. Seed baiting resulted in few germinants, suggesting that mycelial networks produced by the eastern fungal symbiont were rare. Results suggest that eastern pinedrops rarity is influenced by the distribution and rarity of its fungal symbiont.

KELSALL, N., C. HAZARD, AND D. J. LEOPOLD (Department of Environmental and Forest Biology, SUNY College of Environmental Science and Forestry, 1 Forestry Drive Syracuse, New York 13210). Influence of climate factors on demographic changes in the New York populations of the federally-listed Phyllitis scolopendrium (L.) Newm. var. americana J. Torrey Bot. Soc. 131:161-168. 2004.-Approximately 92% of the U.S. population of American hart's-tongue fern (Phyllitis scolopendrium var. americana) is found in Onondaga and Madison counties, New York, in 17 distinct colonies. All New York colonies have been periodically censused since 1916, though rarely in the same year. This study examines census data of the total New York population during and following the severe drought of 1999 and during the summers of 2000 and 2002. Additionally, the long-term (1922-2002) census data of seven colonies are examined relative to climate data for the same period. The total New York population has declined overall since the last previous census in 1995. No climate variables explained more than 23% of variation in population size of any growth phase. The lack of controlling, largescale climate factors suggests that the distribution of hart's-tongue fern in New York is determined mostly by the occurence of habitat capable of buffering climatic fluctuations. Future work should combine small-scale habitat and large-scale climate factors in any analysis.