Impacts of long‐term elevated atmospheric CO2 concentrations on communities of arbuscular mycorrhizal fungi

The ecological impacts of long‐term elevated atmospheric CO₂ (eCO₂) levels on soil microbiota remain largely unknown. This is particularly true for the arbuscular mycorrhizal (AM) fungi, which form mutualistic associations with over two‐thirds of terrestrial plant species and are entirely dependent on their plant hosts for carbon. Here, we use high‐resolution amplicon sequencing (Illumina, HiSeq) to quantify the response of AM fungal communities to the longest running (>15 years) free‐air carbon dioxide enrichment (FACE) experiment in the Northern Hemisphere (GiFACE); providing the first evaluation of these responses from old‐growth (>100 years) semi‐natural grasslands subjected to a 20% increase in atmospheric CO₂. eCO₂ significantly increased AM fungal richness but had a less‐pronounced impact on the composition of their communities. However, while broader changes in community composition were not observed, more subtle responses of specific AM fungal taxa were with populations both increasing and decreasing in abundance in response to eCO₂. Most population‐level responses to eCO₂ were not consistent through time, with a significant interaction between sampling time and eCO₂ treatment being observed. This suggests that the temporal dynamics of AM fungal populations may be disturbed by anthropogenic stressors. As AM fungi are functionally differentiated, with different taxa providing different benefits to host plants, changes in population densities in response to eCO₂ may significantly impact terrestrial plant communities and their productivity. Thus, predictions regarding future terrestrial ecosystems must consider changes both aboveground and belowground, but avoid relying on broad‐scale community‐level responses of soil microbes observed on single occasions.     

The phylogenetic composition and structure of soil microbial communities shifts in response to elevated carbon dioxide

One of the major factors associated with global change is the ever-increasing concentration of atmospheric CO₂. Although the stimulating effects of elevated CO₂ (eCO₂) on plant growth and primary productivity have been established, its impacts on the diversity and function of soil microbial communities are poorly understood. In this study, phylogenetic microarrays (PhyloChip) were used to comprehensively survey the richness, composition and structure of soil microbial communities in a grassland experiment subjected to two CO₂ conditions (ambient, 368 p.p.m., versus elevated, 560 p.p.m.) for 10 years. The richness based on the detected number of operational taxonomic units (OTUs) significantly decreased under eCO₂. PhyloChip detected 2269 OTUs derived from 45 phyla (including two from Archaea), 55 classes, 99 orders, 164 families and 190 subfamilies. Also, the signal intensity of five phyla (Crenarchaeota, Chloroflexi, OP10, OP9/JS1, Verrucomicrobia) significantly decreased at eCO₂, and such significant effects of eCO₂ on microbial composition were also observed at the class or lower taxonomic levels for most abundant phyla, such as Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes and Acidobacteria, suggesting a shift in microbial community composition at eCO₂. Additionally, statistical analyses showed that the overall taxonomic structure of soil microbial communities was altered at eCO₂. Mantel tests indicated that such changes in species richness, composition and structure of soil microbial communities were closely correlated with soil and plant properties. This study provides insights into our understanding of shifts in the richness, composition and structure of soil microbial communities under eCO₂ and environmental factors shaping the microbial community structure.     

Positive climate feedbacks of soil microbial communities in a semi‐arid grassland

Soil microbial communities may be able to rapidly respond to changing environments in ways that change community structure and functioning, which could affect climate–carbon feedbacks. However, detecting microbial feedbacks to elevated CO₂ (eCO₂) or warming is hampered by concurrent changes in substrate availability and plant responses. Whether microbial communities can persistently feed back to climate change is still unknown. We overcame this problem by collecting microbial inocula at subfreezing conditions under eCO₂ and warming treatments in a semi‐arid grassland field experiment. The inoculant was incubated in a sterilised soil medium at constant conditions for 30 days. Microbes from eCO₂ exhibited an increased ability to decompose soil organic matter (SOM) compared with those from ambient CO₂ plots, and microbes from warmed plots exhibited increased thermal sensitivity for respiration. Microbes from the combined eCO₂ and warming plots had consistently enhanced microbial decomposition activity and thermal sensitivity. These persistent positive feedbacks of soil microbial communities to eCO₂ and warming may therefore stimulate soil C loss.     

Effects of elevated CO2, nitrogen deposition, and decreased species diversity on foliar fungal plant disease

Three components of global change, elevated CO₂, nitrogen addition, and decreased plant species richness (‘diversity’), increased the percent leaf area infected by fungi (pathogen load) for much to all of the plant community in one year of a factorial grassland experiment. Decreased plant diversity had the broadest effect, increasing pathogen load across the plant community. Decreased diversity increased pathogen load primarily by allowing remaining plant species to increase in abundance, facilitating spread of foliar fungal pathogens specific to each plant species. Changes in plant species composition also strongly influenced community pathogen load, with communities that lost less disease prone plant species increasing more in pathogen load. Elevated CO₂ increased pathogen load of C₃ grasses, perhaps by decreasing water stress, increasing leaf longevity, and increasing photosynthetic rate, all of which can promote foliar fungal disease. Decreased plant diversity further magnified the increase in C₃ grass pathogen load under elevated CO₂. Nitrogen addition increased pathogen load of C₄ grasses by increasing foliar nitrogen concentration, which can enhance pathogen infection, growth, and reproduction. Because changes in foliar fungal pathogen load can strongly influence grassland ecosystem processes, our study suggests that increased pathogen load can be an important mechanism by which global change affects grassland ecosystems.     

贝加尔针茅草原土壤真菌群落结构对氮素和水分添加的响应

研究氮沉降和降雨变化对土壤真菌群落结构的互作效应,对未来预测多个气候变化因子对草地生态系统的交互作用具有重要意义。以施氮和灌溉模拟氮沉降和降雨增加,采用裂区设计,应用高通量测序技术,研究8个氮添加水平(0、15、30、50、100、150、200、300 kg N hm~(-2)a~(-1))和2个水分添加水平(不灌溉、模拟夏季增雨100 mm灌溉)对土壤真菌群落结构的影响。结果表明,氮素和水分添加后,土壤真菌群落中占优势的门类分别为接合菌门Zygomycota(22.0%—48.9%)、担子菌门Basidiomycota(7.8%—18.5%)、子囊菌门Ascomycota(9.4%—20.1%)、球囊菌门Glomeromycota(0.7%—3.1%)、壶菌门Chytridiomycota(0.1%—1.3%)。常规降雨条件下,随着氮添加水平升高,接合菌门相对丰度呈现出先升高后降低的趋势,N50处理最高;子囊菌门相对丰度在高氮添加时(N100—N300)呈升高趋势。而在氮素和水分同时添加条件下,随着氮添加水平升高,接合菌门相对丰度呈降低趋势,子囊菌门相对丰度变化则不明显。在相同的氮添加水平下,水分添加使接合菌门相对丰度增加,而担子菌门、子囊菌门、球囊菌门和壶菌门的相对丰度降低。在不同氮素和水分添加条件下,有5个土壤真菌门类11个真菌纲相对丰度变化显著。接合菌门的Mortierella属,担子菌门的Entolomataceae科和Geastrum属相对丰度变化极显著,可作为土壤真菌群落结构变化的指示种。PCo A分析结果也表明氮素和水分添加改变了土壤真菌群落结构。植物-土壤-微生物系统的结构方程模型结果表明,植物群落组成及植物物种丰富度的变化是土壤真菌群落结构发生变化的主要影响因素,土壤无机氮及p H的变化主要通过影响植物群落间接影响真菌群落,其对真菌群落的直接影响则较小。综上,氮素和水分添加改变了土壤真菌群落结构,且两者存在明显的互作效应,水分添加可改变氮添加对土壤真菌群落的影响。     

Biomass responses in a temperate European grassland through 17 years of elevated CO2

Future increase in atmospheric CO₂ concentrations will potentially enhance grassland biomass production and shift the functional group composition with consequences for ecosystem functioning. In the “GiFACE” experiment (Giessen Free Air Carbon dioxide Enrichment), fertilized grassland plots were fumigated with elevated CO₂ (eCO₂) year‐round during daylight hours since 1998, at a level of +20% relative to ambient concentrations (in 1998, aCO₂ was 364 ppm and eCO₂ 399 ppm; in 2014, aCO₂ was 397 ppm and eCO₂ 518 ppm). Harvests were conducted twice annually through 23 years including 17 years with eCO₂ (1998 to 2014). Biomass consisted of C3 grasses and forbs, with a small proportion of legumes. The total aboveground biomass (TAB) was significantly increased under eCO₂ (p = .045 and .025, at first and second harvest). The dominant plant functional group grasses responded positively at the start, but for forbs, the effect of eCO₂ started out as a negative response. The increase in TAB in response to eCO₂ was approximately 15% during the period from 2006 to 2014, suggesting that there was no attenuation of eCO₂ effects over time, tentatively a consequence of the fertilization management. Biomass and soil moisture responses were closely linked. The soil moisture surplus (c. 3%) in eCO₂ manifested in the latter years was associated with a positive biomass response of both functional groups. The direction of the biomass response of the functional group forbs changed over the experimental duration, intensified by extreme weather conditions, pointing to the need of long‐term field studies for obtaining reliable responses of perennial ecosystems to eCO₂ and as a basis for model development.     

Controls over Soil Nitrogen Pools in a Semiarid Grassland Under Elevated CO2 and Warming

Long-term responses of terrestrial ecosystems to the combined effects of warming and elevated CO₂ (eCO₂) will likely be regulated by N availability. The stock of soil N determines availability for organisms, but also influences loss to the atmosphere or groundwater. eCO₂ and warming can elicit changes in soil N via direct effects on microbial and plant activity, or indirectly, via soil moisture. Detangling the interplay of direct- and moisture-mediated impacts on soil N and the role of organisms in controlling soil N will improve predictions of ecosystem-level responses. We followed individual soil N pools over two growing seasons in a semiarid temperate grassland, at the Prairie Heating and CO₂ Enrichment experiment. We evaluated relationships of N pools with environmental factors and explored the role of plants by assessing plant biomass, plant N, and plant inputs to soil. We also assessed N forms in plots with and without vegetation to remove plant-mediated effects. Our study demonstrated that the effects of warming and eCO₂ are highly dependent on individual N form and on year. In this water-constrained grassland, eCO₂, warming and their combination appear to impact soil N pools through a complex combination of direct- and moisture-mediated effects. eCO₂ decreased NO₃ ^? but had neutral to positive effects on NH₄ ⁺ and dissolved organic N (DON), particularly in a wet year. Warming increased NO₃ ^? availability due to a combination of indirect drying and direct temperature-driven effects. Warming also increased DON only in vegetated plots, suggesting plant mediation. Our results suggest that impacts of combined eCO₂ and warming are not always equivalent for plant and soil pools; although warming can help offset the decrease in NO₃ ^? availability for plants under eCO₂, the NO₃ ^? pool in soil is mainly driven by the negative effects of eCO₂.     

Fungal biomass production in response to elevated atmospheric CO2 in a Glomus mosseae–Prunus cerasifera model system

Biomass and length of intraradical and extraradical mycorrhizal mycelium under ambient (aCO₂) and elevated (eCO₂ ) atmospheric CO₂ was investigated using a non-destructive in vivo experimental model system. Time-course experiments allowed measurements of intact extraradical mycelium spreading from mycorrhizal roots of Prunus cerasifera micropropagated plants inoculated with the arbuscular mycorrhizal fungus Glomus mosseae, in controlled environmental chambers. The length of extraradical mycelium was significantly increased at the highest CO₂ concentration, ranging from 10.7 to 20.3 m at aCO₂ and eCO₂, respectively. The biochemical determination of mycelial glucosamine content allowed the evaluation of intraradical and extraradical fungal biomass, which were 2 and 3 times larger at eCO₂ than at aCO₂. Present data show that Glomus mosseae responds to increases of CO₂ concentrations producing larger mycorrhizal networks which may potentially represent carbon sink agents in soil ecosystems.     

Impacts of 3 years of elevated atmospheric CO2 on rhizosphere carbon flow and microbial community dynamics

Carbon (C) uptake by terrestrial ecosystems represents an important option for partially mitigating anthropogenic CO₂ emissions. Short‐term atmospheric elevated CO₂ exposure has been shown to create major shifts in C flow routes and diversity of the active soil‐borne microbial community. Long‐term increases in CO₂ have been hypothesized to have subtle effects due to the potential adaptation of soil microorganism to the increased flow of organic C. Here, we studied the effects of prolonged elevated atmospheric CO₂ exposure on microbial C flow and microbial communities in the rhizosphere. Carex arenaria (a nonmycorrhizal plant species) and Festuca rubra (a mycorrhizal plant species) were grown at defined atmospheric conditions differing in CO₂ concentration (350 and 700 ppm) for 3 years. During this period, C flow was assessed repeatedly (after 6 months, 1, 2, and 3 years) by ¹³C pulse‐chase experiments, and label was tracked through the rhizosphere bacterial, general fungal, and arbuscular mycorrhizal fungal (AMF) communities. Fatty acid biomarker analyses and RNA‐stable isotope probing (RNA‐SIP), in combination with real‐time PCR and PCR‐DGGE, were used to examine microbial community dynamics and abundance. Throughout the experiment the influence of elevated CO₂ was highly plant dependent, with the mycorrhizal plant exerting a greater influence on both bacterial and fungal communities. Biomarker data confirmed that rhizodeposited C was first processed by AMF and subsequently transferred to bacterial and fungal communities in the rhizosphere soil. Over the course of 3 years, elevated CO₂ caused a continuous increase in the ¹³C enrichment retained in AMF and an increasing delay in the transfer of C to the bacterial community. These results show that, not only do elevated atmospheric CO₂ conditions induce changes in rhizosphere C flow and dynamics but also continue to develop over multiple seasons, thereby affecting terrestrial ecosystems C utilization processes.     

Long-term elevated precipitation induces grassland soil carbon loss via microbe-plant–soil interplay

Global climate models predict that the frequency and intensity of precipitation events will increase in many regions across the world. However, the biosphere-climate feedback to elevated precipitation (eP) remains elusive. Here, we report a study on one of the longest field experiments assessing the effects of eP, alone or in combination with other climate change drivers such as elevated CO₂ (eCO₂), warming and nitrogen deposition. Soil total carbon (C) decreased after a decade of eP treatment, while plant root production decreased after 2 years. To explain this asynchrony, we found that the relative abundances of fungal genes associated with chitin and protein degradation increased and were positively correlated with bacteriophage genes, suggesting a potential viral shunt in C degradation. In addition, eP increased the relative abundances of microbial stress tolerance genes, which are essential for coping with environmental stressors. Microbial responses to eP were phylogenetically conserved. The effects of eP on soil total C, root production, and microbes were interactively affected by eCO₂. Collectively, we demonstrate that long-term eP induces soil C loss, owing to changes in microbial community composition, functional traits, root production, and soil moisture. Our study unveils an important, previously unknown biosphere-climate feedback in Mediterranean-type water-limited ecosystems, namely how eP induces soil C loss via microbe-plant–soil interplay.     

Elevated carbon dioxide accelerates the spatial turnover of soil microbial communities

Although elevated CO₂ (eCO₂) significantly affects the α‐diversity, composition, function, interaction and dynamics of soil microbial communities at the local scale, little is known about eCO₂ impacts on the geographic distribution of micro‐organisms regionally or globally. Here, we examined the β‐diversity of 110 soil microbial communities across six free air CO₂ enrichment (FACE) experimental sites using a high‐throughput functional gene array. The β‐diversity of soil microbial communities was significantly (P < 0.05) correlated with geographic distance under both CO₂ conditions, but declined significantly (P < 0.05) faster at eCO₂ with a slope of ?0.0250 than at ambient CO₂ (aCO₂) with a slope of ?0.0231 although it varied within each individual site, indicating that the spatial turnover rate of soil microbial communities was accelerated under eCO₂ at a larger geographic scale (e.g. regionally). Both distance and soil properties significantly (P < 0.05) contributed to the observed microbial β‐diversity. This study provides new hypotheses for further understanding their assembly mechanisms that may be especially important as global CO₂ continues to increase.     

Warming and Elevated CO<Subscript>2</Subscript> Interact to Alter Seasonality and Reduce Variability of Soil Water in a Semiarid Grassland

Global changes that alter soil water availability may have profound effects on semiarid ecosystems. Although both elevated CO₂ (eCO₂) and warming can alter water availability, often in opposite ways, few studies have measured their combined influence on the amount, timing, and temporal variability of soil water. Here, we ask how free air CO₂ enrichment (to 600 ppmv) and infrared warming (+?1.5 °C day, +?3 °C night) effects on soil water vary within years and across wet-dry periods in North American mixed-grass prairie. We found that eCO₂ and warming interacted to influence soil water and that those interactions varied by season. In the spring, negative effects of warming on soil water largely offset positive effects of eCO₂. As the growing season progressed, however, warming reduced soil water primarily (summer) or only (autumn) in plots treated with eCO₂. These interactions constrained the combined effect of eCO₂ and warming on soil water, which ranged from neutral in spring to positive in autumn. Within seasons, eCO₂ increased soil water under drier conditions, and warming decreased soil water under wetter conditions. By increasing soil water under dry conditions, eCO₂ also reduced temporal variability in soil water. These temporal patterns explain previously observed plant responses, including reduced leaf area with warming in summer, and delayed senescence with eCO₂ plus warming in autumn. They also suggest that eCO₂ and warming may favor plant species that grow in autumn, including winter annuals and C₃ graminoids, and species able to remain active under the dry conditions moderated by eCO₂.     

Effects of long term CO2 enrichment on microbial community structure in calcareous grassland

Elevated CO₂ generally increases plant productivity, and has been found to alter plant community composition in many ecosystems. Because soil microbes depend on plant-derived C and are often associated with specific plant species, elevated CO₂ has the potential to alter structure and functioning of soil microbial communities. We investigated soil microbial community structure of a species-rich semi-natural calcareous grassland that had been exposed to elevated CO₂ (600 μL L^?1) for 6 growing seasons. We analysed microbial community structure using phospholipid fatty acid (PLFA) profiles and DNA fingerprints obtained by Denaturing Gradient Gel Electrophoresis (DGGE) of 16S rDNA fragments amplified by the Polymerase Chain Reaction (PCR). PLFA profiles were not affected by CO₂ enrichment and the ratio of fungal and bacterial PLFA did not change. Ordination analysis of DNA fingerprints revealed a significant relation between CO₂ enrichment and variation in DNA fingerprints in summer (P=0.01), but not in spring. This variation was due to changes in low-intensity bands, while dominant bands did not differ between CO₂ treatments. Diversity of the bacterial community, as assessed by number of bands in DNA fingerprints and calculation of Shannon diversity indices, was not affected by elevated CO₂. Overall, only minor effects on microbial community structure were detected, corroborating earlier findings that soil carbon inputs did probably change much less than suggested by plant photosynthetic responses.     

Water availability affects seasonal CO2‐induced photosynthetic enhancement in herbaceous species in a periodically dry woodland

Elevated atmospheric CO₂ (eCO₂) is expected to reduce the impacts of drought and increase photosynthetic rates via two key mechanisms: first, through decreased stomatal conductance (g_s) and increased soil water content (V_SWC) and second, through increased leaf internal CO₂ (C_i) and decreased stomatal limitations (S_lim). It is unclear if such findings from temperate grassland studies similarly pertain to warmer ecosystems with periodic water deficits. We tested these mechanisms in three important C₃ herbaceous species in a periodically dry Eucalyptus woodland and investigated how eCO₂‐induced photosynthetic enhancement varied with seasonal water availability, over a 3 year period. Leaf photosynthesis increased by 10%–50% with a 150 μmol mol^?1 increase in atmospheric CO₂ across seasons. This eCO₂‐induced increase in photosynthesis was a function of seasonal water availability, given by recent precipitation and mean daily V_SWC. The highest photosynthetic enhancement by eCO₂ (>30%) was observed during the most water‐limited period, for example, with V_SWC <0.07 in this sandy surface soil. Under eCO₂ there was neither a significant decrease in g_s in the three herbaceous species, nor increases in V_SWC, indicating no “water‐savings effect” of eCO₂. Periods of low V_SWC showed lower g_s (less than ≈ 0.12 mol m^?2 s^?1), higher relative S_lim (>30%) and decreased C_i under the ambient CO₂ concentration (aCO₂), with leaf photosynthesis strongly carboxylation‐limited. The alleviation of S_lim by eCO₂ was facilitated by increasing C_i, thus yielding a larger photosynthetic enhancement during dry periods. We demonstrated that water availability, but not eCO₂, controls g_s and hence the magnitude of photosynthetic enhancement in the understory herbaceous plants. Thus, eCO₂ has the potential to alter vegetation functioning in a periodically dry woodland understory through changes in stomatal limitation to photosynthesis, not by the “water‐savings effect” usually invoked in grasslands.     

Nitrogen input mediates the effect of free-air CO2 enrichment on mycorrhizal fungal abundance

Plots containing Lolium perenne L., Trifolium repens L. or a mixture of both plant species were exposed to elevated atmospheric CO₂ (eCO₂) for 10 consecutive seasons using free‐air CO₂ enrichment technology at ETH Zürich, Switzerland. The CO₂ treatment was crossed with a two‐level nitrogen (N) fertilization treatment. In the tenth year, soil samples were collected on three occasions through the growing season to assess the impact of eCO₂ and N fertilization on mycorrhizal fungal abundance. Soil moisture content, which varied with harvest date, was linked to the vegetation type and was higher under eCO₂. Root weight density was affected by vegetation type: lower for clover, higher for grass. Root weight density was stimulated by eCO₂ and decreased by high N fertilization. The percent root length colonized by mycorrhizal fungi was lowest in the clover plots and highest in the grass plots. High N significantly decreased root length colonized. There was no overall effect of eCO₂ on root length colonized; however, there was a significant eCO₂× N interaction: eCO₂ increased root length colonized at high N, but decreased root length colonized at low N. Extraradical mycorrhizal hyphal density was linked to soil moisture content. Extraradical mycorrhizal hyphal density was not affected by eCO₂ or high N individually, but as for root length colonized, there was a significant eCO₂× N interaction: eCO₂ increased extraradical mycorrhizal hyphal density at low N but not at high N. These environmental effects on root colonization and external mycorrhizal hyphae were independent of soil moisture content and root weight density. This field study demonstrated a significant mediating effect of N fertilization on the response of arbuscular mycorrhizal fungi to eCO₂ irrespective of any change in root biomass.     

Distinct responses of soil microbial communities to elevated CO2 and O3 in a soybean agro-ecosystem

The concentrations of atmospheric carbon dioxide (CO₂) and tropospheric ozone (O₃) have been rising due to human activities. However, little is known about how such increases influence soil microbial communities. We hypothesized that elevated CO₂ (eCO₂) and elevated O₃ (eO₃) would significantly affect the functional composition, structure and metabolic potential of soil microbial communities, and that various functional groups would respond to such atmospheric changes differentially. To test these hypotheses, we analyzed 96 soil samples from a soybean free-air CO₂ enrichment (SoyFACE) experimental site using a comprehensive functional gene microarray (GeoChip 3.0). The results showed the overall functional composition and structure of soil microbial communities shifted under eCO₂, eO₃ or eCO₂+eO₃. Key functional genes involved in carbon fixation and degradation, nitrogen fixation, denitrification and methane metabolism were stimulated under eCO₂, whereas those involved in N fixation, denitrification and N mineralization were suppressed under eO₃, resulting in the fact that the abundance of some eO₃-supressed genes was promoted to ambient, or eCO₂-induced levels by the interaction of eCO₂+eO₃. Such effects appeared distinct for each treatment and significantly correlated with soil properties and soybean yield. Overall, our analysis suggests possible mechanisms of microbial responses to global atmospheric change factors through the stimulation of C and N cycling by eCO₂, the inhibition of N functional processes by eO₃ and the interaction by eCO₂ and eO₃. This study provides new insights into our understanding of microbial functional processes in response to global atmospheric change in soybean agro-ecosystems.     

Plant species richness, elevated CO2, and atmospheric nitrogen deposition alter soil microbial community composition and function

We determined soil microbial community composition and function in a field experiment in which plant communities of increasing species richness were exposed to factorial elevated CO₂ and nitrogen (N) deposition treatments. Because elevated CO₂ and N deposition increased plant productivity to a greater extent in more diverse plant assemblages, it is plausible that heterotrophic microbial communities would experience greater substrate availability, potentially increasing microbial activity, and accelerating soil carbon (C) and N cycling. We, therefore, hypothesized that the response of microbial communities to elevated CO₂ and N deposition is contingent on the species richness of plant communities. Microbial community composition was determined by phospholipid fatty acid analysis, and function was measured using the activity of key extracellular enzymes involved in litter decomposition. Higher plant species richness, as a main effect, fostered greater microbial biomass, cellulolytic and chitinolytic capacity, as well as the abundance of saprophytic and arbuscular mycorrhizal (AM) fungi. Moreover, the effect of plant species richness on microbial communities was significantly modified by elevated CO₂ and N deposition. For instance, microbial biomass and fungal abundance increased with greater species richness, but only under combinations of elevated CO₂ and ambient N, or ambient CO₂ and N deposition. Cellobiohydrolase activity increased with higher plant species richness, and this trend was amplified by elevated CO₂. In most cases, the effect of plant species richness remained significant even after accounting for the influence of plant biomass. Taken together, our results demonstrate that plant species richness can directly regulate microbial activity and community composition, and that plant species richness is a significant determinant of microbial response to elevated CO₂ and N deposition. The strong positive effect of plant species richness on cellulolytic capacity and microbial biomass indicate that the rates of soil C cycling may decline with decreasing plant species richness.     

Microbial assimilation of new photosynthate is altered by plant species richness and nitrogen deposition

To determine how plant species richness impacts microbial assimilation of new photosynthate, and how this may be modified by atmospheric N deposition, we analyzed the microbial assimilation of recent photosynthate in a 6-year-long field experiment in which plant species richness, atmospheric N deposition, and atmospheric CO₂ concentration were manipulated in concert. The depleted δ¹³C of fumigation CO₂ enabled us to investigate the effect of plant species richness and atmospheric N deposition on the metabolism of soil microbial communities in the elevated CO₂ treatment. To accomplish this, we determined the δ¹³C of bacterial, actinobacterial, and fungal phospholipid fatty acids (PLFAs). In the elevated CO₂ conditions of this study, the δ¹³C of bacterial PLFAs (i15:0, i16:0, 16:1ω7c, 16:1ω9c, 10Me16:0, and 10Me18:0) and the fungal PLFA 18:1ω9c was significantly lower in species-rich plant communities than in species-poor plant communities, indicating that microbial incorporation of new C increased with plant species richness. Despite an increase in plant production, total PLFA decreased under N deposition. Moreover, N deposition also decreased fungal relative abundance in species-rich plant communities. In our study, plant species richness directly increased microbial incorporation of new photosynthate, providing a mechanistic link between greater plant detritus production in species-rich plant communities and larger and more active soil microbial community.     

Fungal root colonization responses in natural grasslands after long-term exposure to elevated atmospheric CO2

Arbuscular mycorrhizae, ubiquitous mutualistic symbioses between plant roots and fungi in the order Glomales, are believed to be important controllers of plant responses to global change, in particular to elevated atmospheric CO₂. In order to test if any effects on the symbiosis can persist after long-term treatment, we examined root colonization by arbuscular mycorrhizal (AM) and other fungi of several plant species from two grassland communities after continuous exposure to elevated atmospheric CO₂ for six growing seasons in the field. For plant species from both a sandstone and a serpentine annual grassland there was evidence for changes in fungal root colonization, with changes occurring as a function of plant host species. We documented decreases in percentage nonmycorrhizal fungal root colonization in elevated CO₂ for several plant species. Total AM root colonization (%) only increased significantly for one out of the five plant species in each grassland. However, when dividing AM fungal hyphae into two groups of hyphae (fine endophyte and coarse endophyte), we could document significant responses of AM fungi that were hidden when only total percentage colonization was measured. We also documented changes in elevated CO₂ in the percentage of root colonized by both AM hyphal types simultaneously. Our results demonstrate that changes in fungal root colonization can occur after long-term CO₂ enrichment, and that the level of resolution of the study of AM fungal responses may have to be increased to uncover significant changes to the CO₂ treatment. This study is also one of the first to document compositional changes in the AM fungi colonizing roots of plants grown in elevated CO₂. Although it is difficult to relate the structural data directly to functional changes, possible implications of the observed changes for plant communities are discussed.     

Influence of Elevated CO2 on the Fungal Community in a Coastal Scrub Oak Forest Soil Investigated with Terminal-Restriction Fragment Length Polymorphism Analysis

Sixteen open-top chambers (diameter, 3.66 m) were established in a scrub oak habitat in central Florida where vegetation was removed in a planned burn prior to chamber installation. Eight control chambers have been continuously exposed to ambient air and eight have been continuously exposed to elevated CO₂ at twice-ambient concentration (~700 ppm) for 5 years. Soil cores were collected from each chamber to examine the influence of elevated atmospheric CO₂ on the fungal community in different soil fractions. Each soil sample was physically fractionated into bulk soil, rhizosphere soil, and roots for separate analyses. Changes in relative fungal biomass were estimated by the ergosterol technique. In the bulk soil and root fractions, a significantly increased level of ergosterol was detected in the elevated CO₂ treatments relative to ambient controls. Fungal community composition was determined by terminal-restriction fragment length polymorphism (T-RFLP) analysis of the internal transcribed spacer (ITS) region. The specificities of different ITS primer sets were evaluated against plant and fungal species isolated from the experimental site. Changes in community composition were assessed by principal component analyses of T-RFLP profiles resolved by capillary electrophoresis. Fungal species richness, defined by the total number of terminal restriction fragments, was not significantly affected by either CO₂ treatment or soil fraction.