Microbial communities respond to experimental warming, but site matters

Because microorganisms are sensitive to temperature, ongoing global warming is predicted to influence microbial community structure and function. We used large-scale warming experiments established at two sites near the northern and southern boundaries of US eastern deciduous forests to explore how microbial communities and their function respond to warming at sites with differing climatic regimes. Soil microbial community structure and function responded to warming at the southern but not the northern site. However, changes in microbial community structure and function at the southern site did not result in changes in cellulose decomposition rates. While most global change models rest on the assumption that taxa will respond similarly to warming across sites and their ranges, these results suggest that the responses of microorganisms to warming may be mediated by differences across the geographic boundaries of ecosystems

Fungal community composition and function after long‐term exposure of northern forests to elevated atmospheric CO 2 and tropospheric O 3

The long‐term effects of rising atmospheric carbon dioxide (CO 2 ) and tropospheric O 3 concentrations on fungal communities in soil are not well understood. Here, we examine fungal community composition and the activities of cellobiohydrolase and N ‐acetylglucosaminidase (NAG) after 10 years of exposure to 1.5 times ambient levels of CO 2 and O 3 in aspen and aspen–birch forest ecosystems, and compare these results to earlier studies in the same long‐term experiment. The forest floor community was dominated by saprotrophic fungi, and differed slightly between plant community types, as did NAG activity. Elevated CO 2 and O 3 had small but significant effects on the distribution of fungal genotypes in this horizon, and elevated CO 2 also lead to an increase in the proportion of Sistotrema spp. within the community. Yet, although cellobiohydrolase activity was lower in the forest floor under elevated O 3 , it was not affected by elevated CO 2 . NAG was also unaffected. The soil community was dominated by ectomycorrhizal species. Both CO 2 and O 3 had a minor effect on the distribution of genotypes; however, phylogenetic analysis indicated that under elevated O 3 Cortinarius and Inocybe spp. increased in abundance and Laccaria and Tomentella spp. declined. Although cellobiohydrolase activity in soil was unaffected by either CO 2 or O 3 , NAG was higher (∼29%) under CO 2 in aspen–birch, but lower (∼18%) under aspen. Time series analysis indicated that CO 2 increased cellulolytic enzyme activity during the first 5 years of the experiment, but that the magnitude of this effect diminished over time. NAG activity also showed strong early stimulation by elevated CO 2 , but after 10 years this effect is no longer evident. Elevated O 3 appears to have variable stimulatory and repressive effects depending on the soil horizon and time point examined.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/87135/1/j.1365-2486.2010.02376.x.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/87135/2/GCB_2376_sm_suppinfo_figure-s1-s5.pd

Belowground carbon allocation by trees drives seasonal patterns of extracellular enzyme activities by altering microbial community composition in a beech forest soil

Plant seasonal cycles alter carbon (C) and nitrogen (N) availability for soil microbes, which may affect microbial community composition and thus feed back on microbial decomposition of soil organic material and plant N availability. The temporal dynamics of these plant–soil interactions are, however, unclear.Here, we experimentally manipulated the C and N availability in a beech forest through N fertilization or tree girdling and conducted a detailed analysis of the seasonal pattern of microbial community composition and decomposition processes over 2 yr.We found a strong relationship between microbial community composition and enzyme activities over the seasonal course. Phenoloxidase and peroxidase activities were highest during late summer, whereas cellulase and protease peaked in late autumn. Girdling, and thus loss of mycorrhiza, resulted in an increase in soil organic matter-degrading enzymes and a decrease in cellulase and protease activity.Temporal changes in enzyme activities suggest a switch of the main substrate for decomposition between summer (soil organic matter) and autumn (plant litter). Our results indicate that ectomycorrhizal fungi are possibly involved in autumn cellulase and protease activity. Our study shows that, through belowground C allocation, trees significantly alter soil microbial communities, which may affect seasonal patterns of decomposition processes

Simulated chronic NO 3 − deposition reduces soil respiration in northern hardwood forests

Chronic N additions to forest ecosystems can enhance soil N availability, potentially leading to reduced C allocation to root systems. This in turn could decrease soil CO 2 efflux. We measured soil respiration during the first, fifth, sixth and eighth years of simulated atmospheric NO 3 − deposition (3 g N m −2  yr −1 ) to four sugar maple-dominated northern hardwood forests in Michigan to assess these possibilities. During the first year, soil respiration rates were slightly, but not significantly, higher in the NO 3 − -amended plots. In all subsequent measurement years, soil respiration rates from NO 3 − -amended soils were significantly depressed. Soil temperature and soil matric potential were measured concurrently with soil respiration and used to develop regression relationships for predicting soil respiration rates. Estimates of growing season and annual soil CO 2 efflux made using these relationships indicate that these C fluxes were depressed by 15% in the eighth year of chronic NO 3 − additions. The decrease in soil respiration was not due to reduced C allocation to roots, as root respiration rates, root biomass, and root turnover were not significantly affected by N additions. Aboveground litter also was unchanged by the 8 years of treatment. Of the remaining potential causes for the decline in soil CO 2 efflux, reduced microbial respiration appears to be the most likely possibility. Documented reductions in microbial biomass and the activities of extracellular enzymes used for litter degradation on the NO 3 − -amended plots are consistent with this explanation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72619/1/j.1365-2486.2004.00737.x.pd

Simulated Atmospheric N Deposition Alters Fungal Community Composition and Suppresses Ligninolytic Gene Expression in a Northern Hardwood Forest

High levels of atmospheric nitrogen (N) deposition may result in greater terrestrial carbon (C) storage. In a northern hardwood ecosystem, exposure to over a decade of simulated N deposition increased C storage in soil by slowing litter decay rates, rather than increasing detrital inputs. To understand the mechanisms underlying this response, we focused on the saprotrophic fungal community residing in the forest floor and employed molecular genetic approaches to determine if the slower decomposition rates resulted from down-regulation of the transcription of key lignocellulolytic genes, by a change in fungal community composition, or by a combination of the two mechanisms. Our results indicate that across four Acer-dominated forest stands spanning a 500-km transect, community-scale expression of the cellulolytic gene cbhI under elevated N deposition did not differ significantly from that under ambient levels of N deposition. In contrast, expression of the ligninolytic gene lcc was significantly down-regulated by a factor of 2–4 fold relative to its expression under ambient N deposition. Fungal community composition was examined at the most southerly of the four sites, in which consistently lower levels of cbhI and lcc gene expression were observed over a two-year period. We recovered 19 basidiomycete and 28 ascomycete rDNA 28S operational taxonomic units; Athelia, Sistotrema, Ceratobasidium and Ceratosebacina taxa dominated the basidiomycete assemblage, and Leotiomycetes dominated the ascomycetes. Simulated N deposition increased the proportion of basidiomycete sequences recovered from forest floor, whereas the proportion of ascomycetes in the community was significantly lower under elevated N deposition. Our results suggest that chronic atmospheric N deposition may lower decomposition rates through a combination of reduced expression of ligninolytic genes such as lcc, and compositional changes in the fungal community

Measuring phenol oxidase and peroxidase activities with pyrogallol, l-DOPA, and ABTS: Effect of assay conditions and soil type

Microbial phenol oxidases and peroxidases mediate biogeochemical processes in soils, including microbial acquisition of carbon and nitrogen, lignin degradation, carbon mineralization and sequestration, and dissolved organic carbon export. Measuring oxidative enzyme activities in soils is more problematic than assaying hydrolytic enzyme activities because of the non-specific, free radical nature of the reactions and complex interactions between enzymes, assay substrates, and the soil matrix. We compared three substrates commonly used to assay phenol oxidase and peroxidase in soil: pyrogallol (PYGL, 1,2,3-trihydroxybenzene), l-DOPA (l-3,4-dihydroxyphenylalanine), and ABTS (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). We measured substrate oxidation in three soils across a pH gradient from 3.0 to 10.0 to determine the pH optimum for each substrate. In addition, we compared activities across 17 soils using the three substrates. In general, activities on the substrates followed the trend PYGL>. l-DOPA>ABTS and were inversely related to substrate redox potential. PYGL and ABTS were not suitable substrates at pH>5, and ABTS oxidation often declined with addition of peroxide to the assay. Absolute and relative oxidation rates varied widely among substrates in relation to soil type and assay pH. We also tested whether autoclaved or combusted soils could be used as negative controls for the influence of abiotic factors (e.g., soil mineralogy) on oxidative activity. However, neither autoclaving nor combustion produced reliable negative controls because substrate oxidation still occurred; in some cases, these treatments enhanced substrate oxidation rates. For broad scale studies, we recommend that investigators use all three substrates to assess soil oxidation potentials. For focused studies, we recommend evaluating substrates before choosing a single option, and we recommend assays at both the soil pH and a reference pH (e.g., pH 5.0) to determine the effect of assay pH on oxidase activity. These recommendations should contribute to greater comparability of oxidase potential activities across studies. © 2013 Elsevier Ltd

Models Analyses for Allelopathic Effects of Chicory at Equivalent Coupling of Nitrogen Supply and pH Level on F. arundinacea, T. repens and M. sativa

Alllelopathic potential of chicory was investigated by evaluating its effect on seed germination, soluble sugar, malondialdehyde (MDA) and the chlorophyll content of three target plants species (Festuca arundinacea, Trifolium repens and Medicago sativa). The secretion of allelochemicals was regulated by keeping the donor plant (chicory) separate from the three target plant species and using different pH and nitrogen levels. Leachates from donor pots with different pH levels and nitrogen concentrations continuously irrigated the target pots containing the seedlings. The allelopathic effects of the chicory at equivalent coupling of nitrogen supply and pH level on the three target plants species were explored via models analyses. The results suggested a positive effect of nitrogen supply and pH level on allelochemical secretion from chicory plants. The nitrogen supply and pH level were located at a rectangular area defined by 149 to 168 mg/l nitrogen supply combining 4.95 to 7.0 pH value and point located at nitrogen supply 177 mg/l, pH 6.33 when they were in equivalent coupling effects; whereas the inhibitory effects of equivalent coupling nitrogen supply and pH level were located at rectangular area defined by 125 to 131 mg/l nitrogen supply combining 6.71 to 6.88 pH value and two points respectively located at nitrogen supply 180 mg/l with pH 6.38 and nitrogen supply 166 mg/l with pH 7.59. Aqueous extracts of chicory fleshy roots and leaves accompanied by treatment at different sand pH values and nitrogen concentrations influenced germination, seedling growth, soluble sugar, MDA and chlorophyll of F. arundinacea, T. repens and M. sativa. Additionally, we determined the phenolics contents of root and leaf aqueous extracts, which were 0.104% and 0.044% on average, respectively

Microbial enzymatic responses to drought and to nitrogen addition in a southern California grassland

Microbial enzymes play a fundamental role in ecosystem processes and nutrient mineralization. Therefore understanding enzyme responses to anthropogenic environmental change is important for predicting ecosystem function in the future. In a previous study, we used a reciprocal transplant design to examine the direct and indirect effects of drought and nitrogen (N) fertilization on litter decomposition in a southern California grassland. This work showed direct and indirect negative effects of drought on decomposition, and faster decomposition by N-adapted microbial communities in N-fertilized plots than in non-fertilized plots. Here we measured microbial biomass and the activities of nine extracellular enzymes to examine the microbial and enzymatic mechanisms underlying litter decomposition responses to drought and N. We hypothesized that changes in fungal biomass and potential extracellular enzyme activity (EEA) would relate directly to litter decomposition responses. We also predicted that fungal biomass would dominate the microbial community in our semi-arid study site. However, we found that the microbial community was dominated by bacterial biomass, and that bacteria responded negatively to drought treatment. In contrast to patterns in decomposition, fungal biomass and most potential EEA increased in direct response to drought treatment. Potential EEA was also decoupled from the decomposition response to N treatment. These results suggest that drought and N alter the efficiencies of EEA, defined as the mass of target substrate lost per unit potential EEA. Enzyme efficiencies declined with drought treatment, possibly because reduced water availability increased enzyme immobilization and reduced diffusion rates. In the N experiment, the efficiencies of β-glucosidase, β-xylosidase, and polyphenol oxidase were greater when microbes were transplanted into environments from which they originated. This increase in enzymatic efficiency suggests that microbial enzymes may adapt to their local environment. Overall, our results indicate that drought and N addition may have predictable impacts on the efficiencies of extracellular enzymes, providing a means of linking enzyme potentials with in-situ activities