Long-term impacts of zinc and copper enriched sewage sludge additions on bacterial, archaeal and fungal communities in arable and grassland soils

Long-term impacts of metal contamination derived from sewage sludge on soil microbial communities have been widely evaluated, but confounding effects have made it difficult to draw firm conclusions and thus to advise on safe metal limits. Here we used Multiplex-terminal restriction length fragment polymorphism (M-TRFLP) to assess the long-term impact of sludge-borne Zn and Cu contamination on the structure of bacterial, fungal and archaeal communities across seven different soils at metal levels relevant to current guideline limits. Despite strong effects of site on microbial community structure, analysis of similarity (ANOSIM) demonstrated a small but significant effect of Zn on bacteria (P < 0.001), archaea (P < 0.001), and fungi (P < 0.001). Significant effects of Cu on bacteria (P < 0.001), archaea (P < 0.001) and fungi (P < 0.001) were also observed. Several bacterial and fungal T-RFs were identified as responding to Zn or Cu. For example the bacterial T-RF 72 was negatively correlated with Zn and Cu, and T-RF 259 was positively correlated with Zn. Attempts to identify these bacterial markers of Zn and Cu contamination suggest a negative impact of Cu on Acidobacteria in arable soils. These results demonstrate for the first time, that despite a strong influence of site on microbial community structure, effects of Zn and Cu derived from sewage sludge can be detected as shifts in bacterial, fungal and archaeal communities indicating a common response more than 11 years after sludge addition.     

The influence of soil properties on the structure of bacterial and fungal communities across land-use types

Land-use change can have significant impacts on soil conditions and microbial communities are likely to respond to these changes. However, such responses are poorly characterized as few studies have examined how specific changes in edaphic characteristics do, or do not, influence the composition of soil bacterial and fungal communities across land-use types. Soil samples were collected from four replicated (n = 3) land-use types (hardwood and pine forests, cultivated and livestock pasture lands) in the southeastern US to assess the effects of land-use change on microbial community structure and distribution. We used quantitative PCR to estimate bacterial–fungal ratios and clone libraries targeting small-subunit rRNA genes to independently characterize the bacterial and fungal communities. Although some soil properties (soil texture and nutrient status) did significantly differ across land-use types, other edaphic factors (e.g., pH) did not vary consistently with land-use. Bacterial–fungal ratios were not significantly different across the land-uses and distinct land-use types did not necessarily harbor distinct soil fungal or bacterial communities. Rather, the composition of bacterial and fungal communities was most strongly correlated with specific soil properties. Soil pH was the best predictor of bacterial community composition across this landscape while fungal community composition was most closely associated with changes in soil nutrient status. Together these results suggest that specific changes in edaphic properties, not necessarily land-use type itself, may best predict shifts in microbial community composition across a given landscape. In addition, our results demonstrate the utility of using sequence-based approaches to concurrently analyze bacterial and fungal communities as such analyses provide detailed phylogenetic information on individual communities and permit the robust assessment of the biogeographical patterns exhibited by soil microbial communities.     

Improving soil structure by promoting fungal abundance with organic soil amendments

Building soil structure in agroecosystems is important because it governs soil functions such as air and water movement, soil C stabilization, nutrient availability, and root system development. This study examined, under laboratory conditions, effects of organic amendments comprised of differing proportions of labile and semi-labile C on microbial community structure and macroaggregate formation in three variously textured soils where native structure was destroyed. Three amendment treatments were imposed (in order of increasing C lability): vegetable compost, dairy manure, hairy vetch (Vicia villosa Roth). Formation of water stable macroaggregates and changes in microbial community structure were evaluated over 82 days. Regardless of soil type, formation of large macroaggregates (LMA, >2000 μm diameter) was highest in soils amended with vetch, followed by manure, non-amended control, and compost. Vetch and manure had greater microbially available C and caused an increase in fungal biomarkers in all soils. Regression analysis indicated that LMA formation was most strongly related to the relative abundance of the fungal fatty acid methyl ester (FAME) 18:2ω6c (r = 0.55, p < 0.001), fungal ergosterol (r = 0.58, p < 0.001), and microbial biomass (r = 0.57, p < 0.001). Non-metric multidimensional scaling (NMS) ordination of FAME profiles revealed that vetch and manure drove shifts toward fungal-dominated soil microbial communities and greater LMA formation in these soils. This study demonstrated that, due to their greater amounts of microbially available C, vetch or manure inputs can be used to promote fungal proliferation in order to maintain or improve soil structure.     

Parent material and vegetation influence bacterial community structure and nitrogen functional genes along deep tropical soil profiles at the Luquillo Critical Zone Observatory

Microbial communities mediate every step of the soil nitrogen cycle, yet the structure and associated nitrogen cycle functions of soil microbial communities remain poorly studied in tropical forests. Moreover, tropical forest soils are often many meters deep, but most studies of microbial nitrogen cycling have focused exclusively on surface soils. The objective of our study was to evaluate changes in bacterial community structure and nitrogen functional genes with depth in soils developed on two contrasting geological parent materials and two forest types that occur at different elevations at the Luquillo Critical Zone Observatory in northeast Puerto Rico. We excavated three soil pits to 140 cm at four different sites representing the four soil × forest combinations (n = 12), and collected samples at ten-centimeter increments from the surface to 140 cm. We used bacterial 16S rRNA gene-DGGE (denaturant gradient gel electrophoresis) to fingerprint microbial community structures, and quantitative PCR to measure the abundance of five functional genes involved in various soil nitrogen transformations: nifH (nitrogen fixation), chiA (organic nitrogen decomposition), amoA (ammonia oxidation), nirS (nitrite reduction) and nosZ (nitrous oxide reduction). Multivariate analyses of DGGE fingerprinting patterns revealed differences in bacterial community structure across the four soil × forest types that were strongly correlated with soil pH (r = 0.69, P < 0.01) and nutrient stoichiometry (r² ≥ 0.36, P < 0.05). Across all soil and forest types, nitrogen functional genes declined significantly with soil depth (P < 0.001). Denitrification genes (nirS and nosZ) accounted for the largest proportion of measured nitrogen functional genes. Measured nitrogen functional genes were positively correlated with soil carbon, nitrogen and phosphorus concentrations (P < 0.001) and all genes except amoA were significantly more abundant in the Inceptisol soil type compared with the Oxisol soil type (P < 0.03). Greater abundances and a stronger vertical zonation of nitrogen functional genes in Inceptisols suggest more dynamic nitrogen transformation processes in this soil type. As the first study to examine bacterial nitrogen functional gene abundances below the surface 20 cm in tropical forest soils, our work provides insight into how pedogenically-driven vertical gradients control the nitrogen-cycling capacity of soil microbial communities. While previous studies have shown evidence for redox-driven hotspots in tropical nitrogen cycling on a watershed scale, our study corroborates this finding on a molecular scale.     

Does biochar interfere with standard methods for determining soil microbial biomass and phenotypic community structure?

Due to its high sorption affinity for organic compounds, biochar may interfere with extraction procedures involving such compounds used for microbially-related assays commonly applied to soils. Here we assessed the impact of two biochars (derived from pine bark and produced at 300 and 600 °C) at three concentrations (0, 12.5, and 50 g kg⁻¹) in three distinct arable soils with contrasting textural classes (loamy sand, sandy loam, and clay) on the determination of soil microbial biomass C by fumigation–extraction, fungal biomass by ergosterol analysis, and microbial community structure as defined by phospholipid fatty acid (PLFA) profiling. Biochar did not affect the apparent concentration of soil microbial biomass C and had no significant impact on apparent PLFA profiles. By contrast, the apparent extraction efficiency of ergosterol was affected dependent on soil type, biochar production temperature, and biochar concentration. Nonetheless, ergosterol contents of biochar-amended soils can be accurately estimated by correcting for reduced recovery using an ergosterol spike.     

Habitat selective factors influencing the structural composition and functional capacity of microbial communities in agricultural soils

Key physicochemical factors associated with microbial community composition and functions in Australian agricultural soils were identified. Soils from seven field sites, with varying long-term agricultural management regimes, were characterised physicochemically, on the basis of their bacterial and fungal community structures (using PCR-DGGE), and by assessing potential catabolic functions (MicroResp?). Soil type, rather than agricultural management practice, was the key determinant of microbial community structure and catabolic function (P<0.05). Following multivariate analysis, soil pH was identified as the key habitat-selective physicochemical soil property associated with variation in biological diversity and profiles of organic substrate utilisation. In particular, the capacity of soils to catabolise different C-substrates was closely correlated (ρ=0.604, P=0.001) to pH. With decreasing pH, the catabolism of common low molecular weight organic compounds (especially cysteine and aspartic acid) declined, however catabolism of two others (lysine and arginine) increased. Shifts in the capacity of soil microbiota to cycle common organic compounds have implications for overall geochemical cycling of C and N in acidifying soils. The genetic structure of the bacterial communities in soil strongly correlated with pH (ρ=0.722; P=0.001) and that of soil fungi with pH and % sand (ρ=0.323; P=0.006). Catabolic function was more closely associated with the structure of the bacterial than fungal communities. This work has shown that soil pH is a primary driver of microbial diversity and function in soil. Agricultural management practices thereby act to selectively shift populations and functions against this background.     

Comparison of soil fungal/bacterial ratios in a pH gradient using physiological and PLFA-based techniques

We have compared the total microbial biomass and the fungal/bacterial ratio estimated using substrate-induced respiration (SIR) in combination with the selective inhibition technique and using the phospholipid fatty acid (PLFA) technique in a pH gradient (3.0-7.2) consisting of 53 mature broad-leaved forest soils. A fungal/bacterial biomass index using the PLFA technique was calculated using the PLFA 18:2ω6,9 as an indicator of fungal biomass and the sum of 13 bacterial specific PLFAs as indicator of the bacterial biomass. Good linear correlation (p<0.001) was found between the total microbial biomass estimated with SIR and total PLFAs (totPLFA), indicating that 1 mg biomass-C was equivalent to 130 nmol totPLFA. Both biomass estimates were positively correlated to soil pH. The fungal/bacterial ratio measured using the selective inhibition technique decreased significantly with increasing pH from about 9 at pH 3 to approximately 2 at pH 7, while the fungal/bacterial biomass index using PLFA measurements tended to increase slightly with increasing soil pH. Good correlation between the soil content of ergosterol and of the PLFA 18:2ω6,9 indicated that the lack of congruency between the two methods in estimating fungal/bacterial ratios was not due to PLFA 18:2ω6,9-related non-fungal structures to any significant degree. Several PLFAs were strongly correlated to soil pH (R² values >0.8); for example the PLFAs 16:1ω5 and 16:1ω7c increased with increasing soil pH, while i16:0 and cy19:0 decreased. A principal component analysis of the total PLFA pattern gave a first component that was strongly correlated to soil pH (R²=0.85, p<0.001) indicating that the microbial community composition in these beech/beech-oak forest soils was to a large extent determined by soil pH.     

Vineyard soil bacterial diversity and composition revealed by 16S rRNA genes: Differentiation by geographic features

Here, we examine soil-borne microbial biogeography as a function of the features that define an American Viticultural Area (AVA), a geographically delimited American wine grape-growing region, defined for its distinguishing features of climate, geology, soils, physical features (topography and water), and elevation. In doing so, we lay a foundation upon which to link the terroir of wine back to the soil-borne microbial communities. The objective of this study is to elucidate the hierarchy of drivers of soil bacterial community structure in wine grape vineyards in Napa Valley, California. We measured differences in the soil bacterial and archaeal community composition and diversity by sequencing the fourth variable region of the small subunit ribosomal RNA gene (16S V4 rDNA). Soil bacterial communities were structured with respect to soil properties and AVA, demonstrating the complexity of soil microbial biogeography at the landscape scale and within the single land-use type. Location and edaphic variables that distinguish AVAs were the strongest explanatory factors for soil microbial community structure. Notably, the relationship with TC and TN of the <53 μm and 53–250 μm soil fractions offers support for the role of bacterial community structure rather than individual taxa on fine soil organic matter content. We reason that AVA, climate, and topography each affect soil microbial communities through their suite of impacts on soil properties. The identification of distinctive soil microbial communities associated with a given AVA lends support to the idea that soil microbial communities form a key in linking wine terroir back to the biotic components of the soil environment, suggesting that the relationship between soil microbial communities and wine terroir should be examined further.     

Fungal, bacterial and plant dsDNA contributions to soil total DNA extracted from silty soils under different farming practices: Relationships with chloroform-labile carbon

Twelve differently-managed silty soils from North-Western France were chosen to compare two common methods of quantifying soil microbial biomass: Chloroform fumigation and extraction-labile carbon (CL_C) and microbial double stranded DNA (dsDNA). We also determined the contributions of each of the fungal, bacterial, and plant kingdoms to the total community dsDNA using real-time Polymerase Chain Reaction with kingdom-specific ribosomal primer sets. Regardless of the method, the highest microbial biomasses were associated with long-term untilled plots. Site (locations) specificities could also be detected, especially in conventionally cultivated lands. Regardless of site, a strong linear relationship could be drawn between CL_C and dsDNA in tilled lands (r = 0.91, n = 15, P = 0.01) and in grasslands (r = 0.78, n = 21, P = 0.01). Moreover, we propose a logarithmic model describing all of our silty soils, irrespective of management. In order to explain the non-linearity (log) of this relationship, we tested the hypothesis of a weak plant dsDNA contribution in total dsDNA in comparison with the well-documented root cell contribution to CL_C quantifications. Plant dsDNA never exceeded 2.6% of total dsDNA content for all of the soils studied. Among groups examined, the bacterial dsDNA contribution to the community dsDNA pool was the most site- and/or pedoclimatic-dependent. Fungi constituted a major component of total microbial biomass in grassland or in land with permanent plant cover where their proportion reached almost 50% of total dsDNA. More precisely, fungal dsDNA concentration was highly related to tillage. Our study demonstrated the expediency of the total microbial dsDNA quantification in agricultural silty soils rather than the time-consuming quantification of CL_C. Quantifying the relative contribution of bacterial or fungal biomass in total dsDNA by real-time PCR allows to access to a new level of knowledge of the soil microbial biomass and to reveal the balances between those two kingdoms according to soils or farming practices.     

Composition of the microbial communities in the mineral soil under different types of natural forest

Phospholipid fatty acid (PLFA) patterns were used to describe the composition of the soil microbial communities under 12 natural forest stands including oak and beech, spruce-fir-beech, floodplain and pine forests. In addition to the quantification of total PLFAs, soil microbial biomass was measured by substrate-induced respiration and chloroform fumigation-extraction. The forest stands possess natural vegetation, representing an expression of the natural site factors, and we hypothesised that each forest type would support a specific soil microbial community. Principal component analysis (PCA) of PLFA patterns revealed that the microbial communities were compositionally distinct in the floodplain and pine forests, comprising azonal forest types, and were more similar in the oak, beech and spruce-fir-beech forests, which represent the zonal vegetation types of the region. In the nutrient-rich floodplain forests, the fatty acids 16:1ω5, 17:0cy, a15:0 and a17:0 were the most prevalent and soil pH seemed to be responsible for the discrimination of the soil microbial communities against those of the zonal forest types. The pine forest soils were set apart from the other forest soils by a higher abundance of PLFA 18:2ω6,9, which is typical of fungi and may also indicate ectomycorrhizal fungi associated with pine trees, and high amounts of PLFA 10Me18:0, which is common in actinomycetes. These findings suggest that the occurrence of azonal forest types at sites with specific soil conditions is accompanied by the development of specific soil microbial communities. The study provides information on the microbial communities in undisturbed forest soils which may facilitate interpretation of data derived from managed or even damaged or degraded forests.     

Methyl-mercury affects microbial activity and biomass,bacterial community structure but rarely the fungal community structure

Monomethyl-mercury is one of the most toxic compounds. Methylation of Hg usually appears under anoxic conditions. In Swiss forest soils, methyl-Hg concentrations of up to 3 μg kg⁻¹ soil dw have been observed, but the impact of methyl-Hg on soil microorganisms have rarely been examined so far. In this study, we investigated the effect of increasing concentrations of methyl-Hg (0, 5, 20, 90 μg kg⁻¹ soil dw) on the microbial communities in various forest soils differing in their physico-chemical properties. Experiments were conducted in microcosms under controlled conditions and the basal respiration (BR), the microbial biomass carbon (MBC) and the bacterial and fungal community structures using T-RFLP-profiling were investigated. BR was significantly affected by methyl-Hg. In general, the BR increased with increasing methyl-Hg concentrations, whereas the MBC was significantly reduced. Bacterial communities were more sensitive to methyl-Hg than fungal communities. In five out of seven soils, the bacterial community structures differed significantly between the treatments whereas the fungal communities did not. The impact of methyl-Hg on the soil bacterial communities was site specific. In one soil, a methyl-Hg concentration of already 5 μg kg⁻¹ soil dw significantly affected the relative abundance of 13% bacterial operational taxonomic units (OTU), whereas in other soils concentrations of even 90 μg kg⁻¹ soil dw rarely affected the abundance of OTUs. In this study, for the first time, the impact of methyl-Hg on soil bacterial and fungal communities in forest soils was assessed. We showed that its impact strongly depends on the physico-chemical conditions of the soil and that bacterial communities were more sensitive to methyl-Hg than fungi.     

Changes in soil microbial community composition in response to fertilization of paddy soils in subtropical China

Repeated fertilizer applications to cultivated soils may alter the composition and activities of microbial communities in terrestrial agro-ecosystems. In this study, we investigated the effects of different long term fertilization practices (control (CK), three levels of mineral fertilizer (N₁P₁K₁, N₂P₂K₂, and N₃P₃K₃), and organic manure (OM)) on soil environmental variables and microbial communities by using phospholipid fatty acid (PLFA) biomarkers analysis in subtropical China. Study showed that OM treatment led to increases in soil organic carbon (SOC), total nitrogen (TN) and total phosphorus (TP) contents, while the mineral fertilizer treatment led to increases in dissolved organic carbon (DOC) content. Changes in soil microbial communities (eg. bacteria, actinomycetes) were more noticeable in soils subjected to organic manure applications than in the control soils or those treated with mineral fertilizer applications. Fungal PLFA biomarkers responded differently from the other PLFA groups, the numerical values of fungal PLFA biomarkers were similar for all the OM and mineral fertilizer treatments. PCA analysis showed that the relative abundance of most PLFA biomarkers increased in response to OM treatment, and that increased application rates of the mineral fertilizer changed the composition of one small fungal PLFA biomarker group (namely 18:3ω6c and 16:1ω5c). Further, from the range of soil environmental factors that we examined, SOC, TN and TP were the key determinants affecting soil microbial community. Our results suggest that organic manure should be recommended to improve soil microbial activity in subtropical agricultural ecosystems, while increasing mineral fertilizer applications alone will not increase microbial growth in paddy soils.     

Microbial community composition and rhizodeposit-carbon assimilation in differently managed temperate grassland soils

Rhizodeposit-carbon provides a major energy source for microbial growth in the rhizosphere of grassland soils. However, little is known about the microbial communities that mediate the rhizosphere carbon dynamics, especially how their activity is influenced by changes in soil management. We combined a ¹³CO₂ pulse-labeling experiment with phospholipid fatty acid (PLFA) analysis in differently managed Belgian grasslands to identify the active rhizodeposit-C assimilating microbial communities in these grasslands and to evaluate their response to management practices. Experimental treatments consisted of three mineral N fertilization levels (0, 225 and 450 kg N ha⁻¹ y⁻¹) and two mowing frequencies (3 and 5 times y⁻¹). Phospholipid fatty acids were extracted from surface (0-5 cm) bulk (BU) and root-adhering (RA) soil samples prior to and 24 h after pulse-labeling and were analyzed by gas chromatography-combustion-isotope ratio mass spectrometry (GC-c-IRMS). Soil habitats significantly differed in microbial community structure (as revealed by multivariate analysis of mol% biomarker PLFAs) as well as in gram-positive bacterial rhizodeposit-C uptake (as revealed by greater ¹³C-PLFA enrichment following pulse-labeling in RA compared to BU soil in the 450N/5M treatment). Mowing frequency did not significantly alter the relative abundance (mol%) or activity (¹³C enrichment) of microbial communities. In the non-fertilized treatment, the greatest ¹³C enrichment was seen in all fungal biomarker PLFAs (C16:1ω5, C18:1ω9, C18:2ω6,9 and C18:3ω3,6,9), which demonstrates a prominent contribution of fungi in the processing of new photosynthate-C in non-fertilized grassland soils. In all treatments, the lowest ¹³C enrichment was found in gram-positive bacterial and actinomycetes biomarker PLFAs. Fungal biomarker PLFAs had significantly lower ¹³C enrichment in the fertilized compared to non-fertilized treatments in BU soil (C16:1ω5, C18:1ω9) as well as RA soil (all fungal biomarkers). While these observations clearly indicated a negative effect of N fertilization on fungal assimilation of plant-derived C, the effect of N fertilization on fungal abundance could only be detected for the arbuscular mycorrhizal fungal (AMF) PLFA (C16:1ω5). On the other hand, increases in the relative abundance of gram-positive bacterial PLFAs with N fertilization were found without concomitant increases in ¹³C enrichment following pulse-labeling. We conclude that in situ¹³C pulse-labeling of PLFAs is an effective tool to detect functional changes of those microbial communities that are dominantly involved in the immediate processing of new rhizosphere-C.     

Microbial communities of Lumbricus terrestris L. middens: structure, activity, and changes through time in relation to earthworm presence

Background, aim, and scope  Earthworms make a major contribution to decomposition in ecosystems where they are present, mainly acting in the drilosphere, that is, galleries, burrows, casts, and middens. Earthworm middens are hot-spots of microbial activity and nutrient dynamics and represent a suitable model for studying earthworm-mediated influences on soil microbial communities by alteration of the patch structure of the microbial environment. We studied the structure and activity of the microbial communities in the soil system formed by middens of Lumbricus terrestris and the soil below and surrounding them and the role of earthworms in maintaining these structures through time. Material and methods  We set up an experiment in which middens were either left (control) or removed from their original place (translocated) and left in a nearby area free of earthworm activity for 2 months. After 1 and 2 months we sampled middens, soil below them, and surrounding soil. We analyzed the phospholipid fatty acid (PLFA) profiles and measured respiratory fluxes of CO₂ and CH₄. Results  Microbial communities of middens clearly differed from those of soil below and surrounding soil samples, showing higher bacterial and fungal PLFAs (p < 0.0001 and p < 0.01, respectively); furthermore, changes in microbial communities were stronger in control middens than in translocated middens. Moreover, gram positive and negative bacterial PLFAs were greater in translocated than control middens (p < 0.0001 and p < 0.001, respectively), as well as total organic carbon (p < 0.001). Microbial activity was higher in middens than in soil below and surrounding soil samples both for CO₂ (p < 0.0001) and CH₄ (p < 0.0001). Discussion  Soil bioturbation by the earthworm L. terrestris was strong in their middens, but there was not any effect on soil below and surrounding soil. Microbial communities of middens maintain their biomass and activity when earthworms were not present, whereas they decreased their biomass and increased their activity when earthworms were present. Conclusions  Earthworms strongly enhanced microbial activity measured as CO₂ production in middens, which indicates that there are hot spots for soil microbial dynamics and increasing habitat heterogeneity for soil microorganisms. Moreover, our data strongly support the fact that the impact of this earthworm species in this soil is restricted to their middens and increasing soil heterogeneity. Recommendations and perspectives  Our data indicate that it is not clear if earthworms enhance or depress microbial communities of middens since the microbial activity increased, but did not modify their biomass and this was not dependent on soil organic C content. These results indicate no competence for C pools between this anecic earthworm and microorganisms, which has been found for other earthworm species, mainly endogeics. Conversely, they suggest some type of facilitation due to the release of additional nutrient pools in middens when earthworms are present, through the digestion of middens' material or the addition of casts produced from other food sources.     

Bacterial communities in soil mimic patterns of vegetative succession and ecosystem climax but are resilient to change between seasons

Organism succession during ecosystem development has been researched for aboveground plant communities, however, the associated patterns of change in below-ground microbial communities are less described. In 2008, a study was initiated along a developmental sand-dune soil chronosequence bordering northern Lake Michigan near Wilderness Park (WP). It was hypothesized that soil bacterial communities would follow a pattern of change that is associated with soil, plant, and ecosystem development. This study included 5 replicate sites along 9 soils (n = 45) ranging in age from ∼105 to 4010 years since deposition. Soil bacterial community composition and diversity were studied using bacterial tag-encoded FLX amplicon pyrosequencing of the 16S rRNA gene. Bray–Curtis ordination indicated that bacterial community assembly changed along the developmental soil and plant gradient. The changes were not affected by seasonal differences, despite likely differences in plant root C (e.g. exudates), temperature, and water availability in soil. Soil base cations (Ca, Mg) and pH declined, showing log-linear correlations with soil age (r ∼ 0.83, 0.84 and 0.81; P < 0.01). Bacterial diversity (Simpson's 1/D) declined rapidly during the initial stages of soil development (∼105–450 y) and thereafter (>450 y) did not change. Turnover of plant taxa was also more rapid early during ecosystem development and correlated with bacterial community structural change (P < 0.000001; r = 0.56). It is hypothesized that plants help to drive pedogenic change during early (<450 y) soil development (e.g. pH decline, cation leaching) which drive selection of soil bacterial communities. In mature soils (∼450–4000 y), resilient and stable soil bacterial community structures developed, mimicking steady-state climax communities that were observed during latter stages of primary plant succession. These relationships point to possible feedbacks between plant and bacterial communities during ecosystem development.     

Carbon uptake by a microbial community during 30-day treatment with 13C-glucose of a sandy loam soil fertilized for 20 years with NPK or compost as determined by a GC–C–IRMS analysis of phospholipid fatty acids

To investigate the uptake by the microbial community of easily decomposable exogenous organic C and the proportion of this organic C remaining in soils under long-term fertilization schemes, ¹³C-glucose was supplied to arable soils (aquic inceptisol) following a 20-year (1989–2009) application of compost (CM) or inorganic NPK (NPK), along with a control (no fertilizer). Phospholipid fatty acids (PLFAs) were used as biomarkers for actinobacteria, bacteria and fungi. Gas chromatography–combustion–stable isotope ratio mass spectrometry (GC–C–IRMS) was used to determine the incorporation of ¹³C into individual PLFAs. The concentrations of soil microbial PLFAs significantly (P < 0.05) increased in all three soils after the addition of ¹³C-glucose. Over a 30-day incubation period, the highest PLFA concentrations were on day 7 (control) or day 15 (NPK and CM) for bacteria, and on day 30 for both fungi and actinobacteria. The added ¹³C-glucose was incorporated into bacterial PLFAs first, whilst an increase of ¹³C in fungal and actinobacterial PLFAs was measured on day 7 and 15, respectively. The mean amounts of ¹³C in bacterial, actinobacterial and fungal PLFAs in CM-treated soil during the 30-day incubation period were 0.589, 0.030 and 0.056 μg g⁻¹ soil, respectively, which were significantly (P < 0.05) higher than levels measured in the NPK and control soils. Among the bacterial groups, the amount of ¹³C in Gram-positive (G⁺) bacteria over the entire incubation ranged from 0.326 to 0.440 μg g⁻¹ soil in the CM scheme, which was significantly (P < 0.05) higher than levels detected in the NPK and control regimes. In contrast, ¹³C concentrations in monounsaturated PLFAs (aerobic microorganisms) in the CM-treated soil were 0.030–0.045 μg g⁻¹ soil, which was significantly (P < 0.05) lower than in the NPK schemes. The proportion of glucose-derived ¹³C remaining in soils was ranked as follows: CM (53%) > NPK (41%) > control (28%) after 30 days of incubation. Easily decomposable exogenous organic C was thus more effectively maintained under the CM regime, primarily because, after 20 years, CM had altered the microbial community by reducing the ratio of aerobic to anaerobic microorganisms whilst increasing levels of G⁺ bacteria in soil compared to the control and NPK soils. This study aids our understanding of the transformation and maintenance of easily decomposable organic C in soil over long-term fertilization regimes.     

Methane uptake in soils from Pinus radiata plantations, a reverting shrubland and adjacent pastures: Effects of land-use change, and soil texture, water and mineral nitrogen

Afforestation and reforestation of pastures are key land-use changes in New Zealand that help sequester carbon (C) to offset its carbon dioxide (CO₂) emissions under the Kyoto Protocol. However, relatively little attention has been given so far to associated changes in trace gas fluxes. Here, we measure methane (CH₄) fluxes and CO₂ production, as well as microbial C, nitrogen (N) and mineral-N, in intact, gradually dried (ca. 2 months at 20 °C) cores of a volcanic soil and a heavier textured, non-volcanic soil collected within plantations of Pinus radiata D. Don (pine) and adjacent permanent pastures. CH₄ fluxes and CO₂ production were also measured in cores of another volcanic soil under reverting shrubland (mainly Kunzea var. ericoides (A. Rich) J. Thompson) and an adjacent pasture. CH₄ uptake in the pine and shrubland cores of the volcanic soils at field capacity averaged about 35 and 14 μg CH₄-C m⁻² h⁻¹, respectively, and was significantly higher than in the pasture cores (about 21 and 6 μg CH₄-C m⁻² h⁻¹, respectively). In the non-volcanic soil, however, CH₄-C uptake was similar in most cores of the pine and pasture soils, averaging about 7-9 μg m⁻² h⁻¹, except in very wet samples. In contrast, rates of CO₂ production and microbial C and N concentrations were significantly lower under pine than under pasture. In the air-dry cores, microbial C and N had declined in the volcanic soil, but not in the non-volcanic soil; ammonium-N, and especially nitrate-N, had increased significantly in all samples. CH₄ uptake was, with few exceptions, not significantly influenced by initial concentrations of ammonium-N or nitrate-N, nor by their changes on air-drying. A combination of phospholipid fatty acid (PLFA) and stable isotope probing (SIP) analyses of only the pine and pasture soils showed that different methanotrophic communities were probably active in soils under the different vegetations. The C18 PLFAs (type II methanotrophs) predominated under pine and C16 PLFAs (type I methanotrophs) predominated under pasture. Overall, vegetation, soil texture, and water-filled pore space influenced CH₄-C uptake more than did soil mineral-N concentrations.     

The distribution of nematodes and soil microbial communities across soil aggregate fractions and farm management systems

We hypothesized that nematode and microbial communities vary between soil aggregate fractions due to variations in physical and/or resource constraints associated with each fraction and that this, in turn, contributes to management impacts on whole soil food webs. Nematode and microbial communities were examined within three soil fractions: large macroaggregates (LM; >1000 μm), small macroaggregates (SM; 250-1000 μm) and inter-aggregate soil and space (IS; <250 μm) isolated from soils of four agricultural management systems: conventional tomato (CON), organic tomato (ORG), a minimum till grain-legume intercrop with continuous cover (CC) and an unmanaged riparian corridor (RC). Aggregate fractions appeared to influence nematode assemblages more than did management system. In general the IS and LM fractions contained higher densities of all nematode trophic groups than did SM. Management × fraction interactions for bacterivores and fungivores, however; suggested a non uniform trend across management systems. The IS fraction exhibited stronger trophic links, per the nematode structure index (SI), while the LM and SM fractions had more active fungal decomposition channels as indicated by the channel index (CI). Higher adult to juvenile ratios in the LM and IS than the SM fraction, and a positive correlation between nematode density in the IS fraction and the proportion of macroaggregates in the soil, indicated an association between soil structure and nematode distribution. Microbial communities varied across both aggregate fractions and management systems. Phospholipid fatty acid (PLFA) analysis suggested that the LM fraction contained greater microbial biomass, gram positive bacteria, and eukaryotes than the IS fraction, while SM contained intermediate PLFA associated with these groups. Total PLFA was greater under RC and ORG than under CC or CON. Total PLFA was positively correlated with % C in soil fractions while nematode abundance exhibited no such relationship. Our findings suggest that microbial communities are more limited by resource availability than by habitable pore space or predation, while nematode communities, although clearly resource-dependent, are better associated with habitable pore space for the soil fractions studied here.     

Soil microbial biomass and activity in Chinese tea gardens of varying stand age and productivity

Tea (Camellia sinensis) is a globally important crop and is unusual because it both requires an acid soil and acidifies soil. Tea stands tend to be extremely heavily fertilized in order to improve yield and quality, resulting in a great potential for diffuse pollution. The microbial ecology of tea soils remains poorly understood; an improved understanding is necessary as processes affecting nutrient availability and loss pathways are microbially mediated. We therefore examined the relationships between soil characteristics (pH, organic C, total N, total P, available P, exchangeable Al), the soil microbial biomass (biomass C, biomass ninhydrin-N, ATP, phospholipid fatty acids—PLFAs) and its activities (respiration, net mineralization and nitrification). At the Tea Research Institute, Hangzhou (TRI), we compared fields of different productivity levels (low, medium and high) and at Hongjiashan village (HJS) we compared fields of different stand age (9, 50 and 90 years). At both sites tea soils were compared with adjacent forest soils. At both sites, soil pH was highest in the forest soil and decreased with increasing productivity and age of the tea stand. Soil microbial biomass C and biomass ninhydrin-N were significantly affected by tea production. At TRI, microbial biomass C declined in the order forest>low>high>middle production and at HJS in the order stand age 50>age 9>forest>age 90. Soil pH had a strong influence on the microbial biomass, demonstrated by positive linear correlations with: microbial biomass C, microbial biomass ninhydrin-N, the microbial biomass C:organic C ratio, the microbial biomass ninhydrin-N:total N ratio, the respiration rate and specific respiration rate. Above pH_(KCl) 3.5 there was net N mineralization and nitrification, and below this threshold some samples showed net immobilization of N. A principal component (PC) analysis of PLFA data showed a consistent shift in the community composition with productivity level and stand age. The ratio of fungal:bacterial PLFA biomarkers was negatively and linearly correlated with specific respiration in the soils from HJS (r²=0.93, p=0.03). Our results demonstrate that tea cultivation intensity and duration have a strong impact on the microbial community structure, biomass and its functioning, likely through soil acidification and fertilizer addition.     

Long-term exclusion of plant-inputs to soil reduces the functional capacity of microbial communities to mineralise recalcitrant root-derived carbon sources

Microbial communities in soil are highly species-rich, recognition of which has led to the view that functional redundancy within communities may buffer many impacts of altered community structure on soil functions. In this study we investigated the impact of long-term (>50 years) exclusion of plant-inputs (bare-fallow treatment) on soil microbial community structure and on the ability of the microbial biomass to mineralise tracer additions of ¹³C-labelled plant-derived C-substrates. Exclusion of plant-inputs resulted in depletion of soil organic matter (SOM) and a reduction in microbial biomass size. The microbial community structure was also strongly affected, as indicated by the distinct phospholipid fatty acid (PLFA) profiles in bare-fallow and grassland soils. Mineralisation of labile plant-derived substrates was not perturbed by the bare-fallow treatment. The incorporation of labile plant-derived C into PLFA biomarkers was found to differ between soils, reflecting the distinct community structures of the soils and indicating that these substrates were utilised by a broad range of microbial groups. In contrast, the mineralisation of recalcitrant plant-derived substrates was reduced in bare-fallow soil and the fate of substrate-derived C within PLFA biomarkers was, initially, similar between the soils. These results indicate that utilisation of these recalcitrant substrates was a function restricted to specific groups, and that exclusion of plant-derived inputs to soil had reduced the capacity of bare-fallow microbial communities to utilise this substrate type. Therefore, the study suggests that long-term selective pressure on microbial communities, resulting in altered community structure, may also result in altered functional attributes. This structure-function relationship was apparent for utilisation of recalcitrant plant-derived substrates, but not for the more widely distributed attribute of labile C-substrate utilisation.