Evaluation of microbial methods as potential indicators of soil quality in historical agricultural fields

In agricultural ecosystems that have had consistent cropping histories, standard microbial methods may be used to evaluate past and present practices. Our objective was to evaluate several microbial methods that best indicate cropping histories and soil quality on long-term plots. We selected soil microbial carbon (C), phospholipid analyses, direct counts of total fungal and bacterial biomass, and soil enzymes (phosphatases) to measure direct and indirect microbial activity on the Sanborn Field and Tucker Prairie. The Sanborn Field has been under various cropping and management practices since 1888 and the Tucker Prairie is an uncultivated site. Seven different plots were chosen on the Sanborn Field and random samples were taken in the summit area on the Tucker Prairie, which represented a reference site. Soil microbial biomass C, phospholipids, and enzyme activity were reflective of the cropping and management histories observed on the Sanborn Field. Enzymatic activity was highly correlated to soil organic matter. The direct counts of fungal and bacterial biomass showed that fungal populations dominated these soils, which may be attributed to soil pH. Soil microbial biomass C and enzyme assays seemed to be better potential indicators of cropping histories than the other methods tested in the long-term plots.This paper has been assigned by the Missouri Agricultural Experiment Station to Journal Series no. 12043     

Microbial biomass and activity in pine litter in the presence of Tomocerus minor (Insecta,Collembola)

Summary Laboratory microcosms were used to study microbial populations and biomasses developing in fragmented litter of Pinus nigra Arnold var. nigra (A. et G.). Direct observations (fungal standing crop and fluorescein-stainable mycelia), litter enzyme analyses (cellulase and dehydrogenase), and measurements by physiological methods (microbial CO₂ production and total microbial, fungal, and bacterial viable biomasses) were made at 3-week intervals for 15 weeks. Most variables showed great changes during this period, which were ascribed to a rise in litter moisture content during the initial phase of the experiment, and to substrate depletion towards its final phase. The addition of the collembolan Tomocerus minor (Lubbock) for 1 week enhanced cellulase activities by 4%. When the animals were introduced after 6 weeks, the fungal standing crop was enhanced, and the percentage of fluorescein-stainable mycelia was reduced. Dehydrogenase activity was increased by grazing when the microbial population had been established for 9 weeks or longer. Eucaryotic and procaryotic substrate-induced respiration were positively correlated, which was explained by partial segregation of resources for the two groups. Litter cellulase and dehydrogenase activity showed correlations by other techniques, indicating their suitability as parameters for microbial activity in general, and for the collembolan grazing impact on microbial activity in particular.     

Influence of long-term fertilisation and crop rotation on changes in fungal and bacterial residues in a tropical rice-field soil

Amino sugars, as a microbial residue biomarker, are highly involved in microbial-mediated soil organic matter formation. However, accumulation of microbial biomass and responses of bacterial and fungal residues to the management practices are different and poorly characterized in rice soils. The objectives of this study were to evaluate the effects of mineral fertiliser (MIN), farmyard manure (FYM) and groundnut oil cake (GOC) on crop yield and co-accumulation of microbial residues and microbial biomass under rice-monoculture (RRR) and rice–legume–rice (RLR) systems. In the organic fertiliser treatments and RLR, rice grain yield and stocks of soil and microbial nutrients were significantly higher than those of the MIN treatment and RRR, respectively. The increased presence of saprotrophic fungi in the organic fertiliser treatments and RRR was indicated by significantly increased ergosterol/C_mic ratio and extractable sulphur. In both crop rotation systems, the long-term application of FYM and GOC led to increased bacterial residues as indicated by greater accumulation of muramic acid. In contrast, the higher fungal C/bacterial C ratio and lower ergosterol/C_mic ratio in the MIN treatment, is likely caused by a shift within the fungal community structure towards ergosterol-free arbuscular mycorrhizal fungi (AMF). The organic fertiliser treatments contributed 22 % more microbial residual C to soil organic C compared to the MIN treatment. Our results suggest that the negative relationship between the ratios ergosterol/C_mic and fungal C/bacterial C encourages studying responses of both saprotrophic fungi and AMF when assessing management effects on the soil microbial community.     

Nitrogen dynamics and microbial response in soil amended with either olive pulp or its by-products after biogas production

Olive pulp (OP), the residual material of a two-phase olive oil extraction system, and effluents from hydrogen (EH₂) and methane (ECH₄) production, have been evaluated as soil amendments particularly for their impact on soil mineral nitrogen (N) dynamics, gross N mineralization, and soil microbial biomass N (N_mic). Both N transformation and microbial growth were mainly influenced by the amount and quality of added organic carbon (C). Both OP and EH₂, which contain more carbohydrates and lipids than polyphenolic compounds, stimulated NO₃ ⁻ immobilization during the early incubation period and increased N_mic, saprophytic fungi, and N mineralization. On the contrary, soil amended with ECH₄, which is characterized by the lowest C content but the highest content of polyphenolic compounds, behaved as the control; neither NO₃ ⁻ immobilization nor microbial growth were observed and gross N mineralization was stimulated only at the beginning of the incubation period. Bacterial plate count was significantly correlated with direct bacterial count and fungal count was correlated with N_mic. Therefore, it is suggested that both bacterial and fungal plate counts may be used as indicators of the overall bacterial and fungal populations inhabiting soil, respectively. The knowledge of the impact of these materials on soil N dynamics is crucial for their correct use in agriculture because it has been shown that NO₃ ⁻ availability can be strongly influenced by the addition of different amounts and quality of organic amendment.     

Microbial composition and diversity of an upland red soil under long-term fertilization treatments as revealed by culture-dependent and culture-independent approaches

Background, aim, and scope  Fertilization is an important agricultural practice for increasing crop yields. In order to maintain the soil sustainability, it is important to monitor the effects of fertilizer applications on the shifts of soil microorganisms, which control the cycling of many nutrients in the soil. Here, culture-dependent and culture-independent approaches were used to analyze the soil bacterial and fungal quantities and community structure under seven fertilization treatments, including Control, Manure, Return (harvested peanut straw was returned to the plot), and chemical fertilizers of NPK, NP, NK, and PK. The objective of this study was to examine the effects on soil microbial composition and diversity of long-term organic and chemical fertilizer regimes in a Chinese upland red soil. Materials and methods  Soil samples were collected from a long-term experiment station at Yingtan (28°15′N, 116°55′E), Jiangxi Province of China. The soil samples (0–20 cm) from four individual plots per treatment were collected. The total numbers of culturable bacteria and fungi were determined as colony forming units (CFUs) and selected colonies were identified on agar plates by dilution plate methods. Moreover, soil DNAs were extracted and bacterial 16S rRNA genes and fungal 18S rRNA genes were polymerase chain reaction amplified, and then analyzed by denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing. Results  The organic fertilizers, especially manure, induced the least culturable bacterial CFUs, but the highest bacterial diversity ascertained by DGGE banding patterns. Chemical fertilizers, on the other hand, had less effect on the bacterial composition and diversity, with the NK treatment having the lowest CFUs. For the fungal community, the manure treatment had the largest CFUs but much fewer DGGE bands, also with the NK treatment having the lowest CFUs. The conventional identification of representative bacterial and fungal genera showed that long-term fertilization treatments resulted in differences in soil microbial composition and diversity. In particular, 42.4% of the identified bacterial isolates were classified into members of Arthrobacter. For fungi, Aspergillus, Penicillium, and Mucor were the most prevalent three genera, which accounted for 46.6% of the total identified fungi. The long-term fertilization treatments resulted in different bacterial and fungal compositions ascertained by the culture-dependent and also the culture-independent approaches. Discussion  It was evident that more representative fungal genera appeared in organic treatments than other treatments, indicating that culturable fungi were more sensitive to organic than to chemical fertilizers. A very notable finding was that fungal CFUs appeared maximal in organic manure treatments. This was quite different from the bacterial CFUs in the manure, indicating that bacteria and fungi responded differently to the fertilization. Similar to bacteria, the minimum fungal CFUs were also observed in the NK treatment. This result provided evidence that phosphorus could be a key factor for microorganisms in the soil. Thus, despite the fact that culture-dependent techniques are not ideal for studies of the composition of natural microbial communities when used alone, they provide one of the more useful means of understanding the growth habit, development, and potential function of microorganisms from soil habitats. A combination of culture-dependent and culture-independent approaches is likely to reveal more complete information regarding the composition of soil microbial communities. Conclusions  Long-term fertilization had great effects on the soil bacterial and fungal communities. Organic fertilizer applications induced the least culturable bacterial CFUs but the highest bacterial diversity, while chemical fertilizer applications had less impact on soil bacterial community. The largest fungal CFUs were obtained, but much lower diversity was detected in the manure treatment. The lowest bacterial and also fungal CFUs were observed in the NK treatment. The long-term fertilization treatments resulted in different bacterial and fungal compositions ascertained by the culture-dependent and also the culture-independent approaches. Phosphorus fertilizer could be considered as a key factor to control the microbial CFUs and diversity in this Chinese upland red soil. Recommendations and perspectives  Soil fungi seem to be a more sensitive indicator of soil fertility than soil bacteria. Since the major limitation of molecular methods in soil microbial studies is the lack of discrimination between the living and dead, or active and dormant microorganisms, both culture-dependent and culture-independent methods should be used to appropriately characterize soil microbial diversity.     

Investigating microbial community structure in soils by physiological, biochemical and molecular fingerprinting methods

Several biochemical and molecular methods are used to investigate the microbial diversity and changes in microbial community structure in rhizospheres and bulk soils resulting from changes in management. We have compared the effects of plants on the microbial community, using several methods, in three different types of soils. Pots containing soil from three contrasting sites were planted with Lolium perenne (rye grass). Physiological (Biolog), biochemical (PLFA) and molecular (DGGE and TRFLP) fingerprinting methods were employed to study the change in soil microbial communities caused by the growth of rye grass. Different methods of DNA extraction and nested PCR on TRFLP profiles were examined to investigate whether they gave different views of community structure. Molecular methods were used for both fungal and bacterial diversity. Principal component analysis of Biolog data suggested a significant effect of the plants on the microbial community structure. We found significant effects of both soil type and plants on microbial communities in PLFA data. Data from TRFLP of soil bacterial communities showed large effects of soil type and smaller but significant effects of plants. Effects of plant growth on soil fungal communities were measured by TRFLP and DGGE. Multiple Procrustes analysis suggested that both methods gave similar results, with only soil types having a significant effect on fungal communities. However, TRFLP was more discriminatory as it generated more ribotype fragments for each sample than the number of bands detected by DGGE. Neither methods of DNA extraction nor the nested PCR had any effect on the evaluation of soil microbial community structure. In conclusion, the different methods of microbial fingerprinting gave qualitatively similar results when samples were processed consistently and compatible statistical methods used. However, the molecular methods were more discriminatory than the physiological and biochemical approaches. We believe results obtained from this experiment will have a major impact on soil microbial ecology in general and rhizosphere–microbial interaction studies in particular, as we showed that the different fingerprinting methods for microbial communities gave qualitatively similar results.     

Quantitative assessment of the fungal contribution to microbial tissue in soil

The fungi-to-bacteria ratio in soil ecological concepts and its application to explain the effects of land use changes have gained increasing attention over the past decade. Four different main approaches for quantifying the fungal and bacterial contribution to microbial tissue can be distinguished: (1) microscopic methods, (2) selective inhibition, (3) specific cell membrane components and (4) specific cell wall components. In this review, the different methods were compared and we hypothesized that all these approaches result in similar values for the fungal and bacterial contribution to total microbial biomass, activity, and residues (dead microbial tissue) if these methods are evenly reliable for the estimation of fungal biomass. The fungal contribution to the microbial biomass or respiration varied widely between 2 and 95% in different data sets published over the past three decades. However, the majority of the literature data indicated that fungi dominated microbial biomass, respiration or non-biomass microbial residues, with mean percentages obtained by the different methodological approaches varying between 35 and 76% in different soil groups, i.e. arable, grassland, and forest soils and litter layers. Microscopic methods generally gave the lowest average values, especially in arable and grasslands soils. Very low ratios in fungal biomass C-to-ergosterol obtained by microscopic methods suggest a severe underestimation of fungal biomass by certain stains. Relatively consistent ratios of ergosterol to linoleic acid (18:2ω6,9) indicate that both cell membrane components are useful indicators for saprotrophic and ectomycorrhizal fungi. More quantitative information on the PLFA content of soil bacteria and the 16:1ω5 content of arbuscular mycorrhizal fungi is urgently required to fully exploit the great potential of PLFA measurements. The most consistent results have been obtained from the analysis of fungal glucosamine and bacterial muramic acid in microbial residues. Component-specific δ¹³C analyses of PLFA and amino sugars are a promising prospect for the near future.     

Specific response of fungal and bacterial residues to one-season tillage and repeated slurry application in a permanent grassland soil

The dynamics of fungal and bacterial residues to a one-season tillage event in combination with manure application in a grassland soil are unknown. The objectives of this study were (1) to assess the effects of one-season tillage event in two field trials on the stocks of microbial biomass, fungal biomass, microbial residues, soil organic C (SOC) and total N in comparison with permanent grassland; (2) to determine the effects of repeated manure application to restore negative tillage effects on soil microbial biomass and residues. One trial was started 2 years before sampling and the other 5 years before sampling. Mouldboard ploughing decreased the stocks of SOC, total N, microbial biomass C, and microbial residues (muramic acid and glucosamine), but increased those of the fungal biomarker ergosterol in both trials. Slurry application increased stocks of SOC and total N only in the short-term, whereas the stocks of microbial biomass C, ergosterol and microbial residues were generally increased in both trials, especially in combination with tillage. The ergosterol to microbial biomass C ratio was increased by tillage, and decreased by slurry application in both trials. The fungal C to bacterial C ratio was generally decreased by these two treatments. The metabolic quotient qCO₂ showed a significant negative linear relationship with the microbial biomass C to SOC ratio and a significant positive relationship with the soil C/N ratio. The ergosterol to microbial biomass C ratio revealed a significant positive linear relationship with the fungal C to bacterial C ratio, but a negative one with the SOC content. Our results suggest that slurry application in grassland soil may promote SOC storage without increasing the role of saprotrophic fungi in soil organic matter dynamics relative to that of bacteria.     

Plant and soil microbial biomasses in Agrostis capillaris and Lathyrus pratensis monocultures exposed to elevated O3 and CO2 for three growing seasons

Elevated CO₂ usually promotes plant growth, whereas elevated O₃ often has a negative effect, especially on root biomass. Thus both these gases can indirectly affect the soil microbial community. We exposed Agrostis capillaris and Lathyrus pratensis to realistic levels of O₃ (40-50 ppb) and CO₂ (ambient air + 100 ppm) in open-top chambers during 2002-2004. The experiment shows negative effects of both O₃ and CO₂, especially on the bulk soil of L. pratensis, in terms of the decreased biomasses of total (25% and 31%), actinobacterial (29% and 31%), bacterial (26% and 33%) and mycorrhizal (AM fungal) (31% and 35%) indicator subgroups, analysed by the PLFA (phospholipid fatty acid) method. The fungal:bacterial PLFA biomass ratio decreased in the bulk soil of A. capillaris, especially with elevated CO₂ alone (38%). These longer-term changes are considered to arise mainly from differences between the plant functional types (i.e. grass cf. N₂-fixing legume) in litter quality and soil C:N ratio. The results also point to interactions and multi-trophic feedbacks between elevated O₃, plant, parasitic rust fungi and soil readily available P, accompanied by a shift in N balance in favour of plants rather than soil microorganisms.     

Application of eco-physiological quotients (qCO2 and qD) on microbial biomasses from soils of different cropping histories

Metabolic quotients for CO₂C (qCO₂C) and microbial-C-loss (qD) were studied on soil microbial communities under long-term monoculture (M) or continuous crop rotations (CR). Under defined laboratory conditions the mean qCO₂C (unit CO₂C unit⁻¹ C_mic h⁻¹) of different microbial biomasses from 17 M systems amounted to 1.097 μg CO₂qCO₂CC as compared to 0.645 μg CO₂C of microbial biomasses from 19 CR systems. The 1.7 times higher CO₂C release per unit biomass and time of microbial biomasses from M systems was significantly different at the P =0.001 level.In addition, microbial C-loss in samples from M or CR plots was followed for 5 weeks. Again, mean qD per unit microbial biomass and time was 1.6 times higher (P = 0.01) for microbial biomasses from M systems (0.301 μg C, 14 soils) when compared with CR systems (0.188μg C, 14 soils).These differences were not related to soil texture, C_org or pH of these soils. The effects of environmental influences (soil management) on the microbial pool in terms of a changing energy demand are discussed.     

Humus macro-morphology and soil microbial community changes along a 130-yr-old Fagus sylvatica chronosequence

We aimed to characterize humus macro-morphology and the associated soil microbial community within the unmodified litter (OL), the fragmented and humified layers (FH) and the organo-mineral (A) layer along a beech (Fagus sylvatica L.) forest chronosequence with four stand age-classes (15-, 65-, 95-, 130-yr-old) in Normandy, France. Humus macro-morphology was described with 36 quantitative and semi-quantitative variables. We measured microbial biomass N (N_mic), microbial N quotient (N_mic-to-N_t), fungal ergosterol, bacterial and fungal DNA using 16S and 18S rDNA real-time qPCR and evaluated the potential metabolic profile of heterotrophic bacteria within each soil layer and stand age-class. The log-transform ergosterol/fungal DNA ratio (EFR index) was used as an indicator related to active fungal biomass and the fungal/bacterial (F/B) ratio was calculated from qPCR results. There was a shift from mull (mainly dysmull) to moder humus forms along the chronosequence. While the N_mic did not change significantly, the N_mic-to-N_t decreased along the chronosequence in the OL layer. Ergosterol content increased in FH and A layers and the F/B ratio increased in the FH layer with increasing beech forest age. The EFR index was significantly higher in the OL and A layers of the oldest stands, whereas the highest EFR index in the FH layer occurred in the 15-yr-old stands. The functional diversity of heterotrophic bacteria was greater within OL and FH layers of 130-yr-old stands, but highest in the A layer of 15-yr-old stands while the Average Well Color Development remained stable for all soil layers. We found significant correlations between macro-morphology and microbial variables, especially between FH-based morphology and fungal biomass. Our main results are that beech forest maturation is accompanied by (1) an increase in fungal biomass in the FH layers and, (2) an increase in heterotrophic bacteria functional diversity in the organic layers. We have identified key macro-morphology variables that are good predictors of the structural and functional profile of the soil microbial community during beech forest development.     

Effect of fungal to bacterial biomass ratio on the relationship between CO2 evolution and total soil microbial biomass

The relationship between the fungal: bacterial biomass ratio and the metabolic quotient (qCO₂) was studied in three different soils. In addition, the effect of the fungal: bacterial biomass ratio on the relationship between CO₂ evolution and the size of the soil microbial biomass was examined. Soil samples were collected from three experimental fields amended with various organic materials (Yatsugatake, Ibaraki, and Tochigi fields). The range of the fungal:bacterial biomass ratio in the Yatsugatake and Ibaraki fields was small (1.54–2.24 and 1.11–1.71, respectively), but it was large in the Tochigi field (1.18–3.75). We found a high negative correlation between this ratio and the metabolic quotient (qCO₂=2.10–0.361 (fungal:bacterial biomass ratio), R=–0.851, P<0.01) in the Tochigi field. Therefore, we suggest tha qCO₂ decreases with an increase in the fungal:bacterial biomass ratio, which may be due to a higher efficiency of substrate C use by fungal flora in comparison with bacterial flora. In the Yatsugatake and Ibaraki fields, there was a high positive correlation between CO₂ evolution and total microbial biomass. In contrast, no correlation was observed between these two parameters in the Tochigi field, probably reflecting the wide range of values for the fungal:bacterial biomass ratio. From the results obtained, we suggest that the fungal: bacterial biomass ratio is an important factor regulating the relationship between CO₂ evolution and the size of the microbial biomass.     

水分含量对秸秆还田土壤碳矿化和微生物特性的影响

An 80-d incubation experiment was conducted to investigate straw decomposition,the priming effect and microbial characteristics in a non-fertilized soil(soil 1) and a long-term organic manure-fertilized soil(soil 2) with and without13 C-labeled maize straw amendment under different moisture levels. The soil 2 showed a markedly higher priming effect,microbial biomass C(Cmic),and β-glucosidase activity,and more abundant populations of bacteria and fungi than the soil 1. Also,soil CO2 emission,Cmic,β-glucosidase activity,and bacterial and fungal population sizes were substantially enhanced by straw amendment. In the presence of straw,the amount of straw mineralization and assimilation by microbes in the soil at 55% of water holding capacity(WHC) were significantly higher by 31% and 17%,respectively,compared to those at 25% of WHC. In contrast,β-glucosidase activity and fungal population size were both enhanced as the moisture content decreased. Cmicdecreased as straw availability decreased,which was mainly attributed to the reduction of straw-derived Cmic. Amended soils,except the amended soil 2 at 25% of WHC,had a more abundant fungal population as straw availability decreased,indicating that fungal decomposability of added straw was independent of straw availability. Non-metric multidimensional scaling analysis based on fungal denatured gradient gel electrophoresis band patterns showed that shifts in the fungal community structure occurred as water and straw availability varied. The results indirectly suggest that soil fungi are able to adjust their degradation activity to water and straw availability by regulating their community structure.     

Effect of arsenic contamination on bacterial and fungal biomass and enzyme activities in tropical arsenic-contaminated soils

The importance of assessing the impacts of soil arsenic (As) contamination on microbial properties lay on the fact that microbes are instrumental in nutrient cycling and are therefore indicators of soil quality. In this study, soil chemical extraction methods were used to extract labile and freely exchangeable As (water-soluble As and sodium bicarbonate-extractable As), amorphous/crystalline Fe and Mn oxide-bound As (acid ammonium oxalate-extractable As and hydroxylamine hydrochloride-extractable As), and their impacts on microbial biomass (microbial biomass C, total bacterial and fungal biomass, active bacterial and fungal biomass), enzyme activities representing four major soil biogeochemical cycles, i.e., C (β-glucosidase activity), N (urease activity), P (acid phosphomonoesterase activity), S (acryl-sulfatase activity), and microbial activity (fluorescein diacetate hydrolysis and dehydrogenase activity) were investigated in As-contaminated soils of Ambagarh Chauki block, Chhattisgarh, Central India. The results revealed that the majority of the As in soils resided in the Fe/Mn oxide-bound fraction. The microbial biomass C, total and active fungal biomass, and enzyme activities were significantly inhibited by all the forms of As. However, water-soluble As, even though occupying only a small portion of the total As (0.9–2.9 %), exerted the greatest impact. Interestingly, total and active bacterial biomass was not significantly affected by As toxicity, suggesting their resistance to As. Urease activity was not affected by As pollution.     

Criteria for measurement of microbial growth and activity in soil

Changes in CO₂ evolution, phosphatase and urease activity and ATP contents were related to bacterial and fungal biomass determined microscopically during glucose mineralization at different concentrations of mineral nutrients. Similar results were obtained in a sandy loam and a clay soil except that in the clay the increase in microbial and enzyme activities were delayed. Higher initial rates of CO₂ evolution were noted after the addition of P to a glucose and N amended soil at C:P ratios greater than 30:1. Increases in phosphatase activity coincided with increases in bacterial and fungal populations only in treatments without inorganic P. Peak rates of CO₂ evolution preceded biomass production by 18–24 h, therefore, CO₂ evolution rates did not show a correlation on normal regression analysis with biomass. Soil ATP content was influenced by P concentrations and soil type. ATP was therefore not a specific indicator of biomass in the detailed studies where P concentrations and sequential growth of bacteria and fungi were major factors. Soil urease increased with bacterial and fungal populations. It did not respond to P other than through microbial biomass and was highly correlated with microbial biomass. The results show that no one measurement of microbial biomass or activity is sufficient to interpret microbial growth in the soil system. Each of the criteria measured were sensitive to specific conditions affecting biomass and activity.     

Fungal and bacterial denitrification are differently affected by long-term pH amendment and cultivation of arable soil

Bacteria are considered as playing a predominant role in the production of nitrous oxide (N₂O) in arable soil. Despite the knowledge that fungi are able to denitrify their contribution to denitrifier N₂O production from arable soil is uncertain. Here, we assess the capability of fungi and bacteria to contribute to N₂O emission from arable soil by measuring potential denitrification rates (PDR) as N₂O production, after application of selective inhibitors aimed at distinguishing between fungal and bacterial denitrification, and related PDR to characteristics of the soil microbial community. Soil was sampled from a long-term crop rotation maintained since 1961 at seven different pH levels, ranging in 0.5 increments from pH 4.5 to 7.5, and along a cultivation gradient from freshly ploughed soil to three years under ley grass. Over both pH and cultivation gradients, bacteria contributed up to 54% and fungi contributed to 18% of the PDR. Residual N₂O production that was not targeted by the selective inhibitors and hence could not be attributed to fungi or bacteria might be due to pre-synthesised enzymes or resistant organisms. The PDR of the bacterial community responded positively to increase in soil pH with the lowest PDR at pH 4.2 and the highest around pH 5.9. In contrast, fungal denitrification was not influenced by soil pH. Changes in ester linked fatty acids (ELFA) concentrations showed that whilst total bacterial biomass decreased with increasing pH fungal biomass was not significantly influenced by pH, driving an increase in the ratio of fungal–bacterial biomass. Both fungal biomass and bacterial biomass, and the PDR from the control treatment (no inhibitor application) across the pH gradient were greatest under long-term ley. Concentrations of fatty acids a15:0, 16:1ω7 and 17:1ω8 of microbial origin were positively correlated with the proportion of denitrification activity that was repressed by bacterial inhibitors. This suggests that there is a relationship between organisms that possess the fatty acids a15:0, 16:1ω7 and 17:1ω8, and the function of denitrification. Our results demonstrate that both fungal and bacterial denitrification were occurring in this arable soil. That management for pH and cultivation had differing effects on the potential contribution of fungal and bacterial denitrification to N₂O production has implications for the development of appropriate management practices for mitigation of this greenhouse gas.     

Structure and function of microbial communities in the rhizosphere of Scots pine after tree-felling

We evaluated changes occurring in the rhizosphere microbial communities of Scots pine (Pinus sylvestris L.) due to tree-felling and decrease of the photosynthetic C flow into the soil under field conditions over one growing season. Samples were taken from tree rhizospheres, freshly felled stump rhizospheres and bulk soil. We used culture dependent (CFU counts, community level physiological profiles, CLPPs) and independent methods (fluorogenic MUF-substrates, PLFA pattern and PCR-DGGE) to monitor the microbial communities in soil samples. The numbers of cultivable bacteria and amounts of phosphatase activity in the rhizosphere of trees were significantly higher compared with those in the bulk soil. The organic C consuming community measured by CLPP was stimulated directly after the tree-felling in stump rhizospheres; utilization of the disintegration components of cellulose, hemicellulose and chitin increased. Furthermore, bacterial and fungal biomass as well as chitin decomposers (CFU) increased in the stump rhizosphere. After 11 weeks of tree-felling the stump rhizosphere soluble PO₄-P and NH₄-N as well as amounts of total C and N began to resemble the concentrations measured in the bulk soil. However, the stump rhizosphere community structure detected by PLFA and PCR-DGGE still resembled that of the tree rhizosphere.     

Structural and functional diversity of soil bacterial and fungal communities following woody plant encroachment in the southern Great Plains

In the southern Great Plains (USA), encroachment of grassland ecosystems by Prosopis glandulosa (honey mesquite) is widespread. Mesquite encroachment alters net primary productivity, enhances stores of C and N in plants and soil, and leads to increased levels of soil microbial biomass and activity. While mesquite’s impact on the biogeochemistry of the region is well established, it effects on soil microbial diversity and function are unknown. In this study, soils associated with four plant types (C₃ perennial grasses, C₄ midgrasses, C₄ shortgrasses, and mesquite) from a mesquite-encroached mixed grass prairie were surveyed to in an attempt to characterize the structure, diversity, and functional capacity of their soil microbial communities. rRNA gene cloning and sequencing were used in conjunction with the GeoChip functional gene array to evaluate these potential differences. Mesquite soil supported increased bacterial and fungal diversity and harbored a distinct fungal community relative to other plant types. Despite differences in composition and diversity, few significant differences were detected with respect to the potential functional capacity of the soil microbial communities. These results may suggest that a high level of functional redundancy exists within the bacterial portion of the soil communities; however, given the bias of the GeoChip toward bacterial functional genes, potential functional differences among soil fungi could not be addressed. The results of this study illustrate the linkages shared between above- and belowground communities and demonstrate that soil microbial communities, and in particular soil fungi, may be altered by the process of woody plant encroachment.     

Changes in the composition and diversity of topsoil bacterial,archaeal and fungal communities after 22 years conventional and no-tillage managements in Northern China

A comprehensive comparison about microbial community (bacterial, archaeal and fungal) response to different tillage managements in Northern China remain little studied, in this study we compared no-tillage (NT) versus conventional tillage (CT) management on topsoil microbial community diversity and composition in field experiment. We found that NT practice significantly increased the soil moisture content (SMC), bulk density, stocks of soil organic carbon (SOC), total nitrogen (TN), and microbial biomass carbon and nitrogen (P < 0.05). Moreover, higher levels of bacterial and archaeal alpha diversity were observed in NT relative to CT while unexpectedly, there was no significant difference found in fungal diversity between two treatments. The most pronounced shifts in the composition of the different microbial groups were found for the archaeal community, which followed by bacterial and fungal. NT practice markedly enhanced abundances of Proteobacteria (belongs to bacteria) phyla, Thaumarchaeota phyla (belongs to archaea) and Glomeromycota phyla (belongs to fungi). Redundancy analysis revealed that the factor that most closely correlated with bacterial, archaeal and fungal composition were SMC, TN and SOC, respectively. Considering NT enhanced both microbial composition and C storage in topsoil, we suggest that NT offers significant promise to improve topsoil health in this region.     

Determination of the impact of continuous defoliation of Lolium perenne and Trifolium repens on bacterial and fungal community structure in rhizosphere soil

To determine the effects of defoliation on microbial community structure, rhizosphere soil samples were taken pre-, and post-defoliation from the root tip and mature root regions of Trifolium repens L. and Lolium perenne L. Microbial DNA isolated from samples was used to generate polymerase chain reaction–denaturing gradient gel electrophoresis molecular profiles of bacterial and fungal communities. Bacterial plate counts were also obtained. Neither plant species nor defoliation affected the bacterial and fungal community structures in both the root tip and mature root regions, but there were significant differences in the bacterial and fungal community profiles between the two root regions for each plant. Prior to defoliation, there was no difference between plants for bacterial plate counts of soils from the root tip regions; however, counts were greater in the mature root region of L. perenne than T. repens. Bacterial plate counts for T. repens were higher in the root tip than the mature root region. After defoliation, there was no effect of plant type, position along the root or defoliation status on bacterial plate counts, although there were significant increases in bacterial plate counts with time. The results indicate that a general effect existed during maturation in the root regions of each plant, which had a greater impact on microbial community structure than either plant type or the effect of defoliation. In addition there were no generic consequences with regard to microbial populations in the rhizosphere as a response to plant defoliation.