Agricultural studies of GM maize and the field experimental infrastructure of ECOGEN

Within the ECOGEN project, long-term field experiments with genetically modified maize, Zea mays L. were conducted to study agro-ecological effects on the soil fauna and agro-economic implications of the technology. Here, we describe the study-sites, experimental layout and results of agro-economic relevance. Experiments were conducted during 2002–2005 in Denmark (Foulum), northern France (Varois) and the Midi-Pyrenees region of southern France (Narbons). MON810 Bacillus thuringiensis (Bt)-varieties expressing the Cry1Ab protein, and a T25 glufosinate-ammonium (Basta) tolerant variety expressing the pat-gene encoding phosphinotrinacetyl-transferase were compared with near-isogenic non-Bt varieties, and conventional maize varieties. At Foulum, the maize was harvested for silage. There were no significant differences in yield between Bt-maize and a near-isogenic non-Bt variety, while a small difference in N-concentration of dry matter was detected in 1 year in a range of a measured quality parameters. Similar yield and quality were found in ploughed and reduced tillage treatments in all varieties. At Varois, the maize was harvested at ripeness and no significant differences in grain yield between Bt-maize and near-isogenic non-Bt varieties were found. These results were expected, as only Narbons harbours significant corn-borer populations. At Narbons, the number of Sesamia and Ostrinia corn-borer larvae were significantly lower in the Bt-maize than in a near-isogenic non-Bt variety and for Sesamia even less than in conventional varieties sprayed with pesticides to control corn-borer infestation. Here, Bt-maize produced a higher grain yield and grain size than a near-isogenic non-Bt variety or allowed a significant reduction in pesticide use. Concentrations of Cry1Ab in the Bt-varieties were sufficient to effectively control corn-borer larvae. In soil, Cry1Ab was close to the limit of detection and the protein did not accumulate in the soil year on year.     

Responses by earthworms to reduced tillage in herbicide tolerant maize and Bt maize cropping systems

Genetically modified (GM) crops may affect earthworms either directly through the plant, its root exudates and litter, or indirectly through the agricultural management changes that are associated with GM plant production. In order to investigate such possible effects, we established two field studies of Bacillus thuringiensis (Bt) maize and a glufosinate ammonium tolerant maize and included a reduced tillage (RT) treatment and a conventional tillage (CT) treatment as examples of a likely concomitant change in the agricultural practise. At a French study site at Varois, (Bourgogne), a field grown with the Bt-toxin producing transgenic maize line MON810 was studied for 1 year. At a Danish study site, Foulum (Jutland), 1 year of Bt maize was followed by 2 years of herbicide tolerant (HT) maize. At the French study site, the most prominent effects observed were due to the tillage method where RT significantly reduced the earthworm populations to levels about half of CT. At the Danish study site effects of CT complied with known reduction of anecic earthworms due to this technique and likewise effects of RT were observed for endogeic earthworms. Earthworm populations were diminished with the HT crop, probably due to exposure to the herbicide Basta^® during the two consecutive autumn seasons. This study confirms the importance of including the tillage techniques and pesticide usage when evaluating the environmental effects of new agricultural technologies.     

Evaluation of effects of transgenic Bt maize on microarthropods in a European multi-site experiment

The effects of maize expressing the Bacillus thuringiensis Cry1Ab protein (Bt maize) on soil microarthropods were assessed in the field at four European locations (two in Denmark and two in France) that differ in their climatic conditions or soil properties. Each site was considered as a separate experiment, with separate statistical comparison. Effects of farming practices using Bt maize were compared with conventional farming practices using near-isogenic non-Bt maize and also (at some of the sites) other conventional varieties. Furthermore, at one field site (Foulum, Denmark), the effects of Bt crops were studied in both conventional tillage and reduced tillage contexts. At another field site (Askov, Denmark), Bt maize effects were also compared to the effects of the chemical insecticide dimethoate. Moreover, at three of the field sites (all except Narbons, France), the possibility of a localised Bt effect around the rhizosphere compared to the bulk soil was assessed by sampling within and between maize rows. There were some significant negative effects of Bt maize on microarthropods in soils with a high clay content. Significant differences of the same magnitude also occurred between different conventional varieties of maize, but the effect of dimethoate appeared clearly greater than Bt effects. It is thus debatable if the Bt maize effect is an effect of the Bt toxin or just an effect of the maize variety. Based on the results, it can be concluded that the effect of Bt maize on soil microarthropods was small and within the normal variation expected in conventional agricultural systems.     

The role of laboratory, glasshouse and field scale experiments in understanding the interactions between genetically modified crops and soil ecosystems: A review of the ECOGEN project

The interactions of genetically modified (GM) crops with soil species and ecosystems is complex, requiring both specific and broad spectrum assessments. In the ECOGEN project we undertook experiments at three scales of increasing complexity, using Bt maize expressing the Cry1Ab protein from Bacillus thuringiensis as an example. Test species were selected for laboratory-scale experiments to represent taxonomic groups that we could also monitor at glasshouse and field scales (e.g., nematodes, protozoa, micro-arthropods, earthworms, and snails). In the laboratory, single species were exposed to purified Cry1Ab protein or to Bt maize leaf powder incorporated into simplified diets under controlled conditions. In the glasshouse, multiple test species and soil microbial communities taken from ECOGEN's field sites were exposed to Bt maize plants growing under glasshouse or mesocosm conditions. In the field, evaluations were conducted on our selected indicator groups over multiple sites and growing seasons. Field evaluation included assessment of effects due to the local environment, crop type, seasonal variation and conventional crop management practice (tillage and pesticide use), which cannot be assessed in the glasshouse. No direct effects of Cry1Ab protein or Bt leaf residues were detected on our laboratory test organisms, but some significant effects were detected in the glasshouse. Total nematode and protozoan numbers increased in field soil under Bt maize relative to conventional maize, whilst microbial community structure and activity were unaffected. Field results for the abundance of nematodes and protozoa showed some negative effects of Bt maize, thus contradicting the glasshouse results. However, these negative results were specific to particular field sites and sampling times and therefore were transient. Taking the overall variation found in maize ecosystems at different sites into account, any negative effects of Bt maize at field scale were judged to be indirect and no greater than the impacts of crop type, tillage and pesticide use. Although the ECOGEN results were not predictive between the three experimental scales, we propose that they have value when used with feedback loops between the scales. This holistic approach can used to address questions raised by results from any level of experimentation and also for putting GM crop risk:benefit into context with current agricultural practices in regionally differing agro-ecosystems.     

Specific respiration rates, adenylates, and energy budgets of soil microorganisms after addition of transgenic Bt-maize straw

An arable soil was incubated with straw (stem+leaves) of two transgenic Bt-maize varieties (Novelis: event MON810 and Valmont: event Bt176) and the two corresponding near-isogenic varieties (Nobilis and Prelude). The aim was to evaluate the use of these substrates for microbial growth and maintenance in soil during early decomposition. The addition of Bt-maize straw increased CO₂ production rates and the specific respiration rates CO₂-C/microbial biomass C and CO₂-C/ATP significantly compared with the addition of non-Bt maize straw. This extra energy in the Bt-maize straw could not be used for microbial biomass or ATP and ADP production, and was lost for maintenance. In addition, increased death rates of microbial biomass occurred in the soils treated with the Bt-maize straw from day 3 to 21. Generally, most of the energy was stored in microbial biomass, whereas only 10% of energy was stored in ATP, and only 1-2% in ADP. The AEC (adenylate energy charge: (ATP+0.5×ADP)/(AMP+ADP+ATP)) was not affected by any treatment. The reasons for the lower efficiency of microbial substrate use after adding Bt-maize straw cannot be fully explained by the present experiment. However, a risk assessment has to look at the impact of transgenic plant material on soil microorganisms at different maturity stages.     

Effects of Bt-maize material on the life cycle of the land snail Cantareus aspersus

Insect resistant Bt-maize (MON 810) expresses active Cry1Ab endotoxin derived from Bacillus thuringiensis (Bt). Snails constitute non-target soil species potentially exposed to Bt-toxin through consumption of plant material and soil in fields where transgenic plants have been grown. We studied the effect of the Cry1Ab toxin on survival, growth and egg hatchability of the snail Cantareus aspersus. From the age of 4 to 88 weeks, snails were fed either powdered Bt-maize or non-Bt-maize and exposed to soil samples collected after harvesting either the Bt-maize or non-Bt-maize. We applied four treatments: non-Bt soil + non-Bt-maize (MM); Bt soil + Bt-maize (BB), non-Bt soil + Bt-maize (MB), Bt soil + non-Bt-maize (BM). Eggs laid by snails not exposed to Bt-toxin were also exposed to the two types of soils (Bt and non-Bt soil).At the end of growth (47 weeks of exposure), snails exposed to Bt-toxin in food and soil (BB) had a growth coefficient (GC) 25% lower than unexposed snails (MM). After the first period of reproduction (68 weeks) a significant difference remained for body mass GC between the BB and MM treatments. Differences in body mass were not significant at the end of exposure (88 weeks). For snails not previously exposed to Bt material, hatchability of eggs was similar in the soils tested. The outcome of the experiments indicates that, in growing snails, long-term exposure is needed to reveal an effect of Bt-maize. The hazard analysis of Bt-maize which we performed, based on a worst-case scenario, i.e. snails having no food choice, should now be complemented by other simple measurements, e.g. food intake, to understand the underlying mechanisms involved.     

Microbial response of soils with organic and conventional management history to the cultivation of Bacillus thuringiensis (Bt)-maize under climate chamber conditions

An experiment was carried out in a climate chamber to analyse if Bt-maize may cause particular changes in soils with different levels of microbial biomass and activity due to long-term management history. Among the soils selected, the ones managed organically for 30 years exhibited twice the microbial biomass and 2.6 times the dehydrogenase activity (DHA) of the soil from a field with long-term conventional maize monoculture. Soils were cultivated twice in a row with Bt-maize, its near-isogenic line and a conventional breeding line. We tested the hypotheses that (a) soil microbial biomass and activity are affected by the cultivation of Bt-maize and that (b) the influence of Bt-maize depends on the level of soil microbial biomass and activity. Shoot and root yield and shoot C-content of Bt-maize were higher than the ones of the near-isogenic line. DHA under Bt-maize was 6 % higher, and the metabolic quotient for CO₂ (qCO₂) was 9 % lower than under its near-isogenic line, giving some support to hypothesis (a). No significant interactions of the soils and the varieties used were found in this study, thus hypothesis (b) was not confirmed, and soils with different microbial biomass and activity appear to react in a similar way to the cultivation of Bt-maize.     

Seasonal variation of microbial biomass, activity, and community structure in soil under different tillage and phosphorus management practices

No-tillage systems contribute to physical, chemical and biological changes in the soil. The effects of different tillage practices and phosphorus (P) fertilization on soil microbial biomass, activity, and community structure were studied during the maize growing season in a maize–soybean rotation established for 18 years in eastern Canada. Soil samples were collected at two depths (0–10 and 10–20 cm) under mouldboard plow (MP) and no-till (NT) management and fertilized with 0, 17.5, and 35 kg P ha^?1. Results show that the duration of the growing season had a greater effect on soil microbiota properties than soil tillage or P fertilization at both soil depths. Seasonal fluctuations in soil microbial biomass carbon (SMB-C) and nitrogen (SMB-N), in dehydrogenase and alkaline phosphomonoesterase activities, and in total phospholipids fatty acid (PLFA) level, were greater under NT than MP management. The PLFA biomarkers separated treatments primarily by sampling date and secondly by tillage management, but were not significantly affected by P fertilization. The abundance of arbuscular mycorrhizal fungi (AMF; C16:1ω5) and fungi (C18:2ω6,9) was lower under NT than MP at the 10–20-cm soil depth in July. Phosphorus fertilization increased soil microbial biomass phosphorus (SMB-P) and Mehlich-3 extractable P, but had a limited impact on the other soil properties. In conclusion, soil environmental factors and tillage had a greater effect on microorganisms (biomass and activity) and community structure than P fertilization.     

Exposure and effects assessments of Bt-maize on non-target organisms (gastropods, microarthropods, mycorrhizal fungi) in microcosms

Potential differences between Bt-maize (MEB307 expressing the insecticidal Cry1Ab protein) and a near-isogenic non-Bt variety (Monumental) in their influence on the garden snail (Helix aspersa), soil microarthropods (Collembola, Actinedida, Acaridida, Gamasida and Oribatida) and mycorrhizal fungi were studied. Growing snails were caged in microcosms allowing the development of Bt or non-Bt-maize (Zea mays L.) on a sandy loam soil. After 3 months exposure, survival and growth of snails were similar in both treatments. Cry1Ab protein was detected in the Bt-maize leaves (22–42.2 μg Bt protein g⁻¹ dry wt), in the snail tissues (0.04–0.11 μg Bt-protein g⁻¹ dry wt) and in their faeces (0.034–5 μg Bt-protein g⁻¹ dry wt). Total soil microarthropod abundance and diversity were similar between control (non-Bt-maize) and the genetically modified (GM) Bt-maize microcosms. The mycorrhizal colonization of roots did not differ between Bt and non-Bt-maize (frequency of mycorrhizal roots was 88.7% and 83.3% respectively). The mycorrhizal infectivity of soils, expressed as MI₅₀ (minimum soil dry weight required to colonize 50% of plants) was measured using red clover. MI₅₀ was similar for soils where Bt or non-Bt-maize was cultivated for 4 months. The detection of Cry1Ab protein in the viscera and faeces of H. aspersa exposed to Bt-maize indicates that snails contribute to the transfer of the Bt-protein from plant to soil or snail predators. This may constitute an alternative route of exposure for Bt-protein in soil, but this was without a negative influence on mycorrhizal fungi or microarthropods. Results showed that Bt-maize was not toxic for the selected non-target species exposed for 3 or 4 months. The microcosms and analyses used in this study represent new methods for assessing effects of chronic exposure to GM plants of several diverse, yet ecologically and temporally associated species. As the soil organisms we studied can also be used in standardized ecotoxicological tests (XP X31-205-2 for mycorrhizal fungi, ISO 11267 for Collembola and ISO 15952 for snails), microcosm exposures represent a way to link laboratory and field methods for the ecotoxicological evaluation of GM plants.     

Hierarchical classification of environmental factors and agricultural practices affecting soil fauna under cropping systems using Bt maize

The population dynamics of soil organisms under agricultural field conditions are influenced by many factors, such as pedology and climate, but also farming practices such as crop type, tillage and the use of pesticides. To assess the real effects of farming practices on soil organisms it is necessary to rank the influence of all of these parameters. Bt maize (Zea mays L.), as a crop recently introduced into farming practices, is a genetically modified maize with the Cry1Ab gene which produces a protein toxic to specific lepidopteran insect pests. To assess the effects of Bt maize on non-target soil organisms, we conducted research at a field site in Foulum (Denmark) with a loamy sand soil containing 6.4% organic matter. The study focused on populations of springtails (Collembola) and earthworms (Oligochaeta) from samples taken at the beginning and at the end of the maize crop-growing season during 2 consecutive years. Farming practices, soil parameters, the biological structure of soil communities, and the type and age of the crop at the time of sampling, were used as attributes to predict the total abundance of springtails and biomass of earthworms in general and the abundance or biomass for specific functional groups (epigeic, endogeic and anecic groups for earthworms, and eu-, eu to hemi-, hemi-, hemi to epi- and epiedaphic groups for Collembola). Predictive models were built with data mining tools, such as regression trees that predict the value of a dependent variable from a set of independent variables. Regression trees were constructed with the data mining system M5′. The models were evaluated by qualitative and quantitative measures of performance and two models were selected for further interpretation: anecic worms and hemi-epiedaphic Collembola. The anecic worms (r²=0.83) showed preferences for less clay and more silt soil with medium pH but were not influenced directly by farming practices. The biomass of earthworms was greater in early autumn than in spring or late autumn. Biomass of hemi-epiedaphic Collembola (r²=0.59) increased at the end of the maize growing season, while higher organic matter content and pH tended to increase their biomass in spring. Greater abundance of Collembola was also noted in early autumn if the crop was non-Bt maize. The models assessed by this research did not find any effects of the Bt maize cropping system on functional groups of soil fauna.     

Soil Microbial Community Composition and Diversity in the Rhizosphere of a Chinese Medicinal Plant

Rehmannia glutinosa is an important medicinal plant, but there is a serious problem of decreasing productivity with its continuous cropping on the same land. We hypothesize some relationships between this problem and the disturbed soil ecosystem. In this work, two community‐based microbiological measurements, community‐level physiological profiling (CLPP) using Biolog sole carbon (C) source utilization tests and phospholipid ester–linked fatty acid (PLFA) profiles, were used to evaluate soil microbial community function and composition of different R. glutinosa cropping soils. Field investigation showed that the problems with continuous cropping occurred not only in 2‐year continuous fields but also in 5‐year rotation fields. Soil basal respiration and metabolic quotient were significantly greater in R. glutinosa cropping soils than in the noncropping controls. In contrast, the Shannon index from the Biolog data set was lower in R. glutinosa cropping soils. Both CLPP‐ and PLFA‐based principal component analyses (PCA) showed distinct groupings of soil microbial communities in R. glutinosa rhizosphere, and 11 PLFAs representing different microbes were identified from the principal component scores of PLFAs. Among these, an abundance of PLFA 18:2ω6,9, which is a biomarker of soil fungi, was significantly higher in R. glutinosa cropping soils than control soils. These results suggest an alteration of soil microbial community following R. glutinosa cropping, and this might be an important reason for the constraints associated with continuous cropping.     

Choice of methods for soil microbial community analysis: PLFA maximizes power compared to CLPP and PCR-based approaches

Polyphasic studies that used phospholipid fatty acid analysis (PLFA) in conjunction with community level physiological profiling (CLPP) or PCR-based molecular methods were analyzed in order to evaluate the power of each strategy to detect treatment effects on soil microbial community structure (MCS). We found no studies where CLPP or PCR-based methods differentiated treatments that were not also differentiated by PLFA. In 14 of 32 studies (44%), PLFA differentiated treatments that were not resolved by CLPP analysis. In 5 of 25 studies (20%), PLFA differentiated treatments that were not resolved by PCR-based methods. We discuss PLFA, CLPP, and PCR-based methods with respect to power to discriminate change in MCS versus potential for characterization of underlying population level changes.     

Controls on soil nitrogen cycling and microbial community composition across land use and incubation temperature

We conducted a laboratory incubation of forest (Scots pine (Pinus sylvestris) or beech (Fagus sylvatica)), grassland (Trifolium repens/Lolium perenne) and arable (organic and conventional) soils at 5 and 25 °C. We aimed to clarify the mechanisms of short-term (2-weeks) nitrogen (N) cycling processes and microbial community composition in relation to dissolved organic carbon (DOC) and N (DON) availability and selected soil properties. N cycling was measured by ¹⁵N pool dilution and microbial community composition by denaturing gradient gel electrophoresis (DGGE), phospholipid fatty acid (PLFA) and community level physiological profiles (CLPP). Soil DOC increased in the order of arable<grassland<forest soil while DON and gross N fluxes increased in the order of forest<arable<grassland soil; land use had no affect on respiration rate. Soil DOC was lower, while respiration, DON and gross N fluxes were higher at 25 than 5 °C. Gross N fluxes, respiration and bacterial biomass were all positively correlated with each other. Gross N fluxes were positively correlated with pH and DON, and negatively correlated with organic matter, fungal biomass, DOC and DOC/DON ratio. Respiration rate was positively correlated with bacterial biomass, DON and DOC/DON ratio. Multiple linear modelling indicated that soil pH, organic matter, bacterial biomass, DON and DOC/DON ratio were important in predicting gross N mineralization. Incubation temperature, pH and total-C were important in predicting gross nitrification, while gross N mineralization, gross nitrification and pH were important in predicting gross N immobilization. Permutation multivariate analysis of variance indicated that DGGE, CLPP and PLFA profiles were all significantly (P<0.05) affected by land use and incubation temperature. Multivariate regressions indicated that incubation temperature, pH and organic matter content were important in predicting DGGE, CLPP and PLFA profiles. PLFA and CLPP were also related to DON, DOC, ammonium and nitrate contents. Canonical correlation analysis showed that PLFA and CLPP were related to differences in the rates of gross N mineralization, gross nitrification and soil respiration. Our study indicates that vegetation type and/or management practices which control soil pH and mediate dissolved organic matter availability were important predictors of gross N fluxes and microbial composition in this short-term experiment.     

Long-term no-till management affects microbial biomass but not community composition in Canadian prairie agroecosytems

No-till (NT) management greatly reduces soil physical disturbance and can result in a stratification of nutrients and organic matter in the soil profile due to the retention of crop residues on the soil surface potentially affecting the dynamics of microbial interactions in the soil. Microbial abundance and diversity can be used to assess the relative impact of management on the long-term sustainability of cropping systems. The objective of this study was to assess the impact of long-term NT vs. conventional tillage (CT) management on soil microbial community structure at four different sites on the Canadian prairies using phospholipid fatty acid analysis (PLFA) and DNA fingerprinting. Analysis of 16S and 18S rDNA using denaturing gradient gel electrophoresis revealed high inherent variability within bacterial and fungal community fingerprints among replicate field plots. Differences in bacterial and fungal phylogeny were related to depth in the soil profile but not to tillage management. Abundance of individual PLFA biomarkers were 7 to 86% greater in NT surface soils (0- to 5-cm depth), except at the Ellerslie site in 2005 where biomass was greater in CT. Responses at the 5- to 10-cm and 10- to 15-cm depths were more varied, in some cases with greater biomass in CT than NT soils. Ordination analysis of PLFA profiles showed clear community separation with depth but not tillage. Physiological stress biomarkers were correlated with simple measures of nutrient concentration and indicated that resource availability was likely the main factor determining community structure. It was concluded that tillage disturbance was not an overriding factor in determining microbial community composition in the long-term NT and CT soils studied. Further study of the interaction of cropping frequency with tillage management is needed to understand the effects of tillage disturbance on microbial turnover of plant derived residues.     

Effects of conventionally bred and <Emphasis Type="BoldItalic">Bacillus thuringiensis</Emphasis> (<Emphasis Type="BoldItalic">Bt</Emphasis>) maize varieties on soil microbial biomass and activity

Genetically modified (GM) maize containing genes from the soil bacterium Bacillus thuringiensis (Bt) was cultivated on 29% of the total maize production area worldwide in 2009. Most studies to date compare Bt-maize varieties with their near isogenic lines; however, there is little information on the variability of conventional maize breeding lines and how the effects of Bt varieties are ranked within. In our study on the potential risks of Bt-maize varieties, we analyzed tissue quality and compared the effects of ten conventional and GM maize varieties on soil microbiological properties in a replicated climate chamber experiment. All maize varieties were cultivated twice in the same soil microcosm. Shoot yields and soluble C in leaf tissue of Bt varieties were higher than the ones of non-Bt. Soil dehydrogenase activity was reduced by 5% under Bt varieties compared to non-Bt, while most of the other soil microbial properties (soil microbial biomass, basal respiration) showed no significant differences between Bt and non-Bt varieties. The leaves and roots of one Bt variety were decomposed to a greater extent than the ones of its near isogenic line; the conventional breeding lines also showed higher values. Changes in crop and soil parameters were found when comparing the first and the second crops, but the effects of repeated cropping were the same for all tested varieties. For the studied parameters, the variation among non-Bt-maize varieties was similar to the difference between Bt and non-Bt varieties.     

Effects of sulfamethoxazole on soil microbial communities after adding substrate

The effect of the antibiotic sulfamethoxazole (SMX) on soil bacteria was studied using two methods (leucine incorporation and Biolog plates) of estimating pollution-induced community tolerance (PICT). SMX was added to an agricultural soil in a microcosm setup. The addition of different substrates (manure and alfalfa), and a non-amended soil, were also studied over 5 weeks. PICT measurements were validated by comparison with other measurements. Community structure was assessed using phospholipid fatty acid (PLFA) analysis and community-level physiological profiling (CLPP), and bacterial growth was estimated using leucine incorporation. Increased PICT was found at SMX concentrations of 20 and 500 mg SMX kg⁻¹ soil in samples containing manure and alfalfa, and at 500 mg SMX kg⁻¹ soil in non-amended soil (only concentration tested) using leucine incorporation. No effect was seen at 1 mg SMX kg⁻¹ soil. It was not necessary to add any substrate to increase the microbial activity in order to detect the effects of a bacteriostatic toxicant such as SMX when using measures based on bacterial growth. Direct inhibition of bacterial growth 2 days after SMX addition was correlated to PICT. No major changes in PICT due to SMX addition were found when using Biolog plates. However, there was a tendency towards increased PICT at the higher SMX concentrations in the manure-amended soil. Thus, different methods of detecting PICT have different sensitivities in detecting the toxic effects of SMX. The effects of substrate amendment were reflected by changes in the microbial community, estimated using both PLFA and CLPP. SMX was found to have a clear effect at the two highest levels of SMX in the manure- and alfalfa-amended soils, with an increase in fungal and a decrease in bacterial PLFAs. Little difference in the PLFA composition was found in the non-amended soil. CLPP was only affected at the highest SMX concentration. Although different variables showed different sensitivities to the effects of SMX, the results were consistent with an initial decrease in bacterial growth rates of sensitive species, which eventually transformed into more tolerant species, altering the community composition.     

Seasonal Patterns of Microbial Community Structure and Enzyme Activities in Coastal Saline Soils of Perennial Halophytes

Perennial halophytes are known to be one of the most influential parameters in coastal ecosystem affecting ecosystem processes. The aim of this study was to investigate the changes in soil microbial community structure and enzyme activities in different halophyte‐covered soils (Arthrocnemum indicum , Aeluropus lagopoides , Heleochloa setulosa and Suaeda nudiflora ) with control soil (un‐vegetated) that were collected in three seasons (rainy, winter and summer) from intertidal coastal soils of Gujarat, India. Soil microbial community structure was assessed using phospholipid fatty acid (PLFA) profiling. Halophytes influenced significantly soil micro‐environment by exerting effects on the soil chemical characteristics, enzyme activities and microbial community structure. The activities of β‐glucosidase, urease and alkaline phosphatase were significantly higher in halophyte‐covered soils than in control soil. Among four halophyte‐covered soils, the highest amounts of total, bacterial, actinomycetes and fungal PLFAs were observed in Arthrocnemum soil. The concentrations of total, bacterial, actinomycetes and fungal PLFAs were also significantly higher in summer and winter seasons than in rainy season, whereas enzyme activities also vary with seasons. The non‐metric multidimensional scaling analysis PLFA profiling revealed that the structure of microbial community significantly differed in all seasons as well as between control and halophyte‐covered soils. These shifts in microbial community were due to the higher abundance of Gram‐positive, total bacterial and actinomycetes PLFAs in summer and winter seasons than in rainy season, whereas abundance of fungal biomarker was significantly higher in rainy season than in other seasons. Among halophytes, significantly higher abundance of Gram‐positive, Gram‐negative and total bacteria was observed in Arthrocnemum , Heleochloa and Suaeda whereas the lowest in control soil. Halophytes exhibited improved soil microbial activities, which is important for healthy ecosystem. Copyright © 2017 John Wiley & Sons, Ltd.     

Short-term effect of tillage and crop rotation on microbial community structure and enzyme activities of a clay loam soil

A field study was carried out to analyze the short-term (2 years) effect of tillage and crop rotation on microbial community structure and enzyme activities of a clay loam soil. The experimental design was a split-plot arrangement of treatments, consisting of two tillage treatments—ridge tillage (RT) and no-tillage (NT)—in combination with two crop rotation treatments—corn (Zea mays L.) monoculture and a 2-year corn-soybean (Glycine max L.) rotation. Phospholipid fatty acid (PLFA) profiles were used to assess soil microbial community structure. No-tillage resulted in significantly higher total PLFAs compared to the RT treatment, which was accompanied by higher activities of protease, β-glucosaminidase, and β-glucosidase. This suggests a close link between soil microbial communities and enzyme activities in response to tillage. The increase of total microbial lipid biomass in the NT soils was due to the increase in both fungal and bacterial PLFAs. Crop rotation had little effect on soil bacterial communities and enzyme activities, but it significantly influenced soil fungal communities, particularly arbuscular mycorrhizal fungi. Soils under monoculture corn had higher fungal biomass than soils under corn-soybean rotation regardless of tillage treatment.     

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.     

土壤微生物群落结构与多样性对农田集约化管理程度降低的响应

Using a scheme of agricultural fields with progressively less intensive management （deintensification）, different management practices in six agroecosystems located near Goldsboro, NC, USA were tested in a large-scale experiment, including two cash-grain cropping systems employing either tillage （CT） or no-tillage （NT）, an organic farming system （OR）, an integrated cropping system with animals （IN）, a successional field （SU）, and a plantation woodlot （WO）. Microbial phospholipid fatty acid （PLFA） profiles and substrate utilization patterns （BIOLOG ECO plates） were measured to examine the effects of deintensification on the structure and diversity of soil microbial communities. Principle component analyses of PLFA and BIOLOG data showed that the microbial community structure diverged among the soils of the six systems.Lower microbial diversity was found in lowly managed ecosystem than that in intensive and moderately managed agroecosystems, and both fungal contribution to the total identified PLFAs and the ratio of microbial biomass C/N increased along with agricultural deintensification. Significantly higher ratios of C/N （P 〈 0.05） were found in the WO and SU systems, and for fungal/bacterial PLFAs in the WO system （P 〈 0.05）. There were also significant decreases （P 〈 0.05） along with agricultural deintensification for contributions of total bacterial and gram positive （G＋） bacterial PLFAs.Agricultural deintensification could facilitate the development of microbial communities that favor soil fungi over bacteria.