Methanotrophic communities in a landfill cover soil as revealed by [13C] PLFAs and respiratory quinones: Impact of high methane addition and landfill leachate irrigation

The soil microbial communities of a landfill cover substrate, which was treated with landfill gas (100 l CH₄ m^?2 d^?1) and landfill leachate for 1.5 years, were investigated by phospholipid fatty acid (PLFA), ergosterol and respiratory quinone analyses. The natural ¹³C depletion of methane was used to assess the activity of methanotrophs and carbon turnover in the soil system. Under methane addition, the soil microbial community was dominated by PLFAs (14:0 and 16:1 isomers) and quinones (ubiquinone-8 and 18-methylene-ubiquinone-8) related to type I methanotrophs, and 18:1 PLFAs contained in type II methanotrophs. While type I methanotrophic PLFAs were ¹³C depleted, i.e. type I methanotrophs were actively oxidising and assimilating methane, ¹³C depletion of 18:1 PLFAs was low and inconsistent with their abundance. This, possibly reflects isotopic discrimination, assimilation of carbon derived from type I methanotrophs and a high contribution of non-methanotrophic bacteria to the 18:1 isomers. Landfill leachate irrigation caused the methanotrophic community to shift closer to the soil surface. It also decreased 18:1 PLFAs, while type I methanotrophs were probably stimulated. Gram positive bacteria, but not fungi, were also ¹³C depleted and consequently involved in the secondary turnover of carbon originating from methanotrophic bacteria. Cy17:0 PLFA was ¹³C depleted in deep soil layers, indicating anaerobic methane oxidation.     

Effect of soil moisture and bovine urine on microbial stress

While many studies have examined the cycling of urinary nutrients, few have focused on the effects ruminant urine might have on the soil microbial community. Urine application can cause microbial communities to become stressed, potentially changing community composition and microbial function with subsequent effects on nutrient dynamics. Identification of the factors that stress microbes may assist in explaining ruminant urine effects on nutrient cycling. In this laboratory study bovine urine, with either a high (15.0 g K⁺ l^?1) or low (10.4 g K⁺ l^?1) salt concentration, was added to repacked soil cores maintained at high or low soil moisture contents (70 or 35% water-filled pore space, respectively). Control cores did not receive urine. Microbial stress was measured using phospholipid fatty acid (PLFA) biomarker ratios. Urine addition increased stress as indicated by a decrease in the iso15:0/anteiso15:0 PLFA ratio from >1.35 to <0.95 in both wet and dry soils and by an increase in the 18:1ω9trans/18:1ω9cis PLFA ratio from 1.4 to 1.9 from day 8 onwards in wet soils. Higher stress was indicated by a lower Gram-positive/Gram-negative PLFA ratio in the urine treatments than in the control treatments on day 29 and this may have been a response to the reduction in substrate availability as the experiment progressed. The PLFA biomarkers showed that the salt treatments did not induce stress. Stress induced by urine addition and wet soil treatments was also indicated by principal component analyses and the metabolic quotient for CO₂, respectively. Thus microbial stress was induced by both urine addition and high soil moisture content, but not specifically by increasing the urinary salt concentration.     

Influence of carbon amendments on soil denitrifier abundance in soil microcosms

It is known that carbon (C) amendments increase microbial activity in anoxic soil microcosm studies, however the effects on abundance of total and denitrifier bacterial communities is uncertain. Quantitative PCR was used to target the 16S rRNA gene for the total bacterial community, the nosZ functional gene to reflect a broad denitrifier community, and functional genes from narrow denitrifier communities represented by Pseudomonas mandelii and related species (cnorB_P) and Bosea/Bradyrhizobium/Ensifer spp. (cnorB_B). Repacked soil cores were amended with varying amounts of glucose and red clover plant tissue (0–1000 mg C kg^? 1 of soil) and incubated for 96 h. Carbon amendment significantly increased respiration as measured by cumulative CO₂ emissions. Inputs of red clover or glucose at 1000 mg C kg^? 1 of soil caused increased abundance in the total bacteria under the conditions used. There was about an approximate 2-fold increase in the abundance of bacteria bearing the nosZ gene, but only in treatments receiving 500 or 1000 mg C kg^? 1 of soil of glucose or red clover, respectively. Additions of ≥ 500 mg C kg^? 1 soil of red clover and ≥ 250 mg C kg^? 1 of glucose increased cnorB_P-gene bearing denitrifiers. Changes in abundance of the targeted communities were related to C availability in soil, as indicated by soil respiration, regardless of C source. Applications of C amendments at rates that would occur in agricultural soils not only increase microbial activity, but can also induce changes in abundance of total bacterial and denitrifier communities in studies of anoxic soil microcosms.     

Effects of dazomet on soil organisms and recolonisation of fumigated soil

Fumigation is a common practice to control soil pathogens, but little is known about the impacts of fumigation on other soil biota groups. The purpose of this study was to investigate the effects of fumigation on soil biota, including microorganisms, nematodes, and microarthropods. Bacteria were the most resistant group and some survived following treatment with 2000 mg kg⁻¹ dazomet. Some soil fungi survived 100 mg kg⁻¹ dazomet, although they were mainly Trichoderma. The fungi pathogenic to ginseng were all killed at 100 mg kg⁻¹, and showed both inter- and intra-species variation with respect to dazomet susceptibility. Among the nematodes, Aphelenchus was relatively resistant. The results suggested that susceptibility of soil organisms to dazomet differs between species, and that tolerant organisms may engage in recolonisation. In microcosm experiments, the microbial biomass and community were assessed using phospholipid fatty acid (PLFA) analysis while recolonisation of soil organisms was controlled by mesh size. The bacterial PLFA levels were changed little after fumigation, whereas the fungal PLFA levels gradually increased after fumigation. Principal analysis of the PLFA levels and the ratio of gram-negative to gram-positive bacteria showed that fumigation altered the microbial community. The number of nematodes did not recover even at 12 weeks after fumigation. The increased Collembolan numbers suggest that fumigated soil could be recolonised by specific organisms that have adapted to the conditions. In field experiments, we tested the ability of organic materials to enhance the recolonisation of fumigated soil by soil organisms. Bean powder and rice bran increased the microbial PLFA levels and nematode numbers at 6 weeks and 12 weeks after treatment, and the abundance of nematodes continued to increase 42 weeks after fumigation. The abundance of microarthropods was only slightly affected by the presence of the organic materials. We suggest that treating fumigated soils with organic materials is an effective technique to promote soil organism numbers. In addition, Trichoderma was observed to be relatively resistant to fumigation, and therefore, we propose that the fumigation effect can be improved by using a combination of resistant Trichoderma and dazomet.     

Soil organic matter in soil depth profiles: Distinct carbon preferences of microbial groups during carbon transformation

This study investigates how carbon sources of soil microbial communities vary with soil depth. Microbial phospholipid fatty acids (PLFA) were extracted from 0–20, 20–40 and 40–60 cm depth intervals from agricultural soils and analysed for their stable carbon isotopes (δ¹³C values). The soils had been subjected to a vegetation change from C3 (δ¹³C≈?29.3‰) to C4 plants (δ¹³C≈?12.5‰) 40 years previously, which allowed us to trace the carbon flow from plant-derived input (litter, roots, and root exudates) into microbial PLFA. While bulk soil organic matter (SOM) reflected ≈12% of the C4-derived carbon in top soil (0–20 cm) and 3% in deeper soil (40–60 cm), the PLFA had a much higher contribution of C4 carbon of about 64% in 0–20 cm and 34% in 40–60 cm. This implies a much faster turnover time of carbon in the microbial biomass compared to bulk SOM. The isotopic signature of bulk SOM and PLFA from C4 cultivated soil decreases with increasing soil depth (?23.7‰ to ?25.0‰ for bulk SOM and ?18.3‰ to ?23.3‰ for PLFA), which demonstrates decreasing influence of the isotopic signature of the new C4 vegetation with soil depth. In terms of soil microbial carbon sources this clearly shows a high percentage of C4 labelled and thus young plant carbon as microbial carbon source in topsoils. With increasing soil depth this percentage decreases and SOM is increasingly used as microbial carbon source. Among all PLFA that were associated to different microbial groups it could be observed that (a) depended on availability, Gram-negative and Gram-positive bacteria prefer plant-derived carbon as carbon source, however, (b) Gram-positive bacteria use more SOM-derived carbon sources while Gram-negative bacteria use more plant biomass. This tendency was observed in all three-depth intervals. However, our results also show that microorganisms maintain their preferred carbon sources independent on soil depth with an isotopic shift of 3–4‰ from 0–20 to 40–60 cm soil depth.     

Formation and use of microbial residues after adding sugarcane sucrose to a heated soil devoid of soil organic matter

A 67-day incubation experiment was carried out with a soil initially devoid of any organic matter due to heating, which was amended with sugarcane sucrose (C₄-sucrose with a δ¹³C value of ?10.5‰), inorganic N and an inoculum for recolonisation and subsequently at day 33 with C₃-cellulose (δ¹³C value of ?23.4‰). In this soil, all organic matter is in the microbial biomass or in freshly formed residues, which makes it possible to analyse more clearly the role of microbial residues for decomposition of N-poor substrates. The average δ¹³C value over the whole incubation period was ?10.7‰ in soil total C in the treatments without C₃-cellulose addition. In the CO₂ evolved, the δ¹³C values decreased from ?13.4‰ to ?15.4‰ during incubation. In the microbial biomass, the δ¹³C values increased from ?11.5‰ to ?10.1‰ at days 33 and 38. At day 67, 36% of the C₄-sucrose was left in the treatment without a second amendment. The addition of C₃-cellulose resulted in a further 7% decrease, but 4% of the C₃-cellulose was lost during the second incubation period. Total microbial biomass C declined from 200 μg g^?1 soil at day 5 to 70 μg g^?1 soil at day 67. Fungal ergosterol increased to 1.5 μg g^?1 soil at day 12 and declined more or less linearly to 0.4 μg g^?1 soil at day 67. Bacterial muramic acid declined from a maximum of 35 μg g^?1 soil at day 5 to a constant level of around 16 μg g^?1 soil. Glucosamine showed a peak value at day 12. Galactosamine remained constant throughout the incubation. The fungal C/bacterial C ratio increased more or less linearly from 0.38 at day 5 to 1.1 at day 67 indicating a shift in the microbial community from bacteria to fungi during the incubation. The addition of C₃-cellulose led to a small increase in C₃-derived microbial biomass C, but to a strong increase in C₄-derived microbial biomass C. At days 45 and 67, the addition of N-free C₃-cellulose significantly decreased the C/N ratio of the microbial residues, suggesting that this fraction did not serve as an N-source, but as an energy source.     

Microbial utilization and mineralization of [14C]glucose added in six orders of concentration to soil

The substrate availability for microbial biomass (MB) in soil is crucial for microbial biomass activity. Due to the fast microbial decomposition and the permanent production of easily available substrates in the rooted top soil mainly by plants during photosynthesis, easily available substrates make a very important contribution to many soil processes including soil organic matter turnover, microbial growth and maintenance, aggregate stabilization, CO₂ efflux, etc. Naturally occurring concentrations of easily available substances are low, ranging from 0.1 μM in soils free of roots and plant residues to 80 mM in root cells. We investigated the effect of adding ¹⁴C-labelled glucose at concentrations spanning the 6 orders of magnitude naturally occurring concentrations on glucose uptake and mineralization by microbial biomass. A positive correlation between the amount of added glucose and its portion mineralized to CO₂ was observed: After 22 days, from 26% to 44% of the added 0.0009 to 257 μg glucose C g^?1 soil was mineralized. The dependence of glucose mineralization on its amount can be described with two functions. Up to 2.6 μg glucose C g^?1 soil (corresponds to 0.78% of initial microbial biomass C), glucose mineralization increased with the slope of 1.8% more mineralized glucose C per 1 μg C added, accompanied by an increasing incorporation of glucose C into MB. An increased spatial contact between micro-organisms and glucose molecules with increasing concentration may be responsible for this fast increase in mineralization rates (at glucose additions <2.6 μg C g^?1). At glucose additions higher than 2.6 μg C g^?1 soil, however, the increase of the glucose mineralization per 1 μg added glucose was much smaller as at additions below 2.6 μg C g^?1 soil and was accompanied by decreasing portions of glucose ¹⁴C incorporated into microbial biomass. This supports the hypothesis of decreasing efficiency of glucose utilization by MB in response to increased substrate availability in the range 2.6–257 μg C g^?1 (=0.78–78% of microbial biomass C). At low glucose amounts, it was mainly stored in a chloroform-labile microbial pool, but not readily mineralized to CO₂. The addition of 257 μg glucose C g^?1 soil (0.78 μg C glucose μg^?1 C micro-organisms) caused a lag phase in mineralization of 19 h, indicating that glucose mineralization was not limited by the substrate availability but by the amount of MB which is typical for 2nd order kinetics.     

Impact of the antibiotic sulfadiazine and pig manure on the microbial community structure in agricultural soils

Large amounts of veterinary antibiotics enter soil via manure of treated animals. The effects on soil microbial community structure are not well investigated. In particular, the impact of antibiotics in the presence of manure is poorly understood. In this study, two agricultural soils, a sandy Cambisol (KS) and a loamy Luvisol (ML), were spiked with manure and sulfadiazine (SDZ; 0, 10 and 100 μg g^?1) and incubated for 1, 4, 32 and 61 days. Untreated controls received only water. The microbial community structure was characterised by investigating phospholipid fatty acids (PLFA) and using PCR–denaturing gradient gel electrophoresis (DGGE) of 16S rDNA. The total concentration of PLFA increased with addition of manure and was reduced by both SDZ concentrations at incubation times >4 days. The SDZ addition decreased the bacteria:fungi ratio. The largest stress level, measured as ratio of PLFA (cyc17:0 + cyc19:0)/(16:1ω7c + 18:1ω7c), was found for the controls, followed by the manure treatments and the SDZ treatments. A discriminant analysis of the PLFA clearly separated treatments and incubation times. Both soils differed in total PLFA concentrations and Gram^?:Gram⁺ ratios, but showed similar changes in PLFA pattern upon soil treatment. Effects of manure and SDZ on the bacterial community structure were also revealed by DGGE analysis. Effects on pseudomonads and β-proteobacteria were less pronounced. While community structure remained altered even after two months, the extractable concentrations of SDZ decreased exponentially and the remaining solution concentrations after 32 days were ≤27% of the spiking concentration. Our results demonstrate that a single addition of SDZ has prolonged effects on the microbial community structure in soils.     

Accounting for variability in soil microbial communities of temperate upland grassland ecosystems

This study aimed to determine the factors which regulate soil microbial community organisation and function in temperate upland grassland ecosystems. Soil microbial biomass (C_mic), activity (respiration and potential carbon utilisation) and community structure (phospholipid fatty acid (PLFA) analysis, culturing and community level physiological profiles (CLPP) (Biolog^®)) were measured across a gradient of three upland grassland types; Festuca–Agrostis–Galium grassland (unimproved grassland, National Vegetation Classification (NVC) — U4a); Festuca–Agrostis–Galium grassland, Holcus–Trifolium sub-community (semi-improved grassland, NVC — U4b); Lolium–Cynosurus grassland (improved grassland, NVC — MG6) at three sites in different biogeographic areas of the UK over a period of 1 year. Variation in C_mic was mainly due to grassland type and site (accounting for 55% variance, v, in the data). C_mic was significantly (P<0.001) high in the unimproved grassland at Torridon (237.4 g C m⁻² cf. 81.2 g C m⁻² in semi- and 63.8 g C m⁻² in improved grasslands) and Sourhope (114.6 g C m⁻² cf. in 44.8 g C m⁻² semi- and 68.3 g C m⁻² in improved grasslands) and semi-improved grassland at Abergwyngregyn (76.0 g C m⁻² cf. 41.7 g C m⁻² in un- and 58.3 g C m⁻² in improved grasslands). C_mic showed little temporal variation (v=3.7%). Soil microbial activity, measured as basal respiration was also mainly affected by grassland type and site (n=32%). In contrast to C_mic, respiration was significantly (P<0.001) high in the improved grassland at Sourhope (263.4 l h⁻¹m⁻² cf. 79.6 l h⁻¹m⁻² in semi- and 203.9 l h⁻¹m⁻² unimproved grasslands) and Abergwyngregyn (198.8 l h⁻¹m⁻² cf. 173.7 l h⁻¹m⁻² in semi- and 88.2 l h⁻¹m⁻² unimproved grasslands). Microbial activity, measured as potential carbon utilisation, agreed with the respiration measurements and was significantly (P<0.001) high in the improved grassland at all three sites (A₅₉₀ 0.14 cf. 0.09 in semi- and 0.07 in unimproved grassland). However, date of sampling also had a significant (P<0.001) impact on C utilisation potential (v=24.7%) with samples from April 1997 having highest activity at all three sites. Variation in microbial community structure was due, predominantly, to grassland type (average v=23.6% for bacterial and fungal numbers and PLFA) and date of sampling (average v=39.7% for bacterial and fungal numbers and PLFA). Numbers of culturable bacteria and bacterial PLFA were significantly (P<0.001) high in the improved grassland at all three sites. Fungal populations were significantly (P<0.01) high in the unimproved grassland at Sourhope and Abergwyngregyn. The results demonstrate a shift in soil microbial community structure from one favouring fungi to one favouring bacteria as grassland improvement increased. Numbers of bacteria and fungi were also significantly (P<0.001) higher in August than any other sampling date. Canonical variate analysis (CVA) of the carbon utilisation data significantly (P<0.05) differentiated microbial communities from the three grassland types, mainly due to greater utilisation of sugars and citric acid in the improved grasslands compared to greater utilisation of carboxylic acids, phenolics and neutral amino acids in the unimproved grasslands, possibly reflecting substrate availability in these grasslands. Differences in C_mic, activity and community structure between grassland types were robust over time. In addition, broad scale measures of microbial growth and activity (C_mic and respiration) showed little temporal variation compared to measures of soil microbial community structure, which varied quantitatively with respect to environmental variables (temperature, moisture) and plant productivity, hence substrate supply.     

Effects of vegetation on soil microbial C,N, and P dynamics in a tropical forest and savanna of Central Africa

The forest–savanna transition zone is widely distributed on nutrient-poor oxisols in Central Africa. To reveal and compare the nutrient cycle in relation to soil microbes for forest and savanna vegetation in this area, we evaluated seasonal fluctuations in microbial biomass carbon (MBC), nitrogen (MBN), and phosphorus (MBP) for 13 months as well as soil moisture, temperature, soil pH levels, and nutrients for both vegetation types in eastern Cameroon. Soil pH was significantly lower in forest (4.3) than in savanna (5.6), and soil N availability was greater in forest (87.1 mg N kg⁻¹ soil) than in savanna (32.9 mg N kg⁻¹ soil). We found a significant positive correlation between soil moisture and MBP in forest, indicating the importance of organic P mineralization for MBP, whereas in savanna, we found a significant positive correlation between soil N availability and MBP, indicating N limitation for MBP. These results suggest that for soil microbes, forest is an N-saturated and P-limited ecosystem, whereas savanna is an N-limited ecosystem. Additionally, we observed a significantly lower MBN and larger MB C:N ratio in forest (50.7 mg N kg⁻¹ soil and 8.6, respectively) than in savanna (60.0 mg N kg⁻¹ soil and 6.5, respectively) during the experimental period, despite the rich soil N condition in forest. This may be due to the significantly lower soil pH in forest, which influences the different soil microbial communities (fungi-to-bacteria ratio) in forest versus savanna, and therefore, our results indicate that, in terms of microbial N dynamics, soil pH rather than soil substrate conditions controls the soil microbial communities in this area. Further studies should be focused on soil microbial community, such as PLFA, which was not evaluated in the present study.     

Evaluating effects of sewage sludge and household compost on soil physical,chemical and microbiological properties

Recycling of organic wastes within agriculture may help maintain soil fertility via effects on physical, chemical and biological properties. Efficient use, however, requires an individual assessment of waste products, and effects should be compared with natural variations due to climate and soil type. An 11-month incubation experiment was conducted between April 1998 and March 1999, in which a sandy loam without or with anerobically digested sewage sludge (4.2 t dry matter (DM) ha⁻¹) or household compost (17 t DM ha⁻¹) was incubated under constant laboratory conditions at 10 °C, as well as in the field. The following properties were monitored: wet-stability of soil aggregates, clay dispersibility, hot-water extractable carbohydrates, resin-extractable P_i, inorganic N, biomass C and N, PLFA profiles, FDA hydrolysis activity, β-glucosidase activity and CO₂ evolution. In general, effects of waste amendment were positive, but moderate compared to the dynamics observed in unamended soil, and mainly occurred in the first several weeks after amendment. The temporal dynamics of inorganic N, FDA hydrolysis activity, biomass C and PLFA composition appeared to be faster under the fluctuating climatic conditions in the field. To evaluate accumulated effects of repeated waste applications, soil was also sampled from a field trial, in which the sewage sludge and household compost had been applied at the same rates as in the incubation study for three consecutive years. Sampling took place after the final harvest, i.e. 5 months after the final waste application. Compost amendment had increased potentially mineralizable N by a factor of 1.8, and sludge amendment had increased the amount of resin-extractable P_i by a factor of 1.6. However, there were no accumulated effects of waste amendment on the fraction of soil in wet-stable aggregates, or on the microbiological properties tested, which supported the observation from the incubation study that effects of organic wastes were transient.     

Impact of allelochemical exuded from allelopathic rice on soil microbial community

Allelopathic rice releases allelochemicals from its roots to paddy soils at early growth stages to inhibit neighboring weeds. However, little is currently known about the effects of allelochemicals on soil microbes. In this study, we show that allelopathic rice can have great impact on the population and community structure of soil microbes. Allelopathic rice PI312777 seedlings reduced the culturable microbial population and total PLFA when compared to non-allelopathic rice Liaojing-9. Similar results were observed when, instead of growing seedlings, soils were incubated with plant root exudates. This result demonstrates that the composition of root exudates from the rice varieties tested contributes to the soil microbial community. Further experiments showed that the microbial community was affected by the allelochemical 5,4′-dihydroxy-3′,5′-dimethoxy-7-O-β-glucopyranosylflavone exuded from allelopathic rice roots, through immediately hydrolyzing glucose with stimulation on soil bacteria and aglycone (5,7,4′-trihydroxy-3′,5′-dimethoxyflavone) with inhibition on soil fungi. This result indicates that the flavone O-glycoside can provide carbon and interact with soil microbes. PC analysis of the fatty acid data clearly separated the allelopathic PI312777 and the non-allelopathic Liaojing-9 variety (PC1 = 46.4%, PC2 = 20.3%). Similarly, the first principal component (PC1 = 37.4%) together with the second principal component (PC2 = 17.3%) explained 54.7% of the variation between the allelopathic and non-allelopathic root exudates. Furthermore, the canonical correlation between allelopathic root exudates and the flavone O-glycoside was statistically significant (Canonical R = 0.889, χ² (25) = 69.72, p = 0.0041). Although the data generated in this study were not completely consistent between culturable microbes and PLFA profile, it is a fact that variation in soil microbial populations and community structures could be distinguished by the allelopathic and non-allelopathic rice varieties tested. Our results suggest that individual components of rice root exudates, such as allelochemicals from allelopathic rice, can modify the soil microbial community.     

Evaluation of an optimal extraction method for measuring d-ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) in agricultural soils and its association with soil microbial CO2 assimilation

Assimilating atmospheric carbon (C) into terrestrial ecosystems is recognized as a primary measure to mitigate global warming. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the dominant enzyme by which terrestrial autotrophic bacteria and plants fix CO₂. To investigate the possibility of using RubisCO activity as an indicator of microbial CO₂ fixation potential, a valid and efficient method for extracting soil proteins is needed. We examined three methods commonly used for total soil protein extraction. A simple sonication method for extracting soil protein was more efficient than bead beating or freeze–thaw methods. Total soil protein, RubisCO activity, and microbial fixation of CO₂ in different agricultural soils were quantified in an incubation experiment using ¹⁴C-CO₂ as a tracer. The soil samples showed significant differences in protein content and RubisCO activity, defined as nmol CO₂ fixed g⁻¹ soil min⁻¹. RubisCO activities ranged from 10.68 to 68.07 nmol CO₂ kg⁻¹ soil min⁻¹, which were closely related to the abundance of cbbL genes (r = 0.900, P = 0.0140) and the rates of microbial CO₂ assimilation (r = 0.949, P = 0.0038). This suggests that RubisCO activity can be used as an indicator of soil microbial assimilation of atmospheric CO₂.     

Relationships between soil pH and microbial properties in a UK arable soil

Effects of changing pH along a natural continuous gradient of a UK silty-loam soil were investigated. The site was a 200 m soil transect of the Hoosfield acid strip (Rothamsted Research, UK) which has grown continuous barley for more than 100 years. This experiment provides a remarkably uniform soil pH gradient, ranging from about pH 8.3 to 3.7. Soil total and organic C and the ratio: (soil organic C)/(soil total N) decreased due to decreasing plant C inputs as the soil pH declined. As expected, the CaCO₃ concentration was greatest at very high pH values (pH > 7.5). In contrast, extractable Al concentrations increased linearly (R² = 0.94, p < 0.001) from below about pH 5.4, while extractable Mn concentrations were largest at pH 4.4 and decreased at lower pHs. Biomass C and biomass ninhydrin-N were greatest above pH 7. There were statistically significant relationships between soil pH and biomass C (R² = 0.80, p < 0.001), biomass ninhydrin-N (R² = 0.90, p < 0.001), organic C (R² = 0.83, p < 0.001) and total N (R² = 0.83, p < 0.001), confirming the importance of soil organic matter and pH in stimulating microbial biomass growth. Soil CO₂ evolution increased as pH increased (R² = 0.97, p < 0.001). In contrast, the respiratory quotient (qCO₂) had the greatest values at either end of the pH range. This is almost certainly a response to stress caused by the low p. At the highest pH, both abiotic (from CaCO₃) and biotic Co₂ will be involved so the effects of high pH on biomass activity are confounded. Microbial biomass and microbial activity tended to stabilise at pH values between about 5 and 7 because the differences in organic C, total N and Al concentrations within this pH range were small. This work has established clear relationships between microbial biomass and microbial activity over an extremely wide soil pH range and within a single soil type. In contrast, most other studies have used soils of both different pH and soil type to make similar comparisons. In the latter case, the effects of soil pH on microbial properties are confounded with effects of different soil types, vegetation cover and local climatic conditions.     

Responses of soil microbial biomass and community composition to biological soil crusts in the revegetated areas of the Tengger Desert

As a key component of desert ecosystems, biological soil crusts (BSCs) play an important role in dune fixation and maintaining soil biota. Soil microbial properties associated with the colonization and development of BSCs may indicate soil quality changes, particularly following dune stabilization. However, very little is known about the influence of BSCs on soil microbes in sand dunes. We examined the influence of BSCs on soil microbial biomass and community composition in revegetated areas of the Tengger Desert. BSCs increased soil microbial biomass (biomass C and N), microbial phospholipid fatty acid (PLFA) concentrations and the ratio of fungal to bacterial PLFAs. The effects varied with crust type and crust age. Moss crusts had higher microbial biomass and microbial PLFA concentrations than cyanobacteria-lichen crusts. Crust age was positively correlated with microbial biomass C and N, microbial PLFA concentrations, bacterial PLFA concentrations, fungal PLFA concentrations and the ratio of fungal to bacterial PLFAs. BSCs significantly affected microbial biomass C and N in the 0–20 cm soil layers, showing a significant negative correlation with soil depth. The study demonstrated that the colonization and development of BSCs was beneficial for soil microbial properties and soil quality in the revegetated areas. This can be attributed to BSCs increasing topsoil thickness after dunes have been stabilized, creating suitable habitats and providing an essential food source for soil microbes.     

The effects of fresh and stabilized pruning wastes on the biomass,structure and activity of the soil microbial community in a semiarid climate

The incorporation of organic amendments from pruning waste into soil may help to mitigate soil degradation and to improve soil fertility in semiarid ecosystems. However, the effects of pruning wastes on the biomass, structure and activity of the soil microbial community are not fully known. In this study, we evaluate the response of the microbial community of a semiarid soil to fresh and composted vegetal wastes that were added as organic amendments at different doses (150 and 300 t ha⁻¹) five years ago. The effects on the soil microbial community were evaluated through a suite of different chemical, microbiological and biochemical indicators, including enzyme activities, community-level physiological profiles (CLPPs) and phospholipid fatty acid analysis (PLFA). Our results evidenced a long-term legacy of the added materials in terms of soil microbial biomass and enzyme activity. For instance, cellulase activity reached 633 μg and 283 μg glucose g⁻¹ h⁻¹ in the soils amended with fresh and composted waste, respectively. Similarly, bacterial biomass reached 116 nmol g⁻¹ in the soil treated with a high dose of fresh waste, while it reached just 66 nmol g⁻¹ in the soil amended with a high dose of composted waste. Organic amendments produced a long-term increase in microbiological activity and a change in the structure of the microbial community, which was largely dependent on the stabilization level of the pruning waste but not on the applied dose. Ultimately, the addition of fresh pruning waste was more effective than the application of composted waste for improving the microbiological soil quality in semiarid soils.     

An outline of soil nematode succession on abandoned fields in South Bohemia

Secondary succession of nematodes was studied in 1–48-year-old abandoned fields on cambisols in South Bohemia, Czech Republic, and compared with cultivated field and sub-climax oak forests. Bacterivores were the predominant group in the cultivated field whereas in forests root-fungal feeders (mainly Filenchus) were almost as abundant as bacterivores. The total abundance of nematodes in the cultivated field averaged 868 × 10³ ind m⁻². During the first three years of succession the abundance practically did not change (775 × 10³ ind m⁻²), the fauna was still similar to that in cultivated field but the biomass increased mainly due to Aporcelaimellus. Then the abundance increased up to 3731 × 10³ ind m⁻² in 7–8-year-old abandoned fields, plant parasites (Helicotylenchus) dominated and the fungal-based decomposition channel was activated. Later the abundance stabilised at between 1086 and 1478 × 10³ ind m⁻² in 13–25-year-old successional meadow stages with high population densities of omnivores and predators. The total abundance of nematodes was low in the 12–13-year-old willow shrub stage (594 × 10³ ind m⁻²), increased in the 35–48-year-old birch shrub stage (1760 × 10³ ind m⁻²) and the nematode fauna developed towards a forest community. The diversity and maturity of nematode communities generally increased with the age of abandoned fields but the highest values were in meadow stages (81–113 species, 57–68 genera, MI 2.73–3.30). The development of meadow arrested succession towards forests or diverted succession towards a waterlogged ecosystem. The succession of nematodes was influenced by the method of field abandonment (bare soil vs. legume cover, mowing) that affected the formation of either a shrub or meadow stage, and by the soil water status. The composition of the nematode fauna indicated that the soil food web could recover faster from agricultural disturbance under successive meadows than under shrubs.     

Microbial properties and soil respiration in submontane forests of Venezuelian Guyana: characteristics and response to fertilizer treatments

The distribution of vegetation types in Venezuelan Guyana (in the ‘Canaima’ National Park) represents a transitional stage in a long term process of savannization, a process considered to be conditioned by a combined chemical and intermittent drought stress. All types of woody vegetation in this environment accumulate large amounts of litter and soil organic carbon (SOC). We hypothesized that this accumulation is caused by low microbial activity. During 1 year we measured microbial biomass carbon (Cmic), microbial respiration and soil respiration of stony Oxisols (Acrohumox) at a tall, a medium and a low forest and with three chemical modifications of site conditions by the addition of NO₃⁻, Ca²⁺ and PO₄³⁻ as possible limiting elements. Due to high SOC contents, mean Cmic was 1 mg g soil⁻¹ in the mineral topsoil and 3 mg g soil⁻¹ in the forest floor. Mean microbial respiration in the mineral topsoil and the forest floor were 165 and 192 μg CO₂-C g soil⁻¹ d⁻¹, respectively. We calculated high mean metabolic quotients (qCO₂) of 200 mg CO₂-C g Cmic⁻¹ d⁻¹ in the litter layer and 166 mg CO₂-C g Cmic⁻¹ d⁻¹ in the mineral topsoil, while the Cmic-to-SOC ratios were as low as 1.0% in the litter layer and 0.8% in the mineral topsoil. Annual soil respiration was 9, 12 and 10 Mg CO₂-C ha⁻¹ yr⁻¹ in the tall, medium and low forest, respectively. CO₂ production was significantly increased by CaHPO₄ fertilization, but no consistent effects were caused by Ca²⁺ and NO₃⁻, fertilization. Our findings indicate that Cmic and microbial respiration are reduced by low nutrient concentrations and low litter and SOC quality. Reduced microbial decomposition may have contributed to SOC accumulation in these forests.     

Biochar soil amendment increased bacterial but decreased fungal gene abundance with shifts in community structure in a slightly acid rice paddy from Southwest China

Biochar’s role on greenhouse gas emission and plant growth has been well addressed. However, there have been few studies on changes in soil microbial community and activities with biochar soil amendment (BSA) in croplands. In a field experiment, biochar was amended at rates of 0, 20 and 40 t ha⁻¹ (C0, C1 and C2, respectively) in May 2010 before rice transplantation in a rice paddy from Sichuan, China. Topsoil (0–15 cm) was collected from the rice paddy while rice harvest in late October 2011. Soil physico-chemical properties and microbial biomass carbon (MBC) and nitrogen (MBN) as well as selected soil enzyme activities were determined. Based on 16S rRNA and 18S rRNA gene, bacterial and fungal community structure and abundance were characterized using terminal-restriction fragment length polymorphism (T-RFLP) combined with clone library analysis, denaturing gradient gel electrophoresis (DGGE) and quantitative real-time PCR assay (qPCR). Contents of SOC and total N and soil pH were increased but bulk density decreased significantly. While no changes in MBC and MBN, gene copy numbers of bacterial 16S rRNA was shown significantly increased by 28% and 64% and that of fungal 18S rRNA significantly decreased by 35% and 46% under BSA at 20 and 40 t ha⁻¹ respectively over control. Moreover, there was a significant decrease by 70% in abundance of Methylophilaceae and of Hydrogenophilaceae with an increase by 45% in Anaerolineae abundance under BSA at 40 t ha⁻¹ over control. Whereas, using sequencing DGGE bands of fungal 18S rRNA gene, some bands affiliated with Ascomycota and Glomeromycota were shown inhibited by BSA at rate of 40 t ha⁻¹. Significant increases in activities of dehydrogenase, alkaline phosphatases while decreased β-glucosidase were also observed under BSA. The results here indicated a shift toward a bacterial dominated microbial community in the rice paddy with BSA.     

Soil enzyme activities,microbial community composition and function after 47 years of continuous green manuring

Green manuring practices can influence soil microbial community composition and function and there is a need to investigate the influence compared with other types of organic amendment. This study reports long-term effects of green manure amendments on soil microbial properties, based on a field experiment started in 1956. In the experiment, various organic amendments, including green manure, have been applied at a rate of 4 t C ha⁻¹ every second year. Phospholipid fatty acid analysis (PLFA) indicated that the biomass of bacteria, fungi and total microbial biomass, but not arbuscular mycorrhizal (AM) fungi, generally increased due to green manuring compared with soils receiving no organic amendments. Some differences in abundance of different microbial groups were also found compared with other organic amendments (farmyard manure and sawdust) such as a higher fungal biomass and consequently a higher fungal/bacterial ratio compared with amendment with farmyard manure. The microbial community composition (PLFA profile) in the green manure treatment differed from the other treatments, but there was no effect on microbial substrate-utilization potential, determined using the Biolog EcoPlate. Protease and arylsulphatase activities in the green manure treatment were comparable to a mineral fertilized treatment receiving no additional C, whereas acid phosphatase activity increased. It can be concluded that green manuring had a beneficial impact on soil microbial properties, but differed in some aspects to other organic amendments which might be attributed to differences in quality of the amendments.