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

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

Wood biochar increases nitrogen retention in field settings mainly through abiotic processes

Nitrogen (N) is an essential element associated with crop yield and its availability is largely controlled by microbially-mediated processes. The abundance of microbial functional genes (MFG) involved in N transformations can be influenced by agricultural practices and soil amendments. Biochar may alter microbial functional gene abundances through changing soil properties, thereby affecting N cycling and its availability to crops. The objective of this study was to assess the effects of wood biochar application on N retention and MFG under field settings. This was achieved by characterising soil labile N and their stable isotope compositions and by quantifying the gene abundance of nifH (nitrogen fixation), narG (nitrate reduction), nirS, nirK (nitrite reduction), nosZ (nitrous oxide reduction), and bacterial and archeal amoA (ammonia oxidation). A wood-based biochar was applied to a macadamia orchard soil at rates of 10 t ha⁻¹ (B10) and 30 t ha⁻¹ (B30). The soil was sampled after 6 and 12 months. The abundance of narG in both B10 and B30 was lower than that of control at both sampling months. Canonical Correspondence Analysis showed that soil variables (including dissolved organic C, NO₃⁻–N and NH₄⁺–N) and sampling time influenced MFG, but biochar did not directly impact on MFG. Twelve months after biochar application, NH₄⁺–N concentrations had significantly decreased in both B10 (4.74 μg g⁻¹) and B30 (5.49 μg g⁻¹) compared to C10 (13.9 μg g⁻¹) and C30 (17.9 μg g⁻¹), whereas NO₃⁻–N concentrations increased significantly in B30 (24.7 μg g⁻¹) compared to B10 (12.7 μg g⁻¹) and control plots (6.18 μg g⁻¹ and 7.97 μg g⁻¹ in C10 and C30 respectively). At month 12, significant δ¹⁵N of NO₃⁻–N depletion observed in B30 may have been caused by a marked increase in NO₃⁻–N availability and retention in those plots. Hence, it is probable that the N retention in high rate biochar plots was mediated primarily by abiotic factors.     

Impact of inorganic nitrogen additions on microbes in biological soil crusts

Many studies have shown that changes in nitrogen (N) availability affect the diversity and composition of soil microbial community in a variety of terrestrial systems, but less is known about the responses of microbes specific to biological soil crusts (BSCs) to increasing N additions. After seven years of field experiment, the bacterial diversity in lichen-dominated crusts decreased linearly with increasing inorganic N additions (ambient N deposition; low N addition, 3.5 g N m⁻² y⁻¹; medium N addition, 7.0 g N m⁻² y⁻¹; high N addition, 14.0 g N m⁻² y⁻¹), whereas the fungal diversity exhibited a distinctive pattern, with the low N-added crust containing a higher diversity than the other crusts. Pyrosequencing data revealed that the bacterial community shifted to more Cyanobacteria with modest N additions (low N and medium N) and to more Actinobacteria and Proteobacteria and much less Cyanobacteria with excess N addition (high N). Our results suggest that soil pH, together with soil organic carbon (C), structures the bacterial communities with N additions. Among the fungal communities, the relative abundance of Ascomycota increased with modest N but decreased with excess N. However, increasing N additions favored Basidiomycota, which may be ascribed to increases in substrate availability with low lignin and high cellulose contents under elevated N conditions. Bacteria/fungi ratios were higher in the N-added samples than in the control, suggesting that the bacterial biomass tends to dominate over that of fungi in lichen-dominated crusts after N additions, which is especially evident in the excess N condition. Because bacteria and fungi are important components and important decomposers in BSCs, the alterations of the bacterial and fungal communities may have implications in the formation and persistence of BSCs and the cycling and storage of C in desert ecosystems.     

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.     

Effects of simulated nitrogen deposition on soil respiration components and their temperature sensitivities in a semiarid grassland

Nitrogen (N) deposition to semiarid ecosystems is increasing globally, yet few studies have investigated the ecological consequences of N enrichment in these ecosystems. Furthermore, soil CO₂ flux – including plant root and microbial respiration – is a key feedback to ecosystem carbon (C) cycling that links ecosystem processes to climate, yet few studies have investigated the effects of N enrichment on belowground processes in water-limited ecosystems. In this study, we conducted two-level N addition experiments to investigate the effects of N enrichment on microbial and root respiration in a grassland ecosystem on the Loess Plateau in northwestern China. Two years of high N additions (9.2 g N m⁻² y⁻¹) significantly increased soil CO₂ flux, including both microbial and root respiration, particularly during the warm growing season. Low N additions (2.3 g N m⁻² y⁻¹) increased microbial respiration during the growing season only, but had no significant effects on root respiration. The annual temperature coefficients (Q₁₀) of soil respiration and microbial respiration ranged from 1.86 to 3.00 and 1.86 to 2.72 respectively, and there was a significant decrease in Q₁₀ between the control and the N treatments during the non-growing season but no difference was found during the growing season. Following nitrogen additions, elevated rates of root respiration were significantly and positively related to root N concentrations and biomass, while elevated rates of microbial respiration were related to soil microbial biomass C (SMBC). The microbial respiration tended to respond more sensitively to N addition, while the root respiration did not have similar response. The different mechanisms of N addition impacts on soil respiration and its components and their sensitivity to temperature identified in this study may facilitate the simulation and prediction of C cycling and storage in semiarid grasslands under future scenarios of global change.     

Prolonged drought changes the bacterial growth response to rewetting

Rewetting a dry soil can result in two response patterns of bacterial growth and respiration. In type 1, bacterial growth starts to increase linearly immediately upon rewetting and respiration rates are highest immediately upon rewetting. In type 2, bacterial growth starts to increase exponentially after a lag period with a secondary increase in respiration occurring at the start of the exponential increase in growth. We previously observed that the type 1 response occurred after rewetting 4-day dried soil and type 2 for 1-year dried soil. Here we studied in detail how the duration of drought related to the two types of responses of bacterial growth and respiration to rewetting. Soil was air dried for different time periods from 4 days up to 48 weeks. Upon rewetting, bacterial growth and respiration was measured repeatedly at 17 °C during one week. Drought periods of ≤2 weeks resulted in a type 1 response whereas drought periods of ≥4 weeks resulted in a type 2 response. The lag period increased with drought duration and reached a maximum of ca. 18 h. The bacterial growth response was also affected by incubation of moist soil before drying–rewetting. The lag period increased with duration of moist soil incubation before the 4-day drying–rewetting event and reached also a maximum of ca. 18 h. The exponential growth increase in the type 2 response coincided with a secondary increase in respiration, which increased in magnitude with increasing drought duration. Cumulative respiration increased with drought duration and was ca. 4 times higher after 48 weeks of drought compared to 4 days. Thus, prolonged drought affected the response type of bacterial growth and respiration to rewetting, and also increased lag period, the magnitude of the secondary increase in respiration and total C release. The effect of drought was, however, modified by the lenght of the incubation period of moist soil before drought, suggesting that soil conditions before a drying–rewetting event need consideration when evaluating microbial responses.     

Taxon-specific responses of soil bacteria to the addition of low level C inputs

The addition of small or trace amounts of carbon to soils can result in the release of 2-5 times more C as CO₂ than was added in the original solution. The identity of the microorganisms responsible for these so-called trigger effects remains largely unknown. This paper reports on the response of individual bacterial taxa to the addition of a range of ¹⁴C-glucose concentrations (150, 50 and 15 and 0 μg C g⁻¹ soil) similar to the low levels of labile C found in soil. Taxon-specific responses were identified using a modification of the stable isotope probing (SIP) protocol and the recovery of [¹⁴C] labelled ribosomal RNA using equilibrium density gradient centrifugation. This provided good resolution of the ‘heavy’ fractions ([¹⁴C] labelled RNA) from the ‘light’ fractions ([¹²C] unlabelled RNA). The extent of the separation was verified using autoradiography. The addition of [¹⁴C] glucose at all concentrations was characterised by changes in the relative intensity of particular bands. Canonical correspondence analysis (CCA) showed that the rRNA response in both the ‘heavy’ and ‘light’ fractions differed according to the concentration of glucose added but was most pronounced in soils amended with 150 μg C g⁻¹ soil. In the ‘heavy RNA’ fractions there was a clear separation between soils amended with 150 μg C g⁻¹ soil and those receiving 50 and 15 μg C g⁻¹ soil indicating that at low C inputs the microbial community response is quite distinct from that seen at higher concentrations. To investigate these differences further, bands that changed in relative intensity following amendment were excised from the DGGE gels, reamplified and sequenced. Sequence analysis identified 8 taxa that responded to glucose amendment (Bacillus, Pseudomonas, Burkholderia, Bradyrhizobium, Actinobacteria, Nitrosomonas, Acidobacteria and an uncultured β-proteobacteria). These results show that radioisotope probing (RNA-RIP) can be used successfully to study the fate of labile C substrates, such as glucose, in soil.     

Characteristics of earthworms (Eisenia fetida) in PAHs contaminated soil amended with sewage sludge or vermicompost

Earthworms can be used to remove polycyclic aromatic hydrocarbons (PAHs) from soil, but this might affect their survival and they might accumulate the contaminants. Sterilized and unsterilized soil was contaminated with phenanthrene (Phen), anthracene (Anth) and benzo(a)pyrene (BaP), added with or without Eisenia fetida, sewage sludge or vermicompost. Survival, growth, cocoon formation and concentrations of PAHs in the earthworms were monitored for 70 days. Addition of sewage sludge to sterilized or unsterilized soil maintained the number of earthworms and their survival was 94%. The addition of sludge significantly increased the weight of earthworms 1.3 times compared to those kept in the unamended soil or in soil amended with vermicompost. The weight of earthworms was significantly lower in sterilized than in unsterilized soil. Cocoons were only detected when sewage sludge was added to unsterilized soil. A maximum concentration of 62.3 μg Phen kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost after 7 days and 22.3 μg Phen kg⁻¹ when kept in the unamended unsterilized soil after 14 days. Concentrations of Phen in the earthworms decreased thereafter and ≤2 μg kg⁻¹ after 28 days. A maximum Anth concentration of 82.5 μg kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost and 45.8 μg Anth kg⁻¹ when kept in the unamended unsterilized soil after 14 days. A maximum concentration of 316 μg BaP kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost after 56 days and 311 μg BaP kg⁻¹ when kept in the unsterilized soil amended with vermicompost after 28 days. The amount of BaP in the earthworm was generally largest after 28 days, but after 70 days still 60 μg kg⁻¹ was found in E. fetida when kept in the sterilized soil amended with sewage sludge. It was found that E. fetida survived in PAHs contaminated soil and accumulated only small amounts of the contaminants, but sewage sludge was required as food for its survival and cocoon production.     

Enzymatic activities and microbial communities in an Antarctic dry valley soil: Responses to C and N supplementation

The soils of the Antarctic dry valleys are exposed to extremely dry and cold conditions. Nevertheless, they contain small communities of micro-organisms that contribute to the biogeochemical transformations of the bioelements, albeit at slow rates. We have determined the dehydrogenase, β-glucosidase, acid and alkaline phosphatase and arylsulphatase activities and the rates of respiration (CO₂ production) in laboratory assays of soils collected from a field experiment in an Antarctic dry valley. The objective of the field experiment was to test the responses of the soil microbial community to additions of C and N in simple (glucose and NH₄Cl) and complex forms (glycine and lacustrine detritus from the adjacent lake comprising principally cyanobacterial necromass). The soil samples were taken 3 years after the experimental treatments had been applied. In unamended soil, all enzyme activities and respiration were detected indicating that the enzymatic capacity to mineralize organic C, P and S compounds existed in the soil, despite the very low organic matter content. Relative to the control (unamended soil), respiration was significantly increased by all the experimental additions of C and N except the smallest NH₄Cl addition (1 mg N g⁻¹ soil) and the smallest detritus addition (1.5 mg C g⁻¹ soil and 0.13 mg N g⁻¹ soil). The activities of all enzymes except dehydrogenase were increased by C and combined large C (10 mg C g⁻¹ soil) and N additions, but either unchanged or diminished by addition of either N only or N (up to 10 mg N g⁻¹ soil) with only small C (1 mg C g⁻¹ soil) additions in the form of glucose and NH₄Cl. This suggests that in the presence of a large amount of N, the C supply for enzyme biosynthesis was limited. When normalized with respect to soil respiration, only arylsulphatase per unit of respiration showed a significant increase with C and N additions as glucose and NH₄Cl, consistent with S limitation when C and N limitations have been alleviated. Based on the positive responses of enzyme activity, detritus appeared to provide either conditions or resources which led to a larger biological response than a similar amount of C and more N added in the form of defined compounds (glucose, NH₄Cl or glycine). Assessment of the soil microbial community by ester-linked fatty acid (ELFA) analysis provided no evidence of changes in the community structure as a result of the C and N supplementation treatments. Thus the respiration and enzyme activity responses to supplementation occurred in an apparently structurally stable or unresponsive microbial community.     

Assaying the effects of chemical ameliorants with earthworms and plants exposed to a heavily polluted metalliferous soil

The study examined the effects of chemical ameliorant additions (1% montmorillonite, 1% hydroxylapatite, or 1% ferrous oxide) on the availability of cadmium (Cd), copper (Cu), lead (Pb) and zinc (Zn) to the earthworm, Lumbricus rubellus, exposed for 4 weeks to a circumneutral heavily polluted soil (Cd = 220 μg g^–1; Cu = 35 μg g^–1; Pb = 6070 μg g^–1; Zn = 124500 μg g^–1) in 1:0–1:3 dilutions with a clean soil, under laboratory conditions. Soil type (i.e. the dilution series) had a strong influence on the 1 M ammonium acetate extractable metal fractions in soil and on worm-tissue concentrations of Cd, Pb and Zn. Soil treatments (i.e. amelioration) significantly reduced only the soil Zn extractable fraction; Zn concentrations in worms tended to be lower in amended soils. A second experiment, involving curly cress (Lepidium sativum), grown either directly in the serial soil dilutions with 5% ameliorant additions or in the water-extractable fractions of the soils, indicated that root growth is a more sensitive endpoint of metal availability than chlorophyll assays. It was concluded that: (i) chemical immobilization of metals is probably most effective in soils with low to moderate degrees of metal pollution; (ii) an integrated suite of bioassays incorporating different, ecologically relevant, taxa is to be recommended for monitoring metal bioavailabilities and biological effects.     

Applying an oxygen-based respiratory assay to assess soil microbial responses to substrate and N availability

Documented approaches for measuring soil microbial activities and their controlling factors under field conditions are needed to advance understanding of soil microbial processes for numerous applications. We manipulated field plots with carbon (C) and nitrogen (N) additions to test the capability of a respiratory assay to: (1) measure respiration of endogenous soil C in comparison to field-measured CO₂ fluxes; (2) determine substrate-induced respiratory (SIR) activities that are consistent with substrate availability in the field; and, (3) report N availability in the field based on assay responses with and without added N. The respiratory assay utilizes a microplate containing an oxygen-sensitive fluorescent ruthenium dye. Respiratory activities measured with this approach have previously been shown to occur within short (6–8 h) incubation periods using low substrate concentrations that minimize enrichment during the assay. Field treatments were conducted in a randomized full-factorial design with C substrate (casamino acids, glucose, or none) and inorganic N (±) as the treatment factors. With one exception, we found that respiration of endogenous soil C in the assay responded to the field treatments in a similar manner to CO₂ fluxes measured in the field. Patterns of SIR with low concentrations of added amino acid or carbohydrate substrate (200 μg C g⁻¹ soil) were consistent with field treatments. The ratio (N_ratio) of carbohydrate respiration with added N (25 μg N g⁻¹ soil) to the same without N in the assay was significantly (P < 0.05) decreased by field N amendment. The carbohydrate N_ratio exhibited a logarithmic relationship (r = 0.64, P < 0.05) with extractable inorganic soil nitrate and ammonium concentrations. These data significantly extend and support the capability of this oxygen-based respiratory assay to evaluate in situ soil activities and examine factors that limit these activities.     

Induced N-limitation of bacterial growth in soil: Effect of carbon loading and N status in soil

Application of C-rich plant residues can change the soil system from C-limitation for microbial growth to limitation by other nutrients. However, the initial nutrient status of the soil may interact with the added amount of residues in determining limitation. We studied this interactive effect in soils from the Harvard Forest LTER, where annual addition of N since 1988 has resulted in soils with different N-status: No N (Unfertilized), 50 (Low N) and 150 (High N) kg N ha⁻¹. We hypothesized that adding C-rich substrate would change the soil from being C- to being N-limited for bacterial growth and that the extent of N-limitation would be higher with increasing substrate additions, while becoming less evident in soil with increasing N-status. We compared the effect of adding two C-rich substrates, starch (0, 10, 20, 40 mg g⁻¹ soil) and straw (0, 20, 40, 80 mg g⁻¹), incubating the soils for up to 3 and 4 weeks for starch and straw, respectively. Nutrient limitations were studied by measuring bacterial growth 3 days after adding C as glucose and N as NH₄NO₃ in a full factorial design. Initially bacterial growth in all soils was C-limited. As hypothesized, adding C-rich substrates removed the C-limitation, with lower amounts of starch and straw needed in the unfertilized and Low N soils than in the High N soil. Combinations of different N-status of the soil and amendment levels of starch and straw could be found, where bacterial growth appeared close to co-limited both by available C and N. However, at even higher amendment levels, presumable resulting in N-limitation, bacterial growth still responded less by adding N then C-limited soils by adding C. Thus, in a C-limited soil there appeared to be N available immediate for growth, while in an N-limited soil, easily available C was not immediately available.     

Soil microbial communities response to herbicide 2,4-dichlorophenoxyacetic acid butyl ester

2,4-Dichlorophenoxyacetic acid butyl ester (2,4-D butyl ester) is extensively applied for weed control in cultivation fields in China, but its effect on soil microbial community remains obscure. This study investigated the microbial response to 2,4-D butyl ester application at different concentrations (CK, 10, 100 and 1000 μg g^?1) in the soils with two fertility levels, using soil dilution plate method and phospholipid fatty acid (PLFA) analysis. Culturable microorganisms were affected by the herbicide in both soils, particularly at the higher concentration. After treating soil with 100 μg g^?1 herbicide, culturable bacteria and actinomycetes were significantly higher, compared to other treatments. Treatment of soil with 1000 μg g^?1 2,4-D butyl ester caused a decline in culturable microbial counts, with the exception of fungal numbers, which increased over the incubation time. PLFA profiles showed that fatty acids for Gram-negative (GN) bacteria, Gram-positive (GP) bacteria, total bacteria and total fungi, as well as total PLFAs, varied with herbicide concentration for both soil samples. As herbicide concentration increased, the GN/GP ratio decreased dramatically in the two soils. The higher stress level was in the treatments with high concentrations of herbicide (1000 μg g^?1) for both soils. Principal component analysis of PLFAs showed that the addition of 2,4-D butyl ester significantly shifted the microbial community structure in the two soils. These results showed that the herbicide 2,4-D butyl ester might have substantial effects on microbial population and microbial community structure in agricultural soils. In particular, the effects of 2,4-D butyl ester were greater in soil with low organic matter and fertility level than in soil with high organic matter and fertility level.     

Long-term organic phosphorus mineralization in Spodosols under forests and its relation to carbon and nitrogen mineralization

In forest soils where a large fraction of total phosphorus (P) is in organic forms, soil micro-organisms play a major role in the P cycle and plant availability since they mediate organic P transformations. However, the correct assessment of organic P mineralization is usually a challenging task because mineralized P is rapidly sorbed and most mineralization fluxes are very weak. The objectives of the present work were to quantify in five forest Spodosols at soil depths of 0-15 cm net mineralization of total organic P and the resulting increase in plant available inorganic P and to verify whether net or gross P mineralization could be estimated using the C or N mineralization rates. Net mineralization of total organic P was derived from the net changes in microbial P and gross mineralization of P in dead soil organic matter. We studied very low P-sorbing soils enabling us to use lower extractants to assess the change in total inorganic P as a result of gross mineralization of P in dead soil organic matter. In addition, to enable detection of gross mineralization of P in dead soil organic matter, a long-term incubation (517 days) experiment was carried out. At the beginning of the experiment, total P contents of the soils were very low (19-51 μg g⁻¹) and were essentially present as organic P (17-44 μg g⁻¹, 85-91%) or microbial P (6-14 μg g⁻¹; 24-39%). Conversely, the initial contents of inorganic P were low (2-7 μg g⁻¹; 9-15%). The net changes in the pool size of microbial P during the 517 days of incubation (4-8 μg g⁻¹) and the amounts of P resulting from gross mineralization of dead soil organic matter (0.001-0.018 μg g⁻¹ day⁻¹; 0.4-9.5 μg g⁻¹ for the entire incubation period) were considerable compared to the initial amounts of organic P and also when compared to the initial diffusive iP fraction (<0.3 μg g⁻¹). Diffusive iP corresponds to the phosphate ions that can be transferred from the solid constituents to the soil solution under a gradient of concentration. Net mineralization of organic P induced an important increase in iP in soil solution (0.6-10 μg g⁻¹; 600-5000% increase) and lower increases in diffusive iP fractions (0.3-5 μg g⁻¹; 300-2000% increase), soil solid constituents having an extremely low reactivity relative to iP. Therefore, soil micro-organisms and organic P transformations play a major role in the bioavailability of P in these forest soils. In our study, the dead soil organic matter was defined as a recalcitrant organic fraction. Probably because gross mineralization of P from this recalcitrant organic fraction was mainly driven by the micro-organisms’ needs for energy, the rates of gross mineralization of C, N and P in the recalcitrant organic fraction were similar. Indirect estimation of gross mineralization of P in dead soil organic matter using the gross C mineralization rate seems thus an alternative method for the studied soils. However, additional studies are needed to verify this alternative method in other soils. No relationships were found between microbial P release and microbial C and N releases.     

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

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

DIBOA: Fate in soil and effects on root-knot nematode egg numbers

The benzoxazinoid 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) is produced by rye (Secale cereale) and may contribute to plant-parasitic nematode suppression when rye plants are incorporated as a green manure. We investigated the fate of DIBOA in soil and DIBOA's effects on nematode reproduction. Soil in plastic bags was treated with DIBOA at concentrations ranging from 1.1 to 18 μg g⁻¹ dry soil, and with the root-knot nematode Meloidogyne incognita. Control soils were treated with water or with 0.31% methanol, with or without nematodes. DIBOA concentrations extracted from the soil were measured at selected times for 5 consecutive days. The soil from each bag was then placed into a pot in the greenhouse, and a cucumber seedling was transplanted into each pot. Five weeks later, only the highest DIBOA concentration, 18 μg g⁻¹ soil, reduced nematode egg numbers. At 0 h, DIBOA measured in soil ranged from 19.68 to 35.51% of the initial DIBOA concentration, and was dependent on the concentration added to the soil. DIBOA half-life was from 18 to 22 h, and very little DIBOA was present in soil after 120 h. Identified breakdown products accounted for only 4% at maximum of the initially added DIBOA. The results of our study demonstrate that high soil concentrations of DIBOA are necessary to suppress M. incognita; DIBOA may not be a major factor in nematode suppression by a rye cover crop.     

Interrelationships between microbial and soil properties in young volcanic ash soils of Nicaragua

The activity and biomass of soil microorganisms were measured in soils from 25 different arable sites in the Pacific region of Nicaragua with the objective of elucidating their interrelationship with soil textural and soil chemical properties. All soils developed from recent volcanic deposits but differ in their particle size distribution. Short-term phosphorus fixation capacity varied widely and was, on average, 11% of added P. In contrast, long-term P fixation capacity varied within a small range of around 55%. Mean basal respiration was 8.6 μg CO₂–C d⁻¹ g⁻¹ soil, average contents of biomass C, biomass P, and ergosterol as an indicator of fungal biomass were 116, 1.95, and 0.34 μg g⁻¹ soil, respectively. They were all, except biomass P, significantly lower in the sandy than in the loamy soils. The mean biomass C-to-soil C ratio was 0.69%, the mean metabolic quotient 95 mg CO₂–C d⁻¹ g⁻¹ biomass C, the mean ergosterol-to-biomass C ratio 0.31% and the mean biomass C-to-P ratio 107. The very low ergosterol-to-biomass C ratio indicates that fungi contribute only a relatively small percentage to the microbial biomass. The biomass C-to-P ratio exceeded considerably the soil C-to-total P ratio. Metabolic quotient qCO₂ and ergosterol-to-biomass C were both negatively correlated with biomass C-to-soil C ratio and clay content, indicating positive correlations between qCO₂ and ergosterol-to-biomass C ratio and between biomass C-to-soil C ratio and clay content. Key problems of soil fertility and soil quality in Nicaragua are low availability of soil organic matter and phosphorus to soil microorganisms, which are magnified by a low percentage of fungi, probably reducing the ability of soil to provide nutrients for plant growth.     

Influence of acetamiprid on soil enzymatic activities and respiration

The effect of a new pesticide, acetamiprid, applied at normal field concentration (0.5 mg kg⁻¹ dried soil) and at high concentration (5 and 50 mg kg⁻¹ dried soil), on soil enzyme activities and soil respiration in upland soil was studied. The results showed that acetamiprid had a strong negative influence on soil respiration and phosphatase activity, and the enzyme activities in soil treated with 5 and 50 mg kg⁻¹ dry soil were significantly (P < 0.05) lower than the CK over the course of incubation. The 7-, 14-, and 35-day EC10 for phosphatase were 11, 15, and 11 mg kg⁻¹ dry soil, respectively. The 21-day EC10 and EC50 for soil respiration was 0.005 and 83 mg kg⁻¹ dry soil. The activity of dehydrogenase was enhanced after acetamiprid application for 2 weeks and the enzyme activities in samples treated with 0.5, 5 and 50 mg kg⁻¹ dry soil was about 2.5-, 1.5- and 2-fold to that of the control on sample day 28. Variance of urease and catalase had no distinct relationship with the application concentration. The activity of proteinase was not significantly affected within the first 2 weeks but inhibited from the fourth week after acetamiprid application and was only 0.45-fold to that of the control on sample day 28. Overall, acetamiprid at normal field dose would not pose a toxicological threat to soil enzymes, but a certain potential threat to soil respiration.     

Immobilization and mineralization of nitrogen in a saline and alkaline soil during microbial use of sugarcane filter cake amended with glucose

A 42-day incubation was conducted to study the effect of glucose and ammonium addition adjusted to a C/N ratio of 12.5 on sugarcane filter cake decomposition and on the release of inorganic N from microbial residues formed initially. The CO₂ evolved increased in comparison with the non-amended control from 35% of the added C with pure +5 mg g⁻¹ soil filter cake amendment to 41% with +5 mg g⁻¹ soil filter cake +2.5 mg g⁻¹ soil glucose amendment to 48% with 5 mg g⁻¹ soil filter cake +5 mg g⁻¹ soil glucose amendment. The different amendments increased microbial biomass C and microbial biomass N within 6 h and such an increase persisted. The fungal cell-membrane component ergosterol initially showed a disproportionate increase in relation to microbial biomass C, which completely disappeared by the end of the incubation. The cellulase activity showed a 5-fold increase after filter cake addition, which was not further increased by the additional glucose amendment. The cellulase activity showed an exponential decline to values around 4% of the initial value in all treatments. The amount of inorganic N immobilized from day 0 to day 14 increased with increasing amount of C added, in contrast to the control treatment. After day 14, the immobilized N was re-mineralized at rates between 1.3 and 1.5 μg N g⁻¹ soil d⁻¹ in the treatments being more than twice as high as in the control treatment. This means that the re-mineralization rate is independent of the actual size of the microbial residues pool and also independent of the size of the soil microbial biomass.     

Changes of microbial properties in (near-) rhizosphere soils after phytoextraction by Sedum alfredii H: A rhizobox approach with an artificial Cd-contaminated soil

The ultimate goal of soil remediation is to restore soil health. Soil microbial parameters are considered to be effective indicators of soil health. The aim of this study was to determine the effects of phytoextraction on microbial properties through the measurement of soil microbial biomass carbon, soil basal respiration and enzyme activities. For this purpose, a pre-stratified rhizobox experiment was conducted with the Cd hyperaccumulator Sedum alfredii H. for phytoextraction Cd from an artificial contaminated soil (15.81 mg kg⁻¹) under greenhouse conditions. The plant and soil samples were collected after growing the plant for three and six months with three replications. The results indicated that the ecotype of S. alfredii H. originating from an ancient silver mining site was a Cd-hyperaccumulator as it showed high tolerance to Cd stress, the shoot Cd concentration were as high as 922.6 mg kg⁻¹ and 581.9 mg kg⁻¹ at the two samplings, and it also showed high BF (58.4 and 36.8 after 3 and 6 months growth), and TF (5.8 and 5.1 after 3 and 6 months growth). The amounts of Cd accumulated in the shoots of S. alfredii reached to an average of 1206 μg plant⁻¹ after 6 months growth. Basal respiration, invertase and acid phosphatase activities of the rhizosphere soil separated by the shaking method were significantly higher (P < 0.01) than that of the near-rhizosphere soil and the unplanted soil after 3 months growth, so were microbial biomass carbon, urease, invertase and acid phosphatase activities of the rhizosphere soil after 6 months growth. Acid phosphatase activity of the 0–2 mm sub-layer rhizosphere soil collected by the pre-stratified method after 3 months growth was significantly higher (P < 0.05) than that of other sub-layer rhizosphere soils and bulk soil, and so were microbial biomass carbon, basal respiration, urease, invertase and acid phosphatase activities of the 0–2 mm sub-layer rhizosphere soil after 6 months growth. It was concluded that phytoextraction by S. alfredii could improve soil microbial properties, especially in rhizosphere, and this plant poses a great potential for the remediation of Cd contaminated soil.