The significance of microbial biomass sulphur in soil

The soil microbial biomass S fraction of total organic S in soil is considered to be relatively labile and the most active S pool for S turnover in soil. Its significance has been demonstrated in studies of S deficiency in agronomic situations and in those of S pollution from high atmospheric inputs. The utility of the CHCl₃ fumigation-extraction technique for the measurement of microbial S has been proved for a range of soils and conditions. The various methodologies currently available are discussed, including the need for determination of the conversion (K _s) factor. Microbial S values, summarized from the available literature, ranged from 3 to 300 g S g^-1 dry weight soil. They were generally greater in grassland than in arable systems, though the greatest values were obtained in the few examples from forest and peatland soil systems. Microbial S values showed direct relationships with both microbial C and with total soil organic S. Again, there were significant differences between arable and grassland systems. The effect of factors such as organic and inorganic inputs as well as soil physical conditions on microbial S are described. Microbial S turnover rates were estimated from seasonal, ³⁵S-labelling and modelling studies. These rates varied between an approximately annual turnover rate in undisturbed soils up to 80 year^-1 following the addition of readily available substrates. Prospective future research areas are also outlined.     

Determination of soil microbial biomass phosphorus in acid red soils from southern China

A CHCl₃ fumigation and 0.03 M NH₄F-0.025 M HCl extraction procedure was used to measure microbial biomass P (P_mic) in 11 acid red soils (pH <6.0) from southern China and the results compared to those obtained by the commonly-used CHCl₃ fumigation and 0.5 M NaHCO₃ extraction method. Extraction with NH₄F-HCl was found to be more effective and accurate than NaHCO₃ extraction for detecting the increase of P from microbial biomass P following chloroform fumigation due to its higher efficiency in extracting both native labile phosphate and added phosphate (³²P) in the soils. This was confirmed by the recovery of ³²P from in situ ³²P-labeled soil microbial biomass following fumigation and extraction by the NH₄F-HCl solution. Soil microbial biomass P, measured by the NH₄F-HCl extraction method, was more comparable with soil microbial biomass C (with a more narrow C:P ratio range of 4.3 to 22.3 and a mean of 15.6 in the microbial biomass), than that obtained by NaHCO₃ solution (with a mean C:P ratio of 30.7 and a wide range of 14.9 to 48.9). K_p, the fraction of soil microbial biomass P extracted after CHCl₃ fumigation, by the NH₄F-HCl solution was 0.34. The amount of microbial biomass P determined (using K_p =0.34) was 3–400% (mean 131%) higher than that obtained by the NaHCO₃ extraction (using K_p =0.40) for the 11 red soils studied. The results suggest that the CHCl₃ fumigation and NH₄F-HCl extraction method is more reliable for measuring microbial biomass P than the NaHCO₃ extraction method in acid red soils.     

Hot water extracted organic matter: chemical composition and temporal variations in a long-term field experiment

Hot water-soluble organic matter was extracted from soil samples collected weekly between April and October in untreated and NPK+farmyard manure-fertilized plots in the 88-year-old Static Experiment (Loess Chernozem) at Bad Lauchstädt, Germany. As shown by solid-state ¹³C-nuclear magnetic resonance spectroscopy (¹³C-NMR) combined with pyrolysis-field ionization mass spectrometry this organic matter fraction was largely composed of carbohydrates and N-containing compounds, in particular amino-N species and amides. This composition and the low pyrolysis temperatures (mainly between 300 and 500°C) indicated its origin from soil biomass and root exudates and lysates, and its presence in the soil solution or weakly adsorbed by soil minerals and humic macromolecules. Long-term fertilization with NPK+farmyard manure resulted in larger mean concentrations of hot water-extracted C and N (0.933 and 0.094 g kg^-1) than soil management without fertilization (0.511 and 0.056 g kg^-1). The C and N extracted by hot water were in the range of 3–5% of total soil C and N. In the two treatments distinct temporal changes were observed, which appeared to be related to population dynamics of soil organisms, root growth and decomposition, and climatic influences on soil.     

Estimating microbial biomass N in soils with and without living roots: Limitations of a pre-extraction step

Special net-closed soil containers were used in a pot experiment with low and high plant densities to give soil samples with and without roots. Soils from the containers were analysed either by the fumigation-extraction method or by a modified procedure starting with a pre-extraction and sieving step to remove plant roots from the samples. In the extracts NO ₃ ^- -N, NH ₄ ⁺ -N, organic N, and total N were measured. Microbial biomass N was calculated from the differences in total N in fumigated and unfumigated soils. Different plant densities had almost no influence on the values of the N compounds using either method. In soils with roots, significantly more organic N (and total N) was found by the fumigation-extraction method compared to soils without roots while no differences were obtained using pre-extractions and sieving. Though the organic N content in pre-extracts from soils with roots was significantly higher than from soils without roots, the NO ₃ ^- -N and NH ₄ ⁺ -N content was lower. Significant differences in biomass N in soils with and without roots were found only with the fumigation-extraction method. Biomass N levels calculated using the results after pre-extraction and sieving were about 50% lower than levels detected using fumigation-extraction alone. With the use of special net-closed soil containers, not only were soil samples produced with and without roots, but it was also possible to induce a rhizophere in the soils. A comparison of the two methods using these soils clearly demonstrated that the method used has profound influence on the final biomass N results. While higher biomass levels were found by fumigation-extraction in soils with roots, because root N becomes extractable after fumigation, the use of a pre-extraction and a sieving step may underestimate the total biomass N content due to the pre-extraction of microbial N (especially from rhizosphere microorganisms) from the sample. Nevertheless, pre-extraction and sieving followed by fumigation-extraction does seem to be the preferable method for biomass N measurement in comparative studies, because in most cases only minor errors will occur.     

Activity and biomass of soil microorganisms at different depths

We measured microbial biomass C and soil organic C in soils from one grassland and two arable sites at depths of between 0 and 90 cm. The microbial biomass C content decreased from a maximum of 1147 (0–10 cm layer) to 24 g g^-1 soil (70–90 cm layer) at the grassland site, from 178 (acidic site) and 264 g g^-1 soil (neutral site) at 10–20 cm to values of between 13 and 12 g g^-1 soil (70–90 cm layer) at the two arable sites. No significant depth gradient was observed within the plough layer (0–30 cm depth) for biomass C and soil organic C contents. In general, the microbial biomass C to soil organic C ratio decreased with depth from a maximum of between 1.4 and 2.6% to a minimum of between 0.5 and 0.7% at 70–90 cm in the three soils. Over a 24-week incubation period at 25°C, we examined the survival of microbial biomass in our three soils at depths of between 0 and 90 cm without external substrate. At the end of the incubation experiment, the contents of microbial biomass C at 0–30 cm were significantly lower than the initial values. At depths of between 30 and 90 cm, the microbial biomass C content showed no significant decline in any of the four soils and remained constant up to the end of the experiment. On average, 5.8% of soil organic C was mineralized at 0–30 cm in the three soils and 4.8% at 30–90 cm. Generally, the metabolic quotient qCO₂ values increased with depth and were especially large at 70–90 cm in depth.     

Comparison of chloroform fumigation-extraction, phospholipid fatty acid, and DNA methods to determine microbial biomass in forest humus

Techniques developed to measure microbial biomass in mineral soils may not give reliable results in humus. We evaluated the relationships between three techniques to estimate microbial biomass in forest humus: chloroform fumigation-extraction (CFE), total extractable phospholipid fatty acids (PLFA), and extractable DNA. There was a good relationship between PLFA and CFE (R²=0.96), with a slope slightly different from that previously reported for mineral soils (1 nmol PLFA corresponded to a flush of 3.2 μg C released by fumigation in humus cf. 2.4 μg C in mineral soil). There was no relationship between DNA concentration and the other two measurements of microbial biomass. This may be due, in part, to the high fungal biomass in forest humus, as DNA concentration per unit biomass is much more variable for fungi than bacteria.     

Organic matter status and the size, activity and metabolic diversity of the soil microbial community in the row and inter-row of sugarcane under burning and trash retention

The concentrations of organic C, labile organic fractions and the size and activity of the microbial community were measured to a depth of 30 cm below the plant row and at distances of 30 and 60 cm into the inter-row area under sugarcane under pre-harvest burning or green cane harvesting with retention of a crop residue (trash) mulch. Total root mass was similar under burning and trashing but under trashing there was a redistribution of roots towards the surface 0-10 cm in the inter-row space as roots proliferated beneath the trash mulch. Soil organic C content decreased in response to both increasing distance from the plant row (to a depth of 20 cm) and burning rather than trashing (to a depth of 10 cm). Declines in K₂SO₄-extractable C, light fraction C, microbial biomass C, basal respiration and aggregate stability in response to distance and burning were much more marked than those for organic C and occurred to a depth of 30 cm. Bulk density was greater under burnt than trashed sugarcane and was greater in the inter-row than row, particularly under burning. Heterotrophic functional diversity (measured by analysis of catabolic response profiles to 36 substrates) was also investigated. Principal component analysis of response profiles demonstrated that soils below the row and those under trashing at 30 cm out from this row were separated from the other soils on PC1 and the sample from the inter-row centre (60 cm out) under burning was separated from the others on PC2. Catabolic evenness was least for the latter soil. It was concluded that soil in the inter-row of burnt sugarcane receives few inputs of organic matter and that conversion to green cane harvesting with retention of a trash mulch greatly improves the organic matter, microbial and physical status of the inter-row soil.     

Seasonal variations of soil microbial biomass and activity in warm- and cool-season turfgrass systems

Plant growth can be an important factor regulating seasonal variations of soil microbial biomass and activity. We investigated soil microbial biomass, microbial respiration, net N mineralization, and soil enzyme activity in turfgrass systems of three cool-season species (tall fescue, Festuca arundinacea Schreb., Kentucky bluegrass, Poa pratensis L., and creeping bentgrass, Agrostis palustris L.) and three warm-season species (centipedegrass, Eremochloa ophiuroides (Munro.) Hack, zoysiagrass, Zoysia japonica Steud, and bermudagrass, Cynodon dactylon (L.) Pers.). Microbial biomass and respiration were higher in warm- than the cool-season turfgrass systems, but net N mineralization was generally lower in warm-season turfgrass systems. Soil microbial biomass C and N varied seasonally, being lower in September and higher in May and December, independent of turfgrass physiological types. Seasonal variations in microbial respiration, net N mineralization, and cellulase activity were also similar between warm- and cool-season turfgrass systems. The lower microbial biomass and activity in September were associated with lower soil available N, possibly caused by turfgrass competition for this resource. Microbial biomass and activity (i.e., microbial respiration and net N mineralization determined in a laboratory incubation experiment) increased in soil samples collected during late fall and winter when turfgrasses grew slowly and their competition for soil N was weak. These results suggest that N availability rather than climate is the primary determinant of seasonal dynamics of soil microbial biomass and activity in turfgrass systems, located in the humid and warm region.     

Simultaneous measurement of S, macronutrients, and heavy metals in the soil microbial biomass with CHCl3 fumigation and NH4NO3 extraction

The study was carried out to investigate whether 1 M NH₄NO₃ extraction is a useful alternative to 10 mM CaCl₂ extraction for estimating soil microbial biomass S and whether the data of CHCl₃-labile NH₄NO₃-extractable macronutrients and heavy metals are useful and in agreement with the available data on element concentrations in soil microorganisms. Microbial biomass C was followed by microbial biomass S after CaCl₂ extraction with an average C/S ratio of 82, and by microbial biomass S after NH₄NO₃ extraction with an average C/S ratio of 57. The mean contribution of CHCl₃-labile metals in relation to the NH₄NO₃-extractable fraction from non-fumigated soils ranged from 0.1 to 112% in the order potassium < magnesium < cadmium < sodium < zinc + nickel < manganese < copper. The mean contribution of CHCl₃-labile metals in relation to the microbial biomass C ranged from 0.03 to 22‰ in the order cadmium < nickel < zinc < manganese < magnesium < copper < sodium < potassium. These relative contributions varied within the different metals from a 4-fold (Na⁺) to a more than 200-fold range (Cu²⁺). Significant positive correlations with microbial biomass C were observed for CHCl₃-labile zinc, sodium and especially potassium. The concentration of all elements except copper in relation to microbial biomass C were in the range known from the limited literature on fungi grown on heavy metal contaminated soils.     

An improved and accurate method for determining the dehydrogenase activity of soils with iodonitrotetrazolium chloride

Summary Conditions for a rapid, precise [100 g iodonitrotetrazolium chloride (INT)-formazan ml^-1 assay mixture], and easily reproducible assay of potential soil dehydrogenase activity are described, using 2(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (iodonitrotetrazolium chloride, INT) as the substrate. Reduced iodonitrotetrazolium formazan (INTF) was measured by spectrophotometry (464 nm) after extraction with N,N-dimethylformamide and ethanol. With this method, the coloured complex formed is highly stable. The effects of pH, buffer concentration, temperature, substrate concentration, amount of soil weight, and reaction time on dehydrogenase activity were investigated. The rate of substrate hydrolysis was proportional to soil weight; the optimal INT reduction was achieved with 1 M TRIS buffer (pH 7.0) at 40 °C. It was possible to determine the biotic and abiotic substrate reduction by comparing assays of autoclaved and unsterile soil samples. Different investigations have confirmed that the intracellular enzyme is highly correlated with the microbial biomass, and indicate that this activity is suitable as an indirect parameter of microbial biomass, measurement.     

A method of processing soil core samples for root studies by subsampling

Root studies are generally believed to be very important in ecological research. Soil coring is a valuable approach to root research, but it requires a very large amount of processing time. We present here a method for processing soil cores consisting of the combination and homogenization of several soil cores from a plot, with subsequent subsampling for root extraction. The required subsample size was determined for a topsoil and a subsoil sample from a groundnut field and was found to be 5–10% of the total soil sample. Advantages and limitations of the method are discussed.     

Impact of agricultural practices on the size and activity of the microbial biomass in a long-term field experiment

The Dehérain long-term field experiment was initiated in 1875 to study the impact of fertilization on a wheat-sugarbeet rotation. In 1987, the rotation was stopped to be replaced by continuous maize. Crop residues were soil-incorporated and the mineral fertilization was doubled in some plots. The impact of those changes on the microbial biomass and activity are presented. In spring 1987, the soil was still in a steady-state condition corresponding to the rotation. The microbial biomass was correlated with total organic C and decreased in the order farmyard manure>mineral NPK>unfertilized control. Microbial specific respiratory activity was higher in the unfertilized treatments. The soil biomass was closely related to soil N plant uptake. In 1989, after 2 years of maize and crop residue incorporation, the steady-state condition corresponding to the previous agricultural practices disappeared. So did the relationship between the biomass and total organic C, and the soil N plant uptake. Biomass specific respiratory activity increased because of low efficiency in the use of maize residues by microbes under N stress.     

Interactions between soil organic matter status, cropping history, method of quantification and sample pretreatment and their effects on measured aggregate stability

 The effects of sample pretreatment (field-moist, air-dried or tension rewetted) on aggregate stability measured by wet sieving or turbidimetry were compared for a group of soil samples ranging in organic C content from 20 to 40 g C kg^–1. Concentrations of total N, total and hot-water-extractable carbohydrate and microbial biomass C were linearly related to those of organic C. Aggregate stability measured by wet sieving using air-dried or field-moist samples and that measured by turbidimetry, regardless of sample pretreatment, increased curvilinearly with increasing soil organic C content. However, when tension-rewetted samples were used for wet sieving, aggregate stability was essentially unaffected by soil organic C content. Measurements of aggregate stability (apart from wet sieving using rewetted soils) were closely correlated with one another and with organic C, total and extractable carbohydrate and microbial biomass C content of the soils. The short-term effects of aggregate stability were also studied. Soils from under long-term arable management and those under long-term arable followed by 1 or 3 years under pasture had similar organic C contents, but aggregate stability measured by turbidimetry and by wet sieving using air-dried or field-moist samples increased with increasing years under pasture. Light fraction C, microbial biomass and hot-water-extractable carbohydrate concentrations also increased. It was concluded that both total and labile soil organic C content are important in relation to water-stable aggregation and that the use of tension-rewetted samples to measure stability by wet sieving is unsatisfactory since little separation of values is achieved. Received: 6 January 1999     

河滨缓冲带植物根系和根际微生物特征及其对农业面源污染物去除效果

对河滨缓冲带常见的3种水生植物根系形态特点、活力特征及其土壤微生物群落多样性进行了研究,并对其农业面源污染物的去除效果进行了调查。结果表明,3种水生植物根系形态和活力特征具有显著差异。根系活力表现为水生鸢尾菖蒲千屈菜,与根尖数呈显著相关。同时,3种水生植物具有显著的根际效益,根际土壤微生物生物量显著高于非根际土壤;根际土壤微生物群落数量为细菌放线菌真菌;土壤微生物群落多样性指数为水生鸢尾菖蒲千屈菜,这与3种水生植物根系活力特征表现一致。3种水生植物河滨缓冲区对农业面源污染物TN、TP和CODCr的去除效果表现为水生鸢尾菖蒲千屈菜。说明不同水生植物根系结构导致根系活力不同,由此引起的土壤微生物群落多样性差异对水生植物农业面源污染物去除效果有一定影响。     

Microbial biomass,N mineralization,and the activities of various enzymes in relation to nitrate leaching and root distribution in a slurry-amended grassland

High rates of cattle slurry application induce NO _inf3 ^sup- leaching from grassland soils. Therefore, field and lysimeter trials were conducted at Gumpenstein (Austria) to determine the residual effect of various rates of cattle slurry on microbial biomass, N mineralization, activities of soil enzymes, root densities, and N leaching in a grassland soil profile (Orthic Luvisol, sandy silt, pH 6.6). The cattle slurry applications corresponded to rates of 0, 96, 240, and 480 kg N ha^-1. N leaching was estimated in the lysimeter trial from 1981 to 1991. At a depth of 0.50 m, N leaching was elevated in the plot with the highest slurry application. In October 1991, deeper soil layers (0–10, 10–20, 20–30, 30–40, and 40–50 cm) from control and slurry-amended plots (480 kg N ha^-1) were investigated. Soil biological properties decreased with soil depth. N mineralization, nitrification, and enzymes involved in N cycling (protease, deaminase, and urease) were enhanced significantly (P<0.05) at all soil depths of the slurry-amended grassland. High rates of cattle slurry application reduced the weight of root dry matter and changed the root distribution in the different soil layers. In the slurry-amended plots the roots were mainly located in the topsoil (0–10 cm). As a result of this study, low root densities and high N mineralization rates are held to be the main reasons for NO _inf3 ^sup- leaching after heavy slurry applications on grassland.     

Effect of bamboo harvest on dynamics of nutrient pools,N mineralization,and microbial biomass in soil

The effect of harvesting bamboo savanna on the dynamics of soil nutrient pools, N mineralization, and microbial biomass was examined. In the unharvested bamboo site NO _inf3 ^sup- -N in soil ranged from 0.37 to 3.11 mg kg^-1 soil and in the harvested site from 0.43 to 3.67 mg kg^-1. NaHCO₃-extractable inorganic P ranged from 0.55 to 3.58 mg kg^-1 in the unharvested site and from 1.01 to 4.22 mg kg^-1 in the harvested site. Over two annual cycles, the N mineralization range in the unharvested and harvested sites was 0–19.28 and 0–24.0 mg kg^-1 soil month^-1, respectively. The microbial C, N, and P ranges were 278–587, 28–64, and 12–26 mg kg^-1 soil, respectively, with the harvested site exhibiting higher values. Bamboo harvesting depleted soil organic C by 13% and total N by 20%. Harvesting increased N mineralization, resulting in 10 kg ha^-1 additional mineral N in the first 1st year and 5 kg ha^-1 in the 2nd year following the harvest. Microbial biomass C, N and P increased respectively by 10, 18, and 5% as a result of bamboo harvesting.     

The hormone-like effect of earthworm casts on plant growth

Summary The fertilizing effect of earthworm casts depends on microbial metabolites, mainly growth regulators. The hormone-like effect of earthworm casts is discussed with reference to the literature and ad hoc experiments. When used in plant propagation, earthworm casts promote root initiation and root biomass and increase root percentage. When applied as a casing layer, earthworm casts stimulate carpophore formation in Agaricus bisporus, and N assimilation. When used in horticulture, earthworm casts have a hormone-like effect, influencing the development and precociousness of plants or inhibiting them. These effects are dependent on dose, application time and plant species. In addition, results recorded on dwarfing, stem elongation and precociousness of flowering suggest that the biological effect of earthworm casts is linked to microbial metabolites that influence plant metabolism, growth and development.     

An innovative method for the treatment of rice straw to improve nitrogen uptake efficiency

An innovative method was used to treat rice straw based on a mixed dilute acid treatment followed by neutralization with ammonia water. This treatment decreased the Si content of the rice straw, thus improving its degradation by soil microorganisms. The plant-available N of soil was greatly improved after the application of the treated rice straw with urea. Soil microbial biomass N was about 50 mg kg^–1 in the soil amended with the treated rice straw and urea but only 40 mg kg^–1 in the soil amended with untreated rice straw and urea. Better synchronization of N supply with the plant requirement for N uptake was obtained when treated rice straw was applied with urea. Recovery of urea-N was 61% when soil was amended with treated rice straw and urea, whereas it was only 46% in soil amended with untreated rice straw and urea, and only 30% in soil treated with urea alone.     

Spatial and temporal controls of soil respiration rate in a high-elevation, subalpine forest

We examined soil respiration to determine what measurable environmental variables can be used to predict variation in soil respiration rates, spatially and temporally, at a high-elevation, mixed conifer, subalpine forest site at the Niwot Ridge Ameriflux Site in Colorado. For three summers, soil respiration rates were measured using soil collars and a portable gas-exchange system. Transects of the collars were established to ensure spatial characterization of the litter-repleted areas beneath tree crowns and the litter-depleted open spaces between tree crowns. Soil temperature and soil moisture were both identified as important drivers of soil respiration rate, but were found to confound each other and to function as primary controls at different scales. Soil temperature represents a primary control seasonally, and soil moisture represents a primary control interannually. Spatially, organic layer thickness, ammonium concentration, water content, and the microbial and soil soluble carbon pools were found to predict variation from point to point. Soil microbial biomass strongly correlated to soil respiration rate, whereas root biomass was identified as a weak predictor of respiration rate and only when controlling for other variables. Spatial variation in soil respiration rate is highly determined by the depth of the soil organic horizon, which in this ecosystem varies predictably according to distance from trees. The conclusions that can be drawn from the study provide the foundation for the development of future models of soil respiration driven by fundamental variables of the climate and soil microenvironment.     

Comparison of the difference method and 15N technique for studying the fate of nitrogen from plant residues in soil

We compared the dynamics of net mineralization of nitrogen (N) derived from white clover material (Ndfc) as measured by the difference and the ¹⁵N methods in a pot experiment with a sandy loam (15°C and pF 2.4) planted with Italian ryegrass. On day 22, mineralized Ndfc (soil mineral N plus plant N uptake) was 5.8% and 1.3% of added N for the ¹⁵N and the difference methods, respectively. The discrepancy was reduced on day 43. On day 64, the relationship was reversed, and on day 98 the values given by the two methods were 22.8% and 29.5%, respectively. The results obtained by the two methods were linearly correlated (r = 0.987) and, on average, did not differ significantly. Nevertheless, the different temporal patterns led to appreciably different parameter values as estimated by fitting of a reparameterized Richards model. On day 22, clover amendment reduced mineralized N derived from soil (Ndfs) by 3.4 mg N pot^–1. The reason for this was that the clover amendment led to a reduction in plant growth and uptake of Ndfs, most likely because of allelopathy, while mineral Ndfs did not increase correspondingly. Clover-induced Ndfs in the microbial biomass of 5.1 mg N pot^–1 suggested that the mineral Ndfs not taken up by plants had been reimmobilized. Towards the end of the experiment, clover-induced Ndfs in the biomass declined to 1.5 mg N pot^–1, while mineralized Ndfs due to clover amendment increased to 5.1 mg N pot^–1. The results strongly suggested that this increase was caused by a real stimulation of humus N mineralization by clover amendment rather than by isotope displacement or pool substitution. Received: 5 May 1997