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.     

Turnover of low molecular weight dissolved organic C (DOC) and microbial C exhibit different temperature sensitivities in Arctic tundra soils

Polar ecosystems are currently experiencing some of the fastest rates of climate warming. An increase in soil temperature in High Arctic regions may stimulate soil permafrost melting and microbial activity, thereby accelerating losses of greenhouse gases. It is therefore important to understand the factors regulating the rates of C turnover in polar soils. Consequently, our aims were to: (1) assess the concentration of low molecular weight (MW) dissolved organic carbon (DOC) in soil, (2) to investigate the temperature-dependent turnover of specific low MW compounds, and (3) to analyse the influence of substrate concentration on C cycling. Microbial mineralisation of labile low MW DOC in two High Arctic tundra soils was investigated using soil solutions spiked with either ¹⁴C-labelled glucose or amino acids. Spiked solutions were added to the top- and sub-soil from two ecosystem types (lichen and Carex dominated tundra), maintained at three temperatures (4–20 °C), and their microbial mineralisation kinetics monitored. ¹⁴CO₂ evolution from the tundra soils in response to ¹⁴C-glucose and -amino acid addition could best be described by a double first order exponential kinetic equation with rate constants k₁ and k₂. Both forms of DOC had a short half-life (t_1/2) in the pool of microbial respiratory substrate (t_1/2 = 1.07 ± 0.10 h for glucose and 1.63 ± 0.14 h for amino acids; exponential coefficient k₁ = 0.93 ± 0.07 and 0.64 ± 0.06 h^?1 respectively) whilst the second phase of mineralisation, assumed to be C that had entered the microbial biomass, was much slower (average k₂ = 1.30 × 10^?3 ± 0.49 × 10^?4 h^?1). Temperature had little effect on the rate of mineralisation of ¹⁴C used directly as respiratory substrate. In contrast, the turnover rate of the ¹⁴C immobilized in the microbial biomass prior to mineralisation was temperature sensitive (k₂ values of 0.99 × 10^?3 h^?1 and 1.66 × 10^?3 h^?1 at 4 and 20 °C respectively). Concentration-dependent glucose and amino acid mineralisation kinetics of glucose and amino acids (0–10 mM) were best described using Michaelis–Menten kinetics; there was a low affinity for both C substrates by the microbial community (K_m = 4.07 ± 0.41 mM, V_max = 0.027 ± 0.005 mmol kg^?1 h^?1). In conclusion, our results suggest that in these C limiting environments the flux of labile, low MW DOC through the soil solution is extremely rapid and relatively insensitive to temperature. In contrast, the turnover of C incorporated into higher molecular weight microbial C pools appears to show greater temperature sensitivity.     

Accumulation of As,Pb, Zn,Cd and Cu and arbuscular mycorrhizal status in populations of Cynodon dactylon grown on metal-contaminated soils

Metal(loid) accumulation and arbuscular mycorrhizal (AM) status of the dominant plant species, Cynodon dactylon, growing at four multi-metal(loid)s-contaminated sites and an uncontaminated site of China were investigated. Up to 94.7 As mg kg^?1, 417 Pb mg kg^?1, 498 Zn mg kg^?1, 5.8 Cd mg kg^?1 and 27.7 Cu mg kg^?1 in shoots of C. dactylon were recorded. The plant was colonized consistently by AM fungi (33.0–65.5%) at both uncontaminated site and metal-contaminated sites. Based on morphological characteristics, fourteen species of AM fungi were identified in the rhizosphere of C. dactylon, with one belonging to the genus of Acaulospora and the other thirteen belonging to the genus of Glomus. Glomus etunicatum was the most common species associated with C. dactylon growing at metal-contaminated sites. Spore abundance in the rhizosphere of C. dactylon growing at the metal-contaminated soils (22–82 spores per 25 g soil) was significantly lower than that of the uncontaminated soils (371 spores per 25 g soil). However, AM fungal species diversity in the metal-contaminated soils was significantly higher than that in the uncontaminated soils. This is the first report of AM status in the rhizosphere of C. dactylon, the dominant plant survival in metal-contaminated soils. The investigation also suggests that phytorestoration of metal-contaminated sites might be facilitated using the appropriate plant with the aid of tolerant AM fungi.     

Impact of bovine urine deposition on soil microbial activity,biomass, and community structure

Nitrogen (N) from urine excreted by grazing animals can be transformed into N compounds that have detrimental effects on the environment. These include nitrate, which can cause eutrophication of waterways, and nitrous oxide, which is a greenhouse gas. Soil microbes mediate all of these N transformations, but the impact of urine on microbes and how initial soil conditions and urine chemical composition alter their responses to urine are not well understood. This study aimed to determine how soil inorganic N pools, nitrous oxide fluxes, soil microbial activity, biomass, and the community structure of bacteria containing amoA (nitrifiers), nirK, and nirS (denitrifiers) genes responded to the addition of urine over time. Bovine urine containing either a high (15.0 g K⁺ l^?1) or low salt content (10.4 g K⁺ l^?1) was added to soil cores at either low or high moisture content (hereafter termed dry and wet soil respectively; 35% or 70% water-filled pore space after the addition of urine). Changes in soil conditions, inorganic N pools, nitrous oxide fluxes, and the soil microbial community were then measured 1, 3, 8, 15, 29 and 44 days after urine addition. Urine addition increased soil ammonium concentrations by up to 2 mg g d.w.^?1, soil pH by up to 2.7 units, and electrical conductivity (EC) by 1.0 and 1.6 dS m^?1 in the low and high salt urine treatments respectively. In response, nitrate accumulation and nitrous oxide fluxes were lower in dry compared to wet urine-amended soils and slightly lower in high compared to low salt urine-amended soils. Nitrite concentrations were elevated (>3 μg g d.w.^?1) for at least 15 days after urine addition in wet urine-amended soils, but were only this high in the dry urine-amended soils for 1 day after the addition of urine. Microbial biomass was reduced by up to half in the wet urine-amended soils, but was largely unaffected in the dry urine-amended soils. Urine addition affected the community structure of ammonia-oxidising and nitrite-reducing bacteria; this response was also stronger and more persistent in wet than in dry urine-amended soils. Overall, the changes in soil conditions caused by the addition of urine interacted to influence microbial responses, indicating that the effect of urine on soil microbes is likely to be context-dependent.     

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.     

Production and dissolution rates of earthworm-secreted calcium carbonate

Earthworms secrete granules of calcium carbonate. These are potentially important in soil biogeochemical cycles and are routinely recorded in archaeological studies of Quaternary soils. Production rates of calcium carbonate granules by the earthworm Lumbricus terrestris L. were determined over 27 days in a range of soils with differing chemical properties (pH, organic matter content, water holding capacity, bulk composition, cation exchange capacity and exchangeable cations). Production rate varied between soils, lay in the range $0–0.043{\text{mmol}}_{{\text{CaCO}}_3}$ (0–4.3 mg) earthworm^?1 d^?1 with an average rate of $8\times {10}^{?3}{\text{mmol}}_{{\text{CaCO}}_3}$ (0.8 mg) earthworm^?1 d^?1 and was significantly correlated (r = 0.68, P ≤ 0.01) with soil pH. In a second experiment lasting 315 days earthworms repeatedly (over periods of 39–57 days) produced comparable masses of granules. Converting individual earthworm granule production rates into fluxes expressed on a per hectare of land per year basis depends heavily on estimates of earthworm numbers. Using values of 10–20 L. terrestris m^?2 suggests a rate of $18–3139{\text{mol}}_{{\text{CaCO}}_3}{\text{ha}}^{?1}{\text{yr}}^{?1}$. Data obtained from flow-through dissolution experiments suggest that at near neutral pH, granule geometric surface area-normalised dissolution rates are similar to those for other biogenic and inorganic calcites. Fits of the data to the dissolution relationship r = k(1 ? Ω)ⁿ where r = dissolution rate, k = a rate constant, Ω = relative saturation and n = the reaction order gave values of k = 1.72 × 10^?10 mol cm^?2 s^?1 and n = 1.8 for the geometric surface area-normalised rates and k = 3.51 × 10^?13 mol cm^?2 s^?1 and n = 1.8 for the BET surface area-normalised rates. In 196 day leaching column experiments trends in granule dissolution rate referenced to soil chemistry corresponded to predictions made by the SLIM model for dissolution of limestone in soil. If soil solution approaches saturation with respect to calcium carbonate, granule dissolution will slow or even stop and granules be preserved indefinitely. Granules have the potential to be a small but significant component of the biogeochemical cycling of C and Ca in soil.     

The temperature dependence of dicyandiamide (DCD) degradation in soils: A data synthesis

Dicyandiamide (DCD, C₂H₄N₄) is a nitrification inhibitor that has been studied for more than 80 years. However, there are few papers that have examined the use of DCD on dairy farms where cattle graze pasture and where urine is the primary form of nitrogen (N) deposited onto soils. After DCD was applied (10 kg DCD ha^?1) with bovine urine (700–1200 kg N ha^?1) to five soils throughout New Zealand, the reduction in direct nitrous oxide (N₂O) emissions was significant and remarkably consistent (71 ± 8%, average ± standard error). The application of DCD to these soils occurred in autumn and winter; daily average soil temperature (T) was reported but these data were not further analysed. Perusal of the literature suggested no consensus on the temperature dependence of DCD degradation in soils. Based on published data from controlled-environment studies of soils sampled in four countries, we quantified the relation between T and the time for DCD concentration in soils to decline to half its application value (t_½) as t_½ (T) = 168e^?0.084T with parameter standard errors of ±16 d and ±0.011 d^?1, respectively (n = 16). For example, at 5 °C a 1 °C increase in T reduced t_½ from 110 to 101 d whereas at 25 °C the reduction was 20–19 d. Analysing T data from the New Zealand trials using our t_½ (T) function, over 43–89 d when direct N₂O emissions from treated plots became indistinguishable from the controls, the estimated percentage of applied DCD remaining in the soil averaged 43 ± 10%. These calculations suggested the apparently remaining DCD was ineffective with respect to direct N₂O emissions. In the absence of measurements, explanations for this interpretation included vertical displacement of the DCD and sorption onto organic matter in soils. The consistent DCD efficacy from these trials corresponded with T generally <10 °C, so it is suggested as an application criteria for the reduction of direct N₂O emissions from pastoral soils subjected to urine excretion by grazing cattle.     

Nitrogen mineralization in the ruderal sub-alpine communities in Mount Uludağ, Turkey

Nitrogen mineralization and nitrification in the soil of sub-alpine ruderal community of Mount Uludağ, Bursa, Turkey was measured for 1 year, under field conditions with Verbascum olympicum and Rumex olympicus being the dominant pioneer species under dry and wet sites, respectively. Seasonal fluctuations were observed in N mineralization and nitrification. The net N mineralization and nitrification were high in early summer and winter, due to high moisture. The annual net N mineralization rate (for the 0–15 cm soil layer) was higher under R. olympicus (188 kg N ha⁻¹ yr⁻¹) than under V. olympicum (96 kg N ha⁻¹ yr⁻¹). A significant positive correlation between net N mineralization and soil organic C (r² = 0.166), total N (r² = 0.141) and water content (r² = 0.211) was found. Our results indicate that N mineralization rate is high in soils of ruderal communities on disturbed sites and varies with dominant species and, a difference in net N mineralization rate can be attributed to organic C, total N and moisture content of soils.     

What are the limits of the drilosphere? An incubation experiment using Metaphire posthuma

The near infrared reflectance spectroscopy (NIRS) method was used in the present study to compare earthworm-made soil aggregates to aggregates found in the surrounding bulk soil. After initially assessing the daily cast production of Metaphire posthuma, boxes with soil incubated with M. posthuma and control soils were subjected to wetting in order to reorganize the soil structure. After two months of incubation, soil aggregates produced by earthworms (casts and burrows), soil aggregates that were appeared to be unaffected by earthworms (bulk soil without visible trace of earthworm bioturbation from the earthworm treatment) and soil aggregates that were entirely unaffected by earthworms (control – no earthworm – treatment) were sampled and their chemical signatures analyzed by NIRS. The production of below-ground and surface casts reached 14.9 g soil g worm^?1 d^?1 and 1.4 g soil g worm^?1 d^?1, respectively. Soil aggregates from the control soils had a significantly different NIRS signature from those sampled from boxes with earthworms. However, within the earthworm incubation boxes the NIRS signature was similar between cast and burrow aggregates and soil aggregates from the surrounding bulk soil. We conclude that the high cast production by M. posthuma and the regular reorganization of the soil structure by water flow in and through the soil lead to a relatively homogenous soil structure. Given these results, we question the relevance of considering the bulk soil that has no visible activity of earthworm activity as a control to determine the effect of earthworms on soil functioning.     

Partitioning soil surface CO2 efflux into autotrophic and heterotrophic components,using natural gradients in soil δ13C in an undisturbed savannah soil

We used natural gradients in soil and vegetation δ¹³C signatures in a savannah ecosystem in Texas to partition soil respiration into the autotrophic (Ra) and heterotrophic (Rh) components. We measured soil respiration along short transects from under clusters of C₃ trees into the C₄ dominated grassland. The site chosen for the study was experiencing a prolonged drought, so an irrigation treatment was applied at two positions of each transect. Soil surface CO₂ efflux was measured along transects and CO₂ collected for analysis of the δ¹³C signature in order to: (i) determine how soil respiration rates varied along transects and were affected by localised change in soil moisture and (ii) partition the soil surface CO₂ efflux into Ra and Rh, which required measurement of the δ¹³C signature of root- and soil-derived CO₂ for use in a mass balance model.The soil at the site was unusually dry, with mean volumetric soil water content of 8.2%. Soil respiration rates were fastest in the centre of the tree cluster (1.5 ± 0.18 μmol m^?2 s^?1; mean ± SE) and slowest at the cluster–grassland transition (0.6 ± 0.12 μmol m^?2 s^?1). Irrigation produced a 7–11 fold increase in the soil respiration rate. There were no significant differences (p > 0.5) between the δ¹³C signature of root biomass and respired CO₂, but differences (p < 0.01) were observed between the respired CO₂ and soil when sampled at the edge of the clusters and in the grassland. Therefore, end member values were measured by root and soil incubations, with times kept constant at 30 min for roots and 2 h for soils. The δ¹³C signature of the soil surface CO₂ efflux and the two end member values were used to calculate that, in the irrigated soils, Rh comprised 51 ± 13.5% of the soil surface CO₂ efflux at the mid canopy position and 57 ± 7.4% at the drip line. In non-irrigated soil it was not possible to partition soil respiration, because the δ¹³C signature of the soil surface CO₂ efflux was enriched compared to both the end member values. This was probably due to a combination of the very dry porous soils at our study site (which may have been particularly susceptible to ingress of atmospheric CO₂) and the very slow respiration rates of the non-irrigated soils.     

Rapid estimation of microbial biomass in grassland soils by ultra-violet absorbance

A reliable and simple technique for estimating soil microbial biomass (SMB) is essential if the role of microbes in many soil processes is to be quantified. Conventional techniques are notoriously time-consuming and unreproducible. A technique was investigated that uses the UV absorbance at 280 nm of 0.5 M K₂SO₄ extracts of fumigated and unfumigated soils to estimate the concentrations of carbon, nitrogen and phosphorus in the SMB. The procedure is based on the fact that compounds released after chloroform fumigation from lysed microbial cells absorb in the near UV region. Using 29 UK permanent grassland soils, with a wide range of organic matter (2.9–8.0%) and clay contents (22–68%), it was demonstrated that the increase in UV absorbance at 280 nm after soil fumigation was strongly correlated with the SMB C (r=0.92), SMB N (r=0.90) and SMB P (r=0.89), as determined by conventional methods. The soils contained a wide range of SMB C (412–3412 μg g⁻¹ dry soil), N (57–346 μg g⁻¹ dry soil) and P (31–239 μg g⁻¹ dry soil) concentrations. It was thus confirmed that the UV absorbance technique described was a rapid, simple, precise and relatively inexpensive method of estimating soil microbial biomass.     

Improved effectiveness and efficiency in measuring soil enzymes as universal soil quality indicators using microplate fluorimetry

We tested the relevance of the microplate fluorimetric (F) assay for five enzymes in contrasting land uses, including woodland, grassland, cultivated and contaminated lands, as compared to the standard spectrophotometric (P) method. Enzymatic activity measured by the P method ranged from 0 to 56.04 nmol-pNP g^?1 min^?1 (median = 4) while the F method revealed lower values ranging from 0 to 6.22 nmol-MUB g^?1 dry soil min^?1 (median = 1). The values obtained by the P method were around 8 times higher than those revealed by the F method. However, the F method revealed significant differences in enzyme activity in orchard parcels (land use with low variations in soil properties). We concluded that the F method improves the effectiveness and the efficiency of measuring universal soil quality indicators using enzymes.     

Differences in soil enzyme activities,microbial community structure and short-term nitrogen mineralisation resulting from farm management history and organic matter amendments

Changes in soil microbial biomass, enzyme activities, microbial community structure and nitrogen (N) dynamics resulting from organic matter amendments were determined in soils with different management histories to gain better understanding of the effects of long- and short-term management practices on soil microbial properties and key soil processes. Two soils that had been under either long-term organic or conventional management and that varied in microbial biomass and enzyme activity levels but had similar fertility levels were amended with organic material (dried lupin residue, Lupinus angustifolius L.) at amounts equivalent to 0, 4 and 8 t dry matter lupin ha^?1. Microbial biomass C and N, arginine deaminase activity, fluorescein diacetate hydrolysis, dehydrogenase enzyme activity and gross N mineralisation were measured in intervals over an 81-day period. The community structure of eubacteria and actinomycetes was examined using PCR–DGGE of 16S rDNA fragments. Results suggested that no direct relationships existed between microbial community structure, enzyme activities and N mineralisation. Microbial biomass and activity changed as a result of lupin amendment whereas the microbial community structure was more strongly influenced by farm management history. The addition of 4 t ha^?1 of lupin was sufficient to stimulate the microbial community in both soils, resulting in microbial biomass growth and increased enzyme activities and N mineralisation regardless of past management. Amendment with 8 t lupin ha^?1 did not result in an increase proportional to the extra amount added; levels of soil microbial properties were only 1.1–1.7 times higher than in the 4 t ha^?1 treatment. Microbial community structure differed significantly between the two soils, while no changes were detected in response to lupin amendment at either level during the short-term incubation. Correlation analyses for each treatment separately, however, revealed differences that were inconsistent with results obtained for soil biological properties suggesting that differences might exist in the structure or physiological properties of a microbial component that was not assessed in this study.     

Modeling organic carbon dynamics under no-tillage and plowed systems in tropical soils of Brazil using CQESTR

CQESTR simulates the effect of management practices on soil organic carbon (SOC) stocks. The beta version of the model had been calibrated and validated for temperate regions. Our objective was to evaluate the CQESTR model performance for simulating carbon dynamics as affected by tillage practices in two tropical soils (Ultisol and Oxisol) in southeastern and northeastern Brazil. In the southeast (20.75 S 42.81 W), tillage systems consisted of no tillage (NT); reduced tillage (RT) (one disc plow and one harrow leveling [RT1] or one heavy disc harrow and one harrow leveling [RT2]); and conventional tillage (CT) (two heavy disc harrows followed by one disc plow and two harrow levelings). In the northeast (7.55 S 45.23 W), tillage systems consisted of NT, RT (one chisel plow and one harrow leveling), and CT (one disk plow, two heavy disk harrowings, and two harrow levelings). CQESTR underestimated SOC at both sites, especially under NT systems, indicating that adjustments (e.g., the inclusion of clay mineralogy factor) are necessary for more accurate simulation of SOC in the tropics. In spite of this, measured and simulated values of SOC in the 0–20 cm depth were well correlated (southeast, R² = 0.94, p < 0.01; northeast, R² = 0.88, p < 0.05). With respect to initial conditions (native forest), CQESTR estimated a decrease in SOC stocks in plowed and no-tillage systems. In 2006, in the southeast, SOC stocks were 28.8, 23.7, 23.2, and 22.0 Mg ha^?1 under NT, RT2, RT1, and CT, respectively; in the northeast, stocks were 36.0, 33.8, and 32.5 Mg ha^?1 under NT, RT, and CT, respectively. The model estimated carbon emissions varying from 0.36 (NT) to 1.05 Mg ha^?1 year^?1 (CT) in the southeast and from 0.30 (NT) to 0.82 (CT) Mg ha^?1 year^?1 in the northeast. CQESTR prediction of SOC dynamics illustrates acceptable performance for the two tropical soils of Brazil.     

Cultivation effects on temporal changes of organic carbon and aggregate stability in desert soils of Hexi Corridor region in China

Sustainable agricultural use of cultivated desert soils has become a concern in Hexi Corridor in Gansu Province of China, because loss of topsoil in dust storms has been recently intensified. We chose four desert sites to investigate the effects of cultivation (cropping) on (i) soil organic C and its size fractions and (ii) soil aggregate stability (as a measure of soil erodibility). These parameters are of vital importance for evaluating the sustainability of agricultural practices.Total organic C as well as organic C fractions in soil (coarse organic C, 0.1–2 mm; young organic C, 0.05–0.1 mm; stable organic C, <0.05 mm) generally increased with the duration of the cultivation period from 0 (virgin soil, non-cultivated) to more than 30 years (p < 0.05). Compared to total organic C in virgin soils (2.3–3.5 g kg⁻¹ soil), significantly greater values were found after 10 to >20 years of cultivation (6.2–7.1 g kg⁻¹ soil). The increase in organic C in desert soils following prolonged cultivation was mainly the consequence of an increase in the coarse organic C. The increase in total organic C in soil was also dependent on clay content [total organic C = 0.96 + 0.249 clay content (%) + 0.05 cultivation year, R² = 0.48, n = 27, p < 0.001]. This indicates that clay protected soil organic C from mineralization, and also contributed to the increase in soil organic C as time of cultivation increased.There was a significant positive correlation between aggregate stability and total organic C across all field sites. The water stability of aggregates was low (with water-stable aggregate percentage ∼4% of dry-sieved aggregates of size 1–5 mm). There was no consistent pattern of increase in the soil aggregate stability with time of cultivation at different locations, suggesting that desert soils might remain prone to wind erosion even after 50 years of cultivation. Alternative management options, such as retaining harvested crop residues on soil surface and excluding or minimizing tillage, may permit sustainable agricultural use of desert soils.     

Evapotranspiration and water use efficiency in different-aged Pacific Northwest Douglas-fir stands

We used the eddy-covariance technique to measure evapotranspiration (E) and gross primary production (GPP) in a chronosequence of three coastal Douglas-fir (Pseudotsuga menziesii) stands (7, 19 and 58 years old in 2007, hereafter referred to as HDF00, HDF88 and DF49, respectively) since 1998. Here, we focus on the controls on canopy conductance (g_c), E, GPP and water use efficiency (WUE) and the effect of interannual climate variability at the intermediate-aged stand (DF49) and then analyze the effects of stand age following clearcut harvesting on these characteristics. Daytime dry-foliage Priestley–Taylor α and g_c at DF49 were 0.4–0.8 and 2–6 mm s^?1, respectively, and were linearly correlated (R² = 0.65). Low values of α and g_c at DF49 as well at the other two stands suggested stomatal limitation to transpiration. Monthly E, however, showed strong positive linear correlations to monthly net radiation (R² = 0.94), air temperature (R² = 0.77), and daytime vapour pressure deficit (R² = 0.76). During July–September, monthly E (mm) was linearly correlated to monthly mean soil water content (θ, m³ m^?3) in the 0–60 cm layer (E = 453θ ? 21, R² = 0.69), and GPP was similarly affected. Annual E and GPP of DF49 for the period 1998–2007 varied from 370 to 430 mm and from 1950 to 2390 g C m^?2, respectively. After clearcut harvesting, E dropped to about 70% of that for DF49 while ecosystem evapotranspiration was fully recovered when stand age was ～12 years. This contrasted to GPP, which varied hyperbolically with stand age. Monthly GPP showed a strong positive linear relationship with E irrespective of the stand age. While annual WUE of HDF00 and HDF88 varied with age from 0.5 to 4.1 g C m^?2 kg^?1 and from 2.8 to 4.4 g C m^?2 kg^?1, respectively, it was quite conservative at ～5.3 g C m^?2 kg^?1 for DF49. N-fertilization had little first-year response on E and WUE. This study not only provides important results for a more detailed validation of process-based models but also helps in predicting the influences of climate change and forest management on water vapour and CO₂ fluxes in Douglas-fir forests.     

The effect of organic matter on the structure of soils of different land uses

In this study, the soil structure of two soils (Haplic Chernozem and Eutric Fluvisol) of different land uses (forest, meadow, urban and agro-ecosystem – consisted of four crop rotations) in Slovakia was compared. The soil aggregate stability was determined with a dependence on the chemical composition of plant residues. The quantity and quality of the organic matter was assessed through the parameters of the C and N in size fractions of dry-sieved and water-resistant aggregates. The soil structure of the forest ecosystem was evaluated as the best of all of forms of land use. Differences in the soil structure under the grass vegetation of a meadow (natural conditions) and urban ecosystem were also recorded. The agro-ecosystem was characterised by a higher portion (55.95%) of the most valuable (agronomically) water-resistant aggregate size fraction of 0.5–3 mm. Values of the carbon management index showed that the larger water-resistant aggregates were, the greater were the changes in the organic matter (r = ?0.680, P < 0.05). In addition, a smaller content of dry-sieved aggregates of the 3–5 mm size fraction was observed with higher contents of soil organic carbon (SOC) (r = ?0.728, P < 0.05) and labile carbon (C_L) (r = ?0.760, P < 0.05); there were also greater changes in the soil organic matter and vice versa, higher contents of SOC (r = 0.744, P < 0.05) and C_L (r = 0.806, P < 0.05) greater contents of dry-sieved aggregates of size fraction 0.5–1 mm. The soil structure of agro-ecosystem was superior at a higher content of cellulose (r = ?0.712, P < 0.05) in the plant residues. The higher content of cellulose and hemicellulose in the plant residue of a previous crop was reflected in a smaller C_L content in the water-resistant aggregates (r = ?0.984, P < 0.05). A correlation was observed between a high content of lignin in the plant residue and a smaller SOC content in the water-resistant aggregates (r = ?0.967, P < 0.05). Lastly, a higher content of proteins in the plant residues (r = 0.744, P < 0.05) supported a greater content of dry-sieved aggregates of the 0.5–1 mm size fraction.     

Population size and specific nitrification potential of soil ammonia-oxidizing bacteria under long-term fertilizer management

Population size of soil ammonia-oxidizing bacteria (AOB) was quantified by real-time PCR in a long-term (16 years) field experiment under different fertilizer managements. AOB population sizes in mineral nitrogen-fertilized soils and organic manure-fertilized soil were 10.3 and 3.1 times, respectively, that of the control, while phosphorus and potassium fertilization had no significant effect. On the other hand, the AOB specific nitrification potential (soil nitrification potential per AOB cell) was significantly higher (P < 0.05) in organic manure-fertilized soil than in mineral-fertilized soils and the control, indicating that AOB was likely more metabolically active in organic manure-fertilized soils than in mineral nitrogen-fertilized soils after long-term application.     

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.     

Earthworm-induced N2O emissions in a sandy soil with surface-applied crop residues

Earlier research with endogeic and epigeic earthworm species in loamy arable soil has shown that both earthworm groups can increase nitrous oxide (N₂O) emissions, provided that crop residue placement matches the feeding strategy of the earthworm ecological group(s). However, it is not yet clear whether these effects also occur in sandy soils which typically contain less soil organic matter and have low soil aggregation levels. Here, we aimed to quantify N₂O emissions as affected by endogeic and/or epigeic earthworm species, and to relate changes in N₂O emissions to earthworm-induced changes in soil properties in a sandy soil. A 90 day mesocosm study was conducted with sandy soil and ¹⁵N-labeled radish (Raphanus sativus cv. Adagio L.) residue applied on top. Treatments included: (i) no earthworm addition, (ii) addition of the endogeic species Aporrectodea caliginosa (Savigny), (iii) addition of the epigeic species Lumbricus rubellus (Hoffmeister), and (iv) both species combined. An additional treatment was included without earthworms and with residue manually incorporated into the soil. L. rubellus significantly increased cumulative N₂O emissions from 228 to 859 μg N₂O–N kg^?1 (F_1,12 = 83.12, P < 0.001), whereas A. caliginosa did not affect N₂O emissions. In contrast to earlier studies in loamy soil, no positive interaction between both species with regard to N₂O emissions was found. This was probably related to high competition for organic resources in the relatively poor soil and a low potential for stable soil aggregate formation (and associated anaerobic microsites) by endogeic worms in sandy soil. ¹⁵N isotope analysis revealed that the activity of L. rubellus significantly increased (F_1,12 = 6.20, P = 0.028) the recovery of ¹⁵N in the 250–8000 μm size fraction, indicating incorporation of crop residues into the mineral soil. When residues were manually incorporated, N₂O emissions were significantly (P < 0.008) lower (509 μg N₂O–N kg^?1) than when incorporated by L. rubellus. The high N₂O emissions in the presence of L. rubellus, when compared to manual mixing, suggest a stimulation of microbial activity and/or changes in the microbial community composition. Insights on the earthworm effects on N₂O emission from such soils are discussed.