Microbial community composition shapes enzyme patterns in topsoil and subsoil horizons along a latitudinal transect in Western Siberia

Soil horizons below 30 cm depth contain about 60% of the organic carbon stored in soils. Although insight into the physical and chemical stabilization of soil organic matter (SOM) and into microbial community composition in these horizons is being gained, information on microbial functions of subsoil microbial communities and on associated microbially-mediated processes remains sparse. To identify possible controls on enzyme patterns, we correlated enzyme patterns with biotic and abiotic soil parameters, as well as with microbial community composition, estimated using phospholipid fatty acid profiles. Enzyme patterns (i.e. distance-matrixes calculated from these enzyme activities) were calculated from the activities of six extracellular enzymes (cellobiohydrolase, leucine-amino-peptidase, N-acetylglucosaminidase, chitotriosidase, phosphatase and phenoloxidase), which had been measured in soil samples from organic topsoil horizons, mineral topsoil horizons, and mineral subsoil horizons from seven ecosystems along a 1500 km latitudinal transect in Western Siberia. We found that hydrolytic enzyme activities decreased rapidly with depth, whereas oxidative enzyme activities in mineral horizons were as high as, or higher than in organic topsoil horizons. Enzyme patterns varied more strongly between ecosystems in mineral subsoil horizons than in organic topsoils. The enzyme patterns in topsoil horizons were correlated with SOM content (i.e., C and N content) and microbial community composition. In contrast, the enzyme patterns in mineral subsoil horizons were related to water content, soil pH and microbial community composition. The lack of correlation between enzyme patterns and SOM quantity in the mineral subsoils suggests that SOM chemistry, spatial separation or physical stabilization of SOM rather than SOM content might determine substrate availability for enzymatic breakdown. The correlation of microbial community composition and enzyme patterns in all horizons, suggests that microbial community composition shapes enzyme patterns and might act as a modifier for the usual dependency of decomposition rates on SOM content or C/N ratios.     

The influence of time, storage temperature, and substrate age on potential soil enzyme activity in acidic forest soils using MUB-linked substrates and l-DOPA

The purpose of this experiment was to evaluate whether soil storage and processing methods significantly influence measurements of potential in situ enzyme activity in acidic forest soils. More specifically, the objectives were to determine if: (1) duration and temperature of soil storage; (2) duration of soil slurry in buffer; and (3) age of model substrates significantly influence the activity of six commonly measured soil extracellular enzymes using methylumbelliferone (MUB)-linked substrates and l-dihydroxyphenylalanine (l-DOPA). Soil collected and analyzed for enzyme activity within 2 h was considered the best measure of potential in situ enzyme activity and the benchmark for all statistical comparisons. Sub-samples of the same soil were stored at either 4 °C or −20 °C. In addition to the temperature manipulation, soils experienced two more experimental treatments. First, enzyme activity was analyzed 2, 7, 14, and 21 days after collection. Second, MUB-linked substrate was added immediately (i.e. <20 min) or 2 h after mixing soil with buffer. Enzyme activity of soil stored at 4 °C was not significantly different from soil stored at −20 °C. The duration of soil storage was minimal for β-glucosidase, β-xylosidase, and peroxidase activity. N-acetyl-glucosaminidase (NAGase), phosphatase, and phenol oxidase activity appeared to change the most when compared to fresh soils, but the direction of change varied. Likewise, the activities of these enzymes were most sensitive to extended time in buffer. Fluorometric MUB and MUB-linked substrates generally had a 3-day shelf life before they start to significantly suppress reported activities when kept at 4 °C. These findings suggest that the manner in which acidic forest soils are stored and processed are site and enzyme specific and should not initially be trivialized when conducting enzyme assays focusing on NAGase, phosphatase, and phenol oxidase. The activities of β-glucosidase, β-xylosidase, and peroxidase are insensitive to storage and processing methods.     

我国主要土壤剖面酶活性状况

本世纪五十年代以来,由于科学的发展以及新技术的引入,土壤酶的研究愈来愈为人们所重视。试验研究已证明,土壤酶是土壤的组成分之一。     

Interactions between N fertilization, grass clipping addition and pH in turf ecosystems: Implications for soil enzyme activities and organic matter decomposition

Turf has been acknowledged as an important ecosystem with potential for soil C sequestration. As a major process dictating soil C storage, organic matter decomposition has received little attention in turf systems. Given that soil enzyme-catalyzed biochemical reactions are the rate limiting steps of organic matter decomposition, we examined the activities of oxidative and hydrolytic soil enzymes and their relations with soluble organic compounds and soil C and N mineralization in two turf chronosequences with contrasting soil pH and in response to N fertilization and grass clipping addition. In comparison with turf ecosystems under acidic soil, phenol oxidase activity was about two-fold greater in turf ecosystems under alkaline soil and positively correlated to about two-fold differences in soluble phenolics and dissolved organic C between alkaline and acidic soils. However, the activities of hydrolytic enzymes including cellulase, chitinase, and glucosidase were lower in alkaline soil. It appears that the high concentration of soluble phenolics inhibited the activities of hydrolytic enzymes that in turn limited the decomposition of dissolved organic C and resulted in its accumulation in alkaline soil. Nitrogen mineralization was comparable between alkaline and acidic soils, but CO₂ evolution was about two-fold greater in alkaline soil, possibly due to considerable abiotic carbonate dissolution. We observed that mineral N input at 60 mg N kg⁻¹ soil had very minor negative effects on the activities of both phenol oxidase and hydrolytic enzymes. Grass clipping addition did not affect the activity of phenol oxidase, but increased the activities of soil chitinase, cellulase, glucosidase, and glucosaminidase by up to 20% and also soluble phenolics in soil by about 10%. Our results suggest that soil phenol oxidase might regulate the activities of hydrolytic soil enzymes via its control on soluble phenolics and function as an ‘enzymatic latch’ to hold soil organic C in highly managed turf ecosystems. While soil pH is important to affect phenol oxidase activity and therefore decomposition, management practices, i.e., N fertilization and grass clipping addition may indirectly affect the decomposition through enhancing turfgrass productivity and thus soil C input.     

Vertical distribution of soil properties under short‐rotation forestry in Northern Germany

Short‐rotation forestry (SRF) on arable soils has high potentials for biomass production and leads to long‐term no‐tillage management. In the present study, the vertical distributions of soil chemical and microbial properties after 15 y of SRF with willows and poplar (Salix and Populus spp.) in 3‐ and 6‐year rotations on an arable soil were measured and compared to a pertinent tilled arable site. Two transects at different positions in the relief (upper and lower slope; transect 1 and 2) were investigated. Short‐rotation forestry caused significant changes in the vertical distribution of all investigated soil properties (organic and microbial C, total and microbial N, soil enzyme activities), however, the dimension and location (horizons) of significant effects varied. The rotation periods affected the vertical distribution of the soil properties within the SRF significantly. In transect 1, SRF had higher organic‐C concentrations in the subsoil (Bv horizon), whereas in transect 2, the organic‐C concentrations were increased predominantly in the topsoil (Ah horizon). Sufficient plant supply of P and K in combination with decreased concentrations of these elements in the subsoil under SRF pointed to an effective nutrient mobilization and transfer from the deeper soil horizons even in the long term. In transect 1, the microbial‐C concentrations were higher in the B and C horizons and in transect 2 in the A horizons under SRF than under arable use. The activities of β‐glucosidases and acid phosphatases in the soil were predominantly lower under SRF than under arable use in the topsoil and subsoil. We conclude, that long‐term SRF on arable sites can contribute to increased C sequestration and changes in the vertical distribution of soil microbial biomass and soil enzyme activities in the topsoil and also in the subsoil.     

Carbon structure and enzyme activities in alpine and forest ecosystems

The chemical structure of soil organic matter fractions and its relationship to biological processes remains uncertain. We used pyrolysis-gas chromatography/mass spectrometry to analyze the molecular structure of light and heavy fraction C from soils in the San Juan Mountains, Colorado. The soil samples, each replicated three times, were from two elevations (alpine and low forest) within two geochemically distinct basins (igneous and sedimentary). We also analyzed whether variation in the activity of nine enzymes that mediate soil organic matter turnover and nutrient cycling could explain differences in C structure. We found that, across basins and elevation, light fraction and heavy fraction C had distinct chemistries. The light fraction was characterized by an abundance of plant lignin biomarkers, including phenol, 2-methoxy-4-vinyl-(vinylguaiacol) and phenol, 2-methoxy-(guaiacol); in contrast heavy fraction had very little unaltered lignin but an abundance of polysaccharides, such as furfural, and proteins such as pyrrole. In alpine sites, light fraction was less abundant (4.27 versus 31.79 g kg⁻¹) and had a lower C/N ratio (17.25 versus 32.01) than in forests. The alpine sites also had higher activities of phosphatase, β-d-1,4-cellobiosidase, β-1,4-glucosidase, l-leucine aminopeptidase, and β-1,4-xylosidase. Protein abundance in the heavy fraction was correlated with peptidase, β-1,4-glucosidase, and phosphatase activities; in the light fraction, protein abundance was correlated with peptidase, xylosidase, and β-d-1,4-cellobiosidase activities. β-1,4-N-acetyl-glucosaminidase was negatively correlated with polysaccharides in the light and heavy fractions and positively correlated with lignin in the light fraction. However, there were not always significant correlations between enzymes and substrates. We suggest that this is likely because soil organic matter chemistry reflects long-term decomposition processes while enzyme dynamics fluctuate with current conditions or due to the presence of a pool of sorbed enzymes in the heavy fraction. While alpine and forest ecosystem C distribution and enzyme activities varied, substantial depletion of lignin derivatives in the heavy fraction across sites suggest that these compounds do not persist in stable soil C pools.     

Soil enzyme activity in an old-growth northern hardwood forest: Interactions between soil environment,ectomycorrhizal fungi and plant distribution

Plants and soil microbes produce extracellular enzymes (EE) that catalyze the hydrolysis of nitrogen (N) and phosphorus (P) containing compounds in soil and other enzymes involved in degradation of lignin and cellulose. We explored whether soil enzyme activity involved in carbon (C), N and P cycling were correlated with plant distribution, soil chemical conditions and the identity of fungi colonizing tree roots in an old growth forest remnant. Terminal restriction fragment length polymorphism (TRFLP) was used to determine the presence of root fungi and standard fluorometric analysis was used to determine soil enzyme activities. Soil enzymes were consistently positively correlated with soil C and N, but not CN ratio. Soil P was also correlated with enzyme activity during both June and September sampling. We saw no significant relationships between herbaceous plant cover and enzyme activity in June, but there were significant positive correlations between α-glucosidase and herbaceous plant coverage in September. We also found that some enzymes were significantly correlated with the identity of fungi colonizing tree roots separated from the soil cores. Chitinase and β-glucosidase were positively correlated with the genera Russula and Piloderma while chitinase was negatively correlated with Amanita and Entoloma. In addition, phosphatase was positively correlated with Russula, Meliniomyces and Solenopezia. Our results suggest that enzyme activity in old growth forest soils are affected by a variety of environmental factors, and that herbaceous plants and some root fungi may be associated with sites of elevated or decreased decomposition potential and nutrient cycling.     

Enzyme activities in hill land soils of the Appalachian region

Abstract

The top two horizons of 14 major hill land soils of the Appalachian Region were evaluated for activities of add phospha‐tase (AP), pyrophosphatase (PP1), arylsuifatase (AS) and urease (UR) enzymes. A relationship between enzyme activities and soil properties was examined. Surface horizons contained higher enzyme activities than the subsurface horizons. Overall, enzyme activities were positively correlated with soil moisture content, percent water‐filled porosity, C, N, K, Mn and CEC. Activities of AP and PP1 were negatively correlated with soil pH. The AP and UR activities were positively related to various forms of P. Enzyme activities were positively related to total and organic S. Pot trial was performed 1n green house conditions to evaluate the relationship between enzyme activities and snapbean (Phaseolus vulgaris L.) growth and N, P, and S uptake. With a few exceptions, activities of AP, AS and UR were positively related to snapbean shoot wt. and shoot content of N, P, and S. The PP1 activities of subsurface horizons were positively related to shoot wt. and mineral content. Enzyme activities varied from one soil horizon to another. The effects of various soil properties and their interacting factors on activities of the four enzyme systems are discussed.     

Soil organic matter stabilization in acidic forest soils is preferential and soil type-specific

Minerals with large specific surface areas promote the stabilization of soil organic matter (SOM). We analysed three acidic soils (dystric, skeletic Leptic Cambisol; dystric, laxic Leptic Cambisol; skeletic Leptic Entic Podzol) under Norway spruce (Picea abies) forest with different mineral compositions to determine the effects of soil type on carbon (C) stabilization in soil. The relationship between the amount and chemical composition of soil organic matter (SOM), clay content, oxalate‐extractable Fe and Al (Fe_o; Al_o), and dithionite‐extractable Fe (Fe_d) before and after treatment with 10% hydrofluoric acid (HF) in topsoil and subsoil horizons was analysed. Radiocarbon age, ¹³C CPMAS NMR spectra, lignin phenol content and neutral sugar content in the soils before and after HF‐treatment were determined and compared for bulk soil samples and particle size separates. Changes in the chemical composition of SOM after HF‐treatment were small for the A‐horizons. In contrast, for B‐horizons, HF‐soluble (mineral‐associated) and HF‐resistant (non‐mineral‐associated) SOM showed systematic differences in functional C groups. The non‐mineral associated SOM in the B‐horizons was significantly depleted in microbially‐derived sugars, and the contribution of O/N‐alkyl C to total organic C was less after HF‐treatment. The radiocarbon age of the mineral‐associated SOM was younger than that of the HF‐resistant SOM in subsoil horizons with small amounts of oxalate‐extractable Al and Fe. However, in horizons with large amounts of oxalate‐extractable Al and Fe the HF‐soluble SOM was considerably older than the HF‐resistant SOM. In acid subsoils a specific fraction of the organic C pool (O/N‐alkyl C; microbially‐derived sugars) is preferentially stabilized by association with Fe and Al minerals. Stabilization of SOM with the mineral matrix in soils with large amounts of oxalate‐extractable Al_o and Fe_o results in a particularly stable and relatively old C pool, which is potentially stable for thousands of years.     

三江平原岗地白浆土水解酶活性和动力学特性对不同连作农作物的响应

作物种植会对农田生态系统产生一定的影响。大田试验条件下,在黑龙江省853农场岗地白浆土上连续6年种植玉米、大豆、小麦、水稻,研究了土壤理化性质以及土壤中与碳、氮、磷、硫元素转化相关的9种水解酶活性和动力学特性的响应;同时研究了不同作物种植对土壤脲酶、磷酸单酯酶、磷酸二酯酶、芳基硫酸酯酶及β-葡糖苷酶动力学特性的影响。结果表明,大豆连作土壤的有效氮、总碳、总氮、总磷和总硫含量都稍高;大豆处理土壤pH值略低,但其它三种作物种植下的土壤均呈微酸性,差异不显著。土壤水解酶动力学参数对种植作物的反应与表观活性的反应不一致。玉米连作土壤蛋白酶和磷酸单酯酶活性高于其它处理;小麦处理的磷酸二酯酶和芳基硫酸酯酶活性最高,水稻连作土壤蛋白酶、磷酸二酯酶和磷酸三酯酶活性最低。连年种植小麦处理的土壤脲酶、磷酸二酯酶以及芳基硫酸酯酶Vmax显著高于其它处理,小麦连作土壤β-葡萄糖苷酶、脲酶、磷酸二酯酶和芳基硫酸酯酶的Vmax/Km值显著高于其它处理,可以看出在此处理下土壤酶具有较强的催化潜势。     

Phenol oxidase, peroxidase and organic matter dynamics of soil

Extracellular enzymes mediate the degradation, transformation and mineralization of soil organic matter. The activity of cellulases, phosphatases and other hydrolases has received extensive study and in many cases stoichiometric relationships and responses to disturbances are well established. In contrast, phenol oxidase and peroxidase activities, which are often uncorrelated with hydrolase activities, have been measured in only a small subset of soil enzyme studies. These enzymes are expressed for a variety of purposes including ontogeny, defense and the acquisition of carbon and nitrogen. Through excretion or lysis, these enzymes enter the environment where their aggegrate activity mediates key ecosystem functions of lignin degradation, humification, carbon mineralization and dissolved organic carbon export. Phenol oxidases and peroxidases are less stable in the environment than extracellular hydrolases, especially when associated with organic particles. Activities are also affected, positively and negatively, by interaction with mineral surfaces. High spatiotemporal variation obscures their relationships with environmental variables and ecological process. Across ecosystems, phenol oxidase and peroxidase activities generally increase with soil pH, a finding not predicted from the pH optima of purified enzymes. Activities associated with plant litter and particulate organic matter often correlate with decomposition rates and potential activities generally increase with the lignin and secondary compound content of the material. At the ecosystem scale, nitrogen amendment alters the expression of phenol oxidase and peroxidase enzymes more broadly than culture studies imply and these responses correlate with positive and negative changes in litter decomposition rates and soil organic matter content. At the global scale, N amendment of basidiomycete-dominated soils of temperate and boreal forest ecoystems often leads to losses of oxidative enzyme activity, while activities in grassland soils dominated by glomeromycota and ascomycetes show little net response. Land use that leads to loss of soil organic matter tends to increase oxidative activities. Across ecosystems, soil organic matter content is not correlated with mean potential phenol oxidase and peroxidase activities. A multiple regression model that includes soil pH, mean annual temperature, mean annual precipitation and potential phenol oxidase activity accounts for 37% of the variation in soil organic matter (SOM) content across ecosystems (n = 63); a similar model for peroxidase activity describes 32% of SOM variance (n = 43). Analysis of residual variation suggest that suites of interacting factors create both positive and negative feedbacks on soil organic matter storage. Soils with high oxygen availability, pH and mineral activity tend to be substrate limited: high in situ oxidative activities limit soil organic matter accumulation. Soils with opposing characteristics are activity limited: low in situ oxidative activities promote soil organic matter storage.     

太湖地区典型水稻土FDA水解酶活性的剖面分布特征

本文主要对太湖地区12个典型水稻土剖面的荧光素二乙酸酯(FDA)水解酶活性及其它相关酶活性进行了分析。耕作层的FDA水解酶活性最高,随土层加深活性明显下降,其中部分土壤在犁底层已经很难检测到其活性。部分样点的酶活性在剖面中呈明显的梯度下降趋势。高产水稻土壤表层FDA水解酶活性差异较大,在50～100g/(g.h)范围内,多数为60～80g/(g.h)。土壤酶活性最高与最低间的差异达到近一倍左右。同时FDA水解酶活性与-葡糖苷酶、脲酶、脱氢酶、酸性磷酸酶和芳基硫酸酯酶活性之间有极显著正相关关系(P0.01),与碱性磷酸酶呈显著相关(P0.05);与土壤养分指标如全氮、全磷、速效氮、有机碳之间有极显著相关性,与土壤pH呈显著负相关(P0.01)关系。     

Biochemical characterization of urban soil profiles from Stuttgart, Germany

The knowledge of biochemical properties of urban soils can help to understand nutrient cycling in urban areas and provide a database for urban soil management. Soil samples were taken from 10 soil profiles in the city of Stuttgart, Germany, differing in land use—from an essentially undisturbed garden area to highly disturbed high-density and railway areas. A variety of soil biotic (microbial biomass, enzyme activities) and abiotic properties (total organic C, elemental C, total N) were measured up to 1.9 m depth. Soil organic matter was frequently enriched in the subsoil. Microbial biomass in the top horizons ranged from 0.17 to 1.64 g C kg⁻¹, and from 0.01 to 0.30 g N kg⁻¹, respectively. The deepest soil horizon at 170-190 cm, however, contained 0.12 g C kg⁻¹ and 0.05 kg N kg⁻¹ in the microbial biomass. In general, arylsulphatase and urease activity decreased with depth but in three profiles potentially mineralizable N in the deepest horizons was higher than in soil layers directly overlying. In deeply modified urban soils, subsoil beside topsoil properties have to be included in the evaluation of soil quality. This knowledge is essential because consumption of natural soils for housing and traffic has to be reduced by promoting inner city densification.     

Enzyme activities in apple orchard agroecosystems: How are they affected by management strategy and soil properties

This study under field experimental conditions in apple orchard agroecosystems investigated the effects of pest management strategies (i.e. none, organic, conventional and integrated) on enzyme activities, in relation to soil properties. Enzyme activities chosen are implicated in the major biogeochemical nutrient cycles such as C (cellulase, fluoresceine diacetate hydrolase, β-galactosidase, β-glucosidase, phenol oxidase), N (arylamidase), P (acid and alkaline phosphomonoesterases, phosphodiesterase and phosphotriesterase) and S (arylsulfatase). Redundancy analyses and decomposition of the variances were performed to clarify how enzyme activities are affected by management strategy and soil properties. Results showed that the effects and their proportion attributable to management strategy and soil properties varied considerably depending on enzyme activity. Phenol oxidase activity was the only case where total variance was principally explained by management strategy (i.e. conventional and integrated) rather than by soil properties, and thus it seems to be an attractive potential indicator to assess soil quality in this agrochemical context.     

Responses of soil microbial communities and functions associated with organic carbon mineralization to nitrogen addition in a Tibetan grassland

Alpine grasslands with a high soil organic carbon(SOC)storage on the Tibetan Plateau are experiencing rapid climate warming and anthropogenic nitrogen(N)deposition;this is expected to substantially increase the soil N availability,which may impact carbon(C)cycling.However,little is known regarding how N enrichment influences soil microbial communities and functions relative to C cycling in this region.We conducted a 4-year field experiment on an alpine grassland to evaluate the effects of four different rates of N addition(0,25,50,and 100 kg N ha^-1 year^-1)on the abundance and community structure(phospholipid fatty acids,PLFAs)of microbes,enzyme activities,and community level physiological profiles(CLPP)in soil.We found that N addition increased the microbial biomass C(MBC)and N(MBN),along with an increased abundance of bacterial PLFAs,especially Gram-negative bacterial PLFAs,with a decreasing ratio of Gram-positive to Gram-negative bacteria.The N addition also stimulated the growth of fungi,especially arbuscular mycorrhizal fungi,reducing the ratio of fungi to bacteria.Microbial functional diversity and activity of enzymes involved in C cycling(β-1,4-glucosidase and phenol oxidase)and N cycling(β-1,4-N-acetyl-glucosaminidase and leucine aminopeptidase)increased after N addition,resulting in a loss of SOC.A meta-analysis showed that the soil C/N ratio was a key factor in the response of oxidase activity to N amendment,suggesting that the responses of soil microbial functions,which are linked to C turnover relative to N input,primarily depended upon the soil C/N ratio.Overall,our findings highlight that N addition has a positive influence on microbial communities and their associated functions,which may reduce soil C storage in alpine grasslands under global change scenarios.     

Microbial activities in forest soils exposed to chronic depositions from a lignite power plant

Atmospheric emissions of fly ash and SO₂ from lignite-fired power plants strongly affect large forest areas in Germany. The impact of different deposition loads on the microbial biomass and enzyme activities was studied at three forest sites (Picea abies (L.) Karst.) along an emission gradient of 3, 6, and 15 km downwind of a coal-fired power plant (sites Ia, II, and III, respectively), representing high, moderate and low emission rates. An additional site (site Ib) at a distance of 3 km from the power plant was chosen to study the influence of forest type on microbial parameters in coniferous forest soils under fly ash and SO₂ emissions. Soil microbial biomass C and N, CO₂ evolved and activities of l-asparaginase, l-glutaminase, β -glucosidase, acid phosphatase and arylsulfatase (expressed on dry soil and organic C basis) were determined in the forest floor (L, Of and Oh horizon) and mineral top soil (0-10 cm). The emission-induced increases in ferromagnetic susceptibility, soil pH, concentrations of mobile (NH₄NO₃ extractable) Cd, Cr, and Ni, effective cation exchange capacity and base saturation in the humus layer along the 15 km long transect significantly (P<0.05) reflected the effect of past depositions of alkaline fly ash. Soil microbial and biochemical parameters were significantly (P<0.05) affected by chronic fly ash depositions. The effect of forest type (i.e. comparison of sites Ia and Ib) on the studied parameters was generally dominated by the deposition effect. Alkaline depositions significantly (P<0.05) decreased the microbial biomass C and N, microbial biomass C-to-N ratios and microbial biomass C-to-organic C ratios. Microbial respiration, metabolic quotient (qCO₂) and the activities of l-asparaginase, l-glutaminase, β-glucosidase, acid phosphatase and arylsulfatase were increased by long-term depositions from the power plants. Acid phosphatase had the highest specific (enzyme activities expressed per unit organic C) activity values among the enzymes studied and arylsulfatase the lowest. The responses of the microbial biomass and soil respiration data to different atmospheric deposition loads were mainly controlled by the content of organic C and cation exchange capacity, while those of enzyme activities were governed by the soil pH and concentrations of mobile heavy metals. We concluded that chronic fly ash depositions decrease litter decomposition by influencing specific microbial and enzymatic processes in forest soils.     

Microbial performance under increasing nitrogen availability in a Mediterranean forest soil

In forest ecosystems, the external nitrogen (N) inputs mainly involve wet and dry depositions that potentially alter inorganic N availability in the soil and carbon (C) turnover. This study assesses the effect of a slow increase of inorganic N availability on microbial community activity and functionality in a Mediterranean forest soil. A four-month incubation experiment was performed with soil collected from the organic layer of a forest site and fertilized with a solution of ammonium nitrate. The fertilizer was supplied at an equivalent of 0, 10, 25, 50 and 75 kg N ha⁻¹ (0, 0.3, 0.7, 1.3 and 2 mg N g⁻¹ for control N0 and treatments N1, N2, N3 and N4, respectively). The incubation was carried out under optimal conditions, with the addition of the nutritive solution in small aliquots once a week to mimic the phenomenon of N deposition. In order to isolate the effect of N, the pH of the NH₄NO₃ solutions was adjusted to soil pH, and phosphorus was added in order to prevent any nutrient limitation effect. Inorganic N, C-mineralization, the activity of one oxidative enzyme (o-diphenol oxidase) and 8 hydrolitic enzymes (α-glucosidase, β-glucosidase, N-acetyl-β-d-glucosaminidase, cellulase, leucine amino-peptidase, acid phosphatase, butyric esterase and β-xylosidase) and the community level physiological profile (CLPP) were measured and analyzed during the whole incubation and at the end of the experiment as a proxy for microbial decomposition activity. In the first month, the highest N availability (N4) repressed the microbial respiration activity but stimulated microbial enzymatic activity, suggesting a change of C-pathways from spilling to enzymes and biomass investment. The treatments N1, N2 and N3 had no effect in the same period. Throughout the incubation, a general stress condition affected all the treated soils. As a consequence, treated soils exhibited higher respiration rates than the control. This was accompanied by a loss of functional diversity and an end-detected decline in biomass C. Although at the end of incubation most of the soil features showed a clear correlation with the inorganic N pool, the organic C content was strongly affected by different patterns of microbial activity during the experiment: the highest N treatment (N4) showed a lower C loss than the N3 treatment. Overall, the experiment showed how inorganic N availability can potentially alter the C cycle in a Mediterranean forest soil. The effect is non linear, depending on microbial community dynamics, on the community’s ability to adapt given the time scale of the process, and on N supply amount. Our study also revealed a common pattern in the short-term response to N addition in other, similar ecosystems with different climatic conditions.     

Enzyme activities in agricultural soils fumigated with methyl bromide alternatives

Pre-plant fumigation of agricultural soils with a combination of methyl bromide (MeBr) and chloropicrin (CP) to control nematodes, soil-borne pathogens and weeds has been a common practice in strawberry (Fragaria X ananassa Duchesne) production since the 1960s. MeBr will be phased out by 2005, but little is known about the impacts of alternative fumigants on soil microbial processes. We investigated the response of microbial biomass and enzyme activities in soils fumigated over two years with MeBr+CP and the alternatives propargyl bromide (PrBr), InLine, Midas and CP. Results were compared to control soils, which were not fumigated for the last 4-5 years for Watsonville and Oxnard, respectively, but had a 10 year history of MeBr+CP fumigation (history soils). Soil samples (0-15 cm) were taken from two sites in the coastal areas of California, USA, in Watsonville and Oxnard, at peak strawberry production after two years of repeated application. In addition to the soil enzymes, the activities of purified reference enzymes of β-glucosidase, acid phosphatase and arylsulfatase were assayed before and after fumigation with MeBr+CP and alternative biocides. At the Oxnard site, microbial respiration significantly decreased in soils fumigated with MeBr+CP (P=0.036), while microbial biomass C and N showed no response to fumigation at both sites. These results may indicate that fumigation promotes the growth of resistant species or that soil microorganisms had recovered at the time of sampling. Repeated soil fumigation with MeBr+CP significantly decreased the activities of β-glucosidase and acid phosphatase at the Watsonville site, and dehydrogenase activity at the Oxnard site. Although, enzyme activities in soils fumigated with PrBr, InLine, Midas and CP were lower compared to the control soil, effects were, in general, not significant. Fumigation with MeBr+CP and alternatives reduced the activities of purified reference enzymes by 13, 76 and 28% for acid phosphatase, β-glucosidase and arylsulfatase, respectively. Mean enzyme protein concentrations in fumigated agricultural soils were 2.93, 0.105, and 2.95 mg protein kg⁻¹ soil for acid phosphatase, β-glucosidase and arylsulfatase, respectively, all lower than in control soils. Organic matter turnover and nutrient cycling, and thus, the long-term productivity of agricultural soils seem unaffected in soils repeatedly fumigated with PrBr, InLine, Midas and CP.     

Spatial variability of enzyme activities and microbial biomass in the upper layers of Quercus petraea forest soil

Extracellular lignocellulose-degrading enzymes are responsible for the transformation of organic matter in hardwood forest soils. The spatial variability on a 12 × 12 m plot and vertical distribution (0–8 cm) of the ligninolytic enzymes laccase and Mn-peroxidase, the polysaccharide-specific hydrolytic enzymes endoglucanase, endoxylanase, cellobiohydrolase, 1,4-β-glucosidase, 1,4-β-xylosidase and 1,4-β-N-acetylglucosaminidase and the phosphorus-mineralizing acid phosphatase were studied in a Quercus petraea forest soil profile. Activities of all tested enzymes exhibited high spatial variability in the L and H horizons. Acid phosphatase and 1,4-β-N-acetylglucosaminidase exhibited low variability in both horizons, while the variability of Mn-peroxidase activity in the L horizon, and endoxylanase and cellobiohydrolase activities in the H horizon were very high. The L horizon contained 4× more microbial biomass (based on PLFA) and 7× fungal biomass (based on ergosterol content) than the H horizon. The L horizon also contained relatively more fungi-specific and less actinomycete-specific PLFA. There were no significant correlations between enzyme activities and total microbial biomass. In the L horizon cellulose and hemicellulose-degrading enzymes correlated with each other and also with 1,4-β-N-acetylglucosaminidase and acid phosphatase activities. Laccase, Mn-peroxidase and acid phosphatase activities correlated in the H horizon. The soil profile showed a gradient of pH, organic carbon and humic compound content, microbial biomass and enzyme activities, all decreasing with soil depth. Ligninolytic enzymes showed preferential localization in the upper part of the H horizon. Differences in enzyme activities were accompanied by differences in the microbial community composition where the relative amount of fungal biomass decreased and actinomycete biomass increased with soil depth. The results also showed that the vertical gradients occur at a small scale: the upper and lower parts of the H horizon only 1 cm apart were significantly different with respect to seven out of nine activities, microbial biomass content and community composition.     

Effects of yak excreta on soil organic carbon mineralization and microbial communities in alpine wetlands of southwest of China

Purpose

Improving knowledge of how soil organic carbon (SOC) mineralization responds to excreta application is essential to better understand whether wetland carbon (C) pools will react to grazing. We investigated microbial activity and community structure in the different treatments of excreta addition experiments to examine how soil C mineralization responds to the excreta input in terms of microbial activities and compositions in wetland soils.

Materials and methods

The microcosms of mineralization incubation of excreta addition were established. The structure of the microbial community was described by the fatty acid composition of the phospholipids (PLFA). The methylumbelliferyl-linked substrates (MUB) and l-dihydroxyphenylalanine (L-DOPA) substrates were used to investigate the activities of β-glucosidase (BG), N-acetyl-glucosaminidase (NAG), acid phosphatase (AP), cellobiohydrolase (CBH), and phenol oxidase (PO).

Results and discussion

Excreta addition altered the cumulative C mineralization in swamp meadow (SM) and peatland (PL) soils, but SM was lower than PL. Excreta addition increased the biomass of individual PLFA and the fungi/bacteria ratio, suggesting that microbes are stimulated by nutrients and that the soil microbial community composition is modified by excreta inputs. The hydrolytic enzyme activities were higher in the PL soils than in the SM soils, but the trend was opposite for PO activity. The changes in pH, fungi, actinomycetes (ACT), AP, and CBH after yak fecal input significantly influenced the soil CO₂ efflux. Our findings suggest that yak grazing could influence the rate of C cycling in wetland soils by influencing microbial communities, enzyme activities, and soil pH.

Conclusions

This study suggest that the yak excreta addition increased cumulative C mineralization in SM and PL soils, and the effect of dung addition was more significant than urine addition. The effect of yak excreta addition on SOC mineralization was related with the soil pH, microorganism structure, and enzyme activity which modified by the excreta addition. Soil pH, fungi, AP, and CBH were positively correlated with SOC mineralization, but ACT was negatively correlated with SOC mineralization. In addition, the changes in C and N sources with yak excreta addition play an important role in altering microbial enzyme activities. The input of yak feces into wetlands because of grazing could increase SOC mineralization and thereby promote C emission.