Vertical gradients of potential enzyme activities in soil profiles of European beech,Norway spruce and Scots pine dominated forest sites

Management of forest sites has the potential to modulate soil organic matter decomposition by changing the catalytic properties of soil microorganisms within a soil profile. In this study we examined the impact of forest management intensity and soil physico-chemical properties on the variation of enzyme activities (β-glucosidase, β-xylosidase, α-glucosidase, phenol oxidase, N-acetyl-glucosaminidase, l-leucine aminopeptidase, phosphatase) in the topsoil and two subsoil horizons in three German regions (Schorfheide-Chorin, Hainich-Dün, Schwäbische Alb). The sandy soils in the Schorfheide-Chorin (SCH) showed lower ratios of the activity of carbon (C) acquiring enzymes (β-glucosidase) relative to nitrogen (N) acquiring enzymes (N-acetyl-glucosaminidase + l-leucine aminopeptidase), and activity of C acquiring enzymes relative to phosphorous (P) acquiring enzymes (phosphatase) than the finer textured soils in the Hainich-Dün (HAI) and Schwäbische Alb (ALB), indicating a shift in investment to N and P acquisition in the SCH. All enzyme activities, except phenol oxidase activity, decreased in deeper soil horizons as concentrations of organic C and total N did, while the decrease was much stronger from the topsoil to the first subsoil horizon than from the first subsoil to the second subsoil horizon. In contrast, phenol oxidase activity showed no significant decrease towards deeper soil horizons. Additionally, enzyme activities responsible for the degradation of more recalcitrant C relative to labile C compounds increased in the two subsoil horizons. Subsoil horizons in all regions also indicate a shift to higher N acquisition, while the strength of the shift depended on the soil type. Further, our results clearly showed that soil properties explained most of the total variance of enzyme activities in all soil horizons followed by study region, while forest management intensity had no significant impact on enzyme activities. Among all included soil properties, the clay content was the variable that explained the highest proportion of variance in enzyme activities with higher enzyme activities in clay rich soils. Our results highlight the need for large scale studies including different regions and their environmental conditions in order to derive general conclusions on which factors (anthropogenic or environmental) are most influential on enzyme activities in the whole soil profile in the long term at the regional scale.     

Extracellular enzyme activity in the mycorrhizospheres of a boreal fire chronosequence

Saprotrophic microbes are typically credited with producing extracellular enzymes that recycle organic matter, though roots and mycorrhizal fungi also can contribute and may compete with the saprotrophs. We examined extracellular enzyme activity associated with the mycorrhizospheres of arbuscular mycorrhizal, ectomycorrhizal, dual-colonized (arbuscular and ectomycorrhizal), and ericoid mycorrhizal plants in a fire chronosequence in Alaska. Bulk soil and soil from beneath host plants were gathered in July 2004 and assayed for five enzymes that target organic C, P, and N substrates. Compared to bulk soil, activities of the C-targeting enzymes β-1,4-glucosidase and peroxidase were lower in arbuscular mycorrhizospheres and ericoid mycorrhizospheres, respectively. Moreover, extracellular enzyme activity varied among mycorrhizosphere types. Specifically, N-targeting leucine aminopeptidase was highest in arbuscular mycorrhizospheres, followed by ericoid and ectomycorrhizal/dual-colonized mycorrhizospheres; β-1,4-glucosidase had the reverse pattern. In addition, enzymatic stoichiometry suggested that extracellular enzyme producers invested more in C-acquisition than in N-acquisition in recent fire scars compared to mature forests. These data extend previous findings that roots and mycorrhizal fungi compete with saprotrophs by showing that the strength of this effect varies by mycorrhizal host. As a result the community composition of mycorrhizal host plants might mediate enzymatic activity in boreal soils.     

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.     

Independent roles of ectomycorrhizal and saprotrophic communities in soil organic matter decomposition

The relative roles of ectomycorrhizal (ECM) and saprotrophic communities in controlling the decomposition of soil organic matter remain unclear. We tested the hypothesis that ECM community structure and activity influences the breakdown of nutrient-rich biopolymers in soils, while saprotrophic communities primarily regulate the breakdown of carbon-rich biopolymers. To test this hypothesis, we used high-throughput techniques to measure ECM and saprotrophic community structure, soil resource availability, and extracellular enzyme activity in whole soils and on ECM root tips in a coastal pine forest. We found that ECM and saprotroph richness did not show spatial structure and did not co-vary with any soil resource. However, species richness of ECM fungi explained variation in the activity of enzymes targeting recalcitrant N sources (protease and peroxidase) in bulk soil. Activity of carbohydrate- and organic P- targeting enzymes (e.g. cellobiohydrolase, β-glucosidase, α-glucosidase, hemicellulases, N-acetyl-glucosaminidase, and acid phosphatase) was correlated with saprotroph community structure and soil resource abundance (total soil C, N, and moisture), both of which varied along the soil profile. These observations suggest independent roles of ECM fungi and saprotrophic fungi in the cycling of N-rich, C-rich, and P-rich molecules through soil organic matter. Enzymatic activity on ECM root tips taken from the same soil cores used for bulk enzyme analysis did not correlate with the activity of any enzyme measured in the bulk soil, suggesting that ECM contributions to larger-scale soil C and nutrient cycling may occur primarily via extramatrical hyphae outside the rhizosphere.     

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.     

Areal activities and stratification of hydrolytic enzymes involved in the biochemical cycles of carbon, nitrogen, sulphur and phosphorus in podsolized boreal forest soils

A novel approach allowing on-site high throughput enzyme activity measurements by fluorogenic model substrates was applied to study the functioning of enzymes involved in biochemical cycling of nutrients in boreal forest soil ecosystems. The examined enzymes comprised α-glucosidase, β-glucosidase, β-xylosidase, β-cellobiosidase, N-acetyl-glucosamidase, acetate-esterase, butyrate-esterase, phosphomonoesterase, sulphatase and aminopeptidase, whereby spatial and seasonal variation of their activity was investigated over nine seasons in 2 years. The studied sites of boreal podzolized soil of Pinus sylvestris and Picea abies forest were located in central Finland. Activity of all enzymes except sulphatase was highest in the humus layer in all seasons. Maximum sulphatase activity was located below the humus layer in the soil column. Annual activities of acetate-esterase, butyrate-esterase, β-glucosidase and phosphomonosterase calculated to in situ temperature during the year were 480-700, 690-950, 110-190 and 110-200 mol m⁻² year⁻¹, respectively. They were up to 100 fold higher than the other six measured activities. The overall turnover capacity of the enzymes was >1000 mol of ester linked carbon, >700 mols carbon from different carbohydrates, 100-200 mol of ester linked phosphate, 10-40 mol of ester linked sulphate m⁻² year⁻¹. Winter time (November-April) contributed from 7 to 32% to the annual turnover capacity indicating important enzyme activities also during a cold period of the year. Clear-cutting of the tree stand did not adversely affect enzyme activities related to the cycling of carbon, nitrogen, sulphur and phosphorus during the year. The pH optimum for hemicellulose and cellulose hydrolysing enzymes was pH 3-4 and the pH optimum of phosphomonoesterase, sulphatase, aminopeptidase and N-acetyl-glucosamidase was 4-5. This shows that the hydrolytic activities were adapted to the acid pH-values of the soils. The soil hydrolytic potential was many fold higher as compared to the actual amount of litter it received in the P. sylvestris and P. abies forests.     

Effects of the ecological restoration practices of prescribed burning and mechanical thinning on soil microbial enzyme activities and leaf litter decomposition

The effects of ecological restoration on belowground processes such as decomposition are generally unknown. To assess the immediate effects of prescribed fire and mechanical thinning on belowground processes, we measured the activities of five extracellular enzymes (phosphatase, β-glucosidase, β-N-acetylglucosaminidase, phenol oxidase, and lignin-peroxidase) in soils and on decomposing Quercus falcata leaf litter in unburned, burned, and burned and thinned plots in a mesic forest in northern Mississippi. Decomposition rates of Q. falcata leaf litter were also assessed at each plot. Soil phosphatase activity decreased after a prescribed burn and was related to an increase in soil organic matter in plots that had been burned. Soil β-N-acetylglucosaminidase activity increased after a burn, and was related to a decrease in leaf litter. Leaf litter enzyme activity showed no consistent patterns amongst treatments, or between individual enzymes, while decomposition rates of leaf litter were slightly accelerated in the treatment plots, but not significantly so. Decomposition rates were related to cumulative enzyme activity, with phenol oxidase and lignin-peroxidase having the highest apparent efficiencies in degrading the leaf material. Overall, the microbial degradation of Q. falcata leaf litter was more efficient in plots that were burned and thinned than in the other plots. Increases in the efficiency of litter decomposition coupled with reductions in litter inputs due to canopy thinning likely allows for increased solar penetration to the soil, and could promote the restoration of the shade-intolerant species that once dominated the understory. Post-burn increases in β-N-acetylglucosaminidase activity and decreases in phosphatase activity also suggest a potential shift in the soil community from phosphorus limitation to nitrogen limitation following a fire.     

Enzyme activities as affected by soil properties and land use in a tropical watershed

Enzyme activities play key roles in the biochemical functioning of soils, including soil organic matter formation and degradation, nutrient cycling, and decomposition of xenobiotics. Knowledge of enzyme activities can be used to describe changes in soil quality due to land use management and for understanding soil ecosystem functioning. In this study, we report the activities of the glycosidases (β-glucosidase, α-galactosidase, and β-glucosaminidase), acid phosphatase, and arylsulfatase, involved in C (C and N for β-glucosaminidase), P, and S cycling, respectively, as affected by soil order and land use within a watershed in north-central Puerto Rico (Caribbean). Representative surface soil (0–15 cm) samples were taken from 84.6% of the total land area (45,067 ha) of the watershed using a completely randomized design. The activity of α-galactosidase was greater in soils classified as Oxisols than in soils classified as Ultisols and Inceptisols, and it was not affected by land use. The activity of β-glucosidase was greater in Oxisols compared to the Inceptisols and Ultisols, and it showed this response according to land use: pasture > forest > agriculture. The activity of β-glucosaminidase was higher in Oxisols than the other soil orders, and it was higher under pasture compared to forest and agriculture. Acid phosphatase and arylsulfatase activities were greater in Oxisols and Ultisols than in Inceptisols, and they decreased in this order due to land use: forest = pasture > agriculture. As a group, β-glucosaminidase, β-glucosidase, and acid phosphatase activities separated the sites under forest and pasture from those under agriculture in a three-dimensional plot. Thus, enzyme activities in Inceptisols under agriculture could be increased to levels comparable to other soil orders with conservative practices similar to those under pasture and secondary forest growth. Our findings demonstrate that within this watershed, acid and low fertility soils such as Oxisols and Ultisols have in general higher enzyme activities than less weathered tropical soils of the order Inceptisols, probably due to their higher organic matter content and finer texture; and that the activities of these enzymes respond to management with agricultural practices decreasing key soil biochemical reactions of soil functioning.     

Modelling in situ activities of enzymes as a tool to explain seasonal variation of soil respiration from agro-ecosystems

Understanding in situ enzyme activities could help clarify the fate of soil organic carbon (SOC), one of the largest uncertainties in predicting future climate. Here, we explored the role of soil temperature and moisture on SOM decomposition by using, for the first time, modelled in situ enzyme activities as a proxy to explain seasonal variation in soil respiration. We measured temperature sensitivities (Q₁₀) of three enzymes (β-glucosidase, xylanase and phenoloxidase) and moisture sensitivity of β-glucosidase from agricultural soils in southwest Germany. Significant seasonal variation was found in potential activities of β-glucosidase, xylanase and phenoloxidase and in Q₁₀ for β-glucosidase and phenoloxidase activities but not for xylanase. We measured moisture sensitivity of β-glucosidase activity at four moisture levels (12%–32%), and fitted a saturation function reflecting increasing substrate limitation due to limited substrate diffusion at low water contents. The moisture response function of β-glucosidase activity remained stable throughout the year. Sensitivity of enzymes to temperature and moisture remains one of the greatest uncertainties in C models. We therefore used the response functions to model temperature-based and temperature and moisture-based in situ enzyme activities to characterize seasonal variation in SOC decomposition. We found temperature to be the main factor controlling in situ enzyme activities. To prove the relevance of our modelling approach, we compared the modelled in situ enzyme activities with soil respiration data measured weekly. Temperature-based in situ enzyme activities explained seasonal variability in soil respiration well, with model efficiencies between 0.35 and 0.78. Fitting an exponential response function to in situ soil temperature explained soil respiration to a lesser extent than our enzyme-based approach. Adding soil moisture as a co-factor improved model efficiencies only partly. Our results demonstrate the potential of this new approach to explain seasonal variation of enzyme related processes.     

A method for linking in situ activities of hydrolytic enzymes to associated organisms in forest soils

A root window-based, enzyme-imprinted, membrane system has been modified to enable visualization of the activities of hydrolytic enzymes (acid phosphatase, aminopeptidase, chitinase, and β-glucosidase) in situ in forest soils. The approach can be used to correlate the distribution of enzyme activity with visible features such as roots, mycorrhizas, or mycelial mats. In addition, it enables accurate spatial soil sampling for analysis of microbial communities associated with enzyme activities. The substrates are colorimetric conjugates of napthol, where color develops instantly in the field, or fluorimetric conjugates of 4-methylumbelliferone, whose fluorescent products are detected by a gel-documenting system. The method will allow important questions about the relationship between taxonomic and functional diversity of soil microorganisms to be addressed and identification of enzyme activity hot-spots in soil.     

Reconciling apparent variability in effects of biochar amendment on soil enzyme activities by assay optimization

We studied the effects of a biochar made from fast pyrolysis of switchgrass on four soil enzymes (β-glucosidase, β-N-acetylglucosaminidase, lipase, and leucine aminopeptidase) to determine if biochar would consistently modify soil biological activities. Thus, we conducted a series of enzyme assays on biochar-amended soils. Inconsistent results from enzyme assays of char-amended soils suggested that biochar had variable effects on soil enzyme activities, thus we conducted a second experiment to determine if biochar reacts predictably with either enzyme or substrate in in vitro reactions. Both colorimetric and fluorescent assays were used for β-glucosidase and β-N-acetylglucosaminidase. Seven days after biochar was added to microcosms of 3 different soils, fluorescence-based assays revealed some increased enzyme activities (up to 7-fold for one measure of β-glucosidase in a shrub-steppe soil) and some decreased activities (one-fifth of the unamended control for lipase measured in the same shrub-steppe soil), compared to non-amended soil. In an effort understand the varied effects, purified enzymes or substrates were briefly exposed to biochar and then assayed. In contrast to the soil assays, except for β-N-acetylglucosaminidase, the exposure of substrate to biochar reduced the apparent activity of the enzymes, suggesting that sorption reactions between substrate and biochar impeded enzyme function. Our findings indicate that fluorometric assays are more robust to, or account for, this sorption better than the colorimetric assays used herein. The activity of purified β-N-acetylglucosaminidase increased 50-75% following biochar exposure, suggesting a chemical enhancement of enzyme function. In some cases, biochar stimulates soil enzyme activities, to a much greater degree than soil assays would indicate, given that substrate reactivity can be impeded by biochar exposure. We conclude that the effects of biochar on enzyme activities in soils are highly variable; these effects are likely associated with reactions between biochar and the target substrate.     

Measurements of microbial community activities in individual soil macroaggregates

The functional potential of single soil macroaggregates may provide insights into the localized distribution of microbial activities better than traditional assays conducted on bulk quantities of soil. Thus, we scaled down enzyme assays for β-glucosidase, N-acetyl-β-d-glucosaminidase, lipase, and leucine aminopeptidase to measure of the enzyme potential of individual macroaggregates (250–1000 μm diameter). Across all enzymes, the smallest macroaggregates had the greatest activity and the range of enzyme activities observed in all macroaggregates supports the hypothesis that functional potential in soil may be distributed in a patchy fashion. Paired analyses of ATP as a surrogate for active microbial biomass and β-glucosidase on the same macroaggregates suggest the presence of both extracellular β-glucosidase functioning in macroaggregates with no detectable ATP and also of relatively active microbial communities (high ATP) that have low β-glucosidase potentials. Studying function at a scale more consistent with microbial habitat presents greater opportunity to link microbial community structure to microbial community function.     

Shrub encroachment does not reduce the activity of some soil enzymes in Mediterranean semiarid grasslands

Shrub encroachment is a worldwide phenomenon with implications for desertification and global change. We evaluated its effects on the activities of urease, phosphatase and β-glucosidase in Mediterranean semiarid grasslands dominated by Stipa tenacissima by sampling 12 sites with and without resprouting shrubs along a climatic gradient. The presence of shrubs affected the evaluated enzymes at different spatial scales. Soils under S. tenacissima tussocks and in bare ground areas devoid of vascular plants had higher values of phosphatase and urease when the shrubs were present. For the β-glucosidase, this effect was site-specific. At the scale of whole plots (30 m × 30 m), shrubs increased soil enzyme activities between 2% (β-glucosidase) and 22% (urease), albeit these differences were significant only in the later case. Our results indicate that shrub encroachment does not reduce the activity of extracellular soil enzymes in S. tenacissima grasslands.     

Long-term effect on soil biochemical status of a Vertisol under conservation tillage system in semi-arid Mediterranean conditions

Long-term field experiments are expected to provide important information regarding soil properties affected by conservation management practices. Several studies have shown that soil enzyme activities are sensitive in discriminating among soil management effects. In this study we evaluated the long-term effect of direct drilling (DD) under a crop rotation system (cereals–sunflower–legumes), on the stratification of soil organic matter content and on biochemical properties in a dryland in southwest Spain. The results were compared to those obtained under conventional tillage (CT). Soil biochemical status was evaluated by measuring the enzymatic activities (dehydrogenase, β-glucosidase, alkaline phosphatase and arylsulphatase) during the flowering period of a pea crop. Soil samples were collected in May 2007 at three depths (0–5, 5–10 and 10–20 cm).Total organic carbon (TOC) contents and values of soil enzyme activities were higher in soils subjected to DD than to CT, specifically at 0–5 cm depth. Although a slight decrease of TOC and enzymatic activities with increasing soil depth was observed, no significant differences were found among different depths of the same treatment. This could be related to the high clay content of the soil, a Vertisol. Enzyme activities values showed high correlation coefficients (from r = 0.799 to r = 0.870, p < 0.01) with TOC. Values of activity of the different enzymes were also correlated (p < 0.01).Values of stratification ratios did not show significant differences between tillage practices. The high clay content of the soil is responsible for this lack of differences because of the protection by clay mineral of TOC and soil enzymes activities.Long-term soil conservation management by direct drilling in a dryland farming system improved the quality of a clay soil, especially at the surface, by enhancing its organic matter content and its biological status.     

Enzymatic activity and stabilization of organic matter in soil with different detritus inputs

ABSTRACT

The aim of the study was to assess the effects of different forest stands (Silver fir (Abies alba) and sycamore maple (Acer pseudoplatanus) with common hornbeam (Carpinus betulus)) on the enzymes activities and microbial biomass. The objective was to explore how changes in the detritus inputs affect soil organic matter (SOM) composition. The content of SOM fraction has been compared with soil enzyme activities. The investigation was carried out in the ?wi?tokrzyskie Mountains of central Poland. Twenty investigation plots were selected, including fir stands (10 plots) and maple with hornbeam stands (10 plots). Contents of organic C, N and base cations, pH, hydrolytic acidity, and soil texture were investigated. The content of C and N in the physically separated SOM fractions was distinguished. The study indicated only small changes in soil properties and stabilization of organic matter as a result of different detritus inputs. The maple-hornbeam and fir stands have a similar influence on microbiological processes and the SOM. Acidity of soil is a major factor affecting microbial activity and therefore pH affects enzyme dynamics. Differences in soil pH confirmed the stronger acidifying effects of fir stands compared to maple-hornbeam stands. Additionally, fir stands stimulate β-glucosidase activity, probably through a simultaneous interaction of mycorrhizal fungi in the roots of fir stands.     

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.     

Changes in enzyme activities of spruce (Picea balfouriana) forest soil as related to burning in the eastern Qinghai-Tibetan Plateau

The effect of forest fire on soil enzyme activity of spruce (Picea balfouriana) forest in the eastern Qinghai-Tibetan Plateau was assessed. Six specific enzymes were chosen for investigation: invertase, acid phosphatase, proteinase, catalase, peroxidase and polyphenoloxidase. It was found that the activities of invertase and proteinase were reduced by burning, but the activities of acid phosphatase, polyphenoloxidase and peroxidase increased. Meanwhile, burning significantly (P < 0.05) resulted in the decrease of concentrations of available N and K of 0–20 cm depth layer soil, and significantly (P < 0.05) decreased concentrations of organic matter content, total N and P, as well as available N, P and K in soil at both 20–40 and 40–60 cm depths except for available P at 20–40 cm soil depth. These results illustrated that burning could influence the enzyme activities and chemical properties of soil not only of upper but also lower soil layers. Correlation analysis indicated that invertase activities in 0–20 cm depth layer soil were significantly positively correlated with organic matter, total N and P, as well as available N and P. Furthermore, all six enzymes studied were sensitive to fire disturbance, and thus could be used as indicators of soil quality. Our study also showed that soil enzyme activities were associated with soil depth, decreasing from top to bottom in both burned and unburned spruce forests. The distribution pattern of soil enzyme activities suggested that the rate of organic matter decomposition and nutrient cycling depended on soil depth, which had important structural and functional characteristics in nutrient cycling dynamics and implications in plantation nutrient management. The finding that burning effects on enzyme activities and soil properties between different soil layers were homogenized was attributed to the 8-years’ regeneration of forest after burning.     

不同施肥模式对设施秋冬茬芹菜生育期间土壤酶活性的影响

【目的】利用在天津的日光温室蔬菜不同施肥模式定位试验,研究了不同施肥模式对设施菜田土壤酶活性的影响,为设施蔬菜高效施肥和菜田土壤可持续利用提供依据。【方法】取样调查在第9茬蔬菜(秋冬茬芹菜)进行。定位试验设6个处理,在等氮磷钾条件下,分别为1)全部施用化肥氮(4/4CN),2)3/4化肥氮+1/4猪粪氮(3/4CN+1/4PN),3)2/4化肥氮+2/4猪粪氮(2/4CN+2/4PN),4)1/4化肥氮+3/4猪粪氮(1/4CN+3/4PN),5)2/4化肥氮+1/4猪粪氮+1/4秸秆氮(2/4CN+1/4PN+1/4SN),6)2/4化肥氮+2/4秸秆氮(2/4CN+2/4SN)。在芹菜基肥施用前和定植后30、60、90、110天,采取0—20 cm土壤样品,测定土壤α-葡萄苷酶、β-木糖苷酶、β-葡萄苷酶、β-纤维二糖苷酶、几丁质酶、磷酸酶和脲酶的活性,分析其与土壤微生物量碳氮及土壤可溶性有机碳氮含量之间的关系。【结果】芹菜生育期间不同施肥模式土壤α-葡萄苷酶、β-木糖苷酶、β-葡萄苷酶、β-纤维二糖苷酶、几丁质酶和磷酸酶的活性总体上先增后降,较高土壤酶活性均出现在芹菜定植后60~90 d; 土壤脲酶活性总体上呈逐渐升高的趋势。芹菜季有机无机肥料配施模式土壤α-葡萄苷酶、β-木糖苷酶、β-葡萄苷酶、β-纤维二糖苷酶、几丁质酶、磷酸酶和脲酶的活性较4/4CN模式平均分别增加22.9%~92.0%、20.1%~152.4%、23.1%~145.1%、28.7%~273.8%、9.2%~207.8%、13.7%~86.8%和6.5%~56.5%,其中以配施秸秆模式土壤酶活性相对较高,较4/4CN模式平均分别增加59.9%~92.0%、98.9%~152.4%、90.3%~145.1%、171.6%~273.8%、106.4%~207.8%、68.8%~86.8%和30.7%~56.5%。土壤酶活性与土壤微生物量碳氮、可溶性有机碳氮含量及芹菜产量之间总体上呈显著或极显著正相关关系。【结论】同等养分投入量下,设施菜田土壤酶活性表现为有机无机肥料配合显著高于单施化肥,又以配施秸秆效果更佳; 土壤酶活性与土壤微生物量碳氮、可溶性有机碳氮含量和蔬菜产量之间密切相关。说明有机无机肥配施,特别是配施一定的秸秆可有效提高土壤酶活性,维持较高的菜田土壤肥力,有利于设施蔬菜的可持续和高效生产。     

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.