Response of soil enzymatic activity to pore structure under inversion tillage with organic materials incorporation in a Haplic Chernozem.

Soil pore structure affects microbial survival environmental conditions and thus enzyme activity. The mechanisms underlying returning organic materials on soil pore structure and enzymatic activity, however, remain unclear. We therefore conducted a field experiment in the fall of 2018 in northeastern China with a chernozem soil and four treatments: CT, conventional tillage; SCT, returning maize straw incorporation with conventional tillage; SIT, returning maize straw incorporation with inversion tillage; SMIT, returning maize straw and organic manure with inversion tillage. Soil samples were collected from the 0-15 cm and 15-35 cm layers in the fall of 2021. We used X-ray computed tomography to analyze the characteristics of pore structure and extracellular enzymatic stoichiometry to evaluate the limiting factors for soil microorganisms. Inversion tillage and organic materials incorporation can alter the micromorphological structure of entire soil layer, leading to the rearrangement of soil particles and nutrients, thereby augmenting the physicochemical properties in subsoil layer. SMIT exhibited a substantial increase in the number of macropores, porosity and fractal dimension, compared to SCT and SIT. This led to a significantly increased in soil enzyme activities of carbon and nitrogen-limited in SMIT, with increases ranging from 11.67% to 40.16% and from 8.81% to 21.43%, respectively (P < 0.05). Analysis using structural equation modeling revealed that returning organic material was conducive to the development of soil pore structure, characterized by an increase in macropores and fractal dimension and a decrease in the Euler number, had a positive correlation with soil enzyme activity. This, in turn, led to an alleviation in microbial nitrogen limitation. These results indicate that SMIT could serve as a viable choice in enhancing soil structure and fostering a favorable environment for microbial survival. Moreover, they offer essential insights into the microbial strategies responsible for the breakdown of organic matters in Hapli-Udic Cambisol.

Short-term responses of soil enzyme activities and stoichiometry to litter input in Castanopsis carlesii and Cunninghamia lanceolata plantations.

Litter input triggers the secretion of soil extracellular enzymes and facilitates the release of carbon (C), nitrogen (N), and phosphorus (P) from decomposing litter. However, how soil extracellular enzyme activities were controlled by litter input with various substrates is not fully understood. We examined the activities and stoichiometry of five enzymes including ß-1,4-glucosidase, ß-D-cellobiosidase, ß-1,4-N-acetyl-glucosaminidase, leucine aminopeptidase and acidic phosphatase (AP) with and without litter input in 10-year-old Castanopsis carlesii and Cunninghamia lanceolata plantations monthly during April to August, in October, and in December 2021 by using an in situ microcosm experiment. The results showed that: 1) There was no significant effect of short-term litter input on soil enzyme activity, stoichiometry, and vector properties in C. carlesii plantation. In contrast, short-term litter input significantly increased the AP activity by 1.7% in May and decreased the enzymatic C/N ratio by 3.8% in August, and decreased enzymatic C/P and N/P ratios by 11.7% and 10.3%, respectively, in October in C. lanceolata plantation. Meanwhile, litter input increased the soil enzymatic vector angle to 53.8° in October in C. lanceolata plantations, suggesting a significant P limitation for soil microorganisms. 2) Results from partial least squares regression analyses showed that soil dissolved organic matter and microbial biomass C and N were the primary factors in explaining the responses of soil enzymatic activity to short-term litter input in both plantations. Overall, input of low-quality (high C/N) litter stimulates the secretion of soil extracellular enzymes and accelerates litter decomposition. There is a P limitation for soil microorganisms in the study area.

Arsenic stress on soil microbial nutrient metabolism interpreted by microbial utilization of dissolved organic carbon.

In a 120-day microcosm incubation experiment, we investigated the impact of arsenic contamination on soil microbial nutrient metabolism, focusing on carbon cycling processes. Our study encompassed soil basal respiration, key enzyme activities (particularly, ß-1,4-N-acetylglucosaminidase and phosphatases), microbial biomass, and community structure. Results revealed a substantial increase (1.21-2.81 times) in ß-1,4-N-acetylglucosaminidase activities under arsenic stress, accompanied by a significant decrease (9.86%-45.20%) in phosphatase activities (sum of acid and alkaline phosphatases). Enzymatic stoichiometry analysis demonstrated the mitigation of microbial C and P requirements in response to arsenic stress. The addition of C-sources alleviated microbial C requirements but exacerbated P requirements, with the interference amplitude increasing with the complexity of the C-source. Network analysis unveiled altered microbial nutrient requirements and an increased resistance process of microbes under arsenic stress. Microbial carbon use efficiency (CUE) and basal respiration significantly increased (1.17-1.59 and 1.18-3.56 times, respectively) under heavy arsenic stress (500 mg kg-1). Arsenic stress influenced the relative abundances of microbial taxa, with Gemmatimonadota increasing (5.5-50.5%) and Bacteroidota/ Nitrospirota decreasing (31.4-47.9% and 31.2-63.7%). Application of C-sources enhanced microbial resistance to arsenic, promoting cohesion among microorganisms. These findings deepen our understanding of microbial nutrient dynamics in arsenic-contaminated areas, which is crucial for developing enzyme-based toxicity assessment systems for soil arsenic contamination.

Empirical evidence challenges the effectiveness of the enzymatic stoichiometry of glucosidase and phosphatase as an indicator of microbial C vs P limitation.

The ratio of ß-1,4-glucosidase (BG) to acid/alkaline phosphomonoesterase (AP) (BG:AP) is commonly employed as an indicator to assess the relative microbial limitations of carbon (C) and phosphorus (P), whereby a higher BG:AP ratio suggests stronger C limitations. This approach is based on the assumption that BG and AP can represent enzymes targeting C and P, respectively. Nevertheless, it is crucial to recognize that microbial C and P acquisition involves the participation of other enzymes alongside BG and AP, and thus, the capacity of BG and AP to accurately and comprehensively represent the entire spectrum of C and P acquisition is questionable. Here, analyzing previously published data, I present a piece of empirical evidence that challenges the suitability of the BG:AP ratio as an accurate indicator of microbial limitations concerning C vs P. P fertilization decreased BG:AP in up to 27 % out of the total 109 observations, which represents a clear contradiction, as this outcome is interpreted by the enzymatic stoichiometry approach as indicating an intensified P limitation arising from P fertilization. Furthermore, the effect of P fertilization on the BG:AP ratio did not show significant differences between experimental sites characterized by higher BG:AP ratios (indicative of lesser P limitation) and those with lower BG:AP ratios (indicative of greater P limitation). Consequently, I conclude that the BG:AP ratio inadequately reflects microbial C vs P limitations.

Soil nutrient levels regulate the effect of soil microplastic contamination on microbial element metabolism and carbon use efficiency.

Microplastics (MPs) are emerging environmental contaminants in soil ecosystems that disrupt the soil carbon (C) pool. Therefore, the response of microbial metabolism to MP-contaminated soil is crucial for soil-C stabilization. We undertook factorial experiments in a greenhouse with three types of soil microplastics with three levels of soil nutrients and undertook soil physiochemical analyses after 60 days. The present study revealed how the presence of degradable polylactic acid (PLA) and non-degradable polyethylene (PE) MPs affects soil microbial nutrient limitation and C use efficiency (CUE) at varying nutrient concentrations. The presence of PLA in soil with low nutrient levels led to a significant increase (29%) in the activities of nitrogen (N)-acquiring enzymes. In contrast, the presence of MPs had no effect on C- and N-acquiring enzymes. The occurrence of PE caused a 41% reduction in microbial C limitation in high-nutrient soils, and microbial nutrient metabolism was limited by the occurrence of MPs in soils amended with nutrients. A strong positive correlation between microbial C and nutrient limitation in the soil indicates that addressing C limitation followed by amendment of soil with MPs could potentially intensify microbial N limitation in soils with varying nutrients. In comparison, the microbial CUE increased by 10% with the application of degradable MPs (PLA) to soils with a low nutrient status. These findings highlight the significant influence of both degradable PLA and non-degradable PE MPs on soil microbial processes and C dynamics. In conclusion, PLA enhances metabolic efficiency in nutrient-rich soils, potentially aiding C utilization, whereas PE reduces microbial C limitation, offering promise for soil C sequestration strategies. Our findings underscore the importance of considering MPs in soil ecosystem studies and in broader sustainability efforts.

Fear of predators alters herbivore regulation of soil microbial community function.

Fear of predation can affect important ecosystem processes by altering the prey traits expression that, in turn, regulates the quantity and quality of nutritional inputs to soil. Here, we aimed to assist in bridging a knowledge gap in this cascading chain of events by exploring how risk of spider predation may affect grasshopper prey performances, and the activity of various microbial extracellular enzymes in the soil. Using a mesocosms field-experiment, we found that grasshoppers threatened by spider predation ate less, grew slower, and had a higher body carbon to nitrogen ratio. Herbivory increased activity of all microbial extracellular enzymes examined, likely due to higher availability of root exudates. Predation risk had no effect on C-acquiring enzymes but decreased activity of P-acquiring enzymes. We found contrasting results regarding the effect of predation on the activity of N-acetyl-glucosaminidase and leucine arylamidase N-acquiring enzymes, suggesting that predation risk may alter the composition of N-inputs to soil. Our work highlighted the importance of soil microbial enzymatic activity as a way to predict how changes in the aboveground food-web dynamics may alter key ecosystem processes like nutritional-cycling.

Soil extracellular enzyme stoichiometry reflects microbial metabolic limitations in different desert types of northwestern China.

Soil extracellular enzyme activity (EEA) stoichiometry reflects the dynamic balance between microorganism metabolic demands for resources and nutrient availability. However, variations in metabolic limitations and their driving factors in arid desert areas with oligotrophic environments remain poorly understood. In this study, we investigated sites in different desert types in western China and measured the activities of two C-acquiring enzymes (ß-1,4-glucosidase and ß-D-cellobiohydrolase), two N-acquiring enzymes (ß-1,4-N-acetylglucosaminidase and L-leucine aminopeptidase), and one organic-P-acquiring enzyme (alkaline phosphatase) to quantify and compare the metabolic limitations of soil microorganisms based on their EEA stoichiometry. The ratios of log-transformed C-, N-, and P-acquiring enzyme activities for all deserts combined were 1:1.1:0.9, which is close to the hypothetical global mean EEA stoichiometry (1:1:1). We quantified the microbial nutrient limitation by means of vector analysis using the proportional EEAs, and found that microbial metabolism was co-limited by soil C and N. For different desert types, the microbial N limitation increased in the following order: gravel desert < sand desert < mud desert < salt desert. Overall, the study area's climate explained the largest proportion of the variation in the microbial limitation (17.9 %), followed by soil abiotic factors (6.6 %) and biological factors (5.1 %). Our results confirmed that the EEA stoichiometry method can be used in microbial resource ecology research in a range of desert types, and that the soil microorganisms maintained community-level nutrient element homeostasis by adjusting enzyme production to increase uptake of scarce nutrients even in extremely oligotrophic environments such as deserts.

Microbes drive metabolism, community diversity, and interactions in response to microplastic-induced nutrient imbalance.

The impact of conventional and biodegradable microplastics on soil nutrients (carbon and nitrogen) has been widely examined, and the alteration of nutrient conditions further influences microbial biosynthesis processes. Nonetheless, the influence of microplastic-induced nutrient imbalances on soil microorganisms (from metabolism to community interactions) is still not well understood. We hypothesized that conventional and biodegradable microplastic could alter soil nutrients and microbial processes. To fill this knowledge gap, we conducted soil microcosms with polyethylene (PE, new and aged) and polylactic acid (PLA, new and aged) microplastics to evaluate their effects on the soil enzymatic stoichiometry, co-occurrence interactions, and success patterns of soil bacterial communities. New and aged PLA induced soil N immobilization, which decreased soil mineral N by 91-141 %. The biodegradation of PLA led to a higher bioavailable C and wider bioavailable C:N ratio, which further filtered out specific microbial species. Both new and aged PLA had a higher abundance of copiotrophic members (Proteobacteria, 35-51 % in PLA, 26-34 % in CK/PE treatments) and rrn copy number. The addition of PLA resulted in a lower alpha diversity and reduced network complexity. Conversely, because of the chemically stable hydrocarbon structure of PE polymers, the new and aged PE microplastics had a minor effect on soil mineral N, bacterial community composition, and network complexity, but led to microbial C limitation. Collectively, all microplastics increased soil C-, N-, and P -acquiring enzyme activities and reduced the number of keystone species and the robustness of the co-occurrence network. The PLA treatment enhanced nitrogen fixation and ureolysis, whereas the PE treatment increased the degradation of recalcitrant carbon. Overall, the alteration of soil nutrient conditions by microplastics affected the microbial metabolism and community interactions, although the effects of PE and PLA microplastics were distinct.

Extracellular enzyme stoichiometry and microbial resource limitation following various grassland reestablishment in abandoned cropland.

Grassland restoration in abandoned cropland had great impact on soil enzyme stoichiometry and microbial resource limitation, hence altering carbon (C) sequestration progress in soil depending on soil depth and grassland restoration strategy. It is crucial to understand the microbial resource limitation under various restoration strategies, which could have key implication for optimizing management to improve C sequestration in abandoned cropland. The objective of this study was to examine the changes and key regulators of soil enzyme stoichiometry and microbial resource limitation in different soil depths under different management strategies to restore grassland, namely a) cropland as continuous cropping (CR); b) naturally restored grassland (NR); c) grass-based grassland (GG); d) legume-based grassland (LG); e) grass-legume mixed grassland (MG); and f) grass-based grassland with N fertilization (GF). Results showed that converting cropland into grassland increased absolute soil enzyme activities potential for microbial C, nitrogen (N) and phosphorus (P) acquisition by 5-110 %, 25-132 % and 17-215 %, respectively depending on soil depth and grassland restoration strategy. These enzyme activities increased more in surface soil than subsoil with the conversion of cropland into grassland, especially under LG and GF. The strategies to restore grassland, especially LG and GF, significantly decreased enzymatic C:P and N:P ratios. Microbial C limitation was reduced associated with re-establishment of grassland, exacerbating the P limitation depending on grassland restoration strategies, especially under LG and GF. The shift of relative microbial resource limitation from C to P reduced the microbial C use efficiency, reducing the ecosystem C sequestration potential during the restoration of grassland. It appears that increased biomass input and soil C:P ratio are the key drivers to shift microbial resource limitation from C to P during the restoration of grassland. Thus, a moderate harvest of above-ground biomass with a supplement of P may be necessary for improving the C sequestration potential during the restoration of grasslands, especially in the grass-legume mix or grass-based grassland with N fertilization.

Rice-crayfish co-culture increases microbial necromass' contribution to the soil nitrogen pool.

The rice-crayfish co-culture (RC) is a putative sustainable agricultural system. However, studies on the ecological effects of long-term RC systems were still lacking. Here, we compare enzymatic stoichiometry, microbial necromass, and microbial community between the RC and rice monoculture systems (RM). Soil enzymatic stoichiometry analysis showed that after transformation from RM to RC for about three years, ammonium nitrogen (NH4+-N) availability increased to depress relative N-acquiring enzyme production, especially for leucine aminopeptidase. The contents of microbial necromass increased approximately onefold in the RC system, making microbial necromass' contribution to the soil nitrogen (N) reach up to 46.72%. Elevation in NH4+ decreased N-acquiring enzyme, and a relatively more effective C acquisition likely benefited microbial necromass retention and production in the RC system. This study highlights that the rice-crayfish co-culture could modify the N pool of the surface paddy soil.

Assessment of soil enzymatic resilience in chlorpyrifos contaminated soils by biochar aided Pelargonium graveolens L. plantation.

Chlorpyrifos (CP), a broad-spectrum organophosphorus insecticide, is known for deleterious effects on soil enzymatic activities. Hence, the present study aims to examine the resilience effect of biochar (BC) aided Pelargonium graveolens L. plantation on enzymatic activities of chlorpyrifos contaminated soil. The two chlorpyrifos contaminated agriculture soils (with concentrations: S1: 46.1 and S2: 95.5 mg kg-1) were taken for the pot experiment. The plant biomass, plant growth parameters, soil microbial biomass, and enzymatic activities such as alkaline phosphatase, N-acetyl glucosaminidase, aryl sulphatase, cellulase, ß-glucosidase, dehydrogenase, phenoloxidase, and peroxidase enzymes were  examined. Ecoenzyme activities and their stoichiometry were used to enumerate the different indices including geometric mean, weighted mean, biochemical activity indices, integrated biological response, treated-soil quality index, and vector analysis in all treatments. The results of the study demonstrated that the biochar incorporation enhanced the tolerance of P. graveolens (from 42-45% to 55-67%) in chlorpyrifos contaminated soil and reduced the CP accumulation in plants. A reduction in the inhibitory effect of chlorpyrifos on soil enzymatic activities and plant growth by BC incorporation was observed along with an increase in the activities of ecoenzymes (16.7-18.6%) in soil. The investigation indicated more microbial investments in C and P than that in N acquisition under CP stress. The BC amendment catalyzed the activities of lignin and cellulose-degrading enzymes and enhanced nutrition acquisition. The CP contamination and BC amendment have no significant effect on the oil quality of P. graveolens. The study demonstrated that BC-aided P. graveolens plantation offers sustainable phytotechnology for CP contaminated soil with an economic return.

Oxygen availability regulates the quality of soil dissolved organic matter by mediating microbial metabolism and iron oxidation.

Dissolved organic matter (DOM) plays a vital role in biogeochemical processes and in determining the responses of soil organic matter (SOM) to global change. Although the quantity of soil DOM has been inventoried across diverse spatio-temporal scales, the underlying mechanisms accounting for variability in DOM dynamics remain unclear especially in upland ecosystems. Here, a gradient of SOM storage across 12 croplands in northeast China was used to understand links between DOM dynamics, microbial metabolism, and abiotic conditions. We assessed the composition, biodegradability, and key biodegradable components of DOM. In addition, SOM and mineral-associated organic matter (MAOM) composition, soil enzyme activities, oxygen availability, soil texture, and iron (Fe), Fe-bound organic matter, and nutrient concentrations were quantified to clarify the drivers of DOM quality (composition and biodegradability). The proportion of biodegradable DOM increased exponentially with decreasing initial DOM concentration due to larger fractions of depolymerized DOM that was rich in small-molecular phenols and proteinaceous components. Unexpectedly, the composition of DOM was decoupled from that of SOM or MAOM, but significantly related to enzymatic properties. These results indicate that microbial metabolism exhibited a dominant role in DOM generation. As DOM concentration declined, increased soil oxygen availability regulated DOM composition and enhanced its biodegradability mainly through mediating microbial metabolism and Fe oxidation. The oxygen-induced oxidation of Fe(II) to Fe(III) removed complex DOM compounds with large molecular weight. Moreover, increased oxygen availability stimulated oxidase-catalyzed depolymerization of aromatic substances, and promoted production of protein-like DOM components due to lower enzymatic C/N acquisition ratio. As global changes in temperature and moisture will have large impacts on soil oxygen availability, the role of oxygen in regulating DOM dynamics highlights the importance of integrating soil oxygen supply with microbial metabolism and Fe redox status to improve model predictions of soil carbon under climate change.

Crop rotation stage has a greater effect than fertilisation on soil microbiome assembly and enzymatic stoichiometry.

Agronomic practises, such as fertilisation and crop rotation, affect soil microbial communities and functions. However, limited information is available regarding the relative importance of fertilisation and crop rotation stages in determining the soil microbiome and assembly processes. In addition, insights into the connections between the soil microbiome and enzymatic stoichiometry are scarce. In this study, soil samples were collected from a wheat-rice rotation system that received mineral and organic fertiliser inputs for 6 years to investigate soil microbiome assembly, and the relationship between the soil microbiome and enzymatic stoichiometry. Our results revealed that the crop rotation stage strongly affected the soil microbial community structure, assembly, and enzymatic functions compared to that of the fertilisation regime. Enzymatic stoichiometry results and vector analysis implied that mineral and organic fertilisation could alleviate the microbial N limitation. However, no-manure fertilisation led to microbial P limitation during the wheat stage. The decreases in soil pH mainly drove microbial P limitation due to the acidification induced by the mineral fertilisers. Microbial N/P limitation correlated more strongly with the bacterial assembly than with fungal assembly. Moreover, co-occurrence network analysis showed that ecological relationships between microbial taxa and enzymes were more complex during the wheat stage than that during the rice stage. Microbial nodes linked to acid phosphomonoesterase correlated significantly with the soil pH. Our study highlights the distinct responses of the soil microbiome to fertilisation in different crop-rotation stages, and provides novel insights into connections between microbial assembly and enzymatic stoichiometry.

[Extracellular Enzyme Stoichiometry and Microbial Metabolism Limitation During Vegetation Restoration Process in the Middle of the Qinling Mountains, China].

Clarifying the characteristics of soil microbial nutrient limitation and its driving mechanisms during vegetation restoration after farmland abandonment has important implications for revealing soil nutrient cycling and maintaining ecosystem stability. To determine the limitation of soil microbial nutrients and its relationship with soil properties along a chronosequence of abandoned farmland in the middle of the Qinling Mountains, the soil physicochemical properties and five enzyme activities (ß-1,4-glucosidase (BG), cellobiohydrolase (CBH), ß-1,4-N-acetylglucosaminidase (NAG), leucine aminopeptidase (LAP), and acid phosphatase (AP)) were measured, and models of extracellular enzymatic activity were applied. The results showed that the activities of BG, CBH, NAG, LAP, and AP were significantly increased following farmland abandonment. With the increasing years of abandonment, the ratios of (BG+CBH)/(NAG+LAP) and (BG+CBH)/AP significantly decreased, whereas the ratio of (NAG+LAP)/AP increased. Correlation analysis showed that most soil physicochemical properties were significantly correlated with extracellular enzyme activities and extracellular enzymatic stoichiometry. The vector length of extracellular enzymatic stoichiometry decreased with the increase in abandonment years, indicating that the limitation of soil microorganisms on carbon (C) was reduced. Moreover, the vector angles (>45°) showed a decreasing trend, indicating that microbial metabolisms were limited by phosphorus (P) and gradually decreased. Regression analysis showed that the C and P limitations were significantly related to total nutrients, available nutrients, nutrient ratio, and soil physical properties. Partial least squares path modeling (PLS-PM) revealed that the C and P limitations were directly regulated by nutrient ratio. PLS-PM further showed that soil total nutrients indirectly affected soil microbial C and P limitations by affecting nutrient ratio, and nutrient ratio affected the soil metabolism limitation via available nutrients and pH. Our study suggests that the characteristics of microbial metabolism during the vegetation restoration process reflect the mechanism of microorganism-mediated soil nutrient cycling, which provides a theoretical basis for revealing the community dynamics and stability during the vegetation restoration process and maintaining the regional ecological environment security in the Qinling Mountains.

Effect of high soil C/N ratio and nitrogen limitation caused by the long-term combined organic-inorganic fertilization on the soil microbial community structure and its dominated SOC decomposition.

The application of organic fertilizers, such as straw and manure, is an efficient approach to maintain soil productivity. However, the effect of these organic fertilizers on soil microbial nutrient balance has not yet been established. In this study, the effects of the long-term combined organic-inorganic fertilization on microbial community were investigated by conducting a 30-year-long field test. Overall, the following five fertilizer groups were employed: inorganic NP fertilizer (NP), inorganic NK fertilizer (NK), inorganic NPK fertilizer (NPK), NPK + manure (MNPK), and NPK + straw (SNPK). The results indicated that the mean natural logarithm of the soil C:N:P acquisition enzyme ratio was 1.04:1.11:1.00 under organic-inorganic treatments, which showed a deviation from its overall mean ratio of 1:1:1. This indicates that microbial resources do not have a balance. Vector analysis (vector angle <45°) and threshold elemental ratio analysis (RC:N-TERC:N > 0) further demonstrated that the microbial metabolism was limited by Nitrogen (N) under SNPK and MNPK treatments. N limitation further influenced soil microbial community structure and its dominated SOC decomposition. Specifically, Microbial communities transformed into a more oligotrophic-dominant condition (fungal, Acidobacteria, Chloroflexi) from copiotrophic-dominant (Proteobacteria, Actinobacteria) condition with increasing N limitation. Lysobacter genus and Blastocatellaceae family, in the bacterial communities along with the Mortierella elongata species in fungal communities, were markedly associated with the N limitation, which could be the critical biomarker that represented N limitation. Both correlation analysis and partial least squares path modeling showed significant positive effects of N limitation on the ratio of bacterial functional genes (Cellulase/Amylase), involved in recalcitrant SOC degradation.

Soil extracellular enzyme activities and the abundance of nitrogen-cycling functional genes responded more to N addition than P addition in an Inner Mongolian meadow steppe.

Nitrogen (N) and phosphorus (P) availability in soils commonly limit belowground biological processes in terrestrial ecosystems. Soil extracellular enzyme activities (EEAs) and microbial functional groups play critical roles in soil biological processes and nutrient cycling, yet their response to nutrient addition are poorly understood. To address this issue, we applied six fertilization treatments composed of combinations of N (0, 1.55, 13.95 g N m-2 yr-1) and P (0, 5.24 g P m-2 yr-1) for two years in a meadow steppe of Inner Mongolia. Soils were collected from each plot in July and August and analyzed for abundances of N-cycling genes and EEAs, and their relationships with treatments. The addition of N significantly increased C-acquisition enzyme activity and enzyme C:N and C:P ratios. Enzymatic stoichiometry indicated that N addition alleviated microbial demand for N, while it increased microbial C limitation. Microbial C and N limitation were significantly correlated with NH4+-N in July, yet they were correlated with soil water content (SWC) in August. The abundance of amoA significantly increased with N addition and was positively related to mineral-N accumulation. The abundance of denitrifier genes and gaseous N loss potential were accelerated by N addition in July, while a neutral effect was observed in August. Nitrate leaching potential was significantly increased by N addition, yet it declined with P addition in July. P addition also suppressed amoA abundance of ammonia oxidizing bacteria. Partial least squares path modelling indicated that N addition positively affected microbial-C limitation, soil N-loss potential and negatively affected microbial-N limitation. P addition negatively affected soil N-loss potential. Ultimately, this study highlights the importance of soil N availability in regulating microbial metabolism and soil N-loss potential, and enhances our understanding of the mechanisms responsible for variation in microbial nutrient cycling in meadow steppe soils.

[Effects of Farmland Abandonment on Soil Enzymatic Activity and Enzymatic Stoichiometry in the Loess Hilly Region, China].

Clarifying the characteristic of soil enzymatic activity and stoichiometry variations as well as their influencing factors following farmland abandonment have important implications for understanding soil nutrient availability after revegetation and for illuminating the underlying mechanisms of soil nutrient cycling in ecosystems. To determine microbial nutrient limitations after farmland abandonment and to explore the driving factors of the variations in soil enzymatic activity and stoichiometry along a chronosequence of abandoned farmlands (0-, 10-, 20-, and 30-year-old) in the Loess Hilly Region, China, the potential activities of carbon (C)-, nitrogen (N)-, and phosphorus (P)-acquiring enzymes, soil physicochemical properties, and plant diversity and family composition were measured. The results showed that the activities of ß-1,4-N-acetylglucosaminidase (NAG), leucine aminopeptidase (LAP), and alkaline phosphatase (ALP) increased significantly with the increasing years of land abandonment, whereas the activity of ß-1,4-glucosidase (BG) showed the opposite change trend. Additionally, the ratios of BG:(NAG+LAP) and BG:ALP had the same variation trend with BG activity, which decreased significantly with increasing time, but the ratio of (NAG+LAP):ALP showed an increasing trend and then decreased, with the highest values observed in the 20-year sites. Moreover, the vector length of soil enzymatic stoichiometry decreased significantly as the years of land abandonment inceased, suggesting a reduced microbial C limitation after farmland abandonment. The vector angles <45°were observed at farmlands (0-year sites) and 10-year sites, whereas angles >45°were detected at 20-and 30-year sites, indicating that soil microbial communities were N-limited in the first 10 years of land abandonment and thereafter were P-limited. The redundancy analysis (RDA) reveled that soil organic C content, total N content, the C:N and C:P ratios, soil pH values, and plant diversity had significant effects on soil enzymatic activity and stoichiometry. A variation partitioning analysis (VPA) further demonstrated that edaphic and vegetation factors explained 62.0% of the total variance of soil enzymatic activity and stoichiometry. It should be noted that the interaction between vegetation characteristics and soil physicochemical properties was the major factor affecting soil enzymatic activity and stoichiometry, which explained 37.1% of the variance of the soil enzyme characteristics. Collectively, the application of P fertilizer should be considered to mitigate the deficiency of available P in the ecosystem during farmland abandonment, and these findings may provide a theoretical basis for understanding the mechanisms underlying microbe-mediated biogeochemical cycles as well as guiding soil nutrient management and the sustainable development of the ecological environment.

[Stoichiometry of soil extracellular enzymes and its seasonal variation in natural forests with different altitudes in northern Greater Khingan Mountains, China]. / å¤§å ´å®‰å²­åŒ—éƒ¨ä¸åŒæµ·æ‹”å¤©ç„¶æž—åœŸå£¤èƒžå¤–é ¶åŒ–å­¦è®¡é‡ç‰¹å¾åŠå ¶å­£èŠ‚åŠ¨æ€.

In this study, we examined the characteristics and influence mechanism of soil extracellular enzyme activity (EEA) and enzymatic stoichiometry in different soils in forests at different altitudes (750-1420 m) in Aokelidui Mountains in the north of the Greater Khingan Mountains. The results showed that altitude, season and their interactions significantly affected the activities of ß-glucosidase (BG), ß-1,4-N-acetylglucosaminidase (NAG), L-leucine aminopeptidase (LAP), and acid phosphatase (AP). In May, BG and NAG activities gradually increased with increasing altitude, while AP activities increased first and then decreased with increasing altitude. In July, NAG activity increased with altitude, while AP activity increased first and then decreased. In September, NAG activity changed significantly in different altitudes, with the highest activity at 1420 m (124.22 nmol·h-1·g-1). With the increases of altitude, ln(BG): ln(NAG+LAP) showed a decreasing trend. Except for the altitude of 830 m, stoichiometric ratio in all altitudes was the highest in July. The ratio of logarithmic conversion of soil C, N, and P invertase activity was 1:1.25:0.82. Altitude and soil temperature were the main factors affecting soil extracellular enzyme activities. There was a significant positive correlation between soil temperature and BG, NAG, and AP. Enzymatic stoichiometry ln(BG):ln(NAG+LAP) and ln(NAG+LAP):ln(AP) showed significant positive and negative correlations with soil pH, and had a negative and positive relationship with DOC. The ratio of ln(BG):ln(AP) was greatly affected by soil bulk density.