Enzyme Stoichiometry Research Articles

Variations in soil enzyme stoichiometry and microbial nutrient limitations in Camellia oleifera plantations with different ages

The alteration of stand age instigates modifications in soil properties and microbial communities. Understanding the impacts of stand age on soil enzyme stoichiometry and microbial nutrient limitations in Camellia oleifera plantation is crucial for nutrient management. Taking C. oleifera plantation across four age groups (<10 a, 15-25 a, 30-50 a, >60 a) in a subtropical red soil region as test objects, we examined the response of soil enzyme stoichiometry and microbial nutrient limitations to change in stand age and analyzed the pathways for such responses. The results showed that, compared to that of stand age <10 a, enzyme C:N in the 15-25 a was increased and enzyme N:P was significantly reduced. Microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), and microbial biomass phosphorus (MBP) exhibited a trend of initially decreasing and then increasing with stand age. MBN and MBN:MBP were significantly higher in the <10 a compared to that in the 30-50 a. MBC:MBN was significantly higher in the 30-50 a and >60 a compared to the <10 a and 15-25 a. Results of redundancy analysis revealed that soil nutrients, microbial biomass and their stoichiometry explained 92.4% of the variations in enzyme stoichiometry. Partial least squares path modeling (PLS-PM) results demonstrated that soil organic carbon (SOC) had a positive effect on microbial C limitation; MBN, MBN:MBP, MBC:MBP, SOC, and total nitrogen had a nega-tive overall effect on microbial P limitation, whereas soil C:N had a positive overall effect on microbial P limitation. There was a significant positive correlation between microbial C and P limitations. With increasing stand age, microbial nutrient limitation shifted from N and P limitation (<10 a) to C and P limitation (15-25 a, 30-50 a, >60 a).

Inversion Tillage Combined with Organic Fertilizer Application Increased Maize Yield via Improving Soil Pore Structure and Enzymatic Activity in Haplic Chernozem

Inversion tillage and organic fertilizer application can break the plow pan and improve soil quality. However, the effects of combining these practices on the soil microbial resource limitation and maize yield in Haplic Chernozem are unclear. In this research, a field experiment was established in 2018, and soil samples were collected in 2021 in Longjiang County in Northeast China, which is a Haplic Chernozem region. Four treatments comprising conventional tillage (T15), conventional tillage with organic fertilizer (T15+M), inversion tillage (T35), and inversion tillage with organic fertilizer (T35+M) were randomly arranged with four replications. Compared with T15 and T15+M treatments, soil bulk density significantly decreased by 11.1–16.3% in the 15–35 cm layer under T35 and T35+M treatments, accompanied by the improvement in soil pore structure (e.g., soil porosity, circularity, and Euler number). T15+M treatment significantly increased soil organic carbon and soil nutrient contents by 11.1–16.3% and 3.9–24.5% in the 0–15 cm layer compared with other treatments. However, soil organic carbon, total nitrogen, available phosphorus content, microbial biomass, and enzymatic activities reached the maximum values in the 0–35 cm layer under T35+M treatment. In addition, T35+M treatment had the highest maize yield and sustainable yield index. Extracellular enzymatic stoichiometry suggested that soil microorganisms are generally co-limited by carbon and phosphorus in Haplic Chernozem. However, T35+M treatment significantly reduced soil microbial resource limitation, which was one important factor impacting maize yield and sustainability. Random-forest and partial least-squares path modeling showed that T35+M treatment could reduce soil microbial resource limitation and increase the stability and sustainability of the maize yield by improving soil available nutrients, microbial biomass, and pore structure. Therefore, the incorporation of inversion tillage and organic fertilizer is a suitable soil management practice in view of increasing soil quality and crop yields in a Haplic Chernozem region.

Excessive climate warming exacerbates nitrogen limitation on microbial metabolism in an alpine meadow of the Tibetan Plateau: Evidence from soil ecoenzymatic stoichiometry

Soil ecoenzymatic stoichiometry reflects the dynamic equilibrium between microorganism's nutrient requirements and resource availability. However, uncertainties persist regarding the key determinants of nutrient restriction in relation to microbial metabolism under varying degrees of warming. Our long-term and multi-level warming field experiment (control treatment, +0.42 °C, +1.50 °C, +2.55 °C) in a typical alpine meadow unveiled a decline in carbon (C)- and nitrogen (N)-acquired enzymes with escalating warming magnitudes, while phosphorus (P)-acquired enzymes displayed an opposite trend. Employing enzymatic stoichiometry modeling, we assessed the nutrient limitations of microbial metabolic activity and found that C and N co-limited microbial metabolic activities in the alpine meadow. Remarkably, high-level warming (+2.55 °C) exacerbated microbe N limitation, but alleviate C limitations. The structural equation modeling further indicated that alterations in soil extracellular enzyme characteristics (SES) were more effectively elucidated by microbial characteristics (microbial biomass C, N, P, and their ratios) rather than by soil nutrients (total nutrient contents and their ratios). However, the microbial control over SES diminished with higher levels of warming magnitude. Overall, our results provided novel evidence that the factors driving microbe metabolic limitation may vary with the degree of warming in Tibet alpine grasslands. Changes in nutrient demand for microorganism's metabolism in response to warming should be considered to improve nutrient management in adapting to different future warming scenarios.

Straw return with fertilizer improves soil CO2 emissions by mitigating microbial nitrogen limitation during the winter wheat season

Straw return is the key measure of green agricultural production. Nitrogen (N) and phosphorus (P) play pivotal roles in crop growth and carbon (C) cycling in agroecosystems. However, the mechanism of soil CO2 emissions and enzyme stoichiometry regulation by straw return with N and P application remain unknown. Therefore, this study is aimed to use enzyme stoichiometry to investigate the effects of straw return with N and P fertilizer application on soil CO2 emissions, enzyme activity, and microbial nutrient limitation through a two-year field experiment. A split-plot experimental design was used (straw as the main plot and fertilizer as the sub plot). The results indicated that the addition of N, P (SNP) increased CO2 emissions by 25.54 %, and N application (SN) reduced CO2 emissions by 27.01 %, compared with straw return (SW). Enzyme stoichiometry indicated that straw return with N and P application exhibited significant C and N limitations. In soil nutrients and nutrient stoichiometry, compared with the S0W treatment, straw return with fertilizer (SNP, SN) significantly increased microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), dissolved organic carbon (DOC) and dissolved organic nitrogen (DON); SNP treatment increased β-1,4- n −acetylaminoglucosidase (NAG) enzyme activity by 53.72 %, increased l −leucine aminopeptidase (LAP) enzyme activity by 58.11 %, and increased one P-acquiring enzyme alkaline monoalkaline phosphatase (AKP) enzyme activity by 39.49 % (P < 0.05). Structural equation modeling suggested that straw return and fertilization can influence microbial biomass phosphorus (MBP) and vector length (VL) by affecting MBN, which, in turn, affects CO2 emissions. Soil microbes regulate CO2 emissions using stoichiometric flexibility to maintain a stoichiometric balance of nutrients and microbes. Therefore, straw return with N and P fertilizers to winter wheat soils can meet the requirements of microbial stoichiometry, modulate microbial and extracellular enzyme activities, and effectively regulate CO2 emissions and resource limitations.

Effects of Land-Use Type and Salinity on Soil Carbon Mineralization in Coastal Areas of Northern Jiangsu Province

Sea level rise due to glacier melting caused by climate warming is a major global challenge, but the mechanism of the effect of salinity on soil carbon (C) mineralization in different land types is not clear. The pathways by which salinity indirectly affects soil carbon mineralization rates need to be investigated. Whether or not the response mode is consistent among different land-use types, as well as the intrinsic links and interactions between soil microbial resource limitation, environmental stress, microbial extracellular enzyme activity, and soil carbon mineralization, remain to be demonstrated. In this paper, three typical land-use types (wetland, forest, and agroforestry) were selected, and different salinity levels (0‰, 3‰, 6‰, and 32‰) were designed to conduct a 125-day laboratory incubation experiment to determine the soil CO2 release rate, soil physicochemical properties, and soil enzyme activities, and to correlate C mineralization with biotic and abiotic factors. A correlation analysis of soil physical and chemical properties, extracellular enzyme activities, and carbon mineralization rates was conducted to investigate their intrinsic linkages, and a multiple linear regression of C mineralization at different sites was performed to explore the variability of mineralization among different site types. Structural equation models were established in the pre- and post-incubation stages to study the pathways of soil C mineralization at different incubation times, and the mechanism of mineralization was further verified by enzyme stoichiometry. The results showed that, at the end of 125 days of incubation, the 32‰ salinity addition reduced the cumulative mineralization of forest and agroforestry types by 28.41% and 34.35%, respectively, compared to the 0‰ salinity addition. Soil C mineralization in the three different land-use types was highly correlated with the active C fractions of readily oxidizable C (ROC), dissolved organic C, and microbial biomass C (MBC) in the soil, with the standardized coefficients of multivariate linear regression reaching 0.67 for MBC in the wetland and −0.843 for ROC in the forest. Under long-term salinity additions, increased salinity would reduce the microbial respiratory quotient value by inhibiting β-glucosidase activity, thus indirectly affecting the rate of CO2 release. With added salinity, the mineralization of non-saline soil was more susceptible to the inhibitory effect of salinity, whereas the mineralization of salinized soil was more controlled by soil C pools.

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, β-1,4-N-acetylglucosaminidase and phosphatases), microbial biomass, and community structure. Results revealed a substantial increase (1.21–2.81 times) in β-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.

Soil Carbon Storage, Enzymatic Stoichiometry, and Ecosystem Functions in Indian Himalayan Legume-Diversified Pastures

The influences of legume diversification on soil carbon (C) pools and sequestration, particularly in Himalayan pastureland, remain poorly understood. Moreover, the impact of legume diversification activities and the stoichiometry of soil enzymes in C biogeochemistry at the ecosystem level remains largely overlooked. The purpose of this study is to investigate the influences of legume diversification on activities and the stoichiometry of soil enzymes and their control of C sequestration in pasturelands. Four experimental fertilized species combinations, namely, SG (50% Festuca arundinacea + 50% Dactylis glomerata), SGL1 (25% Festuca arundinacea + 25% Dactylis glomerata + 50% Onobrychis viciifolia), SGL2 (25% Festuca arundinacea + 25% Dactylis glomerata + 50% Trifolium pratense), SGL12 (25% Festuca arundinacea + 25% Dactylis glomerata + 25% Onobrychis viciifolia + 25% Trifolium pratense), and natural pasture (NG) were compared. Soils under SGL1, SGL2, and SG12 had ~18, 36, and 22% greater soil C than SG, respectively. Among the pastures with fertilization, the C mineralization was suppressed by legume diversification. C sequestration under SGL1, SGL2, and SG12 was ~27, 22, and 38% higher than SG, respectively, at the 0–30 cm soil layer. The ratios of DHA are as follows: for PhOX and DHA, PerOX significantly decreased with an increasing grass–legume mixture, suggesting greater C sequestration. PCA analysis revealed that C sequestration under legume diversification and enzymatic stoichiometry had an indirect but substantial impact on C sequestration. The increasing C sequestration under SGL12 was complemented by higher productivity. Data suggested that increasing legumes in pastureland might greatly enhance ecosystem functions such as soil C storage, productivity, ecorestoration efficiency, and biological activity in Indian Himalayan pastureland.

High starch and hemicellulose labile C degradation functional genes increase soil CO2 emissions follow straw return

Straw return is a key method to regulate soil carbon (C) by microbial utilization accumulating and CO2 releasing, which mainly depends on microbial metabolism processes. The present knowledge of the mode and mechanism of soil CO2 emission under straw return is limited. We established four straw return treatments, including straw shallow return (SS, 0–20 cm), straw subsoil burial (SD, 20–40 cm), straw mulching (SM), and control with no straw (NS). Microbial metabolic limitation and the abundance of C degradation functional genes were quantified using enzyme stoichiometry and high-throughput quantitative PCR-based chip technology, respectively. The results revealed that the addition of straw inputs resulted in an increase in soil nutrient contents and soil CO2 emissions. The cumulative CO2 emissions of SS and SM treatments were 21,013.53 and 20,819.26 kg hm−2, respectively, which significantly increased by 4.70 % and 3.73 % compared with SD treatment. Compared with SM and SD treatments, microbial C and P limitations of SS treatment were significantly enhanced, increasing by 1.15 %–1.20 % and 2.73 %–1.21 %, respectively. Straw return increased the absolute abundance of genes engaged in decomposing diverse C source compounds, although with a reduction in microbial C use efficiency. The absolute abundance of amyA, apu, sga, abfA, xylA, and manB labile C degradation genes was higher in SM than in SD treatment, increasing by 1.11 %–25.88 %. On the contrary, the absolute abundance of glx and lig recalcitrant C degradation genes was lower in SM than in SD treatment, reducing by 2.96 %–3.68 %. Straw return directly impacts soil nutrient contents and indirectly influences microbial biomass carbon (MBC), nutrient limitation, and C degradation genes, as revealed by the partial least squares pathway model, ultimately leading to changes in soil CO2 emissions. This finding should contribute to the enhancement of soil C retention and the selection of appropriate methods for straw incorporation in tillage fields.

Restoring Subtropical Forests: Alleviating P Limitation and Introducing C Limitation Using Evergreen Broad-Leaved Tree Species

Determining which species to utilize for the artificial restoration of subtropical secondary forests has become a focal point in forestry and ecology. To compare the effects of the subtropical secondary forest artificial restoration model on soil microbial nutrient acquisition and limitation, we examined secondary forests (CKs), evergreen coniferous forests (GCPs), evergreen coniferous mixed broad-leaved forests (GCBMs), evergreen mixed broad-leaved forests (GBMs), and natural deciduous broad-leaved mixed forests (DBMs) as research subjects. Among them, GCPs, GCBMs, and GBMs were dominated by the species of the early, middle, and climax stages of subtropical forest succession, respectively. The activities and stoichiometry of β-1,4-glucosidase (BG), β-1,4-N-acetylglucosaminidase (NAG), leucine aminopeptidase (LAP), and acid phosphatase (ACP) in the topsoil were analyzed. The results showed that the forest type significantly affects the activities of BG and LAP rather than NAG or ACP. The BG activity in DBMs was the lowest, while the LAP activity in CKs was significantly higher than that in plantations. Furthermore, the nutrient limitation of microbes was quantitatively analyzed by using the vector analysis of enzyme stoichiometry. The soil microbes in the study area were co-limited by C and P, and the nutrient limitation was in the order of C &gt; P &gt; N. Among the forests, the enzyme stoichiometric ratios in GCPs and DBMs were closest to 1:1:1. From CKs to GBMs, the microbial C limitation was increased, while the P limitation was decreased. The C limitation in DBMs was slightly lower than that in CKs. Overall, the P limitation in evergreen plantations was less than that in CKs. The soil bulk density, C/P, and N/P significantly influenced enzyme activities and stoichiometry. These results suggest that the artificial restoration of subtropical forests using evergreen species alleviated P limitation, while using deciduous broad-leaved species offered potential for alleviating microbial C limitation. Compared with evergreen broad-leaved species, employing pioneer and mid-successional or deciduous broad-leaved species can better achieve balanced microbial nutrient requirements.

The life state of smooth vetch regulates the seasonal dynamics of microbial nutrient limitations in citrus orchards

AbstractCover crops improve soil fertility in orchard ecosystems, however, the response of soil microbial metabolism to fruit tree interactions with cover legumes at different stages of the life cycle remains uncertain, thereby limiting our understanding of the mechanisms that regulate nutrient cycling in orchards with cover crop management. Here, the characteristics of microbial energy (carbon, C) and nutrient (nitrogen, N or phosphorus, P) metabolism in the citrus rhizosphere and bulk soil in orchards were analyzed with two managements, that is, smooth vetch (Vicia villosa Roth var. glabresens Koch) cover and clean tillage, using an extracellular enzymatic stoichiometry model. Microbial C limitation was significantly reduced by cover during the vetch growing season owing to the resulting higher plant root C and soil organic C content. Competition for N between plants and microbes in the bulk soil might lead to significant microbial N limitation in covered orchards during the vetch‐decomposition season, presumably owing to a significant increase in citrus root N and a concomitant decrease in bulk soil total N. Partial least squares path modeling showed that citrus root nutrient contents negatively affected microbial C limitation, while soil nutrients negatively affected microbial relative nutrient limitation in covered orchards. Conversely, both effects were reversed by tillage. Smooth vetch cover mitigated microbial C limitation, and the microbial relative N/P limitation was also influenced by the life state of smooth vetch. Our study highlights the importance of changes in microbial metabolism driven by seasonal nutrient dynamics for understanding soil nutrient cycling and optimizing cover‐legumes management in orchards.

Soil Ecoenzymatic Stoichiometry Reveals Microbial Metabolic Limitations in Apple Orchards with Cover Crop and Organic Fertilizer Incorporation

Understanding the stoichiometry of extracellular enzymes in soil, particularly in relation to nutrient acquisition (e.g., carbon, nitrogen, phosphorus), provides valuable insights into microorganisms’ resource requirements. This study investigates the metabolic constraints of soil microorganisms in response to different growth stages of apple trees under various soil management practices. A 14-year long-term experiment with a split-plot design was conducted, where the main plots received different cover crop treatments (bare vs. cover crop), and subplots were subjected to four fertilizer treatments (CK, M, NPK, MNPK). The significant main and interactive effects of cover crops, fertilizer treatment, and growth period on soil nutrients were observed (p &lt; 0.001). Both cover crop and fertilizer treatments significantly increased the soil organic matter content, with implications for orchard resilience to drought. However, the cover factor alone did not notably influence soil carbon–nitrogen ratios or microbial communities. Microbial carbon limitations were driven by soil water dynamics and microbial biomass, while microbial phosphorus limitations were closely linked to total nitrogen levels. The results underscore the combination of cover crops and MNPK fertilizer-enhanced soil nutrient levels and enzyme activities, mitigating microbial carbon and phosphorus limitations. These findings suggest practical strategies for optimizing fertilization practices to improve soil fertility and address nutrient constraints in orchard ecosystems.

Soil stoichiometric imbalances constrain microbial-driven C and N dynamics in grassland

Grassland restoration leads to excessive soils with carbon (C) and nitrogen (N) contents that are inadequate to fulfill the requirements of microorganisms. The differences in the stoichiometric ratios of these elements could limit the activity of microorganisms, which ultimately affects the microbial C, N use efficiencies (CUE, NUE) and the dynamics of soil C and N. The present study was aimed at quantifying the soil microbial nutrient limitation and exploring the mechanisms underlying microbial-induced C and N dynamics in chrono-sequence of restored grasslands. It was revealed that grassland restoration increased microbial C, N content, microbial C, N uptake, and microbial CUE and NUE, while the threshold elemental ratio (the C:N ratio) decreased, which is mainly due to the synergistic effect of the microbial biomass and enzymatic stoichiometry imbalance after grassland restoration. Finally, we present a framework for the nutrient limitation strategies that stoichiometric imbalances constrain microbial-driven C and N dynamics. These results are the direct evidence of causal relations between stoichiometric ratios, microbial responses, and soil C, N cycling.

Coupling and decoupling of soil carbon and nutrients cycles at different salinity levels in a mangrove wetland: Insights from CUE and enzymatic stoichiometry

Soil carbon (C), nitrogen (N), and phosphorus (P) cycling, in conjunction with microbial metabolism, varies significantly with salinity in coastal areas. However, microbial metabolism limitation on salinity levels has received limited attention. Based on soil microbial carbon use efficiency and enzymatic stoichiometry, microbial nutrient limitation characteristics of soil microbial communities in different salinity levels (4.45 mS·cm−1 - 17.25 mS·cm−1) in a subtropical mangrove wetland were investigated. Compared to low-salinity levels, the activity of soil C-acquiring enzyme activities, enzymatic C:N ratios and enzymatic C:P ratios decreased with medium salinity levels and high salinity levels. Soil microbial metabolism was primarily constrained by C and N at different salinity levels. Boosted regression tree analysis revealed that abiotic factors had the greatest influence on C and N limitation of microbial metabolism at different salinity levels. This study underscores the significance of salinity in microbial metabolic processes and enhances our understanding of how future salinity changes induced by rising sea levels will affect soil carbon and nutrient cycling in coastal wetlands.

Hyperosmotic stress alters enzyme stoichiometry of glucosomes

Hyperosmotic stress alters enzyme stoichiometry of glucosomes

Understanding phosphorus mobilization mechanisms in acidic soil amended with calcium-silicon-magnesium-potassium fertilizer

Calcium-silicon-magnesium-potassium fertilizer (CSMP) is usually used as an amendment to counteract soil acidification caused by historical excessive nitrogen (N) applications. However, the impact of CSMP addition on phosphorus (P) mobilization in acidic soils and the related mechanisms are not fully understood. Specifically, a knowledge gap exists with regards to changes in soil extracellular enzymes that contribute to P release. Such a knowledge gap was investigated by an incubation study with four treatments: i) initial soil (Control), ii) urea (60 mg kg−1) addition (U); iii) CSMP (1%) addition (CSMP) and iv) urea (60 mg kg−1) and CSMP (1%) additions (U + CSMP). Phosphorus mobilization induced by different processes was distinguished by biologically based P extraction. The Langmuir equation, K edge X-ray absorption near-edge structure spectroscopy, and ecoenzyme vector analysis according to the extracellular enzyme activity stoichiometry were deployed to investigate soil P sorption intensity, precipitation species, and microbial-driven turnover of organophosphorus. Results showed that CaCl2 extractable P (or citric acid extractable P) content increased by 63.4% (or 39.2%) in the soil with CSMP addition, compared with the study control. The accelerated mobilization of aluminum (Al)/iron (Fe)-bound P after CSMP addition, indicated by the reduction of the sum of FePO4·2H2O and AlPO4 proportion, contributed to this increase. The decrease of P sorption capacity can also be responsible for it. The CSMP addition increased enzyme extractable P in the soil nearly 7-fold and mitigated the limitations of carbon (C) and P for soil microorganisms (indicated by the enzyme stoichiometry and ecoenzyme vector analysis), suggesting that microbial turnover processes also contribute to P mobilization in amended acidic soil. These findings indicate that the P mobilization in CSMP amended acidic soil not only attributed to both decreasing P sorption capacity and dissolving phosphate precipitation, but also to the increase of the microbial turnover of the organophosphorus pool.

Contradiction with enzymatic stoichiometry theory: Persistent low ratios of β-glucosidase to phosphomonoesterase following 10-year continuous phosphorus fertilization in three subtropical forests

The ratio of β-glucosidase (BG) to phosphomonoesterase (PME) activity (BG:PME) is often used to predict the intensity of microbial phosphorus (P) shortage, with lower BG:PME indicating stronger P shortage (enzymatic stoichiometry theory). Here, we demonstrated that 10-year continuous P fertilization as high as 150 kg P ha−1 yr−1 in the form of NaH2PO4 solution did not elevate the BG:PME up to the level of other terrestrial ecosystems. The BG:PME of primary, secondary, and planted forests were 0.094, 0.067, and 0.089, respectively in P-fertilized plots, which were much lower than global average (0.62 ± 0.04), despite the fact that Bray-extracted P contents were substantially elevated (more than 600 times). Thus, the findings of the current study suggest that BG:PME overestimates P shortage in our P-enriched forests, implying that the enzymatic stoichiometry theory may not be universally applicable.

Role of an arbuscular mycorrhizal fungus in vegetation restoration as indicated by bacterial diversity and microbial metabolic limitation in soil underlying moss biocrusts

Arbuscular mycorrhizal (AM) fungi contribute to the revegetation of degraded ecosystems such as coal mining areas and may indirectly affect the development of biological soil crusts (biocrusts). However, bacterial diversity and microbial nutrient cycling in the soil underlying biocrusts are seldom considered when assessing ecological restoration effects. Bacterial community composition was quantified and compared using 16S rRNA gene sequencing, metabolic limitation of microbes via extracellular enzymatic stoichiometry and their association with carbon use efficiency (CUE) in soil underlying moss biocrusts in vegetated areas inoculated with an AM fungus or uninoculated and in uninoculated unvegetated areas. The AM fungal inoculum significantly influenced bacterial community composition and diversity, extracellular enzyme activities and CUE in soil underlying moss biocrusts mainly by affecting soil nutrients. Correlation analysis revealed that Actinobacteria, Armatimonadetes, Bacteroidetes, Chlorobi, Cyanobacteria, Saccharibacteria, and Verrucomicrobia were positively correlated with soil nutrients carbon (C), nitrogen (N) and phosphorus (P). A vector analysis of extracellular enzyme activity indicates that soil microbial communities underlying moss biocrusts were limited by P but microbial communities in areas inoculated with the AM fungus had the lowest relative P limitation. The alleviation of microbial P limitation increased the microbial CUE and bacterial phylum community and alpha diversity, leading to an increase in subsoil C content underlying moss biocrusts. This indicates that microbial communities in soil underlying moss biocrusts under in restoration areas containing the AM fungus were more stable under environmental stress, and the inclusion of AM fungal inoculation may be recommended as the preferred option for ecosystem restoration in arid and semi-arid coal mining areas.

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 β-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 extracellular enzyme stoichiometry reveals the nutrient limitations in soil microbial metabolism under different carbon input manipulations

Changes in the quality and quantity of litter and root inputs due to climate change and human activities can influence below-ground biogeochemical processes in forest ecosystems. However, it is unclear whether and how much aboveground litter and root inputs affect soil microbial metabolism and nutrient limitation mechanisms. In this study, according to a 4-years field manipulation experiment, litter and root manipulations (control (CK), double litter input (DL), no litter (NL), no root (NR), and no inputs (NI)) were set up to analyze the extracellular enzyme activities and stoichiometric ratios characteristics of 0–10 cm and 10–20 cm soils, explore the metabolic limitations of microorganisms, and clarify the main driving factors restricting nutrient limitation. The results showed that the enzyme activities associated with the C cycling (β-1,4-glucosidase (BG), cellulose disaccharide hydrolase (CBH)) and N cycling (β-1,4-N-acetylglucosaminidase (NAG), leucine aminopeptidase (LAP)) in DL treatment were significantly higher than those in NR treatment. Moreover, enzyme activities related to P cycling are significantly higher in comparison to other treatments. The acid phosphatase (AP), which is related to the P cycle, showed the highest activity under NR treatment. In addition, there was no significant difference in soil microbial metabolic limitation by the different carbon inputs, which did not change the original nutrient limitation pattern. The main drivers of microbial nutrient metabolic limitation included soil physicochemical properties, soil total nutrients, and available nutrients, among which soil SWC and pH presented the greatest influence on microbial C limitation and soil total nutrients showed the greatest influence on microbial N limitation. Changes in soil carbon input altered soil extracellular enzyme activities and their stoichiometric ratios by affecting soil physicochemical properties, total nutrients. This study provides data for the understanding of material cycling in forest ecosystems under environmental change.

Impacts of rice terrace abandonment on soil ecosystem multifunctionality: Insights into reduced microbial diversity and exacerbated nitrogen limitation

Soil ecosystem multifunctionality (EMF) reflects the ability of soils to perform multiple ecological functions, and enzyme stoichiometry indicates microbial involvement in ecosystem nutrient cycling and energy balance. However, it remains unclear how the prevalence of terrace abandonment drives soil EMF and its relationship with microbial nutrient limitations. Soil samples were taken from rice terraces (RT), as well as from dry land (DLT) and forest land (FLT) formed after abandonment, and soil profiles were excavated to a depth of 0–120 cm. It was found that the soil EMF decreased significantly by 144.84% and 142.03%, respectively, in the DLT and FLT compared to RT. Soil EMF was not significantly different between DLT and FLT, and individual soil ecosystem functions were more similar. Microbial phosphorus limitation was observed in this study area, and the abandonment of RT resulted in a shift in microbial limitation from phosphorus to nitrogen, weakening microbial elemental homeostasis. Further analysis revealed that soil microbial diversity was the primary driver of decreased soil EMF, followed by enzyme stoichiometry. Therefore, soil microbial diversity may serve as a biological indicator of soil EMF quality. Overall, the abandonment of RT reduces soil EMF, but it is more beneficial to improve it by alleviating microbial nutrient limitation and fostering microbial diversity. Additionally, this study also highlights the significant impact of enzyme stoichiometry on soil EMF. More importantly, reducing the prevalence of abandoned terraces in mountainous and hilly areas contributes to sustainable soil resource management and cleaner agricultural production.