Microbial communities are crucial for water quality and biogeochemical cycling in freshwaters. Microbes secrete extracellular enzymes to decompose organic matter for their needs of nutrients and scarce elements. Yet, there is a lack of knowledge on microbial metabolic limitations in freshwaters, especially in lake sediments. Here, we examined the carbon, nitrogen, and phosphorus-acquiring extracellular enzyme activities and the bacterial and fungal communities of 30 sediments across Xingkai Lake, the largest freshwater lake in Northeast Asia. We further analyzed the microbial metabolic limitations via extracellular enzyme stoichiometry and explored the direct and indirect effects of abiotic and biotic factors on the limitations. We found that microbial metabolisms were primarily limited by phosphorus in Xingkai Lake. For instance, microbial carbon and phosphorus limitations were closely correlated to abiotic factors like water depth, total dissolved solids, sediment total carbon, and conductivity. The metabolic limitations were also affected by biotic factors, such as showing positive relationships with the alpha and beta diversity of bacteria, and with the beta diversity of fungi. In addition, community compositions of bacteria and fungi were mainly correlated to abiotic factors such as total carbon and dissolved organic carbon, respectively. Collectively, microbial metabolic limitations were affected directly or indirectly by abiotic factors and microbial communities. Our findings indicate that microbial metabolic limitations are not only driven by bacteria and fungi but also by abiotic factors such as water depth and total nitrogen, and thus provide empirical evidence for effective management of freshwater lakes under climate warming and intensified human activities.

Karst rocky desertification refers to the process of land degradation caused by various factors such as climate change and human activities including deforestation and agriculture on a fragile karst substrate. Nutrient limitation is common in karst areas. Moss crust grows widely in karst areas. The microorganisms associated with bryophytes are vital to maintaining ecological functions, including climate regulation and nutrient circulation. The synergistic effect of moss crusts and microorganisms may hold great potential for restoring degraded karst ecosystems. However, our understanding of the responses of microbial communities, especially abundant and rare taxa, to nutrient limitations and acquisition in the presence of moss crusts is limited. Different moss habitats exhibit varying patterns of nutrient availability, which also affect microbial diversity and composition. Therefore, in this study, we investigated three habitats of mosses: autochthonal bryophytes under forest, lithophytic bryophytes under forest and on cliff rock. We measured soil physicochemical properties and enzymatic activities. We conducted high-throughput sequencing and analysis of soil microorganisms. Our finding revealed that autochthonal moss crusts under forest had higher nutrient availability and a higher proportion of copiotrophic microbial communities compared to lithophytic moss crusts under forest or on cliff rock. However, enzyme activities were lower in autochthonal moss crusts under forest. Additionally, rare taxa exhibited distinct structures in all three habitats. Analysis of co-occurrence network showed that rare taxa had a relatively high proportion in the main modules. Furthermore, we found that both abundant and rare taxa were primarily assembled by stochastic processes. Soil properties significantly affected the community assembly of the rare taxa, indirectly affecting microbial diversity and complexity and finally nutrient acquisition. These findings highlight the importance of rare taxa under moss crusts for nutrient acquisition. Addressing this knowledge gap is essential for guiding ongoing ecological restoration projects in karst rocky desertification regions.

Aboveground vegetation restoration shapes the soil microbial community structure and affects microbial resource acquisition. However, the changes in soil microbial resource limitation in subsoil during vegetation restoration are still unclear. In this study, the microbial community structure and resource limitation in an alpine meadow soil profile that had undergone natural restoration for short-term (4-year) and long-term (10-year) restoration in response to vegetation restoration were explored through high-throughput sequencing analysis and extracellular enzyme stoichiometry (EES). There was no significant difference in microbial composition and α diversity between short- and long-term restoration soils. Soil microorganisms in this alpine meadow were mainly limited by phosphorus. Carbon limitation of soil microorganisms was significantly decreased in each layer (0-15, 15-30, 30-45, 45-60, and 60-80 cm corresponding to L1, L2, L3, L4, and L5, respectively) of long-term restoration soils when compared to that of the short-term restoration soil layers, while phosphorus limitation of microorganisms in subsoil (60-80 cm) was significantly increased by 17.38%. Soil nutrients, pH, moisture content, and microbial composition are the main drivers of microbial resource limitation in restoration, and their effects on microbial resource limitation were different in short- and long-term restoration. Meanwhile, key microbial taxa have a significant impact on microbial resource limitation, especially in short-term restoration soils. This study suggested that vegetation restoration significantly affected soil microbial resource limitation, and could alleviate microbial resource limitations by adding nutrients, thus accelerating the process of vegetation restoration in alpine ecosystems.

Soil microbial carbon use efficiency (CUE) is a crucial parameter that can be used to evaluate the partitioning of soil carbon (C) between microbial growth and respiration. However, general patterns of microbial CUE among terrestrial ecosystems (e.g., farmland, grassland, and forest) remain controversial. To address this knowledge gap, data from 41 study sites (n = 197 soil samples) including 58 farmlands, 95 forests, and 44 grasslands were collected and analyzed to estimate microbial CUEs using a biogeochemical equilibrium model. We also evaluated the metabolic limitations of microbial growth using an enzyme vector model and the drivers of CUE across different ecosystems. The CUEs obtained from soils of farmland, forest, and grassland ecosystems were significantly different with means of 0.39, 0.33, and 0.42, respectively, illustrating that grassland soils exhibited higher microbial C sequestration potentials (p < .05). Microbial metabolic limitations were also distinct in these ecosystems, and carbon limitation was dominant exhibiting strong negative effects on CUE. Exoenzyme stoichiometry played a greater role in impacting CUE values than soil elemental stoichiometry within each ecosystem. Specifically, soil exoenzymatic ratios of C:phosphorus (P) acquisition activities (EEAC:P ) and the exoenzymatic ratio of C:nitrogen (N) acquisition activities (EEAC:N ) imparted strong negative effects on soil microbial CUE in grassland and forest ecosystems, respectively. But in farmland soils, EEAC:P exhibited greater positive effects, showing that resource constraints could regulate microbial resource allocation with discriminating patterns across terrestrial ecosystems. Furthermore, mean annual temperature (MAT) rather than mean annual precipitation (MAP) was a critical climate factor affecting CUE, and soil pH as a major factor remained positive to drive the changes in microbial CUE within ecosystems. This research illustrates a conceptual framework of microbial CUEs in terrestrial ecosystems and provides the theoretical evidence to improve soil microbial C sequestration capacity in response to global change.

Long-term input of agricultural chemicals such as pesticides into the soil can increase soil pollution, thereby affecting the productivity and quality of black soil. Triazine herbicide atrazine has been shown to have long-lasting residual effects in black soil. The atrazine residues affected soil biochemical properties, further leading to microbial metabolism restriction. It is necessary to explore the strategies to mitigate the limitations on microbial metabolism in atrazine-contaminated soils. Here, we evaluated the effect of the atrazine on microbial nutrient acquisition strategies as indicated by extracellular enzyme stoichiometry (EES) in four black soils. Atrazine degradation in soil followed the first-order kinetics model across various concentrations ranging from 10 to 100 mg kg-1. We found that the atrazine was negatively correlated with the EES for C-, N-, and P-acquisition. Vector lengths and angles decreased and increased significantly with an increase of atrazine concentration in tested black soils except for Lishu soils. Moreover, the vector angles were >45° for tested four black soils, indicating that atrazine residue had the greatest P-limitation on soil microorganisms. Interestingly, microbial C- and P-limitations with different atrazine concentrations showed a strong linear relationship, especially in Qiqihar and Nongan soils. Atrazine treatment significantly negatively affected microbial metabolic limitation. Soil properties and EES interaction explained up to 88.2% for microbial C-/P-limitation. In conclusion, this study confirms the EES as a useful method in evaluating the effects of pesticides on microbial metabolic limitations.

The application of organic fertilizer can improve soil fertility and maintain soil biodiversity. Soil enzyme activity can reflect the relationship between microbial nutrient demand and environmental nutrient availability. The experiment was established with a split-plot design, which included two main plots and two subplots. The main plots were 3 nitrogen levels (0, 150, 300 kg N ha-1). The two subplots were chemical fertilizer alone and combination of manure and inorganic application; the soil enzyme activity and chemical properties of each treatment were measured and analyzed. In the study, ecological enzyme stoichiometry was used to study the limitation of microbial resources in dryland wheat system. The results showed that the combined application of manure and chemical fertilizers did not significantly increase the activities of soil C, N and P cycling-related enzymes but decreased the activities of soil L-leucine aminopeptidase (LAP). Long-term application of organic fertilizer and mineral fertilizer significantly increased the accumulation of soil organic carbon (SOC) and nitrogen (TN) and increased soil microbial biomass (MBC, MBN). Organic fertilizer treatment significantly increased soil available phosphorus (AP) and soil NO3--N contents, and decreased SOC/AP and TN/AP, but had no significant changes under different nitrogen application levels (N0, N1, N2). GHG emissions were increaseed with the amount of nitrogen applied, the addition of manure did not significantly increase the CO2 and N2O emissions, and soil organic matter mineralization and gas emission fluxes decreased at ripen stage. The C-acquiring enzyme was negatively correlated with N-acquiring enzyme but positively correlated with P-acquiring enzyme. The microbial limiting effect of C and P on microbial metabolism becomes increasingly intense as the reproductive period progresses. Redundancy analysis of soil enzyme activities and chemical properties showed that soil TN and MBN could better explain the variation characteristics of soil enzyme activities. Therefore, the study of soil extracellular enzyme stoichiometry and microbial nutrient restriction can give us a more comprehensive understanding of the soil environment. There are more implications can be given under different nitrogen management modes and different growth stages. The results also provided an effective theoretical basis for regulating the changes of soil microbial environment.

The resource acquisition strategy of soil microorganisms can be reflected by soil extracellular enzyme activity (EEA). However, there are few reports on the application of extracellular enzyme stoichiometry (EES) method to study the difference in microbial metabolic nutrient limitation between rhizosphere and bulk soil. Here, we choose the rhizosphere and bulk soils of Pinus sylvestris var. mongolica (P. sylvestris) plantations with five stand ages in the Mu Us sandy land, and analyzed the variation and differences of microbial metabolic limitation between rhizosphere and bulk soils with stand age by EES method. The results showed that the microbial metabolic C-limitation in the rhizosphere and bulk soil gradually increased with stand age. Almost all the vector angles were less than 45°, which indicated that the soil microbial metabolism was relatively limited by N rather than P. Furthermore, the microbial C- and N-limitation in rhizosphere soils were generally stronger than bulk soils. Soil physical properties (59.73%) explained most of the variations in soil EES based on the variation-partitioning analysis, followed by total nutrients (43.00%). The partial least squares path model suggested that the main driving factor for the variation of soil microbial metabolic C-limitation in the rhizosphere and bulk soils was physical properties, while the microbial N-limitation was for total nutrients. In general, the study emphasized the application of EES methods to assess the dynamic equilibrium between soil microbial resource acquisition and nutrient availability in desert ecosystems. These insights provide guidance for formulating afforestation strategies, such as nutrient management of sandy plantations.