Leucine Aminopeptidase Research Articles

Impacts of shifting from single-species pine forests to distinct agroforestry models on soil fertility, exchangeable cations, and microbial functions

The transition from monoculture to mixed-species agroforestry systems affects soil organic matter and microbial activity. However, the specific dynamics of these changes, particularly within medicinal plant-based agroforestry, remain underexplored. This study investigates the impact of monoculture Pine (Pinus massoniana) forests and four agroforestry models: (M1) Pinus massoniana and Alpina oxyphylla, (M2) Pinus massoniana and Ficus simplicissima, (M3) Pinus massoniana and Amomum villosum, and (M4) Pinus massoniana and Curcuma longa on soil properties and microbial activity in rhizosphere and non-rhizosphere environments. Our results showed significantly higher pH (4.80) and total nitrogen (N) content (1.77 g kg−1) in the rhizosphere of model (M4) compared to (CK). Total organic carbon (TOC) and carbon fractions (POC, DOC, and MBC) also differed significantly across monoculture and agroforestry models, with highest TOC concentrations (31.70 g kg⁻1) in rhizosphere of CK. Exchangeable cations, including Ca2⁺, and Mg2⁺ were significantly higher in the rhizosphere of agroforestry models compared to CK, particularly in M4, where Ca2⁺ was recorded at 12.03 cmol kg−1 with the highest percent base saturation (PBS) at 90.17%. Enzymes leucine aminopeptidase and polyphenol oxidase varied significantly, with higher activity in the rhizosphere of agroforestry models and greater activity in non-rhizosphere of monoculture. Soil microbial respiration (MRes) revealed substantial differences, with an average 17% decrease in rhizosphere soil for models M2 and M4 and a 20.83% reduction in non-rhizosphere soil for model M1 compared to CK. Generalized Linear Model (GLM) demonstrated a significant positive correlation between TOC and MRes (R2 = 0.885, p < 0.01), indicating that higher TOC levels are linked with increased MRes. In conclusion, model M4 most effectively enhanced soil fertility and nutrient availability followed by the other agroforestry models tested. This suggests that integrating medicinal plants into agroforestry systems is a viable strategy for improving ecosystem functioning compared to monoculture practices.

Is enzymatic stoichiometry a reliable indicator of microbial limitations in carbon, nitrogen, or phosphorus?

The enzymatic stoichiometry approach is based on the assumption that the metabolic and stoichiometric requirements of soil microorganisms are reflected in the production of enzymes targeting specific nutrient resources, making enzyme activities a useful tool for assessing microbial nutrient limitations. In this approach, β-1,4-glucosidase (BG), β-1,4-N-acetylglucosaminidase (NAG) (occasionally combined with leucine aminopeptidase, LAP), and acid/alkaline phosphatase (AP) are used as proxies for the broader suite of enzymes responsible for catalyzing complex substrates containing carbon (C), nitrogen (N), and phosphorus (P), respectively, because these enzymes catalyze the terminal reactions that release products containing C, N, and P. The fundamental premise of the approach can thus be reformulated as follows: BG:NAG and BG:AP ratios reflect the relative limitations of C versus N and C versus P, respectively, with higher ratios indicating stronger C limitation. Previous meta-analyses have suggested that the enzymatic stoichiometry approach is unreliable, as N and P fertilization often leads to reduction in BG:NAG and BG:AP, respectively, contradicting the predictions of the approach. However, some researchers question the validity of assessing enzymatic stoichiometry after the artificial addition of nutrients via fertilizers. For providing more convincing evidence, this study conducted a meta-analysis to assess the impact of incorporating various C sources, largely representative of those found in natural ecosystems—glucose, cellulose, and plant residues—on the BG:NAG or BG:AP ratios. The results demonstrated that a considerable number of data points exhibited elevated values (i.e., C addition accelerated C limitations), undoubtedly contradicting the fundamental premise of the approach. These findings highlight the unreliability of enzymatic stoichiometry as an indicator of microbial nutrient limitation.

Networks of ion-pairs are responsible for the large differences in the thermal stability of two structurally similar aminopeptidases.

Aminopeptidases are an important class of enzymes for protein metabolism. Leucyl aminopeptidase (PepL) preferably removes leucine from the N-terminus of small peptides. PepL of Lacticaseibacillus casei was observed to be thermally unstable, while a structurally similar aminopeptidase T (AmpT) of Thermus thermophilus is highly stable. To understand the molecular interaction responsible for the large difference in their stability, molecular dynamics simulations were carried out to study the thermal stability of PepL and AmpT at 300 K to 450 K temperature range over 100 ns. PepL sampled a larger conformational space with a rugged free-energy landscape, while AmpT navigated a smoother energy landscape to reach the global minimum. The RMSD, RMSF, radius of gyration and principal component analysis suggested large movements in PepL than in AmpT with an increase in temperature. Analysis of residue-interaction network revealed AmpT possessing a greater number of low, medium and high energy contacts in comparison to PepL. AmpT showed a higher abundance of ion-pair clusters and ionic residues per cluster compared to PepL. Moreover, AmpT retained a greater number of high-energy contacts at elevated temperatures. These findings showed that the inherently lower stability of PepL originates from a comparatively smaller number of contacts and can be pivotal in engineering PepL for higher stability.

The Responses of Soil Extracellular Enzyme Activities and Microbial Nutrients to the Interaction between Nitrogen and Phosphorus Additions and Apoplastic Litter in Broad-Leaved Korean Pine Forests in Northeast China

The impact of nitrogen and phosphorus deposition alternations, as well as apoplastic litter quality and quantity, on soil nutrient cycling and soil carbon pool processes in forest ecosystems is of considerable importance. Soil ecological enzyme chemistry is a powerful tool for elucidating the nutrient limitations of microbial growth and metabolic processes. In order to explore the responding mechanisms of soil ecological enzyme chemistry to the simultaneous changes in apoplast input and nitrogen and phosphorus deposition in temperate coniferous and broad-leaved mixed forests, an outdoor simulating experiment was conducted. The results demonstrate that the treatments involving apoplastic material and nitrogen and phosphorus additions had significantly impacted soil nutrient levels across different forest types. Apoplastic treatments and N-P additions had a significant effect on the soil total organic carbon (TOC), dissolved organic carbon (DOC), soil total soluble nitrogen (TSN), soil available phosphorus (SAP), soil total nitrogen (TN), soil total phosphorus (TP), and microbial biomass carbon (MBC). However, the effects on soil microbial biomass (MBN) and microbial biomass phosphorus (MBP) were insignificant. The apomictic treatments with N and P addition did not result in a statistically significant change in soil C-hydrolase activities (β-1,4-glucosidase BG, β-1,4-xylosidase BX, cellobiohydrolase CBH, phenol oxidase POX, and peroxidase PER), N-hydrolase activities (β-1,4-N-acetylglucosaminidase NAG and L-leucine aminopeptidase LAP), or P-hydrolase activities (Acid phosphatase AP). Although the apomictic treatments did not yield a significant overall impact on carbon hydrolase activity, they influenced the activity of specific enzymes, such as CBH, LAP, and PER, to varying degrees. The effects on BG, BX, CBH, AP, and C-hydrolase activities were significant for different stand types. The impact of apomictic treatments and N-P additions on soil nitrogen hydrolase activities was inconsequential with a minimal interactive effect. The highest correlation between PER, LAP, and N-hydrolase activities was observed in conjunction with elevated levels of nitrogen and phosphorus addition (N3L0, original litter treatment, and high amounts of N and P addition). These findings may provide a theoretical foundation for the management of ecosystem function in broad-leaved Korean pine forests.

Biocompatible Flash Chemiluminescent Assay Enabled by Sterically Hindered Spiro-Strained-Oxetanyl-1,2-Dioxetane.

Chemiluminescence is the emission of light that occurs as a result of a chemical reaction. Depending on the rate of chemiexcitation, light emission can occur as a long-lasting,glow-type reaction or a rapid, highly intense flash-type reaction. Assays using a flash-type mode of action provide enhanced detection sensitivity compared to those using a glow-type mode. Recently, our group discovered that applying spiro-strain to 1,2-dioxetanes significantly increases their chemiexcitation rate. However, further examination of the structure-activity relationships revealed that the spiro-strain severely compromises the chemical stability of the 1,2-dioxetanes. We hypothesized that a combination of spiro-strain, steric hindrance, and an electron-withdrawing effect, will result in a chemically stable spiro-strained dioxetane with an accelerated chemiexcitation rate. Indeed, spiro-fused tetramethyl-oxetanyl exhibited a 128-fold faster chemiexcitation rate compared to adamantyl while maintaining similar chemical stability, with a half-life of over 400 hours in PBS 7.4 buffer at room temperature. Turn-on probes composed of tetramethyl-oxetanyl spiro-dioxetane exhibited significantly improved chemical stability in bacterial and mammalian cell media compared to previously developed dioxetane probes fused to a cyclobutyl unit. The superior chemical stability enables a tetramethyl-oxetanyl dioxetane probe to detect β-galactosidase activity with enhanced sensitivity in E. coliassays and leucine aminopeptidase activity in cancer cells.

Changes in Soil Bacterial and Fungal Community Composition and Functional Groups During the Artificial Restoration of Degraded Grassland of "Black-Soil Mountain".

About 35% of grassland in Sanjiangyuan area of China has degenerated into black-soil mountain. Artificial grassland is considered to be an effective measure to alleviate the severely degraded grassland in the alpine region of the three rivers and has been widely used. However, the pattern, potential function, and changes of carbon and nitrogen contents of soil microorganisms in degraded grassland in Heimushan by planting artificial grassland are still unclear. In this study, mixed-sown artificial alpine grassland (AG) was the focus of our study, whereas degraded black-soil mountain grassland (BG) and natural alpine grassland (NG) served as controls. Illumina 16S and ITS gene sequence analyses were used to analyze the community structure of the soil bacteria and fungi. The functional groups of NG, AG, and BG were predicted using the FAPROTAX and FUNGuild databases. In addition, the levels of soil carbon, nitrogen, and soil enzyme activities were evaluated. The results indicated a significant increase in the aboveground biomass of BG due to the planting artificial grassland. Moreover, the contents of total carbon (TC), total nitrogen (TN), ammonium nitrogen ( ), microbial biomass carbon (MBC), microbial biomass nitrogen (MBN), and leucine aminopeptidase (LAP) increased in the soil. Planting artificial grasslands changed the composition of bacterial and fungal communities. Among these, the bacterial community was more sensitive to planting artificial grasslands. The relative abundance of bacterial functional groups involved in carbon and nitrogen cycling changed significantly, suggesting that bacteria may play a role in regulating nutrient cycling during artificial grassland planting. Soil TC, TN, LAP, and affected the microbial community structure related to carbon and nitrogen. and β-1,4-glucosidase were carbon and nitrogen factors, respectively, that affected functional changes in fungi. These results indicate that planting artificial grasslands can effectively enhance the productivity of degraded black-soil mountain and regulate soil microbial communities and soil physical and chemical properties.

Release of intracellular enzymes increases the total extracellular activities of carbohydrate processing enzymes in marine environment.

Microbial extracellular enzymatic activities (EEAs) produced by microbes to degrade biopolymers are the 'gatekeeper' of carbon cycle in the marine ecosystem. It is usually assumed that these extracellular enzymes are actively secreted by microbes. But biopolymers degrading enzymes also exist in the intracellular space. Cell lysis will passively release these enzymes into the environments and contribute to the total EEAs. However, to what extent the cell lysis can contribute to the total EEAs are still unclear. Here, using extreme cell lysis method, we evaluated the maximum contribution of cell lysis to total EEAs in culturable marine bacteria and coastal seawater. For carbohydrate processing enzymes (β-glucosidase, alginate lyase and chitinase), the release of intracellular enzymes could contribute positively (up to 56.1% increase for β-glucosidase in seawater) to the total EEAs. For protease and leucine aminopeptidase, the cell lysis did not increase and even decreased the total EEAs. For alkaline phosphatase, the intracellular enzymes generally had no contribution to the total EEAs. These results showed that passively released intracellular enzymes could substantially increase the total extracellular activities of carbohydrate processing enzymes, which should be considered in building the link between the EEAs and organic carbon cycle in the ocean.

Study on Single Nucleotide Polymorphism of LAP3 Gene and Its Correlation with Dairy Quality Traits of Gannan Yak.

This study explored the polymorphism of the leucine aminopeptidase (LAP3) gene and its relationship with milk quality characteristics in Gannan yak. A cohort of 162 Gannan yak was genotyped utilizing the Illumina Yak cGPS 7K BeadChip, and the identified single nucleotide polymorphisms (SNPs) were evaluated for their association with milk protein, casein, lactose, and fat concentrations. The results showed that four SNPs (g.4494G > A, g.5919A > G, g.8033G > C, and g.15,615A > G) in the LAP3 gene exhibited polymorphism with information content values of 0.267, 0.267, 0.293, and 0.114, respectively. All four SNPs were in Hardy-Weinberg equilibrium (p > 0.05). The g.4494G > A and g.5919A > G SNPs were significantly associated with protein content (p < 0.05), with homozygous genotypes showing significantly higher protein content than heterozygous genotypes (p < 0.05). The g.8033G > C SNP was significantly associated with casein content, protein content, non-fat solids, and acidity (p < 0.05), with the CC genotype having significantly higher casein, protein, and non-fat solids content than the GG and GC genotypes (p < 0.05). The g.15,615A > G SNP was significantly associated with average fat globule diameter (p < 0.05). In general, the mutations within the LAP3 gene demonstrated a positive impact on milk quality traits in Gannan yak, with mutated genotypes correlating with enhanced milk quality. These results indicate that the LAP3 gene could be a significant or candidate gene affecting milk quality traits in Gannan yak and offer potential genetic markers for molecular breeding programs in this species.

Macrogenome-based study on the mechanism of Bacillus velezensis SX13 in regulating rhizophere environment and cucumber growth under different cultivation environments

Microbial activities are the dynamic core of nutrient cycling in organic substrates, and the exploitation of plant growth-promoting rhizobacteria strains contributes to sustainable agricultural development. This study aimed to investigate the effect and mechanism of Bacillus velezensis SX13 in nutrient cycling and plant promotion under different substrate supply conditions. The effects of reduced substrate amount (sCK) and inoculation of SX13 strain under both substrate supply conditions (Bv and sBv) on rhizosphere microenvironment and plant growth were investigated using conventional substrate amount (CK) as a control. Results showed no significant difference in the α-diversity indexes (Chao1 and Shannon) of the rhizospheric microbial community among the four treatments. However, nonmetric multidimensional scaling analysis and principal coordinate analysis revealed that compared with CK treatment, the inoculation of SX13 strain and reduced substrate supply reshaped the β-diversity structure of microbial communities. Furthermore, inoculation with B. velezensis SX13 under both substrate supply conditions increased the abundance of Proteobacteria (1.64%–2.46 %), Acidobacteria (14.09%–43.07 %), and Firmicutes (179.29%–861.29 %). The results of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the metabolic pathway with the highest abundance of enriched genes was also the pathway with the most enriched differential genes caused by reducing substrate supply or inoculation of B. velezensis SX13. The rhizosphere inoculation of B. velezensis SX13 significantly up-regulated the top genes related to carbohydrate esterases, carbohydrate binding modules, glycoside hydrolases, glycoside transferases, and polysaccharide lyases. As a result, the activities of carbon and nitrogen cycle-related enzymes such as cellobiohydrolase, β-glucosidase, urease, l-leucine amino peptidase, and β-1 4-N-acetylglucosaminidase were increased, which in turn accelerated nutrient cycling. B. velezensis SX13 and its mediated improvement of the rhizospheric microenvironment resulted in the up-regulation of root CsNRT family genes (such as CsNRT1.1, CsNRT1.4a, CsNRT1.4b, CsNRT1.5a, CsNRT1.5b, CsNRT1.5c, and CsNRT1.8), which accelerated nitrogen uptake, accumulation, and utilization efficiency and ultimately improved the yield and quality of cucumber. The effect of SX13 strain was more stable and efficient under conventional substrate supply conditions than under reduced substrate supply conditions.

Biomarkers of oxidative stress, biochemical changes, and the activity of lysosomal enzymes in the livers of rainbow trout (Oncorhynchus mykiss Walbaum) vaccinated against yersiniosis before a Yersinia ruckeri challenge

Abstract Introduction This study aimed to evaluate biomarkers of oxidative stress (2-thiobarbituric acid reactive substances, aldehyde and ketone derivatives of oxidatively modified proteins and total antioxidant capacity), the activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase), that of lysosomal enzymes (alanyl aminopeptidase, leucyl aminopeptidase, β-N-acetylglucosaminidase and acid phosphatase) and changes in biochemical parameters (alanine aminotransferase, aspartate aminotransferase, de Ritis ratio, lactate dehydrogenase activity, lactate and pyruvate levels and their ratio) in the liver tissue of fish that were vaccinated against enteric redmouth disease and challenged with its causative agent, the bacterium Yersinia ruckeri. Material and Methods The vaccine was administered orally to trout, some of which were challenged with Y. ruckeri 61 days later. For comparison, unvaccinated and unchallenged trout and unvaccinated and challenged trout were also evaluated. Results In the unvaccinated fish, infection with Y. ruckeri disrupted the pro-oxidant/antioxidant balance, led to a significant increase in lipid peroxidation and oxidative modification of proteins, decreased total antioxidant capacity and significantly increased the activity of lysosomal enzymes. In vaccinated fish, the Y. ruckeri challenge increased the activity of glutathione-related enzymes and decreased lipid peroxidation, anaerobic metabolism and the activity of lysosomal enzymes in fish livers relative to the unvaccinated and challenged group. In contrast, these parameters increased after the Y. ruckeri challenge in unvaccinated trout relative to those in the untreated group. Conclusion Vaccination exerted a protective effect during the Y. ruckeri challenge and had no adverse effect on fish livers.

Natural restoration has more stable soil leucine aminopeptidase kinetic characteristics than plantations

Litter and roots strongly influence the kinetics of key enzymes involved in nitrogen (N) cycling; however, the influence and mechanisms responsible for such influence remain unclear. Based on eight and nine years of detritus input and removal experiments, the effects of litter and roots on the kinetics of soil leucine aminopeptidase which can better reflect organic N transformation than activities were studied in plots under different forest restoration types. The results showed that: (1) Double litter treatment didn’t significantly influence the leucine aminopeptidase activity at ten substrate concentrations varied from 0 to 400 μ mol·L−1 in the three forest restoration types, or the Vmax of leucine aminopeptidase activity with the range of 0–12.9 nmol·g−1· h−1. (2) No litter, no roots, and no inputs treatments reduced leucine aminopeptidase activity and its kinetic parameters. The reduction effect was more obvious as the removal time increased. (3) The higher the substrate concentration, the closer positive relationship between total nitrogen content and leucine aminopeptidase activity. Total N had positive effects on Vmax and Vmax/Km. The lower the substrate concentration, the closer positive relationship between leucine aminopeptidase activity and soil ammonium N content. (4) Moisture content accounted for the highest variation of Vmax, followed by total N, total C, pH and nitrate N, explaining 50.2 % variation in Vmax. Total N accounted for the highest variation of Vmax/Km, followed by total C, moisture content, Vmax, nitrate N, and pH, explaining 46.8 % variation. From the perspective of leucine aminopeptidase kinetics, soil organic N transformation in naturally restored secondary forests was the most stable, followed by that in native oriental oak plantations, and then introduced black pine plantations.

Clinical fluorescence-guide surgery: Tracking leucine aminopeptidase for assisting head and neck squamous cell carcinoma surgery via a near-infrared fluorescent probe

The complete excision of tumors is critical for surgical treatment and the most used for intraoperative margin control method is frozen section analysis (FSA). However, due to its subjective and time-intensive property, there is still a lack of a rapid and effective means to clearly delineate the boundaries between normal and tumor tissues. In this work, we first demonstrated that leucine aminopeptidase (LAP) is highly expressed in clinical head and neck squamous cell carcinoma (HNSCC) and could be used as a biomarker of HNSCC. Based on this, we reported a LAP-activated fluorescent probe, SiR-LAP, and showed its effectiveness in identifying HNSCC. Importantly, we also demonstrated that the fluorescence imaging results of the probe in human tumor tissues are highly consistent with pathological staining results, highlighting its potential as an adjunct to surgical resection. In addition, intraoperative pathology section detection by fluorescent probe technology took half the time required for H&E staining. This research is poised to advance the application of fluorescent probes in tumor diagnosis and surgical excision assistance.

Composted maize straw under fungi inoculation reduces soil N2O emissions and mitigates the microbial N limitation in a wheat upland

Enhancement of microbial assimilation of inorganic nitrogen (N) by straw addition is believed to be an effective pathway to improve farmland N cycling. However, the effectiveness of differently pretreated straws on soil N2O emissions and soil N-acquiring enzyme activities remains unclear. In this study, a pot experiment with four treatments (I, no addition, CK; II, respective addition of maize straw, S; III, composted maize straw under no fungi inoculation, SC; and IV, composted maize straw under fungi inoculation, SCPA) at the same quantity of carbon (C) input was conducted under the same amount of inorganic N fertilization. Results showed that the seasonal cumulative N2O emissions following the SCPA treatment were the lowest at 4.03 kg N ha−1, representing a significant reduction of 19 % compared with the CK treatment. The S and SC treatments had no significant effects on N2O emissions. The decrease of soil N2O emissions following the SCPA treatment was mainly attributed to the increase of microbial N assimilation and the increased abundance of functional genes related to N2O reductase. The SCPA treatment significantly decreased soil alkaline phosphatase (ALP) activity and increased leucine aminopeptidase (LAP) activity at the basal fertilization, while increased soil ALP and LAP activity, decreased soil N-Acetyl-β-D-Glucosidase (NAG) activity at harvest. Compared with the CK treatment, the S, SC, and SCPA treatment significantly increased soil β-glucosidase (β-GC) activity at harvest. The decrease in the (NAG+LAP)/ALP ratio following the SCPA treatment indicated that the composted maize straw under fungi inoculation alleviated microbial N limitation at harvest. Moreover, PICRUSt analysis also suggested that the SCPA treatment increased the abundance of bacterial genes associated with N assimilation and N2O reduction, whereas the S and SC treatment did not significantly affect the abundance of N2O reduction genes compared with the CK treatment. Our results suggest that the composted maize straw under fungi inoculation would reduce the risk of N2O emissions and effectively mitigate the microbial N limitation in dryland wheat system.

Serum Leucine Aminopeptidase Activity Patterns Across Various Disease States: Potential Implications for Bleeding and Thrombosis Risk.

Disruptions in the pathways for activating and deactivating proteases in the bloodstream can lead to thrombosis and bleeding issues. Leucine aminopeptidases (LAPs), which are exopeptidases essential for regulating protein and peptide activities, are recognized as clinical biomarkers for liver diseases. However, the relationship between serum LAP activity and the risks of bleeding or thrombosis, as well as the identification of the specific tissues or organs that control LAP levels, is not well understood. We performed a retrospective study to evaluate serum LAP activities in 149,360 patients with 47 different diseases and 9,449 healthy individuals. The analysis was conducted using SPSS V2.6, RStudio V.1.3.1073, and libraries in Python 3.8. Our research revealed that 21 of the 47 diseases studied showed increased median serum LAP activities, while 26 diseases were associated with significantly lower activities, especially those related to thrombosis. Furthermore, most diseases were found to have an increased risk of bleeding and thrombosis, indicated by higher Q25 and lower Q75 LAP activities compared to the control group. Receiver operating characteristic curve analysis confirmed the effectiveness of LAP activities as biomarkers for specific conditions like hepatic encephalopathy, liver cancer, pancreatitis, and pancreatic cancer. Diseases were categorized into clusters with similar bleeding or thrombotic tendencies through principal component analysis. This study highlighted regulatory influence of the liver and pancreas on LAP levels. The established link between serum LAP concentrations and the risk of bleeding or thrombosis paved the way for the development of diagnostic and preventative approaches for various medical conditions.

Litter application increases soil multinutrient cycling in alpine meadow ecosystems on the Tibetan Plateau

In soil, multiple nutrients are cycled simultaneously (multinutrient cycling), rather than a single measurable process. While the impact of litter decomposition on individual soil nutrient cycling in alpine meadow ecosystems on the Tibetan Plateau is well-documented, its effects on soil multinutrient cycling remain unclear. We deployed a three-year litter application experiment in an alpine meadow on the Tibetan Plateau to examine the responses of soil microclimate, extracellular enzyme activities, and bacterial and fungal communities to litter applications, as well as the correlations between these vital factors with soil multinutrient cycling induced by litter application. We showed that litter application raised the soil multinutrient cycling index, temperature, and moisture content by an average of 190 %, 1.2 °C, and 12 %, respectively. Litter application increased the activities of soil extracellular enzyme β-1,4-glucosidase, β-1,4-xylosidase, β-D-cellobiosidase, L-leucine aminopeptidase, acid phosphatase, and phenol oxidase. Moreover, litter application increased the richness and diversity of both soil bacterial and fungal communities, and altered their community structure, but with larger effects on bacterial communities (R2 = 0.43 for bacterial community and R2 = 0.19 for fungal community). This indicates that bacterial communities are more responsive to litter application than fungal communities in alpine meadow soils on the Tibetan plateau. Partial least-square path modeling indicated that soil bacterial and fungal communities and extracellular enzyme activities were significantly positively correlated with soil multinutrient cycling after litter application. The relative abundances of bacterial phyla of Proteobacteria, Actinobacteria, and Verrucomicrobiota, and fungal phyla of Basidiomycota were positively related to most of the critical soil nutrients, indicating that those microbial taxa are the main drivers of soil multinutrient cycling. Overall, this study provides explicit evidence that litter application accelerates Tibetan meadow soil multinutrient cycling, which contributes to an enhanced understanding of the role of litter in sustaining the functions and services of alpine meadow ecosystems on the Tibetan Plateau.

Phosphorus addition stimulates overall carbon acquisition enzymes but suppresses overall phosphorus acquisition enzymes: A global meta-analysis

Of the total land worldwide, 43 % is phosphorus (P)-constrained, which is significantly greater than the proportion of nitrogen-constrained land (18 %). However, there are some uncertainties regarding the responses of enzyme activities to P addition on a global scale. We collected data from 99 publications and performed a meta-analysis to assess the responses of 11 extracellular enzymes, including carbon, nitrogen, and phosphorus acquisition enzymes, and oxidases. P addition significantly increased the activities of β-1, 4-glucosidase, sucrase, cellobiohydrolase, and alkaline phosphatase, while lowering the activities of acid phosphatase and phosphodiesterase. The responses of enzyme activities to P addition differed considerably across the three ecosystems (forest, grassland, and cropland). P addition significantly decreased the activities of acid phosphatase, phosphodiesterase, and P-acquisition enzymes in forests, but increased the activities of leucine aminopeptidase, carbon acquisition enzymes, and nitrogen acquisition enzymes in grasslands. Sucrase, β-1,4-glucosidase, N-acetyl-β-glucosaminidase, alkaline phosphatase, and carbon acquisition enzymes were significantly stimulated in croplands. In addition, all P addition rates significantly increased the activity of alkaline phosphatase, a P-acquisition enzyme, in croplands. Ca(H2PO4)2 had a greater effect on enzyme activity than other fertilizer types, possibly because Ca2+ can improve the soil structure and enhance microbial metabolic activities. Furthermore, structural equation models revealed that mean annual precipitation and ecosystem type had significant impacts on the response of P-acquisition enzymes, but no factor could directly influence the response of carbon-acquisition enzymes. Our investigation of enzyme activities in response to P addition complements the biogeochemical model and helps forecast soil nutrient dynamics. We propose that rather than focusing on a single factor, P fertilizer addition programs should take into account the effects of mean annual precipitation, ecosystem type, and nutrient interactions, which could also reduce P pollution.

Effects of leguminous green manure-crop rotation on soil enzyme activities and stoichiometry

Abstract As a crucial strategy for sustainable agricultural production, green manure-crop rotation can regulate soil nutrient cycling and decrease the reliance on nitrogen fertilizers. However, we still lack a comprehensive understanding of the changes in soil eco-enzyme activities, microbial metabolism, and nutrient limitations caused by leguminous green manure crop rotation. Here, we conducted field experiments of leguminous green manure-crop rotation across China to analyze soil extracellular enzyme activities, specifically β-glucosidase (BG), N-acetyl-β-D-glucosaminidase (NAG), leucine aminopeptidase (LAP), and acid phosphatase (AP). The study revealed that long-term green manure-crop rotation increased carbon and nitrogen accumulation in farmland, with a significant average increase of 20.1% and 36.4% in BG, AP enzyme activities in topsoil, while showing a decrease in ln(NAG+LAP):ln(AP) ratios. The ratios of ln(BG):ln(NAG+LAP) and ln(NAG+LAP):ln(AP) in soil across various regions were typically below 1:1, indicating that soil microbial activity is more constrained by nitrogen and phosphorus nutrients rather than by carbon. Precipitation, temperature, soil total carbon, and total nitrogen were identified as key environmental factors for extracellular enzyme activities and stoichiometric ratios. Our study highlights that the green manure-crop rotation alleviates nitrogen limitation while enhancing phosphorus limitation, and is closely related to the accumulation of total carbon and nitrogen in the soil.

Mechanisms of biochar-based organic fertilizers enhancing maize yield on a Chinese Chernozem: Root traits, soil quality and soil microorganisms

The Chinese Chernozems face extreme challenges of soil degradation and crop productivity decline. Biochar-based organic fertilizer (BOF) is effectively applied to improve crop yield and soil quality. Nonetheless, the mechanisms involved in yield enhancement in interaction with root traits, soil quality, and microorganisms after BOF application remain unclear. Herein, a two-year field experiment with three BOF rates [3.0 Mg ha−1 (BOF3), 6.0 Mg ha−1 (BOF6), 9.0 Mg ha−1 (BOF9)] was conducted to evaluate the impact of BOF on root traits, soil quality, and soil microorganisms, with no BOF applied as control treatment (CK). Compared to CK, BOF increased maize yield by 9.1 ∼ 32.0 % in the second year of application, and the highest yield (16.1 Mg ha−1) was obtained with BOF6. BOF increased the plant growth index by 29.7 ∼ 98.9 %, mainly attributed to the increase in root volume (2.4 ∼ 57.4 %), aboveground biomass (31.4 ∼ 44.0 %), and nitrogen (N) uptake (123.2 ∼ 216.0 %). The soil quality index was increased not only in the topsoil (27.7 ∼ 31.1 %) but also in the subsoil (22.7 ∼ 25.1 %) with BOF, particularly with the two highest rates (BOF6 and BOF9). This was mainly ascribed to the notable increase in microbial biomass carbon, total N, NH4+-N, and N-acquiring enzyme activity (normalized by leucine aminopeptidase and alkaline phosphatase). Moreover, BOF increased the abundance of Actinobacteria, Mortierellomycota at 0–40 cm, Chloroflexi at 0–20 cm, Acidobacteriota, and Ascomycota at 20–40 cm, which confirmed their key roles in activating soil nutrient and subsequent enhancing soil quality and stimulating plant growth. Overall, biochar-based organic fertilizer with a rate of 6.0 Mg ha−1 is a promising fertilizer regime to enhance soil quality by regulating soil keystone taxa, thereby boosting maize growth and yield on the Chernozem soil.

Clay soil amendment suppressed microbial enzymatic activities while increasing nitrogen availability in sandy soils

AbstractConservation management practices often produced positive but limited desirable outcomes in US Southeast sandy soils, likely due to their intrinsically low clay contents that constrain the soil's capacity to preserve organic carbon (C) and nutrients. In the field, we tested the effectiveness of a novel approach, that is, clay soil amendment, to improve sandy soils. In October 2017, clay‐rich soils (25% clay) were spread at 25 metric tons ha−1 and tilled onto a sandy soil (1.9% clay) in the field, which was further mixed by light tillage at 0‐ to 15‐cm depth, followed by planting winter cover crop mixtures (cereal rye, crimson clover, and winter pea). The crop rotation was cotton and corn with cover crop mixtures planted in the winter fallow season. Soils (0–15 cm) were collected in August 2021 and subjected to physio‐biochemical analyses. Clay amendment increased soil clay content to 3.4%, which improved nitrogen (N) availability by 51% but inhibited the activities of C (β‐d‐cellubiosidase [CB]; β‐xylosidase [BX]; N‐acetyl‐β‐glucosaminidase [NAG]) and N (leucine aminopeptidase [LAP]) cycling enzymes, resulting in up to 78% reduction in microbial respiration. A follow‐up kinetic study on BG and LAP enzymes suggested that clay addition can have different impacts on enzymes with diverse biological origins through distinct mechanisms. Clay addition can potentially improve sandy soils by stabilizing the organic inputs in soils. However, more research is required to understand its long‐term impacts making this approach practical.

Effects of exogenous copper on microbial metabolic function and carbon use efficiency of Panax notoginseng planting soil.

Soil copper (Cu) pollution is a serious environmental risk in the Panax notoginseng planting area. However, the effect of Cu on soil microbial metabolism and nutrient cycling in this area remains unknown. Therefore, Biolog ECO-plate and enzyme stoichiometry methods were utilized in this study to investigate the impact of exogenous Cu (control: 0 mg·kg-1; Cu100: 100 mg·kg-1; Cu400: 400 mg·kg-1; and Cu600: 600 mg·kg-1) on the metabolic function of soil microbial and nutrient limitation in the P. notoginseng soil. The results indicated that Cu100 significantly increased soil organic carbon (SOC), total phosphorus (TP), soil C:N, microbial biomass carbon (MBC), and microbial biomass nitrogen (MBN) 9.89%, 15.65%, 17.91%, 61.87%, and 90.56% higher than the control, respectively. Moreover, the carbon source utilization ratio of carbohydrates, amino acids, and amphiphilic compounds of Cu100 also increased by 7.16%, 25.47%, and 84.68%, respectively, compared with the control. The activities of β-1,4-glucosidase, cellobiohyrolase, leucine amino peptidase, β-1,4-N-acetylglucosaminidase, and phosphatase significantly decreased with increasing Cu concentration. Soil enzyme stoichiometry showed that all treatments were limited by nitrogen (vector angle < 45°; 19.045-22.081). Cu600 led to the lowest carbon limitation (1.798) and highest carbon use efficiency (CUE:0.267). The PLS-SEM model also showed that MBC, MBN, MBP, and microbial diversity positively affected carbon and nitrogen limitation (0.654 and 0.424). Soil carbon, nitrogen, phosphorus, stoichiometric ratio, MBC, MBN, and MBP positively affected CUE (0.527 and 0.589). The microbial diversity index significantly negatively affected CUE (-1.490). Multiple linear stepwise regression analyses showed that CUE was mainly influenced by MBC, AP, C:P, and LAP. Thus, P. notoginseng soil can benefit soil microbial carbon and nitrogen limitations at low Cu concentrations. Clarifying the metabolic activity and nutritional status of microorganisms under Cu stress can provide some theoretical basis for realizing China's comprehensive and effective management and control policies for environmental risks from metals by 2035.