Rapid, sensitive analysis method for determining the nitrogen stable isotope ratio of total free amino acids in soil.

RATIONALE: The amino acid-nitrogen (AA-N) isotope analysis of naturally abundant or isotope-labeled samples is indispensable for tracing nitrogen transfer in soil nitrogen biogeochemical cycling processes. Despite the usefulness of AA-N isotope analysis, the preparation methods are complex and time-consuming, and necessitate the use of toxic reagents. METHODS: We present an improved, rapid method for AA-N isotope analysis with high precision. At a high pH, AA-N was released and oxidized to N2 O using ClO- under vacuum. Additionally, purge-and-trap isotope ratio mass spectrometry was used to analyze N2 O. Moreover, we investigated the effect of various factors on the N2 O conversion process with glycine and applied the results to seven representative single-N AAs (alanine, serine, cysteine, aspartic acid, glutamic acid, leucine, and phenylalanine) and five poly-N AAs (lysine, arginine, histidine, tryptophan, and asparagine), as well as side-chain analogs, blank reagent, and other N forms. RESULTS: The concentration of ClO- and the pH were determined to be crucial factors for achieving desirable AA-N to N2 O conversion efficiencies. Glycine-N had the highest N2 O yield of 70%, with isotopic results consistent with those of the reference values at a high precision (within 0.5‰ for natural abundance and 0.01 atom% for 15 N-enrichment) at the nanomolar N level. Additionally, the α-NH2 AAs were labile, and the single-N AAs were more easily converted to N2 O than poly-N AAs. With the exception of γ-aminobutyric acid, the N2 O conversion efficiencies of the side-chain N analogs were very low (below 5%). This method was also applicable to the 15 N analysis of the total free AAs in complex soil samples without interference from analytical blanks and other forms of N. CONCLUSIONS: Our method is highly selective for the α-NH2 groups of an amino acid, and the oxidation of the side chain is difficult. In addition, the method is sensitive, rapid, and convenient, and does not require toxic reagents.

Contrasting pathways of carbon sequestration in paddy and upland soils.

Paddy soils make up the largest anthropogenic wetlands on earth, and are characterized by a prominent potential for organic carbon (C) sequestration. By quantifying the plant- and microbial-derived C in soils across four climate zones, we identified that organic C accrual is achieved via contrasting pathways in paddy and upland soils. Paddies are 39%-127% more efficient in soil organic C (SOC) sequestration than their adjacent upland counterparts, with greater differences in warmer than cooler climates. Upland soils are more replenished by microbial-derived C, whereas paddy soils are enriched with a greater proportion of plant-derived C, because of the retarded microbial decomposition under anaerobic conditions induced by the flooding of paddies. Under both land-use types, the maximal contribution of plant residues to SOC is at intermediate mean annual temperature (15-20°C), neutral soil (pH~7.3), and low clay/sand ratio. By contrast, high temperature (~24°C), low soil pH (~5), and large clay/sand ratio are favorable for strengthening the contribution of microbial necromass. The greater contribution of microbial necromass to SOC in waterlogged paddies in warmer climates is likely due to the fast anabolism from bacteria, whereas fungi are unlikely to be involved as they are aerobic. In the scenario of land-use conversion from paddy to upland, a total of 504 Tg C may be lost as CO2 from paddy soils (0-15 cm) solely in eastern China, with 90% released from the less protected plant-derived C. Hence, preserving paddy systems and other anthropogenic wetlands and increasing their C storage through sustainable management are critical for maintaining global soil C stock and mitigating climate change.

Diazotrophic Community Variation Underlies Differences in Nitrogen Fixation Potential in Paddy Soils Across a Climatic Gradient in China.

Biological nitrogen (N2) fixation as a source of new N input into the soil by free-living diazotrophs is important for achieving sustainable rice agriculture. However, the dominant environmental drivers or factors influencing N2 fixation and the functional significance of the diazotroph community structure in paddy soil across a climatic gradient are not yet well understood. Thus, we characterized the diazotroph community and identified the ecological predictors of N2 fixation potential in four different climate zones (mid-temperate, warm-temperate, subtropical, and tropical paddy soils) in eastern China. Comprehensive nifH gene sequencing, functional activity detection, and correlation analysis with environmental factors were estimated. The potential nitrogenase activity (PNA) was highest in warm-temperate regions, where it was 6.2-, 2.9-, and 2.2-fold greater than in the tropical, subtropical, and mid-temperate regions, respectively; nifH gene abundance was significantly higher in warm-temperate and subtropical zones than in the tropical or mid-temperate zones. Diazotroph diversity was significantly higher in the tropical climate zone and significantly lower in the mid-temperate zone. Non-metric multidimensional scaling and canonical correlation analysis indicated that paddy soil diazotroph populations differed significantly among the four climate zones, mainly owing to differences in climate and soil pH. Structural equation models and automatic linear models revealed that climate and nutrients indirectly affected PNA by affecting soil pH and diazotroph community, respectively, while diazotroph community, C/P, and nifH gene abundance directly affected PNA. And C/P ratio, pH, and the diazotroph community structure were the main predictors of PNA in paddy soils. Collectively, the differences in diazotroph community structure have ecological significance, with important implications for the prediction of soil N2-fixing functions under climate change scenarios.

Flooding and straw returning regulates the partitioning of soil phosphorus fractions and phoD-harboring bacterial community in paddy soils.

Flooding and straw returning are effective agricultural practices in promoting phosphorus (P) availability in paddy soils. However, little is known about the effects of these practices and their interaction on the soil P pools and functional microbes responsible for soil P mobilization. Our 4-year paddy field experiment aimed to analyze the responses of soil P fractions and phoD-harboring bacterial communities in a double-rice cropping system to intermittent flooding (IF) and continuous flooding (CF), in plots with (+ S) and without (-S) straw return. Compared to IF, CF significantly increased soil citrate-P and marginally decreased the HCl-P fractions, suggesting that the stable inorganic P pools are transferred to labile inorganic P at lower redox potentials. Compared to the -S treatments, + S treatments significantly increased the labile organic fractions (enzyme-P). Correspondingly, a decreased soil total organic P concentration was observed in + S treatment. Additionally, + S treatment significantly increased the activity of acid phosphomonoesterase and alkaline phosphomonoesterase and the abundance of phoD-harboring bacteria. These results indicated that straw promoted organic P minimization to release orthophosphate. The diversity of the phoD-harboring bacteria and complexity of the co-occurrence network decreased under the CF + S treatment; however, all keystone species of the phoD-harboring bacteria were retained in this oxygen-deficient environment. This study highlights that irrigation regimes mediate the processes of inorganic P mobilization, while straw returns regulate the processes of organic P mineralization. Additionally, flooding could be a more effective agricultural practice than straw returning to promote soil P availability in paddy soils. KEY POINTS: •Soil P pools and phoD-harboring bacteria communities were assessed. •Straw return mainly affects the mineralization of organic P. •Continuous flooding mainly affects the mobilization of inorganic P.

Value of serum miR-922 and miR-506 expression levels in the diagnosis and prognostic assessment of childhood acute lymphoblastic leukemia. / è¡€æ¸ miR-922及miR-506å¯¹å„¿ç«¥æ€¥æ€§æ·‹å·´ç»†èƒžç™½è¡€ç— è¯Šæ–­åŠé¢„åŽåˆ¤æ–­çš„ä»·å€¼.

OBJECTIVES: To study the value of serum miR-922 and miR-506 expression levels in the diagnosis and prognostic assessment of childhood acute lymphoblastic leukemia (ALL). METHODS: A total of 132 children with ALL (ALL group) and 80 healthy children (healthy control group) were prospectively selected in this study. Quantitative real-time polymerase chain reaction was used to measure the expression levels of serum miR-922 and miR-506 in both groups. Receiver operating characteristic (ROC) curves were plotted to analyze the diagnostic value of miR-922 and miR-506 for childhood ALL. The Kaplan-Meier method was used to plot survival curves, and multivariate COX regression models were used to analyze the risk factors for poor prognosis in children with ALL. RESULTS: The ALL group had significantly higher expression levels of serum miR-922 and miR-506 than the control group (P<0.001). The ROC curve analysis showed that the optimal cut-off values of miR-922 and miR-506 for the diagnosis of childhood ALL were 1.46 and 2.17, respectively. The high miR-922 expression (≥1.46) group and high miR-506 expression (≥2.17) group had significantly higher incidence rates of lymph node enlargement, leukocyte count ≥50×109/L, medium-high risk stratification, mixed-lineage leukemia (MLL) gene rearrangement, and karyotype abnormality than the low miR-922 expression group and low miR-506 expression group (P<0.05). The Kaplan-Meier analysis showed that high expression of miR-922 and miR-506 was associated with short survival time in children with ALL (P<0.05). The multivariate COX regression analysis showed that leukocyte count ≥50×109/L, medium-high risk stratification, MLL gene rearrangement, miR-922≥1.46, and miR-506≥2.17 could indicate poor prognosis in children with ALL (P<0.05). CONCLUSIONS: The expression levels of miR-922 and miR-506 are of good value in the diagnosis and prognostic assessment of childhood ALL.

Correction to: Effects of long-term straw incorporation on lignin accumulation and its association with bacterial laccase-like genes in arable soils.

The first funding No. should be 41671298 instead of 41301298.

Effects of long-term straw incorporation on lignin accumulation and its association with bacterial laccase-like genes in arable soils.

In this study, we aimed to investigate lignin accumulation and its relationship with the composition of bacterial laccase-like genes in three arable lands (i.e., upland limestone soil (UL), upland red soil (UR), and upland-paddy rotation red soil (UPR)), which are subjected to long-term straw incorporation. After 9-13 years of straw incorporation, the lignin content significantly increased from 337.1, 414.5, and 201.6 mg/kg soil to 2096.5, 2092.4, and 1972.2 mg/kg soil in UL, UR, and UPR, respectively. The dominant lignin monomer changed from vanillyl (V)-type to cinnamyl (C)-type in UR. Both V- and C-types were the dominant monomers in UPR, and V-type monomer remained the dominant monomer in UL. Compared with the treatment without straw, straw incorporation significantly promoted the activity of laccase enzyme and the abundance of bacterial laccase-like genes in all soils. The redundancy analysis showed that the main influencing factors on lignin accumulation patterns with straw incorporation were the laccase enzyme activity, nitrogen availability, and some specific bacterial communities possessing the laccase-like genes (e.g., Thermotogae and Acidobacteria). The variation partitioning analysis confirmed that the strongest influencing factor on lignin accumulation was the composition of bacterial laccase-like genes (explained 31.4% of variance). The present study provides novel insights into the importance of bacterial laccase-like genes in shaping lignin monomer accumulation with straw incorporation in arable soils.

Modified method for the extraction of mRNA from paddy soils.

OBJECTIVE: Paddy soils contain relatively greater organic matter and water contents than other soils thereby limiting effective mRNA extraction. A modification of the conventional mRNA soil extraction method specific to paddy soils is described. RESULTS: Two main modifications for co-extraction of DNA and RNA are: (1) addition of 20 % (w/v) sodium dodecyl sulphate to 10 % (w/v) hexadecyltrimethylammonium bromide extraction buffer, and (2) fresh soil, initially frozen at -80 °C, is immediately immersed in extraction buffer. The high-quality total RNA extracted can be directly used in downstream analyses without an additional step to remove humic acid. RNA purification was conducted to remove 5S rRNA, and the mRNA was enriched by selectively digesting rRNA. cDNA synthesised by reverse transcriptase was not contaminated by the reagents or genomic DNA. CONCLUSION: The modified method for mRNA extraction from paddy soil is suitable for analysing the expression of microbial genes from fresh paddy soil.

Abundance and Diversity of CO2-Assimilating Bacteria and Algae Within Red Agricultural Soils Are Modulated by Changing Management Practice.

Elucidating the biodiversity of CO(2)-assimilating bacterial and algal communities in soils is important for obtaining a mechanistic view of terrestrial carbon sinks operating at global scales. "Red" acidic soils (Orthic Acrisols) cover large geographic areas and are subject to a range of management practices, which may alter the balance between carbon dioxide production and assimilation through changes in microbial CO(2)-assimilating populations. Here, we determined the abundance and diversity of CO(2)-assimilating bacteria and algae in acidic soils using quantitative PCR and terminal restriction fragment length polymorphism (T-RFLP) of the cbbL gene, which encodes the key CO(2) assimilation enzyme (ribulose-1,5-bisphosphate carboxylase/oxygenase) in the Calvin cycle. Within the framework of a long-term experiment (Taoyuan Agro-ecosystem, subtropical China), paddy rice fields were converted in 1995 to four alternative land management regimes: natural forest (NF), paddy rice (PR), maize crops (CL), and tea plantations (TP). In 2012 (17 years after land use transformation), we collected and analyzed the soils from fields under the original and converted land management regimes. Our results indicated that fields under the PR soil management system harbored the greatest abundance of cbbL copies (4.33 × 10(8) copies g(-1) soil). More than a decade after converting PR soils to natural, rotation, and perennial management systems, a decline in both the diversity and abundance of cbbL-harboring bacteria and algae was recorded. The lowest abundance of bacteria (0.98 × 10(8) copies g(-1) soil) and algae (0.23 × 10(6) copies g(-1) soil) was observed for TP soils. When converting PR soil management to alternative management systems (i.e., NF, CL, and TP), soil edaphic factors (soil organic carbon and total nitrogen content) were the major determinants of bacterial autotrophic cbbL gene diversity. In contrast, soil phosphorus concentration was the major regulator of algal cbbL community composition. Our results provide new insights into the diversity, abundance, and modulation of organisms responsible for microbial autotrophic CO(2) fixation in red acidic soils subjected to changing management regimes.

Laccase activity is proportional to the abundance of bacterial laccase-like genes in soil from subtropical arable land.

Laccase enzymes produced by both soil bacteria and fungi play important roles in refractory organic matter turnover in terrestrial ecosystems. We investigated the abundance and diversity of fungal laccase genes and bacterial laccase-like genes in soil from subtropical arable lands, and identified which microbial group was associated with laccase activity. Compared with fungal laccase genes, the bacterial laccase-like genes had greater abundance, richness and Shannon-Wiener diversity. More importantly, laccase activity can be explained almost exclusively by the bacterial laccase-like genes, and their abundance had significant linear relationship with laccase activity. Thus, bacterial laccase-like gene has great potential to be used as a sensitive indicator of laccase enzyme for refractory organic matter turnover in subtropical arable lands.

Community structure analysis of soil ammonia oxidizers during vegetation restoration in southwest China.

Soil ammonia oxidizers play a critical role in nitrogen cycling and ecological restoration. The composition and structure of soil ammonia oxidizers and their impacting factors were studied in four typical ecosystem soils, tussock (T), shrub (S), secondary forest (SF), and primary forest (PF), during vegetation restoration in the Karst region of Southwest China. The composition and structure of the ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) communities were characterized by sequencing the amoA and arch-amoA genes, respectively. The diversity of soil ammonia oxidizers (except in S) and plant Shannon diversity index gradually increased with vegetation restoration, and the ammonia oxidizer communities differed significantly (p < 0.001). Amplicons of AOA from the Nitrososphaera cluster dominated all four ecosystem soils. AOB Nitrosospira cluster 3b only appeared in PF and SF soils, while Nitrosospira cluster 3a species were found in all soils. Changes in AOB paralleled the changes in soil ammonium content that occurred with vegetation restoration. Redundancy analysis showed that the distribution of dominant AOB species was linked to pH, soil urease activity, and soil C/N ratio, whereas the distribution of dominant AOA species was mainly influenced by litter nitrogen content and C/N ratio. These results suggested that the composition and structure of the AOB community were more sensitive to changes in vegetation and soil ammonium content, and may be an important indicator of nitrogen availability in Karst ecosystem soils.

Intensive N2 fixation accelerates microbial turnover in cropland soils.

Biological nitrogen fixation (BNF) is strongly affected by the carbon (C) and nitrogen (N) stoichiometry in soil and depends on the input of organic C. Due to the high metabolic costs of nitrogenase activity, however, the response of BNF to organic C input and its impact on microbial turnover remain unclear. To address this knowledge gap, we combined 15N2 tracing with high-throughput sequencing by adding glucose or glucose plus mineral N fertilizer for a 12-day incubation in three cropland soils. Glucose addition alone strongly changed the BNF activity (0.76-2.51 mg N kg-1 d-1), while BNF was completely absent after mineral N fertilization. This switch-on of BNF by glucose addition supported equally high rates of microbial growth and organic C mineralization compared with the direct mineral N assimilation by microorganisms. Glucose-induced BNF was predominantly catalyzed by Azotobacter-affiliated free-living diazotrophs (>50 % of the total nifH genes), which increased with diverse nondiazotrophs such as Nitrososphaera, Bacillus and Pseudoxanthomonas. Structural equation models (SEMs) and random forest (RF) analyses consistently revealed that the soil C:N ratio and Azotobacter-affiliated diazotrophic abundances were the key factors affecting glucose-induced BNF. Our findings emphasize the importance of free-living diazotrophs for microbial turnover of organic C in soil.

[Response of Organic Carbon Mineralization to Nitrogen Addition in Micro-aerobic and Anaerobic Layers of Paddy Soil].

In field conditions, a micro-aerobic layer with 1 cm thickness exists on the surface layer of paddy soil owing to the diffusion of dissolved oxygen via flooding water. However, the particularity of carbon and nitrogen transformation in this specific soil layer is not clear. A typical subtropical paddy soil was collected and incubated with13C-labelled rice straw for 100 days. The responses of exogenous fresh organic carbon(13C-rice straw) and original soil organic carbon mineralization to nitrogen fertilizer addition[(NH4)2SO4]in the micro-aerobic layer(0-1 cm) and anaerobic layer(1-5 cm) of paddy soil and their microbial processes were analyzed based on the analysis of 13C incorporation into phospholipid fatty acid(13C-PLFAs). Nitrogen addition promoted the total CO2 and 13C-CO2 emission from paddy soil by 11.4% and 12.3%, respectively. At the end of incubation, with the addition of nitrogen, the total soil organic carbon (SOC) and13C-recovery rate from rice straw in the anaerobic layer were 2.4% and 9.2% lower than those in the corresponding micro-aerobic layer, respectively. At the early stage(5 days), nitrogen addition increased the total microbial PLFAs in the anaerobic layer with a consistent response of bacterial and fungal PLFAs. However, there was no significant effect from nitrogen on microbial abundance in the micro-aerobic layer. Nitrogen addition had no significant impact on the abundance of total 13C-PLFAs in the micro-aerobic and anaerobic layers, but the abundance of 13C-PLFAs for bacteria and fungi in the micro-aerobic layer was decreased dramatically. At the late stage(100 days), the effect of nitrogen addition on microbial PLFAs was consistent with that at the early stage. The abundances of total, bacterial, and fungal 13C-PLFAs were remarkably increased in the anaerobic layer. However, the abundance of 13C-PLFAs in the micro-aerobic layer showed no significant response to nitrogen addition. During the incubation, the content of NH4+-N in the anaerobic soil layer was higher than that in the micro-aerobic soil layer. This indicates that nitrogen addition increased microbial activity in the anaerobic soil layer caused by the higher NH4+-N concentration, as majority of microorganisms preferred to use NH4+-N. Consequently, the microbial utilization and decomposition of organic carbon in the anaerobic soil layer were accelerated. By contrast, richer available N existed in the form of NO3--N in the micro-aerobic soil layer owing to the ammoxidation process. Thus, the shortage of NO3--N preference microorganisms in the paddy soil environment prohibited the microbial metabolism of organic carbon in the micro-aerobic layer. As a whole, nitrogen fertilization enhanced organic carbon loss via microbial mineralization in paddy soil with a weaker effect in the micro-aerobic layer than that in the anaerobic layer, indicating the limited microbial metabolic activity in the surface micro-aerobic layer could protect the organic carbon stabilization in paddy soil. This study emphasizes the heterogeneity of paddy soil and its significant particularity of carbon and nitrogen transformation in micro-aerobic layers. Consequently, this study has implications for optimizing the forms and method for the application of nitrogen fertilizer in paddy cropping systems.

Land reclamation and short-term cultivation change soil microbial communities and bacterial metabolic profiles.

BACKGROUND: Soil microbes play an important role in many critical ecosystem processes, but little is known about the effects of land reclamation and short-term cultivation on microbial communities in red soil. In this study, soil microbial communities under five land use patterns-artificial pine forest (Fp), tussock and shrub (TS), shrubbery (Sh), sugarcane (Su) and maize and cassava rotation (Ma)-were characterised by DNA fingerprinting and metabolic profiling to reveal how land reclamation and cultivation affect the underlying diversity and function of soil microbial communities in southwestern China. RESULTS: Eight years of reclamation and cultivation significantly affected population size, composition and structure, bacterial metabolic profiles and diversity values (Shannon-Wiener index) of soil microbial communities. Soil organic carbon and pH were the most important factors shaping the underlying microbial communities; however, with significant correlations between soil carbon/nitrogen ratio and bacterial taxonomic and metabolic diversities, soil total nitrogen was a potentially important factor for soil microbial composition and function, as well as soil moisture, cation exchange capacity and physical structure to a lesser extent. In addition, the lowest pH, lower nutrient availability and the most compact soil in pine forest resulted in the lowest microbial taxonomic and metabolic diversities among the five land use patterns studied. CONCLUSION: Soil organic carbon, nitrogen and pH appeared to be the most important factors influencing microbial biomass, composition and function in red soil of southwestern China. The study suggests that measures to lessen the impact of changes in this edaphic environment should be taken to avoid an imbalance of microbial function and improve ecological sustainability in southwestern China.

Soil bacterial community composition and diversity respond to cultivation in Karst ecosystems.

Soil microorganisms play vital roles in recovering and maintaining the health of ecosystems, particularly in fragile Karst ecosystems that are easily degraded after cultivation. We investigated the composition and diversity of soil bacterial communities, based on RFLP and 16S rDNA sequencing, in a cropland, a naturally revegetated land with former cultivation disturbance and a primeval forest in the subtropical Karst of southwest China. Our results illustrated that Proteobacteria accounted for 44.8% of the 600 tested clones, making it the most dominant phylum observed. This phylum was followed by Acidobacteria and Planctomycetes for the three Karst soils analyzed. Compared with the primeval forest soil, the proportions of Proteobacteria were decreased by 30.2 and 37.9%, while Acidobacteria increased by 93.9 and 87.9%, and the Shannon-Wiener diversity indices and the physicochemical parameters declined in the cropland and the revegetated land, respectively. Among the three soils, the proportion of dominant bacterial phyla and the diversity indices in the revegetated land were similar to the cropland, implying the bacterial community in the cropland was relatively stable, and the after-effects of cultivation were difficult to eliminate. However, similar distributions of the four Proteobacteria subphyla were observed between the revegetated land and the primeval forest soil. Furthermore, the proportion of Rhizobiales belonging to α-Proteobacteria was sharply decreased with cultivation compared to the primeval forest soil, while a small cluster of Rhizobiales recurred with vegetation recovery. These results indicated that although the subphyla of the dominant bacterial phylum had some positive responses to 20 years of vegetation recovery, it is a slow process. Our results suggest that priority should be given to conserve the primeval forest and inoculation of functional microorganisms on the basis of vegetation recovery may be more effective for the restoration of Karst ecosystems after cultivation.

[Effects of Organic Materials on Phosphorus Fractions and phoD-harboring Bacterial Community in Karst Soil].

Efficient utilization of organic materials based on the rich resources in the karst region can promote soil fertility. Microorganisms have a crucial influence on soil phosphorus availability. phoD is considered to be the encoding phosphatase gene that can reflect the hydrolysis of organophosphorus compounds for the soil bacterial community. Molecular analysis of the phoD-harboring bacterial gene provides insight into promoting soil phosphorus availability under different fertilization managements. However, the effects of organic materials on soil phosphorus fractions associated with phoD-harboring bacterial communities are poorly understood. This study comprehensively investigated the effects of organic materials on soil phosphorus availability and explored environmental drivers of phoD-harboring bacteria in the Karst region. Here, six treatments were designed in the field as follows:non-fertilized control (CK), inorganic fertilization (NPK), inorganic fertilization combined with straw (NPKS), inorganic fertilization combined with manure (NPKM), inorganic fertilization combined with sludge (NPKL), and inorganic fertilization combined with sugarcane ash (NPKA). The phoD-harboring bacterial community in Karst region soil was analyzed using high-throughput sequencing. The results showed that the content of total P (TP), Olsen-P, and Ca2-P increased with the years after organic material application, whereas the content of CaCl2-P first decreased and then increased. Compared to that under the CK treatment, organic material application, especially NPKL treatment, significantly increased soil total nitrogen (TN), TP, Olsen-P, CaCl2-P, and Ca2-P contents, followed by those in the NPKA and NPKM treatments. Correlation analysis showed that the contents of CaCl2-P, Ca2-P, and Olsen-P were significantly positively correlated with soil exchangeable calcium (Ca-ex) content. Redundancy analysis (RDA) showed that TN, Ca-ex, soil organic carbon (SOC), and total potassium (TK) contents were the key factors affecting soil P fractions. Using high-throughput sequencing, we found that only NPKS increased the richness of phoD-harboring bacteria compared to that under the control treatment. No significant difference was observed in the phoD-harboring bacterial community among all treatments. The RDA model selected the Ca-ex, TK, Olsen-P, pH, and SOC as the key environmental predictors for the phoD-harboring bacterial community. In summary, soil phosphorus availability can be improved through the input of organic materials and inorganic fertilizer combined with manure, sludge, and ash. These additions were suitable for nutrient management and sustainable development in farmland soil in the Karst region of Guangxi.

[Characteristics of Microbial Utilization for Crop Residue-Derived C in Paddy and Upland Soils].

Two typical subtropical agricultural soils, a flooded paddy soil and its adjacent upland, were collected and then incubated with or without 13C-labeled crop residue (maize straw) for 40 days. During the incubation, the mineralization rate of the crop residue was monitored, and the 13C incorporated into fungal and bacterial phospholipid fatty acid (PLFA) was quantified. At the early stage (0.25-1 days), the mineralization rate of crop residue was faster in paddy soil than that in upland soil, whereas the opposite trend was observed from 2 to 20 days. At the late stage (21-40 days), the mineralization rate was similar in both soils. At the end of incubation, 11% of the total crop residue was mineralized in paddy soil, which was about half of that in upland soil (20%). Although paddy soil had a higher amount of microbial biomass (indicated by total PLFA), the total amounts of 13C-PLFA were comparable in both soils, and the enrichment ratio (proportion of 13C to total C in PLFA) was lower in paddy soil than that in upland soil. This indicated that the microbial community in paddy soil was less active in the uptake of crop residue C than that in upland soil. During the incubation, the residue-derived 13C was mainly distributed in bacterial PLFA (up to 86% of total 13C-PLFA, including 59% in gram-positive and 27% in gram-negative bacteria) in paddy soil, and up to 75% of total 13C-PLFA distributed in fungal PLFAs was in upland soil. Thus, bacteria dominated the utilization of crop residue in paddy soil versus fungi in upland soil. Compared with that in upland soil, the microbial activity was suppressed in the anaerobic condition caused by flooding in paddy soil, with a stronger inhibition of fungi than bacteria. Considering the discrepancies of life strategies and necromass turnover between bacteria and fungi, the different dominant microbial groups in the utilization of crop residue in water-logged and well-drained conditions could lead to the distinct accumulation and stabilization of microbial-derived organic matter in paddy and upland soils.

Soil Microbial Resource Limitations and Community Assembly Along a Camellia oleifera Plantation Chronosequence.

Understanding soil microbial element limitation and its relation with the microbial community can help in elucidating the soil fertility status and improving nutrient management of planted forest ecosystems. The stand age of a planted forest determines the aboveground forest biomass and structure and underground microbial function and diversity. In this study, we investigated 30 plantations of Camellia oleifera distributed across the subtropical region of China that we classified into four stand ages (planted <9 years, 9-20 years, 21-60 years, and >60 years age). Enzymatic stoichiometry analysis showed that microbial metabolism in the forests was mainly limited by C and P. P limitation significantly decreased and C limitation slightly increased along the stand age gradient. The alpha diversity of the soil microbiota remained steady along stand age, while microbial communities gradually converged from scattered to clustered, which was accompanied by a decrease in network complexity. The soil bacterial community assembly shifted from stochastic to deterministic processes, which probably contributed to a decrease in soil pH along stand age. Our findings emphasize that the stand age regulated the soil microbial metabolism limitation and community assembly, which provides new insight into the improvement of C and P management in subtropical planted forest.

Biochar protects hydrophilic dissolved organic matter against mineralization and enhances its microbial carbon use efficiency.

A combination of biochar with exogenous organic material in soils is often used in practical farmland management. The objective of this study was to determine how biochar affects organic matter decomposition by studying the decomposition of 13C-labelled hydrophilic (Hi-) and hydrophobic (Ho-) dissolved organic matter (DOM) in acid and neutral soils during a 60-day incubation experiment. The proportions of carbon (C) mineralization in Hi-DOM with or without biochar addition were 32.6% or 34.5% in acid soil (P > 0.05) and 15.4% or 22.3% in neutral soil (P < 0.05), respectively. In contrast, those proportions of Ho-DOM-C mineralization with or without biochar addition were 20.0% or 21.4% in acid soil and 19.0% or 20.5% in neutral soil (P > 0.05), respectively. These results showed that biochar could protect Hi-DOM against mineralization in neutral soil but exhibited less effect on Ho-DOM mineralization in both acid and neutral soils. Additionally, biochar did not affect microbial incorporation of Hi- or Ho-DOM in acid and neutral soils. However, biochar notably improved the microbial carbon use efficiency (CUE) of Hi-DOM while it significantly reduced the CUE of Ho-DOM in neutral soil (P < 0.05), indicating that the effect of biochar on microbial CUE was related to organic matter type and soil pH. This study suggests that Hi-DOM can outperform Ho-DOM to decrease C loss and improve microbial CUE in neutral soil with biochar addition. This phenomenon could be due mainly to the different chemical compositions of Hi-DOM and Ho-DOM and their distinct microbial preference. These findings can provide references for biochar's ability to regulate the decomposition of organic matter.

[Characteristics of soil nitrogen cycle and mechanisms underlying the increase in rice yield with partial substitution of mineral fertilizers with organic manure in a paddy ecosystem: A review]. / 稻作系统有机肥替代部分化肥的土壤氮循环特征及增产机制.

Partial substitution of mineral fertilizers with organic manure is a key strategy for stable and increase crop yield accompanying with zero growth of mineral fertilizers. Based on recent stu-dies, we reviewed the effects of partial substitution of mineral fertilizers with organic manure on rice yield, nitrogen utilization efficiency, soil nitrogen fractions, and microbial nitrogen fixation, ammonification, nitrification, and denitrification in rice paddy ecosystems. We further compared the cha-racteristics of soil nitrogen cycle of mineral fertilizers alone and partial substitution of mineral fertili-zers with organic manure. The partial substitution altered key processes of nitrogen cycling, including enhancement of ammonification, mediation of nitrification and denitrification, reduction of ammonia volatilization and nitrogen loss, improved the status of nitrogen supplements (enriching the supplement of low-molecular-weight organic nitrogen, adjusting the distribution of inorganic nitrogen components, increasing the amount of microbial biomass nitrogen, and decreasing the loss of total nitrogen), improved soil nitrogen supply (increasing supply of small molecule organic nitrogen, coordinating inorganic nitrogen components and proportions, and increasing soil microbial biomass nitrogen and total nitrogen fixation), which promoted nitrogen uptake and regulated nitrogen allocation in rice plant to realize stability and enhancement of rice yield.