Novel molecular insights into pyroptosis in triple-negative breast cancer prognosis and immunotherapy.

BACKGROUND: Patients with triple-negative breast cancer (TNBC) often have a poor prognostic outcome. Current treatment strategies cannot benefit all TNBC patients. Previous findings suggested pyroptosis as a novel target for suppressing cancer development, although the relationship between TNBC and pyroptosis-related genes (PRGs) was still unclear. METHODS: Gene expression data and clinical follow-up of TNBC patients were collected from the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) and Gene Expression Omnibus (GEO). PRGs were screened using weighted gene co-expression network analysis. Cox regression analysis and the least absolute shrinkage and selection operator (i.e. LASSO) technique were applied to construct a pyroptosis-related prognostic risk score (PPRS) model, which was further combined with the clinicopathological characteristics of TNBC patients to develop a survival decision tree and a nomogram. The model was used to calculate the PPRS, and then the overall survival, immune infiltration, immunotherapy response and drug sensitivity of TNBC patients were analyzed based on the PPRS. RESULTS: The PPRS model was closely related to clinicopathological features and can independently and accurately predict the prognosis of TNBC. According to normalized PPRS, patients in different cohorts were divided into two groups. Compared with the high-PPRS group, the low-PPRS group had significantly higher ESTIMATE (i.e. Estimation of STromal and Immune cells in MAlignantTumours using Expression data) score, immune score and stromal score, and it also had overexpressed immune checkpoints and significantly reduced Tumor Immune Dysfunction and Exclusion (TIDE) score, as well as higher sensitivity to paclitaxel, veliparib, olaparib and talazoparib. A decision tree and nomogram based on PPRS and clinical characteristics can improve the prognosis stratification and survival prediction for TNBC patients. CONCLUSIONS: A PPRS model was developed to predict TNBC patients' immune characteristics and response to immunotherapy, chemotherapy and targeted therapy, as well as their survival outcomes.

Comparison of Different Isolation Methods for Plasma-Derived Extracellular Vesicles in Patients with Hyperlipidemia.

Extracellular vesicles are commonly found in human body fluids and can reflect current physiological conditions of human body and act as biomarkers of disease. The quality of isolated extracellular vesicles facilitates the early diagnosis of various diseases accompanied by hyperlipidemia. Nonetheless, there are no reports on which special methods are suitable for isolating extracellular vesicles from the plasma of patients with hyperlipidemia. Thus, this study compared three different research-based extracellular vesicle isolation approaches, namely ultracentrifugation (UC), polyethylene glycol (PEG) precipitation, and size exclusion chromatography (SEC), and determined which of them was the most effective method. We selected blood samples from 12 patients with clinically diagnosed hyperlipidemia and isolated plasma-derived extracellular vesicles using three methods. The morphology of the isolated extracellular vesicles was observed using transmission electron microscopy, while the concentration was detected by asymmetric flow field-flow fractionation and multi-angle light scattering. Marker proteins were identified by Western blotting, and protein composition was evaluated by silver staining. Both determined the contaminations in the extracellular vesicle samples. The results showed that the three methods can be successfully used for the isolation of extracellular vesicles. The extracellular vesicles isolated by UC were larger in size, and the yield was much lower. Although the yield of extracellular vesicles isolated by PEG precipitation was greatly improved, the contamination was increased. Of the three methods, only the SEC-isolated extracellular vesicles were characterized by high yield and low contamination. Therefore, our data suggested that the SEC was a more ideal method for isolating extracellular vesicles from the plasma of patients with hyperlipidemia.

[Effect of Long-term Straw Returning on the Mineralization and Priming Effect of Rice Root-carbon].

Long-term straw returning to the field changes the environmental conditions of rice paddy soil, which affects the mineralization and priming effect of residual rice roots in the soil, but the direction and intensity of its influence is not clear. Therefore, based on a long-term fertilization field experiment, 13C-CO2 isotopic labeling technology and laboratorial incubation were used to analyze the characteristics of mineralization of rice roots and native soil organic carbon, the intensity and direction of the priming effect, and the source partitioning of CO2 emissions in three treatments, consisting of no fertilization (CK), chemical fertilizer (CF), and straw returning with chemical fertilizer (CFS). The results showed that after 120 days of flooding incubation, the root residue (R) increased the cumulative CO2 emissions by 617.41-726.27 mg·kg-1. The cumulative CO2 emissions from roots and root mineralized proportions in the CFS+R and CF+R treatments were 470.82 and 444.04 mg·kg-1, respectively, and 18.8% and 17.8%, respectively. These were significantly higher than those in the CK+R treatment (384.19 mg·kg-1, 15.4%). There was no significant difference in the cumulative CO2 emissions from native soil organic carbon among the three treatments. However, the mineralized proportion of native soil organic carbon in the CFS+R treatment (4.2%) was significantly lower than that in the CF+R and CK+R treatments (5.4% and 5.8%). The priming effect in the CFS+R treatment was 29.6%, which was significantly lower than that in the CK+R treatment (42.5%) and higher than that in the CF+R treatment (14.4%). A total of 23.47% to 27.59% of the cumulative CO2 emission of the flooded paddy soil was from the roots, and the remainder was from the soil. In addition, the proportion of CO2 emission caused by the priming effect was smaller in the CFS+R treatment than that in the CK+R treatment and larger than that in the CF+R treatment. In summary, the long-term straw returning in the flooded paddy soil will increase the mineralization potential of rice roots, but it is more conducive to the stability of the native soil organic carbon.

[Characteristics of Paddy Soil Organic Carbon Mineralization and Influencing Factors Under Different Water Conditions and Microbial Biomass Levels].

Paddy soil often undergoes frequent dry-wet alternation. The change in water status not only affects the physical and chemical properties of the soil, but also changes the structure and diversity of the soil microbial communities, which in turn determines the rate of soil organic carbon mineralization. However, the effects of different water conditions and soil microbial biomass levels on the process of soil organic carbon mineralization and its mechanisms are still unclear. Therefore, this study took typical subtropical paddy soil as the research object, applied a laboratory incubation experiment with two water treatments of dry-wet and continuous flooding, and reduced the soil microbial biomass through chloroform fumigation, thereby obtaining high and low soil microbial biomass carbon contents, to elucidate the influencing mechanisms of microbial biomass and water conditions on organic carbon mineralization in paddy soil. The results showed that during the first 30 d of incubation, the dry-wet treatment was in a non-flooded stage and its cumulative CO2 emissions were significantly lower than those of the continuous flooded treatment. After 30 d, the dry-wet treatment entered the flooded stage. The difference in the cumulative CO2 emissions of the soils with a high microbial biomass carbon content between the dry-wet and continuous flooding treatments gradually decreased, and there was no significant difference on day 78. In the soil with a low microbial biomass carbon content, the cumulative CO2 emissions of the dry-wet treatment on day 78 was still significantly lower than that of the continuous flooded treatment. The soils with a low microbial biomass carbon content showed a faster CO2 emission rate at the beginning of the incubation period (first 20 d), which was 1.1-6.1 times greater than that of the high microbial biomass carbon soils owing to their high soil dissolved organic carbon (DOC) content, and the CO2 emission rate then gradually decreased until it was below that of the soil with a high microbial biomass carbon content. The soil organic carbon mineralization rate became stable later in the incubation period (days 45-78). The stable mineralization rate of the high microbial biomass carbon soil was 20%-30% higher than that of the low microbial biomass carbon soil. The multiple regression analysis results showed that the decrease in the soil DOC content (ΔDOC) and the increase in the Fe2+ content (ΔFe2+) significantly affected the change in cumulative CO2 emissions (ΔCO2) under continuous flooding conditions, but had no effect on ΔCO2 during the flooding stage of the dry-wet treatment. The correlation analysis showed that the daily CO2 emission rate of soils with high microbial biomass carbon was significantly positively correlated with glucosidase activity under dry-wet treatment and significantly negatively correlated with acetylglucosaminidase (NAG) and peroxidase activities under continuous flooding treatment. In the low microbial biomass carbon soils, the daily CO2 emission rate of the continuous flooding treatment was negatively correlated with the NAG activity, but showed no correlation with enzyme activities under dry-wet management. In summary, the cumulative CO2 emissions of dry-wet treatment were lower than those of continuous flooding treatment, and the difference was significant in soils with low microbial biomass carbon. The size of the soil microbial biomass determined the level of the stable soil organic carbon mineralization rate. The amount of soluble organic carbon and iron reduction affected the soil CO2 emissions under continuous flooding conditions, and the soil water conditions affected the daily CO2 emission rate and its key influencing enzymes. This study provides data and theoretical support for the carbon cycle and carbon sequestration potential in paddy soil.

[Effect of Water Management on Rice Growth and Rhizosphere Priming Effect in Paddy Soils].

The rhizosphere priming effect (RPE) caused by carbon inputs from crop rhizodeposits plays a key role in regulating the carbon emission flux and carbon balance of farmland soils. Due to frequent alternations between dry and wet conditions, CO2 and CH4 emissions and the RPE in paddy field ecosystems are significantly different to those of other ecosystems. Therefore, it is of great significance to determine the direction and intensity of the rice RPE under alternations of dry and wet to limit greenhouse gas emissions. In this study, using a 13C-CO2 continuous labeling method combined with a pot-based experiment, the response of rice growth and the RPE under alternating dry and wet and continuous flooding conditions was examined. The results showed that, compared with the continuous flooding treatment, the alternating dry and wet treatments significantly increased aboveground and root biomass and the root-to-root ratio, and also increased soil microbial biomass. Under continuous flooding conditions, fluxes of 13CO2 and 13CH4 increased with rice growth from 10.2 µg·(kg·h)-1 and 2.8 µg·(kg·h)-1 (63 d) to 16.0 µg·(kg·h)-1 and 3.2 µg·(kg·h)-1 (75 d), respectively. During the 12-day drying process, the emissions of 13CO2 and 13CH4 derived from rhizosphere deposited C decreased by 57.5% and 88.1%. Under continuous flooding conditions, the RPE for CO2 and CH4 were positive and increased with the growth of rice. Under the alternating dry and wet treatment, after 12 days of drying, the RPE for CO2 and CH4 was reduced from 0.29 mg·(kg·h)-1 and 12.3 µg·(kg·h)-1 (63 d) to -0.39 mg·(kg·h)-1 and 0.07 µg·(kg·h)-1 (75 d). Thus, alternating wet and dry treatment can effectively promote rice growth and reduce the cumulative emissions of CH4. Therefore, adopting appropriate field water management is of great significance for increasing rice yields and mitigating greenhouse gas emissions.

Metagenomic and 14 C tracing evidence for autotrophic microbial CO2 fixation in paddy soils.

Autotrophic carbon dioxide (CO2 ) fixation by microbes is ubiquitous in the environment and potentially contributes to the soil organic carbon (SOC) pool. However, the multiple autotrophic pathways of microbial carbon assimilation and fixation in paddy soils remain poorly characterized. In this study, we combine metagenomic analysis with 14 C-labelling to investigate all known autotrophic pathways and CO2 assimilation mechanisms in five typical paddy soils from southern China. Marker genes of six autotrophic pathways are detected in all soil samples, which are dominated by the cbbL genes (67%-82%) coding the ribulose-bisphosphate carboxylase large chain in the Calvin cycle. These marker genes are associated with a broad range of phototrophic and chemotrophic genera. Significant amounts of 14 C-CO2 are assimilated into SOC (74.3-175.8 mg 14 C kg-1 ) and microbial biomass (5.2-24.1 mg 14 C kg-1 ) after 45 days incubation, where more than 70% of 14 C-SOC was concentrated in the relatively stable humin fractions. These results show that paddy soil microbes contain the genetic potential for autotrophic carbon fixation spreading over broad taxonomic ranges, and can incorporate atmospheric carbon into organic components, which ultimately contribute to the stable SOC pool.

Unremitting progresses for phosphoprotein synthesis.

Phosphorylation, one of the important protein post-translational modifications, is involved in many essential cellular processes. Site-specifical and homogeneous phosphoproteins can be used as probes for elucidating the protein phosphorylation network and as potential therapeutics for interfering their involved biological events. However, the generation of phosphoproteins has been challenging owing to the limitation of chemical synthesis and protein expression systems. Despite the pioneering discoveries in phosphoprotein synthesis, over the past decade, great progresses in this field have also been made to promote the biofunctional exploration of protein phosphorylation largely. Therefore, in this review, we mainly summarize recent advances in phosphoprotein synthesis, which includes five sections: 1) synthesis of the nonhydrolyzable phosphorylated amino acid mimetic building blocks, 2) chemical total and semisynthesis strategy, 3) in-cell and in vitro genetic code expansion strategy, 4) the late-stage modification strategy, 5) nonoxygen phosphoprotein synthesis.

[Response of Extracellular Enzyme Activities to Substrate Availability in Paddy Soil with Long-term Fertilizer Management].

The availability of carbon (C), nitrogen (N), and other substrates in soil determines the growth and metabolism of microorganisms and affects the activity of extracellular enzymes. To study the activities of ß-1,4-glucosidase (BG) and ß-1,4-N-acetylglucosaminidase (NAG) in response to C and N availability, samples that underwent four treatments-non-fertilization (CK), chemical fertilizer (NPK), combination of organic manure and chemical fertilizer (OM), and mixture of straw and chemical fertilizer (ST)-were collected from long-term fertilization paddy soil and incubated for 0, 4, 8, and 12 months to obtain soil with different C and N availability gradients. The results showed that the dissolved organic carbon(DOC) content of OM and ST treatment samples was 2-3 times higher than that of CK and NPK treatment samples. With the increase of DOC and ammonium (NH4+-N) contents, the activities of BG and NAG and the contents of microbial biomass C (MBC) and N (MBN) showed no increase during incubation within each treatment. Fertilization treatments, incubation time, and their interaction are crucial factors varying the contents of DOC, NH4+-N, MBC, and MBN among different fertilization treatments (P<0.01). There was a positive correlation between MBC/MBN and DOC/NH4+-N of OM treatment (P<0.05) and a negative relationship between ln(BG)/ln(NAG) and DOC/NH4+-N of ST treatment (P<0.01), indicating that the availability of substrates played a key role in the potential activity of extracellular enzymes in paddy soil, and the carbon-nitrogen ratio of microbial biomass was controlled by the C/N stoichiometry of substrates in soil. The results have a certain guiding significance for further study on the variation of extracellular enzyme activity in paddy soil, regulating the balance of carbon and nitrogen, and improving the fertility of paddy soil.

Antibacterial Peptide BSN-37 Kills Extra- and Intra-Cellular Salmonella enterica Serovar Typhimurium by a Nonlytic Mode of Action.

The increasing rates of resistance to traditional anti-Salmonella agents have made the treatment of invasive salmonellosis more problematic, which necessitates the search for new antimicrobial compounds. In this study, the action mode of BSN-37, a novel antibacterial peptide (AMP) from bovine spleen neutrophils, was investigated against Salmonella enterica serovar Typhimurium (S. Typhimurium). Minimum inhibitory concentrations (MICs) and time-kill kinetics of BSN-37 were determined. The cell membrane changes of S. Typhimurium CVCC541 (ST) treated with BSN-37 were investigated by testing the fluorescence intensity of membrane probes and the release of cytoplasmic ß-galactosidase activity. Likewise, cell morphological and ultrastructural changes were also observed using scanning and transmission electron microscopes. Furthermore, the cytotoxicity of BSN-37 was detected by a CCK-8 kit and real-time cell assay. The proliferation inhibition of BSN-37 against intracellular S. Typhimurium was performed in Madin-Darby canine kidney (MDCK) cells. The results demonstrated that BSN-37 exhibited strong antibacterial activity against ST (MICs, 16.67 µg/ml), which was not remarkably affected by the serum salts at a physiological concentration. However, the presence of CaCl2 led to an increase in MIC of BSN-37 by about 4-fold compared to that of ST. BSN-37 at the concentration of 100 µg/ml could completely kill ST after co-incubation for 6 h. Likewise, BSN-37 at different concentrations (50, 100, and 200 µg/ml) could increase the outer membrane permeability of ST but not impair its inner membrane integrity. Moreover, no broken and ruptured cells were found in the figures of scanning and transmission electron microscopes. These results demonstrate that BSN-37 exerts its antibacterial activity against S. Typhimurium by a non-lytic mode of action. Importantly, BSN-37 had no toxicity to the tested eukaryotic cells, even at a concentration of 800 µg/ml. BSN-37 could significantly inhibit the proliferation of intracellular S. Typhimurium.

[Allocation and Stabilization Responses of Rice Photosynthetic Carbon in the Plant-Soil System to Phosphorus Application].

This research studied the response of the input and allocation of photosynthetic carbon (C) to phosphorus (P) in paddy soils. Two treatments were conducted in this experiment:no P application (P0) and the application of 80 mg·kg-1 of P (P80). The rice cultivar was the indica Zhongzao 39. The 13C-CO2 continuous labeling technique was used to identify the photosynthetic C distribution of the rice. The results showed that the application of P80 significantly increased the photosynthates allocation in the rice aboveground, but reduced their allocation in the rhizosphere soil (P<0.05). At the jointing stage, P80 application increased the photosynthetic C content of the rice by 70%, but the root dry weight decreased 31%. Compared with P0, the total C content of the aboveground rice was increased 0.31 g·pot-1 by P80. The ratio of rice roots to shoots decreased with the P80 treatment. Moreover, P80 application led to an increase in the photosynthetic microbial biomass in the non-rhizosphere soil C (13C-MBC) of 0.03 mg·kg-1, but still decreased its allocation in the rhizosphere soil. The allocation of photosynthetic C to the particulate organic matter fraction (POC) and mineral fraction (MOC) in the non-rhizosphere soil showed no significant differences between P0 and P80. Additionally, the P80 fertilization treatment significantly lowered the content of POC in the rhizosphere soil. In summary, P application increased the allocation of photosynthetic C in the soil-rice system, but reduced the accumulation of photosynthetic C in the soil. This research provided a theoretical basis and data supporting the rational application of P fertilizer, and was also of great significance as a study of the transportation and allocation of photosynthetic C and its sequestration potential response to the application of P to the rice soil.

[Characterization of Soil Organic Carbon Mineralization Under Different Gradient Carbon Loading in Paddy Soil].

Available carbon is the most active part of the soil carbon pool. It is also the main carbon source of soil microbes and plays an important role in the processes of soil organic carbon mineralization and accumulation. However, the mechanisms are still not clear how soil organic carbon mineralization and its priming effect (PE) are affected by different input levels of readily available carbon, based on the growth requirements of microbes in paddy soil. In this study, an incubation experiment was conducted by adding different levels (0.5, 1, 3, and 5 times of MBC) of exogenous source organic carbon (13C-glucose) to the soil. The mineralization dynamics of labile organic carbon and its priming effect was investigated. The mineralization rate of glucose-C increased significantly with the increasing carbon loading level. The distribution of glucose-C into rapid and slow C pools was also exponentially correlated with the carbon loading (R2=0.99, P<0.05 and R2=0.99, P<0.05, respectively). Negative PE was observed at high carbon loading (3×MBC and 5×MBC); while positive PE was induced by low carbon loading (0.5×MBC and 1×MBC). The cumulative PE was 160.0 mg·kg-1 and 325.1 mg·kg-1, respectively, at the end of the incubation. Redundancy analysis showed that the main factors affecting the cumulative PE were MBC, MBN, and DOC at the initial glucose mineralization stage, while ß-glucosidase, chitinase, and ammonium nitrogen were the main factors at later stages. Therefore, the readily available carbon loading has an important effect on the organic carbon mineralization and PE in paddy soil. Higher carbon loading was good for the accumulation of organic carbon sequestration in paddy soil. This study is of great scientific significance for revealing the activity of organic carbon in paddy fields and for its contribution to the development of sustainable agriculture.

Identification of a Novel Bcl-2 Inhibitor by Ligand-Based Screening and Investigation of Its Anti-cancer Effect on Human Breast Cancer Cells.

Bcl-2 family protein is an important factor in regulating apoptosis and is associated with cancer. The anti-apoptotic proteins of Bcl-2 family, such as Bcl-2, are overexpression in numerous tumors, and contribute to cancer formation, development, and therapy resistance. Therefore, Bcl-2 is a promising target for drug development, and several Bcl-2 inhibitors are currently undergoing clinical trials. In this study, we carried out a QSAR-based virtual screening approach to develop potential Bcl-2 inhibitors from the SPECS database. Surface plasmon resonance (SPR) binding assay was performed to examine the interaction between Bcl-2 protein and the screened inhibitors. After that, we measured the anti-tumor activities of the 8 candidate compounds, and found that compound M1 has significant cytotoxic effect on breast cancer cells. We further proved that compound M1 downregulated Bcl-2 expression and activated apoptosis by inducing mitochondrial dysfunction. In conclusion, we identified a novel Bcl-2 inhibitor by QSAR screening, which exerted significant cytotoxic activity in breast cancer cells through inducing mitochondria-mediated apoptosis.

[Transformation and Distribution of Soil Organic Carbon and the Microbial Characteristics in Response to Different Exogenous Carbon Input Levels in Paddy Soil].

The turnover of soil organic carbon (SOC) and the activity of soil microbes can be influenced by exogenous carbon. However, microbial response characteristics of the transformation and distribution of available organic carbon under different levels remain unclear in paddy soils. 13C-labeled glucose was used as a typical available exogenous carbon to simulate indoor culture experiments added at different levels of soil microbial biomass carbon (MBC) (0×MBC, 0.5×MBC, 1×MBC, 3×MBC, and 5×MBC) to reveal the process of C-transformation and distribution. The characteristics of microbial response in the process of exogenous carbon turnover was also monitored. The 96-well microplate fluorescence analysis was adopted to determine the activities of cellobiose hydrolase (CBH) and ß-glucosidase (ß-Glu). The results showed that, in 2 d of incubation, the ratio of labeled glucose carbon to dissolved organic carbon (13C-DOC/DOC) or to SOC (13C-SOC/SOC) was positively correlated with the amount of glucose added. The incorporation of glucose C (13C) into MBC reached the highest value (18.96 mg·kg-1) at 3×MBC treatment but decreased thereafter. The 13C allocation rate was mainly positively correlated with MBC, Olsen-P, and DOC. At 60 d, 13C-DOC, 13C-MBC, and 13C-SOC decreased significantly to less than 0.02 mg·kg-1, 2 mg·kg-1, and 10 mg·kg-1 in soil, and it was positively correlated with the amount of glucose added. Compared with CK, CBH enzyme activity increased significantly after the addition of glucose, and for the 3×MBC treatment it was increased by 22.6 times, which was significantly higher than those of other treatments (P<0.05). However, ß-Glu enzyme activity increased only in the 3×MBC and 5×MBC treatments, wherein it decreased with increasing amounts of added glucose. NH4+-N, pH, ß-Glu, and CBH were the primary factors affecting the distribution rate of 13C. In conclusion, the conversion of exogenous carbon to SOC increased with increased amounts of added organic carbon. This changed the activity of soil enzymes; however, microbial utilization of exogenous carbon may have a saturation threshold. Within the saturation threshold, the conversion rate of organic matter was directly proportional to the amount of added organic matter. When the saturation threshold was exceeded, the conversion rate of organic matter decreased. Therefore, the appropriate addition of exogenous carbon is beneficial, as it can increase SOC in rice fields and improve the quality of the crop growth environment.

Three-Level Hepatotoxicity Prediction System Based on Adverse Hepatic Effects.

Hepatotoxicity is a major cause of drug withdrawal from the market. To reduce the drug attrition induced by hepatotoxicity, an accurate and efficient hepatotoxicity prediction system must be constructed. In the present study, we constructed a three-level hepatotoxicity prediction system based on different levels of adverse hepatic effects (AHEs) combined with machine learning, using (1) an end point, hepatotoxicity; (2) four hepatotoxicity severity degrees; and (3) specific AHEs. After collecting and curing 15 873 compound-AHE pairs associated with 2017 compounds and 403 AHEs, we constructed 27 models with three end point levels with the random forest algorithm, and obtained accuracies ranging from 67.0 to 78.2% and the area under receiver operating characteristic curves (AUCs) of 0.715-0.875. The 27 models were fully integrated into a tiered hepatotoxicity prediction system. The existence of hepatotoxicity existence, severity degree, and potential AHEs for a given compound could be inferred simultaneously and systematically. Thus, the tiered hepatotoxicity prediction system allows researchers to have significant confidence in confirming compound hepatotoxicity, analyzing hepatotoxicity from multiple perspectives, obtaining warnings for the potential hepatotoxicity severity, and even rapidly selecting the proper in vitro experiments for hepatotoxicity verification. We also applied three external sets (11 drugs or candidates that failed in clinical trials or were withdrawn from the market, the PharmGKB (offsides) database, and an herbal hepatotoxicity data set) to test and validate the prediction ability of our system. Furthermore, the hepatotoxicity prediction system was adapted into a flow framework based on the Konstanz Information Miner, which was made available for researchers.

[Dynamics of Rice Photosynthesized Carbon Input and Its Response to Nitrogen Fertilization at the Jointing Stage:13 C-CO2 Pulse-labeling].

Photosynthesized carbon (C) is an important source of soil organic C in paddy fields, and its input and distribution are affected by rice growth and soil fertility. Fertilizer application plays an important role in rice growth. The 13C pulse-labeling method was used to quantify the dynamics and distribution of input photosynthesized C in the rice-(rhizosphere-and bulk-) soil system and its response to nitrogen fertilizer (N) application. The results suggested that N fertilization significantly increased the rice aboveground and the root biomass and decreased the rice biomass root/shoot ratio. The amount of assimilated 13C gradually decreased in the rice plants but gradually decreased over 0-6 days and increased over 6-26 days in the rhizosphere and bulk soil during rice growth. N fertilization significantly increased the amount of assimilated 13C in the rhizosphere soil by 9.5%-32.6% compared with the control. In comparison to the unfertilized treatment, the application of N fertilization resulted in higher photosynthetic13C in rice aboveground and in the root by 24.5%-134.7% and 9.1%-106%, respectively. With the N fertilized and unfertilized treatments, 85.5%-93.2% and 91.3%-95.7%, respectively, of input photosynthetic 13C was distributed in the rice plants. The results suggested that N fertilization significantly affected the distribution of photosynthesized C in the rice-soil system (P<0.01). After 26 days of pulse labeling, the distribution of photosynthetic 13C into rice aboveground was increased by 13.4%, while the distribution into the rhizosphere and bulk soil were decreased by 21.9% and 52.2%, respectively, in the N fertilized treatments compared with the unfertilized treatments. Therefore, the N application increased the distribution of photosynthesized carbon in the soil-rice system but decreased the accumulation in the rhizosphere and bulk soil. The findings of this study provided a theoretical basis for our understanding of the dynamic of photosynthetic C in the plant-soil system and the assimilation of the soil organic matter pool in the paddy soil ecosystem.

[Effects of Long-term Fertilization on Enzyme Activities in Profile of Paddy Soil Profiles].

The enzyme activity, which is closely related to soil material cycling (mineralization, transformation, etc.), can reflect soil quality and nutrient status. In order to explore the effect of long-term fertilization on the enzyme activity in paddy soil profile (0-40 cm), soils with organic fertilizer and inorganic fertilizer, and non-fertilized soils were selected, and the carbon and nitrogen contents, and the activities of ß-1,4-glucosidase (BG), and ß-1,4-N-acetylglucosaminidase (NAG) in 10cm depths of soil were analyzed. The results showed that the activities of BG and NAG in the soils treated with inorganic fertilizer and organic fertilizer increased by 0.73-47.87 nmol·(g·h)-1 and 1.33-128.81 nmol·(g·h)-1, and 0.19-9.72 nmol·(g·h)-1 and 0.92-57.66 nmol·(g·h)-1, respectively, compared to those for non-fertilized soil. Soil enzyme activity decreased with increasing soil depth. Soil enzyme activity in soil from 0-20 cm was significantly higher than that of soil from 20-40 cm. Soil enzyme activities were significantly affected by long term fertilization at different soil depths. RDA analysis showed that soil carbon and nitrogen contents had significant positive relationships with the activities of BG and NAG in the 0-20 cm soil profiles, however, negative relationships were observed in the 20-40 cm soil profiles. The long-term application of organic fertilizer significantly increased soil biomass and enzyme activity, both of which decreased with the increase in soil depth. Long-term fertilization could increase soil nutrient contents, microbial biomass, and extracellular enzyme activities, which has important theoretical significance for optimizing farmland fertilizer management and improving soil productivity.

[Allocation of rice photosynthates in plant-soil system in response to elevated CO2 and nitrogen fertilization.] / æ°´ç¨»å ‰åˆç¢³åœ¨æ¤ç‰©-åœŸå£¤ç³»ç»Ÿä¸­çš„åˆ†é åŠå ¶å¯¹CO2升高和施氮的响应.

To examine the allocation of rice photosynthates and its response to the elevated CO2 (800 µL·L-1) and N fertilization (100 mg·kg-1) at both tillering stage and booting stage in plant-soil system, rice was continually labelled with 13CO2. The results showed that the rice root biomass at the tillering stage and the shoot biomass at the booting stage were significantly increased under elevated CO2. Elevated CO2 increased the rice biomass and root-shoot ratio at tillering stage, but reduced it at booting stage. Under elevated CO2, N fertilization promoted shoot biomass during rice growth, but significantly decreased the root biomass at booting stage. Elevated CO2 significantly increased the allocation of assimilated 13C to the soil at the booting stage. N fertilization did not promote the elevated CO2-induced stimulation of assimilated 13C allocated to the soil, and it even decreased the proportion of assimilated 13C in the soil. In summary, elevated CO2 increased the photosynthetic C allocation into soil and promoted the turnover of soil organic carbon in paddy soil. N fertilization enhanced rice shoot biomass but decreased the belowground allocation of photosynthetic C.

[Responses of organic carbon mineralization and priming effect to phosphorus addition in paddy soils]. / ç¨»ç”°åœŸå£¤æœ‰æœºç¢³çŸ¿åŒ–åŠå ¶æ¿€å‘æ•ˆåº”å¯¹ç£·æ·»åŠ çš„å“åº”.

To understand the coupled controlling of carbon (C) and phosphorus (P) on the minera-lization of soil organic carbon and amended substrates in paddy soil, we investigated the effects of P addition on the decomposition of organic carbon and its induced priming effect by using 13C isotope probing technique in microcosm. The results showed that P addition accelerated the release of CO2 but inhibited the release of CH4, leading to 53.1% reduction of total accumulated CH4 and 70.5% reduction of the 13CH4 derived from exotic glucose-13C. P addition altered the carbon distribution during the microbial turnover progress, with 3.6% of glucose-13C being transferred into the labile carbon pool, therein significantly increased potential of the mineralization rate of exogenous C. A transient negative priming effect was observed in the early stage of incubation. With time prolonging, the priming effect on CO2 emission (PECO2) generally increased and then decreased after a peak. The priming effect on CH4 emission (PECH4) kept increasing and finally fluctuated at a relative stable value until the end of the experiment (100 days). P addition increased PECO2 by 32.3% but reduced PECH4 by 93.4%. Results from the RDA and Pearson analysis showed that electric conductivity, oxidation-reduction potential and dissolved organic carbon significantly affected soil C mineralization. There were significantly negative correlations between available phosphorus (Olsen-P) and 13CH4, and between Olsen-P and PECH4. In conclusion, with the addition of exogenous organic matter, P application could reduce CH4 emissions and inhibit its priming effect, acce-lerate the mineralization of SOC, probably improve the nutrient supply, and thus enhance the avai-lability of organic C and promote C cycling in paddy soil.

[Effect of Phosphorus Addition on the Abundance of Autotrophic CO2-Fixation Microorganisms in Rhizospheric Soil from a Phosphorus-Limited Paddy Field].

A rice pot experiment was conducted to investigate the effect of phosphorus addition on the abundance of autotrophic CO2-fixation microorganisms using phosphorus-limited paddy soil from the Changsha Observation and Research Station for the Agricultural Environment. Rice seedlings were transplanted in the paddy soil with or without phosphorus addition, corresponding to P-treated-pot (P) or control pot (CK), respectively. Rhizosphere soils were collected from the P and CK treatments during the tillering and shooting stages. The physical and chemical soil properties were measured and the abundance of autotrophic CO2-fixation microorganisms was quantified with a real-time PCR technique based on four functional genes (cbbL, cbbM, accA, and aclB) involved in three CO2-fixation pathways (CBB cycle, rTCA cycle, and 3-hydroxypropionate/4-hydroxybutyrate cycle). The results show that phosphorus addition improves the concentrations of DOC and Olsen-P and the pH value, whereas negative effects on the MBC and NH4+-N concentrations are revealed during the tillering stage. The effect of phosphorus addition on the NO3--N concentration in the tillering and shooting stages differs. Phosphorus addition significantly increases the abundances of the cbbL, cbbM, accA, and aclB genes, which are 156%, 99%, 110%, and 193% higher than those of the CK treatment in the tillering stage. However, this positive effect is not notable for the cbbL, accA, and aclB genes during the shooting stage. Redundancy analysis (RDA) shows that Olsen-P is the environmental factor that most significantly affects the abundance of autotrophic CO2-fixation microorganisms.