Electrocatalysis-Enabled Defluorinative Cross-Coupling of gem-Difluoroalkenes with Aldehydes and Ketones.

An electrochemical defluorinative cross-coupling of gem-difluoroalkenes with carbonyl compounds was described, by which highly stereoselective monofluoroalkene allyl alcohols were synthesized. The reaction tolerates a broad range of functional groups and has successfully been applied to synthesize complex molecules. Mechanistic studies indicate that the reaction starts from electron reduction of gem-difluoroalkenes to generate radical negative ions, which undergo ß-fluoride elimination and subsequent reduction to form anions. These anions are subsequently trapped by carbonyl compounds to furnish target products.

Invasive Spartina alterniflora accelerates the increase in microbial nitrogen fixation over nitrogen removal in coastal wetlands of China.

Salt marsh plants play a vital role in mediating nitrogen (N) biogeochemical cycle in estuarine and coastal ecosystems. However, the effects of invasive Spartina alterniflora on N fixation and removal, as well as how these two processes balance to determine the N budget, remain unclear. Here, simultaneous quantifications of N fixation and removal via 15N tracing experiment with native Phragmites australis, invasive S. alterniflora, and bare flats as well as corresponding functional gene abundance by qPCR were carried out to explore the response of N dynamics to S. alterniflora invasion. Our results showed that N fixation and removal rates ranged from 0.77 ± 0.08 to 16.12 ± 1.13 nmol/(g·h) and from 1.42 ± 0.14 to 16.35 ± 1.10 nmol/(g·h), respectively, and invasive S. alterniflora generally facilitated the two processes rates. Based on the difference between N removal and fixation rates, net N2 fluxes were estimated in the range of -0.39 ± 0.14 to 8.24 ± 2.23 nmol/(g·h). Estimated net N2 fluxes in S. alterniflora stands were lower than those in bare flats and P. australis stands, indicating that the increase in N removal caused by S. alterniflora invasion may be more than offset by N fixation process. Random forest analysis revealed that functional microorganisms were the most important factor associated with the corresponding N transformation process. Overall, our results highlight the importance of N fixation in evaluating N budget of estuarine and coastal wetlands, providing valuable insights into the ecological effect of S. alterniflora invasion.

Flagellar motors of swimming bacteria contain an incomplete set of stator units to ensure robust motility.

Flagellated bacteria, like Escherichia coli, swim by rotating helical flagellar filaments powered by rotary flagellar motors at their base. Motor dynamics are sensitive to the load it drives. It was previously thought that motor load was high when driving filament rotation in free liquid environments. However, torque measurements from swimming bacteria revealed substantially lower values compared to single-motor studies. We addressed this inconsistency through motor resurrection experiments, abruptly attaching a 1-micrometer-diameter bead to the filament to ensure high load. Unexpectedly, we found that the motor works with only half the complement of stator units when driving filament rotation. This suggests that the motor is not under high load during bacterial swimming, which we confirmed by measuring the torque-speed relationship by varying media viscosity. Therefore, the motor operates in an intermediate-load region, adaptively regulating its stator number on the basis of external load conditions. This ensures the robustness of bacterial motility when swimming in diverse load conditions and varying flagella numbers.

Active pathways of anaerobic methane oxidization in deep-sea cold seeps of the South China Sea.

IMPORTANCE: Cold seeps occur in continental margins worldwide and are deep-sea oases. Anaerobic oxidation of methane is an important microbial process in the cold seeps and plays an important role in regulating methane content. This study elucidates the diversity and potential activities of major microbial groups in dependent anaerobic methane oxidation and sulfate-dependent anaerobic methane oxidation processes and provides direct evidence for the occurrence of nitrate-/nitrite-dependent anaerobic methane oxidation (Nr-/N-DAMO) as a previously overlooked microbial methane sink in the hydrate-bearing sediments of the South China Sea. This study provides direct evidence for occurrence of Nr-/N-DAMO as an important methane sink in the deep-sea cold seeps.

Integrative Transcriptomics and Proteomics Analysis Reveals Immune Response Process in Bovine Viral Diarrhea Virus-1-Infected Peripheral Blood Mononuclear Cells.

Bovine viral diarrhea virus (BVDV) causes bovine viral diarrhea-mucosal disease, inflicting substantial economic losses upon the global cattle industry. Peripheral blood mononuclear cells (PBMCs) are the central hub for immune responses during host-virus infection and have been recognized as crucial targets for BVDV infection. In order to elucidate the dynamics of host-BVDV-1 interaction, this study harnessed RNA-seq and iTRAQ methods to acquire an extensive dataset of transcriptomics and proteomics data from samples of BVDV-1-infected PBMCs at the 12-h post-infection mark. When compared to mock-infected PBMCs, we identified 344 differentially expressed genes (DEGs: a total of 234 genes with downregulated expression and 110 genes with upregulated expression) and 446 differentially expressed proteins (DEPs: a total of 224 proteins with downregulated expression and 222 proteins with upregulated expression). Selected DEGs and DEPs were validated through quantitative reverse transcriptase-polymerase chain reaction and parallel reaction monitoring. Gene ontology annotation and KEGG enrichment analysis underscored the significant enrichment of DEGs and DEPs in various immunity-related signaling pathways, including antigen processing and presentation, complement and coagulation cascades, cytokine-cytokine receptor interaction, and the NOD-like receptor signaling pathway, among others. Further analysis unveiled that those DEGs and DEPs with downregulated expression were predominantly associated with pathways such as complement and coagulation cascades, the interleukin-17 signaling pathway, cytokine-cytokine receptor interaction, the PI3K-Akt signaling pathway, the tumor necrosis factor signaling pathway, and the NOD-like receptor signaling pathway. Conversely, upregulated DEGs and DEPs were chiefly linked to metabolic pathways, oxidative phosphorylation, complement and coagulation cascades, and the RIG-I-like receptor signaling pathway. These altered genes and proteins shed light on the intense host-virus conflict within the immune realm. Our transcriptomics and proteomics data constitute a significant foundation for delving further into the interaction mechanism between BVDV and its host.

Tidal dynamics regulates potential coupling of carbon­nitrogen­sulfur cycling microbes in intertidal flats.

Tide-driven hydrodynamic process causes significant geochemical gradients that influence biogeochemical cycling and ecological functioning of estuarine and coastal ecosystems. However, the effects of tidal dynamics on microbial communities, particularly at the functional gene level, remain unclear even though microorganisms play critical roles in biogeochemical carbon (C), nitrogen (N) and sulfur (S) cycling. Here, we used 16S rRNA gene amplicon sequencing and microarray-based approach to reveal the stratification of microorganisms related to C, N and S cycles along vertical redox gradients in intertidal wetlands. Alpha-diversity of bacteria and archaea was generally higher at the deep groundwater-sediment interface. Microbial compositions were markedly altered along the sediment profile, and these shifts were largely due to changes in nutrient availability and redox potential. Furthermore, functional genes exhibited redox partitioning between interfaces and transition layer, with abundant genes involved in C decomposition, methanogenesis, heterotrophic denitrification, sulfite reduction and sulfide oxidation existed in the middle anoxic zone. The influence of tidal dynamics on sediment function was highly associated with redox state, sediment texture, and substrates availability, leading to distinct distribution pattern of metabolic coupling of microbes involved in energy flux and elemental cycling in intertidal wetlands. These results indicate that tidal cycles are critical in determining microbial community and functional structure, and they provide new insights into sediment microbe-mediated biogeochemical cycling in intertidal habitats.

Non-native Brachiaria humidicola with biological nitrification inhibition capacity stimulates in situ grassland N2O emissions.

Introduction: Brachiaria humidicola, a tropical grass, could release root exudates with biological nitrification inhibition (BNI) capacity and reduce soil nitrous oxide (N2O) emissions from grasslands. However, evidence of the reduction effect in situ in tropical grasslands in China is lacking. Methods: To evaluate the potential effects of B. humidicola on soil N2O emissions, a 2-year (2015-2017) field experiment was established in a Latosol and included eight treatments, consisting of two pastures, non-native B. humidicola and a native grass, Eremochloa ophiuroide, with four nitrogen (N) application rates. The annual urea application rates were 0, 150, 300, and 450 kg N ha-1. Results: The average 2-year E. ophiuroides biomass with and without N fertilization were 9.07-11.45 and 7.34 t ha-1, respectively, and corresponding values for B. humidicola increased to 31.97-39.07 and 29.54 t ha-1, respectively. The N-use efficiencies under E. ophiuroide and B. humidicola cultivation were 9.3-12.0 and 35.5-39.4%, respectively. Annual N2O emissions in the E. ophiuroides and B. humidicola fields were 1.37 and 2.83 kg N2O-N ha-1, respectively, under no N fertilization, and 1.54-3.46 and 4.30-7.19 kg N2O-N ha-1, respectively, under N fertilization. Discussions: According to the results, B. humidicola cultivation increased soil N2O emissions, especially under N fertilization. This is because B. humidicola exhibited the more effective stimulation effect on N2O production via denitrification primarily due to increased soil organic carbon and exudates than the inhibition effect on N2O production via autotrophic nitrification. Annual yield-scaled N2O emissions in the B. humidicola treatment were 93.02-183.12 mg N2O-N kg-1 biomass, which were significantly lower than those in the E. ophiuroides treatment. Overall, our results suggest that cultivation of the non-native grass, B. humidicola with BNI capacity, increased soil N2O emissions, while decreasing yield-scaled N2O emissions, when compared with native grass cultivation.

Spatiotemporal Dynamics of Bacterial Taxonomic and Functional Profiles in Estuarine Intertidal Soils of China Coastal Zone.

Bacteria play an important role in regulating carbon (C), nitrogen (N), and sulfur (S) in estuarine intertidal wetlands. To gain insights into the ecological and metabolic modes possessed by bacteria in estuarine intertidal wetlands, a total of 78 surface soil samples were collected from China's coastal intertidal wetlands to examine the spatial and seasonal variations of bacterial taxonomic composition, assembly processes, and ecological system functions through shotgun metagenomic and 16S rRNA gene sequencing. Obvious spatiotemporal dynamic patterns in the bacterial community structure were identified, with more pronounced seasonal rather than spatial variations. Dispersion limitation was observed to act as a critical factor affecting community assembly, explaining approximately half of the total variation in the bacterial community. Functional bacterial community structure exhibited a more significant latitudinal change than seasonal variability, highlighting that functional stability of the bacterial communities differed with their taxonomic variability. Identification of biogeochemically related links between C, N, and S cycles in the soils showed the adaptive routed metabolism of the bacterial communities and the strong interactions between coupled metabolic pathways. Our study broadens the insights into the taxonomic and functional profiles of bacteria in China's estuarine intertidal soils and helps us understand the effects exerted by environmental factors on the ecological health and microbial diversity of estuarine intertidal flats.

Effects of periodic drying-wetting on microbial dynamics and activity of nitrite/nitrate-dependent anaerobic methane oxidizers in intertidal wetland sediments.

Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) plays an important role in methane (CH4) consumption in intertidal wetlands. However, little is known about the responses of n-DAMO in intertidal wetlands to periodic drying-wetting caused by tidal cycling. Here, comparative experiments (waterlogged, desiccated, reflooded) with the Yangtze estuarine intertidal sediments were performed to examine the effects of periodic tidal changes on n-DAMO microbial communities, abundances, and potential activities. Functional gene sequencing indicated the coexistence of n-DAMO bacteria and archaea in the tide-fluctuating environments and generally higher biodiversity under reflooded conditions than consecutive inundation or emersion. The n-DAMO microbial abundance and associated activity varied significantly during alternative exposure and inundation, with higher abundance and activity under the waterlogged than desiccated conditions. Reflooding of intertidal wetlands might intensify n-DAMO activities, indicating the resilience of n-DAMO microbial metabolisms to the wetting-drying events. Structural equation modeling and correlation analysis showed that n-DAMO activity was highly related to n-DAMO microbial abundance and substrate availability under inundation, whereas salt accumulation in sediment was the primary factor restraining n-DAMO activity under the desiccation. Overall, this study reveals tidal-induced shifts of n-DAMO activity and associated contribution to mitigating CH4, which may help accurately project CH4 emission from intertidal wetlands under different tidal scenarios.

High level expression of recombinant human serum albumin in CHO cells and optimization of its culture technology / 中国生物制品学杂志

@#ObjectiveTo achieve efficient expression of human serum albumin(HSA)in Chinese hamster ovary(CHO)cells and optimize its culture technology,so as to lay a foundation of the large-scale production of HSA.MethodsThe eukaryotic expression vector of HSA was constructed by gene recombination technology,and then electrotransfected into fully suspended CHO cells. The monoclonal cell lines with stable and high expression of HSA were screened by G418 and limited dilution method. By adding glucose,sodium butyrate and supplementalmedium to the basal medium,the cell culture process was optimized to improve the expression of HSA. Finally,the scale-up culture verification was carried out in a 5 L bioreactor.ResultsThe recombinant expression vector pcDNA3.1-HSA was successfully constructed and expressed in fully suspended CHO cells. After two monoclonal screening,the secondary monoclonal cell lines CHO-rHSA-7H2A9 and CHOrHSA-7H2D12 were obtained with high HSA expression of 29. 37 mg/L and 25. 26 mg/L respectively. The HSA expression level reached about 100. 00 mg/L by optimizing the culture process and wasfinally increased to 166. 16 mg/L in the 5 L bioreactor,which was about 30 times higher than that in the supernatant of the first monoclonal cells.Conclusion The high level expression of HSA in CHO cells was achieved,which laid a foundation of the further large-scale production of HSA in the field of biological products and solving the market supply problems.

HSP27 Attenuates cGAS-Mediated IFN-ß Signaling through Ubiquitination of cGAS and Promotes PRV Infection.

Pseudorabies (PR) is a domestic and wild animal infectious disease caused by the pseudorabies virus (PRV) and is one of the major infectious diseases that endanger the global swine industry. Studies have reported that PRV may achieve cross-species transmission from pigs to humans in recent years. Therefore, in-depth exploration of the relationship between PRV and host proteins is of great significance for elucidating the pathogenic mechanism of PRV and anti-PRV infection. Here, we report that heat shock protein 27 (HSP27) ubiquitinates and degrades cyclic GMP-AMP synthase (cGAS) and attenuates cGAS-mediated antiviral responses, thereby promoting PRV infection. Overexpression of HSP27 promoted PRV proliferation in vitro, while knockdown of HSP27 inhibited PRV infection. Importantly, we found that HSP27 inhibited PRV infection or poly(dA:dT)-activated IFN-ß expression. Further studies found that HSP27 may inhibit cGAS-STING-mediated IFN-ß expression through targeting cGAS. In addition, we found that HSP27 can suppress the expression of endogenous cGAS in different cells at both gene transcription and protein expression levels, and that HSP27 interacts with and ubiquitinates cGAS. In conclusion, we reveal for the first time that HSP27 is a novel negative regulator of the cGAS-STING signaling pathway induced by PRV infection or poly(dA:dT) activation and demonstrate that HSP27 plays a crucial role in PRV infection.

High importance of coupled nitrification-denitrification for nitrogen removal in a large periodically low-oxygen estuary.

The coupling between nitrification and denitrification/anammox (nitrate/nitrite used in denitrification/anammox derives from nitrification) is a significant process of reactive nitrogen (N) removal that has attracted much attention. However, the dynamics of coupled nitrification-denitrification/anammox in the periodically low-oxygen estuaries and coasts remain unclear. Here, continuous-flow experiments combined with isotope tracing techniques were conducted in periodically low-oxygen areas of the Yangtze Estuary to reveal the changes in benthic sediment denitrification and anammox as well as their coupling with nitrification. Our results showed that denitrification increased but anammox decreased during low-oxygen summer. The occurrence of low oxygen also promoted coupled nitrification-denitrification but decreased coupled nitrification-anammox. These results implied that decreased dissolved oxygen in summer did not largely restrict nitrification activity, and anaerobic denitrification/anammox regulated the magnitude of coupled nitrification-denitrification/anammox rates. Denitrification (74.95-100 %) was the dominant process in total N removal, while coupled nitrification-denitrification accounted for a higher proportion (45.68-97.05 %) of denitrification, indicating that coupling between nitrification and denitrification played a dominant role in N removal. In addition to dissolved oxygen levels, carbon and N substrate availabilities were also important variables to regulate N transformations. Overall, this study advanced our knowledge of the distribution patterns and controlling factors of N removal processes and highlighted that coupled nitrification-denitrification might have a significant but neglected role in N removal from periodically low-oxygen estuaries.

Long-Term Compost Amendment Spurs Cellulose Decomposition by Driving Shifts in Fungal Community Composition and Promoting Fungal Diversity and Phylogenetic Relatedness.

Cellulose is the most abundant polysaccharide in plant biomass and an important precursor of soil organic matter formation. Fungi play a key role in carbon cycling dynamics because they tend to decompose recalcitrant materials. Here, we applied [12C]cellulose and [13C]cellulose to distinguish the effects of application of compost, nitrogen-phosphorus-potassium (NPK) fertilizer, and no fertilizer (control) for 27 years upon cellulose decomposition via RNA-based stable isotope probing (RNA-SIP). The loss ratio of added cellulose C in compost soil was 67.6 to 106.7% higher than in NPK and control soils during their 20-day incubation. Dothideomycetes (mainly members of the genus Cryptococcus) dominated cellulose utilization in compost soil, whereas the copiotrophic Sordariomycetes were more abundant in NPK and unfertilized soils. Compared with NPK and control soils, compost application increased the diversity of 13C-assimilating fungi. The 13C-labeled fungal communities in compost soil were more phylogenetically clustered and exhibited greater species relatedness than those in NPK and control soils, perhaps because of stringent filtering of narrow-spectrum organic resources and biological invasion originating from added compost. These changes led to an augmented decomposition capacity of fungal species for cellulose-rich substrates and reduced cellulose C sequestration efficiency. The RNA-SIP technique is more sensitive to responses of fungi to altered soil resource availability than DNA-SIP. Overall, long-term compost application modified fungal community composition and promoted fungal diversity and phylogenetic relatedness, accelerating the decomposition of substrate cellulose in soil. This work also highlights the RNA-SIP technique's value for comprehensively assessing the contributions of active fungi to the substrate decomposition process. IMPORTANCE Cellulose is a very rich component in plant biomass and an important precursor of soil organic matter formation. Fungal communities are known to be important drivers of organic carbon accumulation in arable soils. However, current understanding of responses of fungal species to cellulose amendment and the contributions of active fungi to substrate decomposition process is still very superficial. Here, we established a [13C]cellulose microcosm experiment with soils subjected to long-term application of compost, nitrogen-phosphorus-potassium (NPK) fertilizer, and no fertilizer (control). The novel 13C-RNA-SIP technique with subsequent high-throughput sequencing was used to investigate the linkages between active fungal taxa and cellulose decomposition. Our study demonstrated that Dothideomycetes dominated cellulose utilization in compost soil, whereas the copiotrophic Sordariomycetes were more enriched in both NPK and unfertilized soils. We also found that the compost amendment promoted fungal diversity and phylogenetic relatedness and strengthened the decomposition capacity of fungi for cellulose-rich substrates by enhancing synergistic interactions, thereby reducing cellulose C sequestration efficiency. Overall, our research has implications for our understanding of the role of active fungi in cellulose C transformation in soils undergoing different types of long-term nutrient management.

Fungal key players of cellulose utilization: Microbial networks in aggregates of long-term fertilized soils disentangled using 13C-DNA-stable isotope probing.

Long-term compost application accelerates organic carbon (C) accumulation and macroaggregate formation in soil. Stable aggregates and high soil organic C (SOC) content are supposed to increase microbiota activity and promote transformation of litter compounds (i.e., cellulose) into SOC. Here, we used 13C-DNA-stable isotope probing with subsequent high-throughput sequencing to characterize fungal succession and co-occurrence trends during 13C-cellulose decomposition in aggregate size classes in soils subjected to no fertilizer (control), nitrogen-phosphorus­potassium (NPK) fertilizers, and compost (Compost) application for 27 years. Ascomycota (mostly saprotrophic fungi) were always highly competitive for cellulose in all aggregate size classes at the early stages of cellulose decomposition (20 days). Compost-treated soil was enriched with Ascomycota compared to the control soil, wherein Sordariomycetes, the majority, strongly dominated the cellulose utilization (13C incorporation in DNA). 13C-labeled fungal communities converged in the Compost soil, with lower abundance and diversity compared with the NPK and control soils. Such convergence led to greater cellulose decomposition, indicating that compost amendment increased the capacity of a few dominant fungal taxa to decompose litter. Compost soil had more 13C-labeled fungal decomposers in microaggregates and lower fungal decomposers in macroaggregates when compared with the levels in the NPK and control soils. This implies that compost application facilitates fungal colonization towards smaller aggregates. Fungal interactions were reinforced in microaggregates (<250 µm), with more positive associations than those in macroaggregates (>250 µm), indicating greater fungal synergism for recalcitrant resource utilization in microaggregates. The keystone taxa in the co-occurrence networks were not related to cellulose decomposition in microaggregates, but did in macroaggregates. The findings advance a process-based understanding of cellulose utilization by fungal key players based on C and energy availability and the regulation of microbial activity at the aggregate level.

N2O emission dynamics along an intertidal elevation gradient in a subtropical estuary: Importance of N2O consumption.

Studying nitrous oxide (N2O) production and consumption processes along an intertidal elevation gradient can improve the understanding of N2O dynamics among coastal wetlands. A natural-abundance isotope technique was applied to characterize the processes responsible for N2O emission in high, middle and low intertidal zones in the Yangtze Estuary. The results showed that N2O emission rates in high tidal zones (0.84 ± 0.35 nmol g-1 h-1) were significantly higher than those in middle (0.21 ± 0.04 nmol g-1 h-1) and low tidal zones (0.26 ± 0.05 nmol g-1 h-1). Gross N2O production and consumption rates were greater in high and low tidal zones than in middle tidal zones, whereas N2O consumption proportions generally increased from high to low tidal zones. N2O consumption was quite pronounced, implying that N2O emission in estuarine wetlands accounts for only a small fraction of the total production. Higher degrees of N2O consumption were the pivotal driver of less N2O emission in low tidal zones. Bacterial denitrification (>84%) was the dominant pathway, although hydroxylamine (NH2OH) oxidation/fungal denitrification contributed substantially to N2O production in high tidal flats. The contribution to N2O production exhibited a decrease in NH2OH oxidation/fungal denitrification and an increase in bacterial denitrification with decreasing elevation. Changes in N2O dynamics along the elevation gradient were affected by carbon and nitrogen substrate availabilities as well as the redox environments. Overall, our findings highlight the importance of N2O consumption in controlling N2O emission in intertidal wetlands, especially with higher inundation frequencies and durations.

Microbial dynamics and activity of denitrifying anaerobic methane oxidizers in China's estuarine and coastal wetlands.

Estuarine and coastal wetlands, which act as large sources of methane (CH4) and undergo substantial loading of anthropogenic nitrogen (N), provide ideal conditions for denitrifying anaerobic methane oxidation (DAMO) to occur. Yet the microbial mechanisms governing DAMO and the main driving factors in estuarine and coastal ecosystems remain unclear. This study investigated the spatiotemporal distribution and associated activity of DAMO microorganisms along a wide swath of China's coastline (latitudinal range: 22-41°N) using molecular assays and isotope tracing techniques. We uncovered significant spatial and seasonal variation in DAMO bacterial community structure, whereas DAMO archaeal community structure exhibited no seasonal differences. The abundance of DAMO bacterial pmoA gene (2.2 × 105-1.0 × 107 copies g-1) was almost one order of magnitude higher than that of DAMO archaeal mcrA gene (8.7 × 104 -1.8 × 106 copies g-1). A significant positive correlation between pmoA and mcrA gene abundances (p < 0.01) was observed, indicating that DAMO bacteria and archaea may cooperate closely and thus complete nitrate elimination. Potential DAMO rates, in the range of 0.09-23.4 nmol 13CO2 g-1 day-1 for nitrite-DAMO and 0.03-43.7 nmol 13CO2 g-1 day-1 for nitrate-DAMO, tended to be greater in the relatively warmer low-latitudes. Potential DAMO rates were weakly positively correlated with gene abundances, suggesting that DAMO microbial activity could not be predicted directly by gene abundance alone. The heterogeneous variability of DAMO was shaped by interactions among key environmental characteristics (sediment texture, N availability, TOC, Fe3+, salinity of water, and temperature). On a broader continental scale, potential N removal rates of 0.1-11.2 g N m-2 yr-1 were estimated via nitrite-DAMO activity in China's coastal wetlands. Overall, our results highlight the widespread distribution of DAMO microbes and their potential role in eliminating excess N inputs and reducing CH4 emissions in estuarine and coastal ecosystems, which could help mitigate global warming.

HSP27 Protein Dampens Encephalomyocarditis Virus Replication by Stabilizing Melanoma Differentiation-Associated Gene 5.

Heat shock proteins (HSPs) are a protein family that respond to physiological stress, such as heat, starvation, and infection. As cellular protein chaperones, they play an important role in protein folding, assembly, and degradation. Though it is well known that HSP27 is involved in a range of viral infections, its role during an encephalomyocarditis virus (EMCV) infection is not known. Here, we report that EMCV degrades HSP27 and that EMCV proteins 2Cpro and 3Apro are primarily responsible for its degradation. Consequently, loss of cellular HSP27 augmented EMCV proliferation, an effect that could be reversed upon HSP27 overexpression. Importantly, we found that HSP27 positively regulated EMCV-triggered type I interferon (IFN) production. Moreover, overexpression of 2Cpro and 3Apro significantly blocked type I IFN production. We also found for the first time that HSP27, as a molecular chaperone, can specifically interact with MDA5 and stabilize the expression of MDA5. Collectively, this study shows that HSP27 dampens EMCV infectivity by positively regulating EMCV-triggered retinoic acid-inducible gene (RIG)-I-like receptor (RLR)/melanoma differentiation-associated gene 5 (MDA5) signal pathway, while EMCV proteins 2Cpro and 3Apro interact with HSP27 and degrade HSP27 protein expression to allow EMCV proliferation. Our findings provide further mechanistic evidence for EMCV partaking in immune escape mechanisms, and that 2Cpro and 3Apro could serve as potential antiviral targets.

Host protein, HSP90ß, antagonizes IFN-ß signaling pathway and facilitates the proliferation of encephalomyocarditis virus in vitro.

Encephalomyocarditis virus (EMCV) is a small, non-enveloped, single stranded RNA virus which infects a wide variety of mammalian species, and has zoonotic importance. Many host proteins are known to regulate EMCV proliferation by interacting with its structural or nonstructural proteins, but the regulatory role and mechanism of heat shock protein 90ß (HSP90ß), in EMCV infection has not been reported yet. Here, we report that overexpression of HSP90ß significantly promotes the growth and proliferation of EMCV in vitro. On the contrary, down-regulation of HSP90ß by RNAi or geldanamycin inhibits EMCV replication. HSP90ß suppresses IFN-ß responses in the RLRs pathway by targeting the expression of the key adaptor molecules MAVS, TBK1, and IRF3, but not MDA5. This study demonstrates the firsthand information that HSP90ß plays a positive role in viral proliferation by inhibiting EMCV induced IFN-ß production. Collectively, the results reveal new insights into HSP90ß-assisted progression of EMCV infection.

Effect of field-aged biochar on fertilizer N retention and N2O emissions: A field microplot experiment with 15N-labeled urea.

Biochar application is thought to improve crop yield and reduce N leaching and gas emissions; however, little is known about how field-aged biochar affects fertilizer N retention and N2O emissions. Here, a field microplot experiment is established in the North China Plain at maize season by applying 15N-labeled urea to the sandy loam soil both with (Biochar) and without (Control) application of 3-year field-aged biochar at 12 t ha-1. Overall, 25.6-26.2% of the urea N was taken up by maize aboveground biomass, field-aged biochar did not affect yield or fertilizer N recovery efficiency. After maize harvest, the residual ratio of applied N in the soil profile (0-40 cm) was 21.6 and 20.3% under Control and Biochar treatment, respectively, with an increase of 10.2% in the topsoil (0-20 cm) and decrease of 37.2% in the subsoil (20-40 cm) following biochar amendment, probably due to reduced NO3- leaching. Cumulative N2O emissions and urea N-induced N2O emissions under Control treatment were 2.06 and 0.78 kg N ha-1, and significantly decreased to 1.89 and 0.74 kg N ha-1 after Biochar treatment, respectively. N2O emissions derived from the applied N accounted for 38.0 and 39.4% of the total emissions under Control and Biochar treatment, respectively. N2O emissions from decomposition of soil organic N induced by the priming effect of the applied N was 0.69 and 0.56 kg N ha-1 under Control and Biochar treatment, respectively, contributing 33.7 and 29.7% of the total emissions. Overall, our results suggest that field-aged biochar increased the retention of fertilizer N in the topsoil by reducing NO3- leaching, while effectively reduced N2O emissions from fertilizer N and mineralization of organic N in the sandy loam soil.