Effects of glucose on the uptake and metabolism of glycine in pakchoi (Brassica chinensis L.) exposed to various nitrogen sources.

BACKGROUND: Plants can absorb amino acids as a nitrogen (N) source, and glucose is an important part of root rhizodeposition and the soil sugar pool, which participates in the regulation of plant growth and uptake. In pakchoi, the effect of glucose concentration on the glycine N uptake from a nutrient mixture composed of glycine, ammonium, and nitrate, or from a single N solution of glycine alone was studied using specific substrate 15N-labeling and 15N-gas chromatography mass spectrometry. RESULTS: The optimal glucose concentration for plant growth was 4.5 µM or 25 µM when supplied with glycine alone or the N mixture, respectively, and resulted in a >25% increase in seedling biomass. The addition of glucose affected the relative contribution from organic or inorganic sources to overall N uptake. When glucose was added at optimal concentrations, glycine was preferentially used as an N source, while the relative contribution from nitrate was reduced. The limiting step for glycine N contribution was active uptake in the roots in high glucose and single-N-source conditions; however, root metabolism of glycine to serine was limiting in high-glucose and mixed-N-source conditions. CONCLUSIONS: The addition of low concentrations of glucose increased the relative uptake of organic nitrogen and reduced the uptake of nitrate, suggesting a feasible way to decrease nitrate content and increase the edible quality of vegetables.

Hexavalent chromium stress enhances the uptake of nitrate but reduces the uptake of ammonium and glycine in pak choi (Brassica chinensis L.).

Chromium (Cr) pollution affects plant growth and biochemical processes, so, the relative uptake of glycine, nitrate, and ammonium by pak choi (Brassica chinensis) seedlings in treatments with 0mgL-1 and 10mgL-1 Cr (VI) were detected by substrate-specific 15N-labelling in a sterile environment. The short-term uptake of 15N-labelled sources and 15N-enriched amino acids were detected by gas chromatography mass spectrometry to explore the mechanism by which Cr stress affects glycine uptake and metabolism, which showing that Cr stress hindered the uptake of ammonium and glycine but increased significantly the uptake of nitrate. Cr stress did not decrease the active or passive uptake of glycine, but it inhibited the conversion of glycine to serine in pak choi roots, indicating that the metabolism of glycine to serine in roots, rather than the root uptake, was the limiting step in glycine contribution to total N uptake in pak choi. Since Cr affects the relative uptake of different N sources, a feasible way to reduce Cr-induced stress is application of selective fertilization, in particular nitrate, in pak choi cultivation on Cr-polluted soil.

Broadband Light-Harvesting Molecular Triads with High FRET Efficiency Based on the Coumarin-Rhodamine-BODIPY Platform.

Broadband capturing and FRET-based light-harvesting molecular triads, CRBs, based on the coumarin-rhodamine-BODIPY platform were rationally designed and synthesized. The absorption band of CRBs starts from blue-green to yellow-orange regions (330-610 nm), covering the strong radiation scope of sunlight. The peripheral coumarin and BODIPY chromophore energy could transfer to the central acceptor rhodamine by a one-step direct way. The energy of the coumarin moiety could also transfer to the BODIPY unit, subsequently transferring to the rhodamine core by two-step sequential ways. Both the efficiencies of the coumarin moiety and the BODIPY unit to the rhodamine core in CRBs, determined by two different ways, are very high.

Drying-wetting cycle enhances stress resistance of Escherichia coli O157:H7 in a model soil.

Outbreaks of Escherichia coli (E. coli) O157:H7 in farms are often triggered by heavy rains and flooding. Most cells die with the decreasing of soil moisture, while few cells enter a dormant state and then resuscitate after rewetting. The resistance of dormant cells to stress has been extensively studied, whereas the molecular mechanisms of the cross-resistance development of the resuscitated cells are poorly known. We performed a comparative proteomic analysis on O157:H7 before and after undergoing soil dry-wet alternation. A differential expression of 820 proteins was identified in resuscitated cells compared to exponential-phase cells, as determined by proteomics analysis. The GO and KEGG pathway enrichment analyses revealed that up-regulated proteins were associated with oxidative phosphorylation, glycolysis/gluconeogenesis, the citrate cycle (TCA cycle), aminoacyl-tRNA biosynthesis, ribosome activity, and transmembrane transporters, indicating increased energy production and protein synthesis in resuscitated O157:H7. Moreover, proteins related to acid, osmotic, heat, oxidative, antibiotic stress and horizontal gene transfer efficiency were up-regulated, suggesting a potential improvement in stress resistance. Subsequent validation experiments demonstrated that the survival rates of the resuscitated cells were 476.54 and 7786.34 times higher than the exponential-phase cells, with pH levels of 1.5 and 2.5, respectively. Similarly, resuscitated cells showed higher survival rates under osmotic stress, with 7.5%, 15%, and 30% NaCl resulting in survival rates that were 460.58, 1974.55, and 3475.31 times higher. Resuscitated cells also exhibited increased resistance to heat stress, with survival rates 69.64 and 139.72 times higher at 55 °C and 90 °C, respectively. Furthermore, the horizontal gene transfer (HGT) efficiency of resuscitated cells was significantly higher (153.12-fold) compared to exponential phase cells. This study provides new insights into bacteria behavior under changing soil moisture and this may explain O157:H7 outbreaks following rainfall and flooding, as the dry-wet cycle promotes stress cross-resistance development.

Multi-scale simulations of the mechanical behaviors of the W-Cu joint interface with a diffusion layer.

CONTEXT: At the interface of W-Cu after direct jointing, diffusion layers with a thickness of approximately 22 nm are present but often overlooked in simulations of mechanical properties. In this study, an interface model with a W-Cu diffusion layer is developed using molecular dynamics (MD). The effects of the diffusion layers on the elastic-plastic behaviors, dissipation mechanisms, and fracture properties of the interface are analyzed under mode-I (perpendicular to the interface) and mode-II (parallel to the interface). The results demonstrate that the interface model with a diffusion layer exhibits superior mechanical properties under mode-I and mode-II loading compared to the model without a diffusion layer. Furthermore, a multi-scale method based on the classical Paris law is proposed, combining MD and finite element methods to investigate the fatigue crack propagation of W-Cu bimetallic composites under cyclic loading and predict their fatigue life. The findings of this study are meaningful for improving the mechanical properties of W-Cu interface materials, predicting the material's lifespan, and guiding related engineering applications. METHODS: In this study, the molecular dynamics simulations have been carried out by using the Large-scale Atomic/Molecular Massively Parallel Simulator (LAMMPS). The visualization of results is performed using the Open Visualization Tool (OVITO). Common neighbor analysis (CNA) and dislocation analysis (DXA) in OVITO have been employed to capture the structural evolution. Finite element method simulations are performed in Ansys Workbench.

Single fluorescent probe responds to H2O2, NO, and H2O2/NO with three different sets of fluorescence signals.

Hydrogen peroxide (H(2)O(2)) acts as a signaling molecule in a wide variety of signaling transduction processes and an oxidative stress marker in aging and disease. However, excessive H(2)O(2) production is implicated with various diseases. Nitric oxide (NO) serves as a secondary messenger inducing vascular smooth muscle relaxation. However, mis-regulation of NO production is associated with various disorders. To disentangle the complicated inter-relationship between H(2)O(2) and NO in the signal transduction and oxidative pathways, fluorescent reporters that are able to display distinct signals to H(2)O(2), NO, and H(2)O(2)/NO are highly valuable. Herein, we present the rational design, synthesis, spectral properties, and living cell imaging studies of FP-H(2)O(2)-NO, the first single-fluorescent molecule, that can respond to H(2)O(2), NO, and H(2)O(2)/NO with three different sets of fluorescence signals. FP-H(2)O(2)-NO senses H(2)O(2), NO, and H(2)O(2)/NO with a fluorescence signal pattern of blue-black-black, black-black-red, and black-red-red, respectively. Significantly, we have further demonstrated that FP-H(2)O(2)-NO, a single fluorescent probe, is capable of simultaneously monitoring endogenously produced NO and H(2)O(2) in living macrophage cells in multicolor imaging. We envision that FP-H(2)O(2)-NO will be a unique molecular tool to investigate the interplaying roles of H(2)O(2) and NO in the complex interaction networks of the signal transduction and oxidative pathways. In addition, this work establishes a robust strategy for monitoring the multiple ROS and RNS species (H(2)O(2), NO, and H(2)O(2)/NO) using a single fluorescent probe, and the modularity of the strategy may allow it to be extended for other types of biomolecules.

A native-chemical-ligation-mechanism-based ratiometric fluorescent probe for aminothiols.

Thiol-containing amino acids (aminothiols) such as cysteine (Cys) and homocysteine (Hcy) play a key role in various biological processes including maintaining the homeostasis of biological thiols. However, abnormal levels of aminothiols are associated with a variety of diseases. The native chemical ligation (NCL) reaction has attracted great attention in the fields of chemistry and biology. NCL of peptide segments involves cascade reactions between a peptide-α-thioester and an N-terminal cysteine peptide. In this work, we employed the NCL reaction mechanism to formulate a Förster resonance energy transfer (FRET) strategy for the design of ratiometric fluorescent probes that were selective toward aminothiols. On the basis of this new strategy, the ratiometric fluorescent probe 1 for aminothiols was judiciously designed. The new probe is highly selective toward aminothiols over other thiols and exhibits a very large variation (up to 160-fold) in its fluorescence ratio (I(458)/I(603)). The new fluorescent probe is capable of ratiometric detection of aminothiols in newborn calf and human serum samples and is also suitable for ratiometric fluorescent imaging of aminothiols in living cells.

Fluorescent detection of hypochlorous acid from turn-on to FRET-based ratiometry by a HOCl-mediated cyclization reaction.

Hypochlorous acid (HOCl), a reactive oxygen species (ROS), plays a significant biological role in living systems. However, abnormal levels of HOCl are implicated in many inflammation-associated diseases. Therefore, the detection of HOCl is of great importance. In this work, we describe the HOCl-promoted cyclization of rhodamine-thiosemicarbazides to rhodamine-oxadiazoles, which is then exploited as a novel design strategy for the development of a new fluorescence turn-on HOCl probe 2. On the basis of the fluorescence resonance energy transfer (FRET) signaling mechanism, 2 was further converted into 1a and 1b, which represent the first paradigm of FRET-based ratiometric fluorescent HOCl probes. The outstanding features of 1a and 1b include well-resolved emission peaks, high sensitivity, high selectivity, good functionality at physiological pH, rapid response, low cytotoxicity, and good cell-membrane permeability. Furthermore, these excellent attributes enable us to demonstrate, for the first time, the ratiometric imaging of endogenously produced HOCl in living cells by using these novel ratiometric probes. We expect that 1a and 1b will be useful molecular tools for studies of HOCl biology. In addition, the HOCl-promoted cyclization reaction of rhodamine-thiosemicarbazides to rhodamine-oxadiazoles should be widely applicable for the development of different types of fluorescent HOCl probes.

Crop rotation stage has a greater effect than fertilisation on soil microbiome assembly and enzymatic stoichiometry.

Agronomic practises, such as fertilisation and crop rotation, affect soil microbial communities and functions. However, limited information is available regarding the relative importance of fertilisation and crop rotation stages in determining the soil microbiome and assembly processes. In addition, insights into the connections between the soil microbiome and enzymatic stoichiometry are scarce. In this study, soil samples were collected from a wheat-rice rotation system that received mineral and organic fertiliser inputs for 6 years to investigate soil microbiome assembly, and the relationship between the soil microbiome and enzymatic stoichiometry. Our results revealed that the crop rotation stage strongly affected the soil microbial community structure, assembly, and enzymatic functions compared to that of the fertilisation regime. Enzymatic stoichiometry results and vector analysis implied that mineral and organic fertilisation could alleviate the microbial N limitation. However, no-manure fertilisation led to microbial P limitation during the wheat stage. The decreases in soil pH mainly drove microbial P limitation due to the acidification induced by the mineral fertilisers. Microbial N/P limitation correlated more strongly with the bacterial assembly than with fungal assembly. Moreover, co-occurrence network analysis showed that ecological relationships between microbial taxa and enzymes were more complex during the wheat stage than that during the rice stage. Microbial nodes linked to acid phosphomonoesterase correlated significantly with the soil pH. Our study highlights the distinct responses of the soil microbiome to fertilisation in different crop-rotation stages, and provides novel insights into connections between microbial assembly and enzymatic stoichiometry.

The inhibition effect of tea polyphenols on soil nitrification is greater than denitrification in tea garden soil.

Tea polyphenols are the most widely distributed class of secondary metabolites (Camellia sinensis) and account for a considerable proportion of the pruning residues of tea. A large amount of tea polyphenols have fallen down over soil with prunning residues every year. However, the effect of tea polyphenols on soil nitrogen cycle, especially the denitrification process and its related microbial communities, remains unclear. Epigallocatechin gallate (EGCG), the most abundant component of tea polyphenols, was selected to simulate the effects of tea polyphenols on soil nitrification, denitrification, related functional genes and microbial community. The results indicated that addition of EGCG can significantly (p < 0.05) inhibit soil nitrification. Copy numbers of bacterial and archaeal ammonia monooxygenase genes (amoA) decreased as EGCG concentration increased. Further, the ammonia oxidisers exhibited a significantly (p < 0.05) greater niche differentiation under the effect of EGCG compared with the control treatment (no EGCG addition). However, the inhibition effect of EGCG over soil denitrification was most significant at 34 and 36 day of incubation period, and such inhibitory effect was more apparent on nitrification compared with denitrification. EGCG addition increased the diversity of bacterial community. The composition of bacterial community was significantly altered and community variation was primary explained by EGCG, NH4+-N, NO3--N, soil organic carbon contents and potential denitrification rates. EGCG addition significantly increased relative abundance of Proteobacteria and Bacteroidetes phyla whereas decreased Actinobacteria. Overall, tea polyphenols can inhibit soil nitrification to a larger extent than denitrification by reducing the abundance of microorganisms carrying the related functional genes. Our results can serve as important basis of reducing the nitrogen pollution risk in tea orchards and could be considered as a powerful natural nitrification inhibitor to reduce the environmental risks caused by unreasonable nitrogen fertiliser adaptation.

Development of FRET-based ratiometric fluorescent Cu2+ chemodosimeters and the applications for living cell imaging.

Coumarin-rhodamine-based compounds 1a,b were rationally designed and synthesized as novel FRET ratiometric fluorescent chemodosimeters. Ratiometric chemodosimeters 1a,b exhibit several favorable features, including a large variation in the emission ratio, well-resolved emission peaks, high sensitivity, high selectivity, low cytotoxicity, and good cell membrane permeability. Importantly, these excellent attributes enable us to demonstrate ratiometric imaging of Cu(2+) in living cells by using these novel ratiometric fluorescent chemodosimeters.

Development of a ratiometric fluorescent sensor for ratiometric imaging of endogenously produced nitric oxide in macrophage cells.

We described the development of Cou-Rho-NO as the first small-molecule suitable for ratiometric fluorescent imaging of endogenously produced nitric oxide in macrophage cells.