Metabolic pathway engineering of high-salinity-induced overproduction of L-proline improves high-salinity stress tolerance of an ectoine-deficient Halomonas elongata.

Halophilic bacteria have adapted to survive in high-salinity environments by accumulating amino acids and their derivatives as organic osmolytes. L-Proline (Pro) is one such osmolyte that is also being used as a feed stimulant in the aquaculture industry. Halomonas elongata OUT30018 is a moderately halophilic bacterium that accumulates ectoine (Ect), but not Pro, as an osmolyte. Due to its ability to utilize diverse biomass-derived carbon and nitrogen sources for growth, H. elongata OUT30018 is used in this work to create a strain that overproduces Pro, which could be used as a sustainable Pro-rich feed additive. To achieve this, we replaced the coding region of H. elongata OUT30018's Ect biosynthetic operon with the artificial self-cloned proBm1AC gene cluster that encodes the Pro biosynthetic enzymes: feedback-inhibition insensitive mutant γ-glutamate kinase (γ-GKD118N/D119N), γ-glutamyl phosphate reductase, and pyrroline-5-carboxylate reductase. Additionally, the putA gene, which encodes the key enzyme of Pro catabolism, was deleted from the genome to generate H. elongata HN6. While the Ect-deficient H. elongata KA1 could not grow in minimal media containing more than 4% NaCl, H. elongata HN6 thrived in the medium containing 8% NaCl by accumulating Pro in the cell instead of Ect, reaching a concentration of 353.1 ± 40.5 µmol/g cell fresh weight, comparable to the Ect accumulated in H. elongata OUT30018 in response to salt stress. With its genetic background, H. elongata HN6 has the potential to be developed into a Pro-rich cell factory for upcycling biomass waste into single-cell feed additives, contributing to a more sustainable aquaculture industry.IMPORTANCEWe report here the evidence for de novo biosynthesis of Pro to be used as a major osmolyte in an ectoine-deficient Halomonas elongata. Remarkably, the concentration of Pro accumulated in H. elongata HN6 (∆ectABC::mCherry-proBm1AC ∆putA) is comparable to that of ectoine accumulated in H. elongata OUT30018 in response to high-salinity stress. We also found that among the two γ-glutamate kinase mutants (γ-GKD118N/D119N and γ-GKD154A/E155A) designed to resemble the two known Escherichia coli feedback-inhibition insensitive γ-GKD107N and γ-GKE143A, the γ-GKD118N/D119N mutant is the only one that became insensitive to feedback inhibition by Pro in H. elongata. As Pro is one of the essential feed additives for the poultry and aquaculture industries, the genetic makeup of the engineered H. elongata HN6 would allow for the sustainable upcycling of high-salinity waste biomass into a Pro-rich single-cell eco-feed.

Internal Electric Field Facilitates Facet-Dependent Photocatalytic Cl- Utilization on BiOCl in High-Salinity Wastewater for Ammonium Removal.

High Cl- concentration in saline wastewater (e.g., landfill leachate) limits wastewater purification. Catalytic Cl- conversion into reactive chlorine species (RCS) arises as a sustainable strategy, making the salinity profitable for efficient wastewater treatment. Herein, aiming to reveal the structure-property relationship in Cl- utilization, bismuth oxychloride (BiOCl) photocatalysts with coexposed {001} and {110} facets are synthesized. With an increasing {001} ratio, the RCS production efficiency increases from 75.64 to 96.89 µg L-1 min-1. Mechanism investigation demonstrates the fast release of lattice Cl- as an RCS and the compensation of ambient Cl-. Correlation analysis between the internal electric field (IEF, parallel to [001]) and normalized efficiency on {110} (kRCS/S{110}, perpendicular to [001]) displays a coefficient of 0.86, validating that the promoted carrier dynamics eventually affects Cl- conversion on the open layered structure. The BiOCl photocatalyst is well behaved in ammonium (NH4+-N) degradation ranging from 20 to 800 mg N L-1 with different chlorinity (3-12 g L-1 NaCl). The sustainable Cl- conversion into RCS also realizes 85.4% of NH4+-N removal in the treatment of realistic landfill leachate (662 mg of N L-1 NH4+-N). The structure-property relationship provides insights into the design of efficient catalysts for environment remediation using ambient Cl-.

Expression of an engineered salt-inducible proline biosynthetic operon in a Glutamic acid Over-Producing mutant, Halomonas elongata GOP, confers increased proline yield due to enhanced growth under high-salinity conditions.

L-Proline (Pro) is an essential amino acid additive in livestock and aquaculture feeds. Previously, we created a Pro overproducing Halomonas elongata HN6 by introducing an engineered salt-inducible Pro biosynthetic mCherry-proBm1AC operon and deleting a putA gene that encoded a Pro catabolic enzyme in the genome of H. elongata OUT30018. Here, we report a generation of a novel Pro overproducing H. elongata HN10 strain with improved salt tolerance and higher Pro yield by expressing the mCherry-proBm1AC operon and deleting the putA gene in the genome of a spontaneous mutant H. elongata GOP, which overproduces glutamic acid (Glu) that is a precursor for Pro biosynthesis. The optimal salt concentration for growth of H. elongata HN10 was found to be 7% to 8% w/v NaCl, and the average Pro yield of 166 mg/L was achieved when H. elongata HN10 was cultivated in M63 minimal medium containing 4% w/v glucose and 8% w/v NaCl.

Facilitating nitrification and biofilm formation of Vibrio sp. by N-acyl-homoserine lactones in high salinity environment.

The N-acyl-homoserine lactones (AHLs)-mediated quorum-sensing (QS) system played a crucial role in regulating biological nitrogen removal and biofilm formation. However, the regulatory role of AHLs on nitrogen removal bacteria in high salinity environment has remained unclear. This study evaluated the roles and release patterns of AHLs in Vibrio sp. LV-Q1 under high salinity condition. Results showed that Vibrio sp. primarily secretes five AHLs, and the AHLs activity is strongly correlated with the bacterial density. Exogenous C10-HSL and 3OC10-HSL were found to significantly enhance ammonium removal, while making a minor contribution to the growth rate. Both the C10-HSL and 3OC10-HSL promoted the biofilm formation of Vibrio sp. with an enhancement of 1.64 and 1.78 times, respectively. The scanning electron microscope (SEM) and confocal laser scanning microscope (CLSM) observations confirmed the biofilm-enhancing effect of AHLs. Further analysis revealed that AHLs significantly improved bacterial self-aggregation and motility, as well as the level of extracellular polymeric substances (EPS). These findings provide significant guidance on construction of nitrification system at high salinity.

Overexpression of a Fragaria vesca NAM, ATAF, and CUC (NAC) Transcription Factor Gene (FvNAC29) Increases Salt and Cold Tolerance in Arabidopsis thaliana.

The NAC (NAM, ATAF1/2, CUC2) family of transcription factors (TFs) is a vital transcription factor family of plants. It controls multiple parts of plant development, tissue formation, and abiotic stress response. We cloned the FvNAC29 gene from Fragaria vesca (a diploid strawberry) for this research. There is a conserved NAM structural domain in the FvNAC29 protein. The highest homology between FvNAC29 and PaNAC1 was found by phylogenetic tree analysis. Subcellular localization revealed that FvNAC29 is localized onto the nucleus. Compared to other tissues, the expression level of FvNAC29 was higher in young leaves and roots. In addition, Arabidopsis plants overexpressing FvNAC29 had higher cold and high-salinity tolerance than the wild type (WT) and unloaded line with empty vector (UL). The proline and chlorophyll contents of transgenic Arabidopsis plants, along with the activities of the antioxidant enzymes like catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD) under 200 mM NaCl treatment or -8 °C treatment, were higher than those activities of the control. Meanwhile, malondialdehyde (MDA) and the reactive oxygen species (ROS) content were higher in the WT and UL lines. FvNAC29 improves transgenic plant resistance to cold and salt stress by regulating the expression levels of AtRD29a, AtCCA1, AtP5CS1, and AtSnRK2.4. It also improves the potential to tolerate cold stress by positively regulating the expression levels of AtCBF1, AtCBF4, AtCOR15a, and AtCOR47. These findings suggest that FvNAC29 may be related to the processes and the molecular mechanisms of F. vesca response to high-salinity stress and LT stress, providing a comprehensive understanding of the NAC TFs.

Offshore produced water treatment by a biofilm reactor on the seabed: The effect of temperature and matrix characteristics.

In many industrial processes a large amount of water with high salinity is co-produced whose treatment poses considerable challenges to the available technologies. The produced water (PW) from offshore operations is currently being discharged to sea without treatment for dissolved pollutants due to space limitations. A biofilter on the seabed adjacent to a production platform would negate all size restrictions, thus reducing the environmental impact of oil and gas production offshore. The moving bed biofilm reactor (MBBR) was investigated for PW treatment from different oilfields in the North Sea at 10 °C and 40 °C, corresponding to the sea and PW temperature, respectively. The six PW samples in study were characterized by high salinity and chemical oxygen demand with ecotoxic effects on marine algae S. pseudocostatum (0.4%

Influence of Organic Impurities on Fractional Crystallization of NaCl and Na2SO4 from High-Salinity Coal Chemical Wastewater: Thermodynamics and Nucleation Kinetics Analysis.

It is a valid path to realize the zero discharge of coal chemical wastewater by using the fractional crystallization method to recycle the miscellaneous salt in high-salinity wastewater. In this study, the thermodynamics and nucleation kinetics of sodium chloride (NaCl) and sodium sulfate (Na2SO4) crystallization in coal chemical wastewater were systematically studied. Through analyses of solubility, metastable zone width, and induction period, it was found that the impurity dimethoxymethane would increase the solid-liquid interface energy and critical crystal size during the nucleation of Na2SO4. Ternary phase diagrams of the pseudo-ternary Na2SO4-NaCl-H2O systems in simulated wastewater were plotted in the temperature range of 303.15 to 333.15 K, indicating that a co-ionization effect existed between NaCl and Na2SO4, and NaCl had a strong salting out effect on Na2SO4. Finally, the nucleation rate and growth rate of Na2SO4 crystals under simulated wastewater conditions were determined by the intermittent dynamic method, and the crystallization kinetic models of Na2SO4 were established. The crystallization nucleation of Na2SO4 crystals was found to be secondary nucleation controlled by surface reactions. The basic theoretical research of crystallization in this study is expected to fundamentally promote the application of fractional crystallization to realize the resource utilization of high-salinity wastewater in the coal chemical industry.

Efficient bioremediation of multiple steroid hormones by halotolerant 17ß-hydroxysteroid dehydrogenase derived from moderately halophilic Pontibacillus chungwhensis HN14.

Steroid hormones exhibit potent endocrine disrupting activity and have been shown to disrupt the equilibrium of aquatic ecosystems and pose a threat to public health through their persistent and carcinogenic effects. Pontibacillus chungwhensis HN14, a moderately halophilic bacterium with the capacity to effectively degrade various polycyclic aromatic hydrocarbons and other organic pollutants, was previously isolated. Additionally, the strain HN14 showed strong environmental adaptability under various environmental stress conditions. In this study, the steroid degradation by strain HN14 was studied for the first time. We demonstrated that strain HN14 could degrade estradiol (E2) to maintain the growth of the strain and could convert E2 to estrone. Additionally, the efficient substrate degradation efficiency of P. chungwhensis HN14 under high salinity and high substrate concentration conditions was demonstrated. Furthermore, a 17ß-hydroxysteroid dehydrogenase, 17ß-HSD(HN14), was identified in strain HN14. Comparative analysis reveals that 17ß-HSD(HN14) shares approximately 38% sequence identity with 17ß-HSDx from Rhodococcus sp. P14. In addition, 100 µg of purified 17ß-HSD(HN14) could effectively convert about 40% of 0.25 mM of E2 within 1 h period, with an enzyme activity of 17.5 U/mg, and catalyze the dehydrogenation of E2 and testosterone at the C-17 position. The characterization of purified enzyme properties reveals that 17ß-HSD(HN14) exhibits exceptional structural robustness and enzymatic efficacy even under high salinity conditions of up to 20%. Overall, this study enhances our comprehension of steroid biodegradation in strain HN14 and contributes novel ideas and theoretical underpinnings for advancing bioremediation technologies targeting steroid pollution in high-saline environments.

Salt and drought stress-mitigating approaches in sugar beet (Beta vulgaris L.) to improve its performance and yield.

MAIN CONCLUSION: Although sugar beet is a salt- and drought-tolerant crop, high salinity, and water deprivation significantly reduce its yield and growth. Several reports have demonstrated stress tolerance enhancement through stress-mitigating strategies including the exogenous application of osmolytes or metabolites, nanoparticles, seed treatments, breeding salt/drought-tolerant varieties. These approaches would assist in achieving sustainable yields despite global climatic changes. Sugar beet (Beta vulgaris L.) is an economically vital crop for ~ 30% of world sugar production. They also provide essential raw materials for bioethanol, animal fodder, pulp, pectin, and functional food-related industries. Due to fewer irrigation water requirements and shorter regeneration time than sugarcane, beet cultivation is spreading to subtropical climates from temperate climates. However, beet varieties from different geographical locations display different stress tolerance levels. Although sugar beet can endure moderate exposure to various abiotic stresses, including high salinity and drought, prolonged exposure to salt and drought stress causes a significant decrease in crop yield and production. Hence, plant biologists and agronomists have devised several strategies to mitigate the stress-induced damage to sugar beet cultivation. Recently, several studies substantiated that the exogenous application of osmolytes or metabolite substances can help plants overcome injuries induced by salt or drought stress. Furthermore, these compounds likely elicit different physio-biochemical impacts, including improving nutrient/ionic homeostasis, photosynthetic efficiency, strengthening defense response, and water status improvement under various abiotic stress conditions. In the current review, we compiled different stress-mitigating agricultural strategies, prospects, and future experiments that can secure sustainable yields for sugar beets despite high saline or drought conditions.

Unexpectedly Enhanced Organics Removal in Persulfate Oxidation with High Concentration of Sulfate: The Origin and the Selectivity.

Massive anions in high saline wastewater are primary factors that restricted the efficiency of pollutant degradation in advanced oxidation processes (AOPs). Herein, we reported the influence laws of different anions at high concentration on the electron-transfer process in the activation of persulfate, and especially, the sulfate anion exhibited the excellent promotion effect. Depending on the ionic charge, polarizability, and size, the anions exerted diverse effects on the dispersed phase and zeta potential of carbonaceous catalysts, which further embodied in the removal of pollutants. Based on the differences of reaction rate constant in water solution and high saline solution, the order was ClO4- < NO3- < Cl- < SO42- < CO32-, obeying the Hofmeister series. The enhancement of the sulfate anion was widely confirmed with different carbonaceous catalysts and pollutants with various structures. It could be attributed to the higher oxidation capacity, the faster interfacial electron transfer, and the better catalyst dispersion in the high sulfate environment. On the other hand, the decrease of zeta potential of the catalyst induced by sulfate reinforced the electrostatic attraction or repulsion with pollutants, which caused the selectivity of the sulfate promotion effect. Overall, this study provides new insights into the mechanism of influence of anions on AOPs, which refreshed the cognition of the role of sulfate on pollutant degradation, and helps guide the treatment design of high salinity wastewater.

Water Self-Purification with Zero External Consumption by Livestock Manure Resource Utilization.

Improper disposal of waste biomass and an increasing number of emerging contaminants (ECs) in water environment are universal threats to the global environment. Here, we creatively propose a sustainable strategy for the direct resource transformation of livestock manure (LM) into an innovative catalyst (Fe-CCM) for water self-purification with zero external consumption. ECs can be rapidly degraded in this self-purification system at ambient temperature and atmospheric pressure, without any external oxidants or energy input, accompanied by H2O and dissolved oxygen (DO) activation. The performance of the self-purification system is not affected by various types of salinity in the wastewater, and the corresponding second-order kinetic constant is improved 7 times. The enhanced water self-purification mechanism reveales that intermolecular forces between anions and pollutants reinforce electron exchange between pollutants and metal sites on the catalyst, further inducing the utilization of the intrinsic energy of contaminants, H2O, and DO through the interfacial reaction. This work provides new insights into the rapid removal of ECs in complicated water systems with zero external consumption and is expected to advance the resource utilization of livestock waste.

Adaptive laboratory evolution empowers lipids and biomass overproduction in Chlorella vulgaris for environmental applications.

Microalgal strain improvement with commercial features is needed to generate green biological feedstock to produce lipids for bioenergy. Hence, improving algal strain with enhanced lipid content without hindering cellular physiological parameters is pivotal for commercial applications of microalgae. In this report, we demonstrated the adaptive laboratory evolution (ALE) by hypersaline conditions to improve the algal strains for increasing the lipid overproduction capacity of Chlorella vulgaris for environmental applications. The evolved strains (namely E2 and E2.5) without notable impairment in general physiological parameters were scrutinized after 35 cycles. Conventional gravimetric lipid analysis showed that total lipid accumulation was hiked by 2.2-fold in the ALE strains compared to the parental strains. Confocal observation of algal cells stained with Nile-red showed that the abundance of lipid droplets was higher in the evolved strains without any apparent morphological aberrations. Furthermore, evolved strains displayed notable antioxidant potential than the control cells. Interestingly, carbohydrates and protein content were significantly decreased in the evolved cells, indicating that carbon flux was redirected into lipogenesis in the evolved cells. Altogether, our findings demonstrated a potential and feasible strategy for microalgal strain improvement for simultaneous lipids and biomass hyperaccumulation.

Impact of salinity on anaerobic ceramic membrane bioreactor for textile wastewater treatment: Process performance, membrane fouling and machine learning models.

Anaerobic membrane bioreactor (AnMBR) shows great potential for textile wastewater treatment, but high salinity in the influent may undermine its performance. This study evaluated the impact of salinity on the treatment performance of an upflow anaerobic sludge blanket (UASB) configured AnMBR using a flat sheet ceramic membrane. The salinity was stepwise increased (0, 5, 10 and 20 g/L) in four phases of the AnMBR operation. Results indicated that increased salinity jeopardized the COD removal efficiency of AnMBR from 92% to 73%, but had a marginal effect on dye removal efficacy (90-96%). Low salinity (5 g/L) boosted the biogas production whilst high salinity (>10 g/L) had a negative impact. Additionally, the increase of salinity resulted in the soluble microbial production (SMP) concentration soar and membrane fouling rate increase, peaking at a salinity of 10 g/L (Phase III) and recovering back to a lower level at a salinity of 20 g/L (Phase IV). This indicated a transition occurrence at a salinity of 10 g/L (Phase III). The microbial diversity analyses further suggested a transition from salinity-sensitive microbes (Aminiphilus, Caldatribacterium, Mesotoga, Methanobrevibacter, Methanobacterium, Methanosaeta) to salinity-tolerant microbes (Longilinea, Ignavibacterium, Rhodovarius, Bosea and Flexilinea). This transition can be associated with the increase SMP concentration and more severe membrane fouling in Phase III, which were mitigated after a new equilibrium was reached when the microbial consortium acclimatized to the high salinity. Finally, a machine learning model of the Adaboost algorithm was established to predict COD removal under different salinities. Importantly, this study revealed that AnMBR process performance and membrane operation can be maintained for high salinity textile wastewater treatment with a halophilic microbial community growth under high-salinity selection pressure.

Bioaugmentation performance for moving bed biofilm reactor (MBBR) treating mariculture wastewater by an isolated novel halophilic heterotrophic nitrification aerobic denitrification (HNAD) strain (Zobellella B307).

Moving bed biofilm reactor (MBBR) demonstrates weak nitrogen removal for mariculture wastewater treatment under high salinity environment. An isolated novel halophilic heterotrophic nitrification aerobic denitrification (HNAD) strain (Zobellella B307) was applied in MBBR process to enhance nitrogen removal. Results showed that strain Zobellella B307 could remove 90.9% ammonia nitrogen (NH4+-N) and 97.1% nitrate nitrogen (NO3--N) after 10 h cultivation, and strong resistance to salinity variation (high growth and nitrogen removal efficiency with salinity of 65‰) was observed. Besides, the chemical oxygen demand (COD), NH4+-N and NO3--N removal reached 95.6%, 94.4% and 85.7% with the strain added into MBBR process. In addition, microbial community structure analysis reflected that the strain Zobellella B307 successfully proliferated (the relative abundance increased to 2.33%). The HNAD bacteria abundance increased and dominated during the nitrogen removal process with the strain inoculation. A microbial functional analysis revealed that the main dominant functional categories (carbohydrate metabolism and amino acid metabolism) increased with the bioaugmentation of strain Zobellella B307, thus improving the nitrogen removal.

Fracturing Fluid Polymer Thickener with Superior Temperature, Salt and Shear Resistance Properties from the Synergistic Effect of Double-Tail Hydrophobic Monomer and Nonionic Polymerizable Surfactant.

To develop high-salinity, high-temperature reservoirs, two hydrophobically associating polymers as fracturing fluid thickener were respectively synthesized through aqueous solution polymerization with acrylamide (AM), acrylic acid (AA), 2-acrylamido-2-methylpropanesulfonic acid (AMPS), nonionic polymerizable surfactant (NPS) and double-tail hydrophobic monomer (DHM). The thickener ASDM (AM/AA/AMPS/NPS/DHM) and thickener ASD (AM/AA/AMPS/DHM) were compared in terms of properties of water dissolution, thickening ability, rheological behavior and sand-carrying. The results showed that ASDM could be quickly diluted in water within 6 min, 66.7% less than that of ASD. ASDM exhibited salt-thickening performance, and the apparent viscosity of 0.5 wt% ASDM reached 175.9 mPa·s in 100,000 mg/L brine, 100.6% higher than that of ASD. The viscosity of 0.5 wt% ASDM was 85.9 mPa·s after shearing for 120 min at 120 °C and at 170 s-1, 46.6% higher than that of ASD. ASDM exhibited better performance in thickening ability, viscoelasticity, shear recovery, thixotropy and sand-carrying than ASD. The synergistic effect of hydrophobic association and linear entanglement greatly enhancing the performance of ASDM and the compactness of the spatial network structure of the ASDM was enhanced. In general, ASDM exhibited great potential for application in extreme environmental conditions with high salt and high temperatures.

Microbial degradation of petroleum characteristic pollutants in hypersaline environment, emphasizing n-hexadecane and 2,4 di-tert-butylphenol.

In this paper, nine strains of salt-tolerant petroleum-degrading bacteria were applied to an biological aerated filter. Simulating the degradation of high-salinity petroleum wastewater with n-hexadecane and 2,4-ditert-butylphenol as the primary pollutants and analyzing the structure of the biofilm at various salt concentrations. According to the results, when the salinity was 4%, the COD removal efficiency reached 74.34%. Various halotolerant microorganisms have adapted to various salt concentrations. At a salinity of 3%, n-hexadecane exhibited the best degradation effect, with a rate of 83.21%. Shewanella, Acinetobacter, and Marinobacter were the predominant bacterial groups at the time. At 4% salinity, Acinetobacter and Pseudomonas were the predominant bacteria, and the average 2,4-ditert-butylphenol degradation rate was the highest at 63.02%. This study provided an experimental basis for further studying the biological treatment of high-salinity petroleum wastewater.

Functional Insights of Salinity Stress-Related Pathways in Metagenome-Resolved Methanothrix Genomes.

Recently, methanogenic archaea belonging to the genus Methanothrix were reported to have a fundamental role in maintaining stable ecosystem functioning in anaerobic bioreactors under different configurations/conditions. In this study, we reconstructed three Methanothrix metagenome-assembled genomes (MAGs) from granular sludge collected from saline upflow anaerobic sludge blanket (UASB) reactors, where Methanothrix harundinacea was previously implicated with the formation of compact and stable granules under elevated salinity levels (up to 20 g/L Na+). Genome annotation and pathway analysis of the Methanothrix MAGs revealed a genetic repertoire supporting their growth under high salinity. Specifically, the most dominant Methanothrix (MAG_279), classified as a subspecies of Methanothrix_A harundinacea_D, had the potential to augment its salinity resistance through the production of different glycoconjugates via the N-glycosylation process, and via the production of compatible solutes as Nε-acetyl-ß-lysine and ectoine. The stabilization and reinforcement of the cell membrane via the production of isoprenoids was identified as an additional stress-related pathway in this microorganism. The improved understanding of the salinity stress-related mechanisms of M. harundinacea highlights its ecological niche in extreme conditions, opening new perspectives for high-efficiency methanisation of organic waste at high salinities, as well as the possible persistence of this methanogen in highly-saline natural anaerobic environments. IMPORTANCE Using genome-centric metagenomics, we discovered a new Methanothrix harundinacea subspecies that appears to be a halotolerant acetoclastic methanogen with the flexibility for adaptation in the anaerobic digestion process both at low (5 g/L Na+) and high salinity conditions (20 g/L Na+). Annotation of the recovered M. harundinacea genome revealed salinity stress-related functions, including the modification of EPS glycoconjugates and the production of compatible solutes. This is the first study reporting these genomic features within a Methanothrix sp., a milestone further supporting previous studies that identified M. harundinacea as a key-driver in anaerobic granulation under high salinity stress.

ECAP is a key negative regulator mediating different pathways to modulate salt stress-induced anthocyanin biosynthesis in Arabidopsis.

Anthocyanins are a subgroup of plant flavonoids with antioxidant activities and are often induced by various biotic and abiotic stresses in plants, probably to efficiently scavenge free radicals and reactive oxygen species. However, the regulatory mechanisms of salt stress-induced anthocyanin biosynthesis remain unclear. Using molecular and genetic techniques we demonstrated key roles of ECAP in differential salt-responsive anthocyanin biosynthesis pathways in Arabidopsis thaliana. ECAP, JAZ6/8 and TPR2 are known to form a transcriptional repressor complex, and negatively regulate jasmonate (JA)-responsive anthocyanin accumulation. In this study, we demonstrated that under moderate salt stress, the accumulation of anthocyanins is partially dependent on JA signaling, which degrades JAZ proteins but not ECAP. More interestingly, we found that high salinity rather than moderate salinity induces the degradation of ECAP through the 26S proteasome pathway, and this process is independent of JA signaling. Further analysis revealed that ECAP interacts with MYB75 (a transcription factor activating anthocyanin biosynthetic genes) and represses its transcriptional activity in the absence of high salinity. Our results indicated that plants adopt different strategies for fine-tuning anthocyanin accumulation under different levels of salt stress, and further elucidated the complex regulation of anthocyanin biosynthesis during plant development and responses to environmental stresses.

Karyopherin Kap114p-mediated trans-repression controls ribosomal gene expression under saline stress.

Nuclear accessibility of transcription factors controls gene expression, co-regulated by Ran-dependent nuclear localization and a competitive regulatory network. Here, we reveal that nuclear import factor-facilitated transcriptional repression attenuates ribosome biogenesis under chronic salt stress. Kap114p, one of the karyopherin-ßs (Kap-ßs) that mediates nuclear import of yeast TATA-binding protein (yTBP), exhibits a yTBP-binding affinity four orders of magnitude greater than its counterparts and suppresses binding of yTBP with DNA. Our crystal structure of Kap114p reveals an extensively negatively charged concave surface, accounting for high-affinity basic-protein binding. KAP114 knockout in yeast leads to a high-salt growth defect, with transcriptomic analyses revealing that Kap114p modulates expression of genes associated with ribosomal biogenesis by suppressing yTBP binding to target promoters, a trans-repression mechanism we attribute to reduced nuclear Ran levels under salinity stress. Our findings reveal that Ran integrates the nuclear transport pathway and transcription regulatory network, allowing yeast to respond to environmental stresses.

Fabrication of nitrogen-doped graphene nanosheets anchored with carbon nanotubes for the degradation of tetracycline in saline water.

The treatment of wastewater with high salinity is still a challenge because of the quenching effect of various anions on radical processes. The nonradical process may be a more promising pathway. Herein, a 3D structured nitrogen-doped graphene nanosheet anchored with carbon nanotubes (N-GS-CNTs) was prepared by direct pyrolysis of K3Fe(CN)6. The as-prepared catalyst can effectively activate peroxymonosulfate (PMS) for mineralization of tetracycline (TC) over a wide pH range (from 3 to 11) and even in high saline water (500 mM Cl-, HCO3-, etc.). The degradation mechanism was elucidated by both experimental characterizations and DFT calculations. The high catalytic efficiency was attributed to accelerated electron transfer from donor (TC) to acceptor (PMS) in the presence of the catalyst, which acts as electron shuttle mediators to promote a nonradical process. At the same time, the catalyst also enhances the production of singlet oxygen (1O2), hence further increasing the degradation rate. This study not only provides a simple method for synthesizing N-GS-CNT catalysts but also provides new insights into the electron transfer pathway for the removal of organic pollutants under high salinity conditions.