Osmotic Conditions Research Articles

Cell volume maintenance capacity of the sea anemone Bunodosoma cangicum: the effect of copper.

Cell volume regulation is an essential strategy for the maintenance of life under unfavorable osmotic conditions. Mechanisms aimed at minimizing the physiological challenges caused by environmental changes are crucial in anisosmotic environments. However, aquatic ecosystems experience multiple stressors, including variations in salinity and heavy metal pollution. The accumulation of heavy metals in aquatic ecosystems has a significant effect on the biota, leading to impaired function. The aim of this study was to investigate the capacity of volume regulation in isolated cells of the sea anemone Bunodosoma cangicum exposed to nominal copper (Cu) concentrations of 5 and 50µg L-1, associated or not with hypoosmotic (15‰) or hyperosmotic (45‰) shock for 15min. In the absence of the metal, our results showed volume maintenance in all osmotic conditions. Our results showed that cell volume was maintained under all osmotic conditions in the absence of Cu. Similarly, no significant differences were observed in cell volumes under isosmotic and hyperosmotic conditions in the presence of both Cu concentrations. A similar homeostatic response was observed under the hypoosmotic condition with 5µg L-1 Cu. Our results showed an increase in cell volume with exposure of the cells to the hypoosmotic condition and 50µg L-1 Cu. The response could be associated with the increased bioavailability of Cu, reduced ability to resist multixenobiotics and their efflux pathways, and the impairment of water efflux in specialized transmembrane proteins. Therefore, B. cangicum pedal disk cells can tolerate osmotic variations in aquatic ecosystems. However, the capacity to regulate cell volume under hypoosmotic conditions can be affected by the presence of a metal contaminant (50µg L-1 Cu), which could be due to the inhibition of water channels.

Physiological and morphological responses of ‘Irukandji’ polyps to thermal and osmotic conditions: consequences for niche profiling

The Irukandji jellyfish (Carukia barnesi) is a medically important species. While the medusa stage of this species is well known, due to its highly venomous sting, the benthic polyp has core roles in regulating both the timing and abundance of medusa making it a research priority. However, due to their small size, Carukia barnesi polyps have never been found in situ and, basic ecological knowledge surrounding this life stage is limited. In this study we adopt a lab-based approach, utilizing physiological tolerance as a functional tool, to gain new insights into the in situ location for Carukia barnesi polyps. The physiological tolerance of Carukia barnesi polyps was characterized by measuring the oxygen consumption rates of polyps exposed to different salinity/temperature combinations. A total of nine salinities and seven temperatures were investigated, ranging from 11 °C/16‰ to 34 °C/42.5‰, encompassing the spectrum of environments experienced on the Great Barrier Reef. Polyps were also monitored for morphological changes such as asexual reproduction, polyp deterioration, and mortality. Salinity did not have a significant effect on oxygen consumption rates, with Carukia barnesi polyps displaying a significant tolerance to a wide range of salinities. The effect of temperature, however, was statistically significant with oxygen °consumption rates increasing alongside water temperature. There was no statistical evidence to support an interactive effect between salinity and temperature. Based on these results, we conclude that the polyp stage of this species is likely located in an environment with stable temperatures and fluctuating salinities and, consequently, future endeavors aimed at locating this life stage should expand targeted survey areas outside stable oceanic environments, typical of medusa, and encompass dynamic environments such as estuaries and submarine freshwater upwellings.

Grapevine bZIP transcription factor bZIP45 regulates VvANN1 and confers drought tolerance in Arabidopsis.

Drought is a severe environmental condition that restricts the vegetative growth and reduces the yield of grapevine (Vitis vinifera L.). However, the mechanisms underlying grapevine response and adaptation to drought stress remain unclear. In the present study, we characterized an ANNEXIN gene, VvANN1, which plays a positive role in the drought stress response. The results indicated that VvANN1 was significantly induced by osmotic stress. Expression of VvANN1 in Arabidopsis thaliana enhanced osmotic and drought tolerance through modulating the level of MDA, H2O2, and O2 ·- at the seedling stage, implying that VvANN1 might be involved in the process of ROS homeostasis under drought or osmotic stress conditions. Moreover, we used yeast one-hybridization and chromatin immunoprecipitation assays to show that VvbZIP45 could regulate VvANN1 expression by directly binding to the promoter region of VvANN1 in response to drought stress. We also generated transgenic Arabidopsis that constitutively expressed the VvbZIP45 gene (35S::VvbZIP45) and further produced VvANN1Pro::GUS/35S::VvbZIP45 Arabidopsis plants via crossing. The genetic analysis results subsequently indicated that VvbZIP45 could enhance GUS expression in vivo under drought stress. Our findings suggest that VvbZIP45 may modulate VvANN1 expression in response to drought stress and reduce the impact of drought on fruit quality and yield.

How Does Stomatal Density and Residual Transpiration Contribute to Osmotic Stress Tolerance?

Osmotic stress that is induced by salinity and drought affects plant growth and development, resulting in significant losses to global crop production. Consequently, there is a strong need to develop stress-tolerant crops with a higher water use efficiency through breeding programs. Water use efficiency could be improved by decreasing stomatal transpiration without causing a reduction in CO2 uptake under osmotic stress conditions. The genetic manipulation of stomatal density could be one of the most promising strategies for breeders to achieve this goal. On the other hand, a substantial amount of water loss occurs across the cuticle without any contribution to carbon gain when the stomata are closed and under osmotic stress. The minimization of cuticular (otherwise known as residual) transpiration also determines the fitness and survival capacity of the plant under the conditions of a water deficit. The deposition of cuticular wax on the leaf epidermis acts as a limiting barrier for residual transpiration. However, the causal relationship between the frequency of stomatal density and plant osmotic stress tolerance and the link between residual transpiration and cuticular wax is not always straightforward, with controversial reports available in the literature. In this review, we focus on these controversies and explore the potential physiological and molecular aspects of controlling stomatal and residual transpiration water loss for improving water use efficiency under osmotic stress conditions via a comparative analysis of the performance of domesticated crops and their wild relatives.

NMR-based analysis of pore water state of unsaturated compacted bentonite considering the saline effect

Understanding the saline effect on the pore water state of unsaturated compacted bentonite at the microscale is critical for evaluating the performance of barrier materials in nuclear waste geological repositories. In this paper, the pore water redistribution and pore water state of unsaturated compacted bentonite considering the saline effect is investigated via nuclear magnetic resonance (NMR) technique. It is found that, with increasing pore water salinity, the pore water content decreases and pore water is distributed in smaller pores, which is consistent with the swelling deformation behavior of bentonite at the macroscale and the diffuse double layers (DDL) theory at the microscale. In addition, the saline effect on pore water distribution is weakened with the increase in matric suction due to the shrinkage deformation of bentonite during drying. Then, the saline effect on the pore water state of unsaturated compacted bentonite is discussed via the concept of effective activation energy. It is shown that the drainage of pore water and increased pore water salt concentration (or the increase of matric suction and osmotic suction) both elevate the effective activation energy of bentonite, decreasing pore water mobility. The saline effect on the permeability coefficient of bentonite is also discussed. For bentonite under larger matric suction and osmotic suction conditions, the permeability coefficient is smaller.

Synergic mitigation of saline-alkaline stress in wheat plant by silicon and Enterobacter sp. FN0603.

Although microorganisms and silicon are well documented as factors that mitigate salt stress, their effect mitigating saline-alkaline stress in plants remains unknown. In this study, wheat plant seeds were treated with silicon, Enterobacter sp. FN0603 alone and in combination of both. Wheat seeds were soaked in silicon and bacterial solutions and sown in pots containing artificial saline-alkaline soils to compare the effects among all treatments. The results showed that the treatments with silicon and FN0603 alone significantly changed plant morphology, enhanced the rhizosphere soil nutrient content and enzyme activities, improved some important antioxidant enzyme activities (e.g., superoxide dismutase) and the contents of small molecules (e.g., proline) that affected osmotic conditions in the top second leaves. However, treatment with silicon and FN0603 in combination significantly further increased these stress tolerance indexes and eventually promoted the plant growth dramatically compared to the treatments with silicon or FN0603 alone (p < 0.01), indicating a synergic plant growth-promoting effect. High relative abundance of strain FN0603 was detected in the treated plants roots, and silicon further improved the colonization of FN0603 in stressed wheat roots. Strain FN0603 particularly when present in combination with silicon changed the root endophytic bacterial and fungal communities rather than the rhizosphere communities. Bipartite network analysis, variation partitioning analysis and structure equation model further showed that strain FN0603 indirectly shaped root endophytic bacterial and fungal communities and improved plant physiology, rhizosphere soil properties and plant growth through significantly and positively directing FN0603-specific biomarkers (p < 0.05). This synergetic effect of silicon and plant growth-promoting microorganism in the mitigation of saline-alkaline stress in plants via shaping root endophyte community may provide a promising approach for sustainable agriculture in saline-alkaline soils.

Submicron-Sized In Situ Osmotic Pressure Sensors for In Vitro Applications in Biology.

Physical forces are important cues in determining the development and the normal function of biological tissues. While forces generated by molecular motors have been widely studied, forces resulting from osmotic gradients have been less considered in this context. A possible reason is the lack of direct in situ measurement methods that can be applied to cell and organ culture systems. Herein, novel kinds of resonance energy transfer(FRET)-based liposomal sensors are developed, so that their sensing range and sensitivity can be adjusted to satisfy physiological osmotic conditions. Several types of sensors are prepared, either based on polyethylene glycol- (PEG)ylated liposomes with steric stabilization and stealth property or on crosslinked liposomes capable of enduring relatively harsh environments for liposomes (e.g., in the presence of biosurfactants). The sensors are demonstrated to be effective in the measurement of osmotic pressures in pre-osteoblastic in vitro cell culture systems by means of FRET microscopy. This development paves the way toward the in situ sensing of osmotic pressures in biological culture systems.

Molecular Links between Flowering and Abiotic Stress Response: A Focus on Poaceae.

Extreme temperatures, drought, salinity and soil pollution are the most common types of abiotic stresses crops can encounter in fields; these variations represent a general warning to plant productivity and survival, being more harmful when in combination. Plant response to such conditions involves the activation of several molecular mechanisms, starting from perception to signaling, transcriptional reprogramming and protein modifications. This can influence the plant's life cycle and development to different extents. Flowering developmental transition is very sensitive to environmental stresses, being critical to reproduction and to agricultural profitability for crops. The Poacee family contains some of the most widespread domesticated plants, such as wheat, barley and rice, which are commonly referred to as cereals and represent a primary food source. In cultivated Poaceae, stress-induced modifications of flowering time and development cause important yield losses by directly affecting seed production. At the molecular level, this reflects important changes in gene expression and protein activity. Here, we present a comprehensive overview on the latest research investigating the molecular pathways linking flowering control to osmotic and temperature extreme conditions in agronomically relevant monocotyledons. This aims to provide hints for biotechnological strategies that can ensure agricultural stability in ever-changing climatic conditions.

In Silico Dissection of Regulatory Regions of PHT Genes from Saccharum spp. Hybrid and Sorghum bicolor and Expression Analysis of PHT Promoters under Osmotic Stress Conditions in Tobacco

Phosphorus (P) is the second-most essential macronutrient required for the growth and development of plants. It is involved in a number of cellular processes that contribute to the plant’s growth and development. This study investigated Saccharum spp. hybrid and Sorghum bicolor promoter regions of Phosphate transporters (PHT), viz., PHT1, PHT2, PHT3, PHT4, and PHO1, through in silico analysis. The transcription start sites (TSS), conserved motifs, and CpG islands were studied using various computational techniques. The distribution of TSSs indicated the highest promoter prediction scores (1.0). MSh2 and MSb4 were recognized as the common promoter motifs for PHT promoters, found in with 85 to 100% percentage of distribution. The CpG analysis revealed that the promoter regions of most PHT genes had low CpG density, indicating a possible tissue-specific expression. The PHT promoters were investigated for the presence of biotic- and abiotic-stress-associated transcription factor binding sites (TFbs) that revealed the presence of binding motifs for major transcription factors (TFs), namely, AP2/ERF, bHLH, bZIP, MYB, NAC, and WRKY. Therefore, the in-silico analysis of the promoter regions helps us to understand the regulation mechanism of phosphate transporter promoters and gene expression under stress management. The 5′ regulatory region of the EaPHT gene was isolated from Erianthus, a wild relative of the genus Saccharum. The promoter construct was prepared and transformed in tobacco wherein the promoter drove the expression of GUS. Analysis of GUS expression in transgenic tobacco revealed enhanced expression of GUS under salt-stress conditions. This is the first report of the isolation and characterization of a phosphate transporter gene promoter from Erianthus and is expected to be useful for the development of salt-stress transgenic crop plants.

PQBP5/NOL10 maintains and anchors the nucleolus under physiological and osmotic stress conditions

Polyglutamine binding protein 5 (PQBP5), also called nucleolar protein 10 (NOL10), binds to polyglutamine tract sequences and is expressed in the nucleolus. Using dynamic imaging of high-speed atomic force microscopy, we show that PQBP5/NOL10 is an intrinsically disordered protein. Super-resolution microscopy and correlative light and electron microscopy method show that PQBP5/NOL10 makes up the skeletal structure of the nucleolus, constituting the granule meshwork in the granular component area, which is distinct from other nucleolar substructures, such as the fibrillar center and dense fibrillar component. In contrast to other nucleolar proteins, which disperse to the nucleoplasm under osmotic stress conditions, PQBP5/NOL10 remains in the nucleolus and functions as an anchor for reassembly of other nucleolar proteins. Droplet and thermal shift assays show that the biophysical features of PQBP5/NOL10 remain stable under stress conditions, explaining the spatial role of this protein. PQBP5/NOL10 can be functionally depleted by sequestration with polyglutamine disease proteins in vitro and in vivo, leading to the pathological deformity or disappearance of the nucleolus. Taken together, these findings indicate that PQBP5/NOL10 is an essential protein needed to maintain the structure of the nucleolus.

Bilayer lipid membrane formation on surface assemblies with sparsely distributed tethers.

A combined computational and experimental study of small unilamellar vesicle (SUV) fusion on mixed self-assembled monolayers (SAMs) terminated with different deuterated tether moieties (-(CD2)7CD3 or -(CD2)15CD3) is reported. Tethered bilayer lipid membrane (tBLM) formation of synthetic 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine was initially probed on SAMs with controlled tether (d-alkyl tail) surface densities and lateral molecular packing using quartz crystal microbalance with dissipation monitoring (QCM-D). Long time-scale coarse-grained molecular dynamics (MD) simulations were then employed to elucidate the mechanisms behind the interaction between the SUVs and the different phases formed by the -(CD2)7CD3 and -(CD2)15CD3 tethers. Furthermore, a series of real time kinetics was recorded under different osmotic conditions using QCM-D to determine the accumulated lipid mass and for probing the fusion process. It is shown that the key factors driving the SUV fusion and tBLM formation on this type of surfaces involve tether insertion into the SUVs along with vesicle deformation. It is also evident that surface densities of the tethers as small as a few mol% are sufficient to obtain stable tBLMs with a high reproducibility. The described "sparsely tethered" tBLM system can be advantageous in studying different biophysical phenomena, such as membrane protein insertion, effects of receptor clustering, and raft formation.

Germination niche of a neotropical dry forest species: seed osmotic stress and recovery

Abstract: Erythrina velutina Willd is a native Fabaceae with wide occurrence, mainly in the Caatinga Domain with multiple uses. This study aimed to compare the vulnerability/tolerance of seeds of four accessions of E. velutina to osmotic stresses. Four replications were conducted with 25 seeds, germinated in osmotic solutions of NaCl or PEG up to -1.73 MPa. Seeds that did not germinate after 14 days were recovered in distilled water. Obtained data were fitted in Boltzmann curves and t50 and synchrony were evaluated. A probit analysis was performed, and hydrotime and halotime models were constructed. In distilled water, the germination curve was similar for all accessions. When the concentration of salt or PEG in the substrate increased, the accessions differentiated into two groups. Two accessions - Jutaí 2012 and Jutaí 2015 - maintained high germination up to -0.86 MPa NaCl, while Caboclo 2008 and Caiçara 2011 accessions presented a lower germination speed. The hydrotime and halotime analyses separated the germination response of accessions according to their physiological quality and tolerance to osmotic and saline conditions. Seeds viability was maintained at both stresses, since germination was reestablished at distilled water, thus constituting a crucial response for this species’ survival and conservation.

Polyethylene Glycol Mediated Osmotic Stress Impacts on Growth and Biochemical Aspects of Wheat Under Artificial Osmotic Stress Condition

Polyethylene Glycol Mediated Osmotic Stress Impacts on Growth and Biochemical Aspects of Wheat Under Artificial Osmotic Stress Condition

Development of a diluent storage media and effect of cryoprotectants on semen collected from white spotted bamboo sharks (Chiloscyllium plagiosum) and cooled to 5 ºC

Three experiments were performed to evaluate the effects of osmolarity, temperature, media and cryoprotectants on white spotted bamboo shark (Chiloscyllium plagiosum) spermatozoa. Semen was collected from 7 different males using a massage technique with the animals held in tonic immobilization with gills submerged. A Computer Assisted Sperm Analysis (CASA) system was used to assess sperm motility. Experiment one evaluated the effects of Hank’s balanced salt solution (HBSS) with different osmolarities (200, 400, 800 and 1000 mOsm Kg1) and incubation temperatures (5 °C and 24 °C [room temperature]) over time (6 h, 12 h and 24 h). Membrane integrity and morphology were degraded (P < 0.05) over time; however, sperm extended with HBSS 1000 and HBSS 800 were least affected. Higher total motility (TM) and average path velocity (VAP, P > 0.05) sperm parameters were observed with HBSS 1000 and HBSS 800, respectively. In addition, TM, VAP, and curvilinear velocity (VCL) were all reduced (P < 0.05) compared to sperm held at 5 °C. Experiment two compared the effects of sperm diluted in crystal reef water (CRW), elasmobranch ringers (ER) or raw held at 5 °C. Results indicated that CRW had improved viability, TM, VAP, VSL, VCL, and straightness (STR) when compared to sperm diluted in ER or Raw at 5 °C. The final experiment compared different cryoprotectants (dimethyl sulfoxide [DMSO] 10%, DMSO 20%, Ethylene-glycol [EG] 10%, EG 20%, and Trehalose [Tre] 10%, Tre 20%) during incubation at 5 °C. Across all time points, sperm incubated in Tre (10 or 20%) had increased (P < 0.05) membrane integrity and normal morphology compared to all other media and raw. Across all time points, sperm exposed to Tre (10% and 20%) and 10% DMSO had significantly improved TM (P < 0.05) compared to EG (10% and 20%) and 20% DMSO which were similar to raw. Although sperm collected from WSBS are robust and appear to tolerate a wide range of osmotic conditions and media, media made at 1000 mOsm Kg1 should provide the best in-vitro osmotic pressure, while storing at cooler temperatures should improve storage survival. For future cryopreservation studies, media using TRE and DMSO as cryoprotectants should be evaluated.

Isolation and Characterization of Yeasts from Locally Available Foods

A study was conducted to identify different isolates of yeasts which are prospective to be utilized in various industries from locally available foods. Altogether 24 yeast isolates were obtained from fermented fruits and vegetables (banana, cabbage, grapes, lime and mango), pudding, bee honey, toddy and fermented fish samples using the pour plate method. Characterization of the yeasts using several biochemical tests (urease, catalase, liquid carbon and nitrogen assimilation and sugar fermentation tests) revealed that this pool was composed with yeast strains belong to genera of Saccharomyces, Kluvyeromyces, Candida, Pseudozyma, Cryptococcus, Rhodotorula and Debaryomyces. The most effective lactose fermenters were identified as viable candidates for bioethanol production and for manufacturing fermented dairy products for lactose intolerant people. Yeasts with the highest biomass production were suggested as the best viable candidates for industrial single cell protein production using whey, the major byproduct of the dairy industry. The five thermo-tolerant yeasts (Y55, Y57, Y58, Y59 and Y70) and Y069 which was optimally active under 10C were recognized as suitable for industrial applications. The isolates tolerant for high osmotic pressure conditions were identified as potential isolates to be used in highly concentrated food products. Sugarcane juice was recognized as a possible medium for the cultivation of these yeasts in industrial settings. The beneficial yeasts forecasting in this study are expected to screen using molecular biological methods to utilize them in industrial applications.

Deciphering the global roles of Cold shock proteins in Listeria monocytogenes nutrient metabolism and stress tolerance.

Listeria monocytogenes (Lm) accounts for serious public health and food safety problems owing to its stress resilience and pathogenicity. Based on their regulatory involvement in global gene expression events, cold-shock domain family proteins (Csps) are crucial in expression of various stress fitness and virulence phenotypes in bacteria. Lm possesses three Csps (CspA, CspB, and CspD) whose regulatory roles in the context of the genetic diversity of this bacterium are not yet fully understood. We examined the impacts of Csps deficiency on Lm nutrient metabolism and stress tolerance using a set of csp deletion mutants generated in different genetic backgrounds. Phenotype microarrays (PM) analysis showed that the absence of Csps in ∆cspABD reduces carbon (C-) source utilization capacity and increases Lm sensitivity to osmotic, pH, various chemical, and antimicrobial stress conditions. Single and double csp deletion mutants in different Lm genetic backgrounds were used to further dissect the roles of individual Csps in these phenotypes. Selected PM-based observations were further corroborated through targeted phenotypic assays, confirming that Csps are crucial in Lm for optimal utilization of various C-sources including rhamnose and glucose as well as tolerance against NaCl, β-phenyethylamine (PEA), and food relevant detergent stress conditions. Strain and genetic lineage background-based differences, division of labour, epistasis, and functional redundancies among the Csps were uncovered with respect to their roles in various processes including C-source utilization, cold, and PEA stress resistance. Finally, targeted transcriptome analysis was performed, revealing the activation of csp gene expression under defined stress conditions and the impact of Csps on expression regulation of selected rhamnose utilization genes. Overall, our study shows that Csps play important roles in nutrient utilization and stress responses in Lm strains, contributing to traits that are central to the public health and food safety impacts of this pathogen.

Antibacterial Effects of ZnO Nanodisks: Shape Effect of the Nanostructure on the Lethality in Escherichia coli.

The role of the shape of the nanostructure on the antibacterial effects of ZnO nanodisks has been investigated by detailed mass spectrometry-based proteomics along with other spectroscopic and microscopic studies on E. coli. The primary interaction study of the E. coli cells in the presence of ZnO nanodisks showed rigorous cell surface damage disrupting the cell wall/membrane components detected by microscopic and ATR-FTIR studies. Protein profiling of whole-cell extracts in the presence and absence of ZnO nanodisks identified several proteins that are upregulated and downregulated under the stress of the nanodisks. This suggests that the bacterial response to the primary stress leads to a secondary impact of ZnO nanodisk toxicity via regulation of the expression of specific proteins. Results showed that the ZnO nanodisks lead to the over-expression of peptidyl-dipeptidase Dcp, Transketolase-1, etc., which are important to maintaining the osmotic balance in the cell. The abrupt change in osmotic pressure leads to mechanical injury to the membrane, and nutritional starvation conditions, which is revealed from the expression of the key proteins involved in membrane-protein assembly, maintaining membrane integrity, cell division processes, etc. Thus, indicating a deleterious effect of ZnO nanodisk on the protective layer of E. coli. ZnO nanodisks seem to primarily affect the protective membrane layer, inducing cell death via the development of osmotic shock conditions, as one of the possible reasons for cell death. These results unravel a unique behavior of the disk-shaped ZnO nanostructure in executing lethality in E. coli, which has not been reported for other known shapes or morphologies of ZnO nanoforms.

ThASR3 confers salt and osmotic stress tolerances in transgenic Tamarix and Arabidopsis

BackgroundASR (abscisic acid-, stress-, and ripening-induced) gene family plays a crucial role in responding to abiotic stresses in plants. However, the roles of ASR genes protecting plants against high salt and drought stresses remain unknown in Tamarix hispida.ResultsIn this study, a salt and drought-induced ASR gene, ThASR3, was isolated from Tamarix hispida. Transgenic Arabidopsis overexpressing ThASR3 exhibited stimulating root growth and increasing fresh weight compared with wild-type (WT) plants under both salt and water deficit stresses. To further analyze the gain- and loss-of-function of ThASR3, the transgenic T. hispida plants overexpressing or RNA interference (RNAi)-silencing ThASR3 were generated using transient transformation. The overexpression of ThASR3 in Tamarix and Arabidopsis plants displayed enhanced reactive oxygen species (ROS) scavenging capability under high salt and osmotic stress conditions, including increasing the activities of antioxidant enzymes and the contents of proline and betaine, and reducing malondialdehyde (MDA) content and electrolyte leakage rates.ConclusionOur results indicate that ThASR3 functions as a positive regulator in Tamarix responses to salt and osmotic stresses and confers multiple abiotic stress tolerances in transgenic plants, which may have an important application value in the genetic improvement of forest tree resistance.

The two-component system histidine kinase EnvZ contributes to Avian pathogenic Escherichia coli pathogenicity by regulating biofilm formation and stress responses

EnvZ, the histidine kinase (HK) of OmpR/EnvZ, transduces osmotic signals in Escherichia coli K12 and affects the pathogenicity of Shigella flexneri and Vibrio cholera. Avian pathogenic E. coli (APEC) is an extra-intestinal pathogenic E. coli (ExPEC), causing acute and sudden death in poultry and leading to severe economic losses to the global poultry industry. How the functions of EnvZ correlate with APEC pathogenicity was still unknown. In this study, we successfully constructed the envZ mutant strain AE17ΔenvZ and the inactivation of envZ significantly reduced biofilms and altered red, dry, and rough (rdar) morphology. In addition, AE17ΔenvZ was significantly less resistant to acid, alkali, osmotic, and oxidative stress conditions. Deletion of envZ significantly enhanced sensitivity to specific pathogen-free (SPF) chicken serum and increased adhesion to chicken embryonic fibroblast DF-1 cells and elevated inflammatory cytokine IL-1β, IL6, and IL8 expression levels. Also, when compared with the WT strain, AE17ΔenvZ attenuated APEC pathogenicity in chickens. To explore the molecular mechanisms underpinning envZ in APEC17, we compared the WT and envZ-deletion strains using transcriptome analyses. RNA-Seq results identified 711 differentially expressed genes (DEGs) in the envZ mutant strain and DEGs were mainly enriched in outer membrane proteins, stress response systems, and TCSs. Quantitative real-time reverse transcription PCR (RT-qPCR) showed that EnvZ influenced the expression of biofilms and stress responses genes, including ompC, ompT, mlrA, basR, hdeA, hdeB, adiY, and uspB. We provided compelling evidence showing EnvZ contributed to APEC pathogenicity by regulating biofilms and stress response expression.

Pumping Ratio of the Na+/K+ Pump — A Further View

Our current knowledge about the Na+/K+ pump is mainly of the structure and function of the pump itself. The knowledge of the relationship between the function of the pump and other cellular functions is inadequate. This is because we are unaware of what the pumping process is and why the pump takes 3 Na+ to 2 K+ as the pumping ratio. In recent years, I proposed a dipole-charging model to simulate the pumping process of the Na+/K+ pump and showed by this model that this pumping ratio of the Na+/K+ pump is determined by the ratio of the ionic mobilities of K+ to Na+ ions. In this article, I further propose an osmotic balance condition that the potassium ion concentration inside the cell and the sodium ion concentration outside the cell are equal. With this condition, the dipole-charging model is further extended to compare different pumping ratios and it shows that only the pumping ratio 3 Na+ to 2 K+ can be selected during evolution. This result also reveals the relationships between the 3 Na+ to 2 K+ pumping ratio, the membrane resting potential, and the osmotic balance condition of the cell. Furthermore, I discuss the osmotic balance condition, the stability of the dipole-charging model and the dipole-charging process, the electrogenic contribution by the Na+/K+ pump to the resting membrane potential, and the effectiveness and the prospect of the dipole-charging model. The dipole-charging model is capable of explaining all the phenomena related to the pumping process of the Na+/K+ pump and is especially suitable for study in situations under pathologic conditions, such as ischemia or hypoxia. Finally, I present analyses of the free energy change of the coupled overall reaction for the Na+/K+ pump and of some reports on different pumping ratios of the Na+/K+ pump. The exception of the pumping ratio in the brine shrimp Artemia is also discussed.