Variation Characteristics of Root Traits of Different Alfalfa Cultivars under Saline-Alkaline Stress and their Relationship with Soil Environmental Factors

Soil salinization is the main factor that threatens the growth and development of plants and limits the increase of yield.It is of great significance to study the key soil environmental factors affecting plant root traits to reveal the adaptation strategies of plants to saline-alkaline-stressed soil environments.In this study,the root biomass,root morphological parameters and root mineral nutrient content of two alfalfa cultivars with different sensitivities to alkaline stress were analyzed with black soil as the control group and the mixed saline-alkaline soil with a ratio of 7:3 between black soil and saline-alkaline soil as the saline-alkaline treatment group.At the same time,the correlation analysis of soil salinity indexes,soil nutrient indexes and the activities of key enzymes involved in soil carbon,nitrogen and phosphorus cycles was carried out.The results showed that compared with the control group,the pH,EC,and urease(URE)of the soil surrounding the roots of two alfalfa cultivars were significantly increased,while soil total nitrogen(TN),total phosphorus(TP),organic carbon(SOC),andα-glucosidase activity(AGC)were significantly decreased under saline-alkaline stress.There was no significant difference in root biomass and root morphological parameters of saline-alkaline tolerant cultivar GN under saline-alkaline stress.The number of root tips(RT),root surface area(RS)and root volume(RV)of AG were reduced by 61.16%,44.54%,and 45.31%,respectively,compared with control group.The ratios of K^(+)/Na^(+),Ca^(2+)/Na^(+)and Mg^(2+)/Na^(+)of GN were significantly higher than those of AG(p<0.05).The root fresh weight(RFW)and dry weight(RDW),root length(RL),RV and RT of alfalfa were positively regulated by soil SOC and TN,but negatively regulated by soil pH,EC,and URE(p<0.01).Root Ca^(2+)/Na+ratio was significantly positively correlated with soil TN,TP and SOC(p<0.01).The absorption of Mg and Ca ions in roots is significantly negatively regulated by soilβ-glucosidase activity(BGC)and acid phosphatase activity(APC)(p<0.05).This study improved knowledge of the relationship between root traits and soil environmental factors and offered a theoretical framework for elucidating how plant roots adapt to saline-alkaline stressed soil environments.

Grain Yield,Biomass Accumulation,and Leaf Photosynthetic Characteristics of Rice under Combined Salinity-Drought Stress

Simultaneous stresses of salinity and drought often coincide during rice-growing seasons in saline lands,primarily due to insufficient water resources and inadequate irrigation facilities.Consequently,combined salinity-drought stress poses a major threat to rice production.In this study,two salinity levels(NS,non-salinity;HS,high salinity)along with three drought treatments(CC,control condition;DJ,drought stress imposed at jointing;DH,drought stress imposed at heading)were performed to investigate their combined influences on leaf photosynthetic characteristics,biomass accumulation,and rice yield formation.Salinity,drought,and their combination led to a shortened growth period from heading to maturity,resulting in a reduced overall growth duration.Grain yield was reduced under both salinity and drought stress,with a more substantial reduction under the combined salinity-drought stress.The combined stress imposed at heading caused greater yield losses in rice compared with the stress imposed at jointing.Additionally,the combined salinity-drought stress induced greater decreases in shoot biomass accumulation from heading to maturity,as well as in shoot biomass and nonstructural carbohydrate(NSC)content in the stem at heading and maturity.However,it increased the harvest index and NSC remobilization reserve.Salinity and drought reduced the leaf area index and SPAD value of flag leaves and weakened the leaf photosynthetic characteristics as indicated by lower photosynthetic rates,transpiration rates,and stomatal conductance.These reductions were more pronounced under the combined stress.Salinity,drought,and especially their combination,decreased the activities of ascorbate peroxidase,catalase,and superoxide dismutase,while increasing the contents of malondialdehyde,hydrogen peroxide,and superoxide radical.Our results indicated a more significant yield loss in rice when subjected to combined salinity-drought stress.The individual and combined stresses of salinity and drought diminished antioxidant enzyme activities,inhibited leaf photosynthetic functions,accelerated leaf senescence,and subsequently lowered assimilate accumulation and grain yield.

Ferroptosis and endoplasmic reticulum stress in ischemic stroke

Ferroptosis is a form of non-apoptotic programmed cell death,and its mechanisms mainly involve the accumulation of lipid peroxides,imbalance in the amino acid antioxidant system,and disordered iron metabolism.The primary organelle responsible for coordinating external challenges and internal cell demands is the endoplasmic reticulum,and the progression of inflammatory diseases can trigger endoplasmic reticulum stress.Evidence has suggested that ferroptosis may share pathways or interact with endoplasmic reticulum stress in many diseases and plays a role in cell survival.Ferroptosis and endoplasmic reticulum stress may occur after ischemic stroke.However,there are few reports on the interactions of ferroptosis and endoplasmic reticulum stress with ischemic stroke.This review summarized the recent research on the relationships between ferroptosis and endoplasmic reticulum stress and ischemic stroke,aiming to provide a reference for developing treatments for ischemic stroke.

Overexpression of the transcription factor MdWRKY115 improves drought and osmotic stress tolerance by directly binding to the MdRD22 promoter in apple

Abiotic stress reduces plant yield and quality.WRKY transcription factors play key roles in abiotic stress responses in plants,but the molecular mechanisms by which WRKY transcription factors mediate responses to drought and osmotic stresses in apple(Malus×domestica Borkh.)remain unclear.Here,we functionally characterized the apple GroupⅢWRKY gene MdWRKY115.qRT-PCR analysis showed that MdWRKY115 expression was up-regulated by drought and osmotic stresses.GUS activity analysis revealed that the promoter activity of MdWRKY115 was enhanced under osmotic stress.Subcellular localization and transactivation assays indicated that MdWRKY115 was localized to the nucleus and had a transcriptional activity domain at the N-terminal region.Transgenic analysis revealed that the overexpression of MdWRKY115 in Arabidopsis plants and in apple callus markedly enhanced their tolerance to drought and osmotic stresses.DNA affinity purification sequencing showed that MdWRKY115 binds to the promoter of the stress-related gene MdRD22.This binding was further verified by an electrophoretic mobility shift assay.Collectively,these findings suggest that MdWRKY115 is an important regulator of osmotic and drought stress tolerance in apple.

Piezo1 channel exaggerates ferroptosis of nucleus pulposus cells by mediating mechanical stress-induced iron influx

To date,several molecules have been found to facilitate iron influx,while the types of iron influx channels remain to be elucidated.Here,Piezo1 channel was identified as a key iron transporter in response to mechanical stress.Piezo1-mediated iron overload disturbed iron metabolism and exaggerated ferroptosis in nucleus pulposus cells(NPCs).Importantly,Piezo1-induced iron influx was independent of the transferrin receptor(TFRC),a well-recognized iron gatekeeper.Furthermore,pharmacological inactivation of Piezo1 profoundly reduced iron accumulation,alleviated mitochondrial ROS,and suppressed ferroptotic alterations in stimulation of mechanical stress.Moreover,conditional knockout of Piezo1(Col2a1-CreERT Piezo1^(flox/flox))attenuated the mechanical injury-induced intervertebral disc degeneration(IVDD).Notably,the protective effect of Piezo1 deficiency in IVDD was dampened in Piezo1/Gpx4 conditional double knockout(cDKO)mice(Col2a1-CreERT Piezo1^(flox/flox)/Gpx4^(flox/flox)).These findings suggest that Piezo1 is a potential determinant of iron influx,indicating that the Piezo1-iron-ferroptosis axis might shed light on the treatment of mechanical stress-induced diseases.

Synergistic effects of carbon cycle metabolism and photosynthesis in Chinese cabbage under salt stress

Chinese cabbage(Brassica rapa ssp. pekinensis) has a long cultivation history and is one of the vegetable crops with the largest cultivation area in China. However, salt stress severely damages photosynthesis and hormone metabolism, nutritional balances, and results in ion toxicity in plants. To better understand the mechanisms of salt-induced growth inhibition in Chinese cabbage, RNA-seq and physiological index determination were conducted to explore the impacts of salt stress on carbon cycle metabolism and photosynthesis in Chinese cabbage. Here, we found that the number of thylakoids and grana lamellae and the content of starch granules and chlorophyll in the leaves of Chinese cabbage under salt stress showed a time-dependent response, first increasing and then decreasing. Chinese cabbage increased the transcript levels of genes related to the photosynthetic apparatus and carbon metabolism under salt stress, probably in an attempt to alleviate damage to the photosynthetic system and enhance CO_(2) fixation and energy metabolism. The transcription of genes related to starch and sucrose synthesis and degradation were also enhanced;this might have been an attempt to maintain intracellular osmotic pressure by increasing soluble sugar concentrations. Soluble sugars could also be used as potential reactive oxygen species(ROS) scavengers, in concert with peroxidase(POD)enzymes, to eliminate ROS that accumulate during metabolic processes. Our study characterizes the synergistic response network of carbon metabolism and photosynthesis under salt stress.

Pre‑hatch thermal manipulation of embryos and post‑hatch baicalein supplementation mitigated heat stress in broiler chickens

Background High environmental temperatures induce heat stress in broiler chickens,affecting their health and pro-duction performance.Several dietary,managerial,and genetics strategies have been tested with some success in mitigating heat stress(HS)in broilers.Developing novel HS mitigation strategies for sustaining broiler production is critically needed.This study investigated the effects of pre-hatch thermal manipulation(TM)and post-hatch baica-lein supplementation on growth performance and health parameters in heat-stressed broilers.Results Six hundred fertile Cobb 500 eggs were incubated for 21 d.After candling on embryonic day(ED)10,238 eggs were thermally manipulated at 38.5℃ with 55%relative humidity(RH)from ED 12 to 18,then transferred to the hatcher(ED 19 to 21,standard temperature)and 236 eggs were incubated at a controlled temperature(37.5℃)till hatch.After hatch,180-day-old chicks from both groups were raised in 36 pens(n=10 birds/pen,6 replicates per treatment).The treatments were:1)Control,2)TM,3)control heat stress(CHS),4)thermal manipulation heat stress(TMHS),5)control heat stress supplement(CHSS),and 6)thermal manipulation heat stress supplement(TMHSS).All birds were raised under the standard environment for 21 d,followed by chronic heat stress from d 22 to 35(32–33℃ for 8 h)in the CHS,TMHS,CHSS,and TMHSS groups.A thermoneutral(22–24℃)environment was maintained in the Control and TM groups.RH was constant(50%±5%)throughout the trial.All the data were analyzed using one-way ANOVA in R and GraphPad software at P<0.05 and are presented as mean±SEM.Heat stress significantly decreased(P<0.05)the final body weight and ADG in CHS and TMHS groups compared to the other groups.Embryonic TM significantly increased(P<0.05)the expression of heat shock protein-related genes(HSP70,HSP90,and HSPH1)and antioxidant-related genes(GPX1 and TXN).TMHS birds showed a significant increment(P<0.05)in total cecal volatile fatty acid(VFA)concentration compared to the CHS birds.The cecal microbial analysis showed significant enrichment(P<0.05)in alpha and beta diversity and Coprococcus in the TMHSS group.Conclusions Pre-hatch TM and post-hatch baicalein supplementation in heat-stressed birds mitigate the detrimental effects of heat stress on chickens’growth performance,upregulate favorable gene expression,increase VFA produc-tion,and promote gut health by increasing beneficial microbial communities.

A semi-analytical model for coupled flow in stress-sensitive multi-scale shale reservoirs with fractal characteristics

A large number of nanopores and complex fracture structures in shale reservoirs results in multi-scale flow of oil. With the development of shale oil reservoirs, the permeability of multi-scale media undergoes changes due to stress sensitivity, which plays a crucial role in controlling pressure propagation and oil flow. This paper proposes a multi-scale coupled flow mathematical model of matrix nanopores, induced fractures, and hydraulic fractures. In this model, the micro-scale effects of shale oil flow in fractal nanopores, fractal induced fracture network, and stress sensitivity of multi-scale media are considered. We solved the model iteratively using Pedrosa transform, semi-analytic Segmented Bessel function, Laplace transform. The results of this model exhibit good agreement with the numerical solution and field production data, confirming the high accuracy of the model. As well, the influence of stress sensitivity on permeability, pressure and production is analyzed. It is shown that the permeability and production decrease significantly when induced fractures are weakly supported. Closed induced fractures can inhibit interporosity flow in the stimulated reservoir volume (SRV). It has been shown in sensitivity analysis that hydraulic fractures are beneficial to early production, and induced fractures in SRV are beneficial to middle production. The model can characterize multi-scale flow characteristics of shale oil, providing theoretical guidance for rapid productivity evaluation.

Adverse effects of early-life stress:focus on the rodent neuroendocrine system

Early-life stress is associated with a high prevalence of mental illnesses such as post-traumatic stress disorders,attention-deficit/hyperactivity disorder,schizophrenia,and anxiety or depressive behavior,which constitute major public health problems.In the early stages of brain development after birth,events such as synaptogenesis,neuron maturation,and glial differentiation occur in a highly orchestrated manner,and external stress can cause adverse long-term effects throughout life.Our body utilizes multifaceted mechanisms,including neuroendocrine and neurotransmitter signaling pathways,to appropriately process external stress.Newborn individuals first exposed to early-life stress deploy neurogenesis as a stress-defense mechanism;however,in adulthood,early-life stress induces apoptosis of mature neurons,activation of immune responses,and reduction of neurotrophic factors,leading to anxiety,depression,and cognitive and memory dysfunction.This process involves the hypothalamus-pituitary-adrenal axis and neurotransmitters secreted by the central nervous system,including norepinephrine,dopamine,and serotonin.The rodent early-life stress model is generally used to experimentally assess the effects of stress during neurodevelopment.This paper reviews the use of the early-life stress model and stress response mechanisms of the body and discusses the experimental results regarding how early-life stress mediates stress-related pathways at a high vulnerability of psychiatric disorder in adulthood.

Evolution of microstructure and properties of a novel Ni-based superalloy during stress relief annealing

We discussed the decrease in residual stress,precipitation evolution,and mechanical properties of GH4151 alloy in different annealing temperatures,which were studied by the scanning electron microscope(SEM),high-resolution transmission electron microscopy(HRTEM),and electron backscatter diffraction(EBSD).The findings reveal that annealing processing has a significant impact on diminishing residual stresses.As the annealing temperature rose from 950 to 1150℃,the majority of the residual stresses were relieved from 60.1 MPa down to 10.9 MPa.Moreover,the stress relaxation mechanism transitioned from being mainly controlled by dislocation slip to a combination of dislocation slip and grain boundary migration.Meanwhile,the annealing treatment promotes the decomposition of the Laves,accompanied by the precipitation ofμ-(Mo_(6)Co_(7))starting at 950℃ and reaching a maximum value at 1050℃.The tensile strength and plasticity of the annealing alloy at 1150℃ reached the maximum(1394 MPa,56.1%)which was 131%,200%fold than those of the as-cast alloy(1060 MPa,26.6%),but the oxidation process in the alloy was accelerated at 1150℃.The enhancement in durability and flexibility is primarily due to the dissolution of the brittle phase,along with the shape and dispersal of theγ′phase.

U-Net Models for Representing Wind Stress Anomalies over the Tropical Pacific and Their Integrations with an Intermediate Coupled Model for ENSO Studies

El Niño-Southern Oscillation(ENSO)is the strongest interannual climate mode influencing the coupled ocean-atmosphere system in the tropical Pacific,and numerous dynamical and statistical models have been developed to simulate and predict it.In some simplified coupled ocean-atmosphere models,the relationship between sea surface temperature(SST)anomalies and wind stress(τ)anomalies can be constructed by statistical methods,such as singular value decomposition(SVD).In recent years,the applications of artificial intelligence(AI)to climate modeling have shown promising prospects,and the integrations of AI-based models with dynamical models are active areas of research.This study constructs U-Net models for representing the relationship between SSTAs andτanomalies in the tropical Pacific;the UNet-derivedτmodel,denoted asτUNet,is then used to replace the original SVD-basedτmodel of an intermediate coupled model(ICM),forming a newly AI-integrated ICM,referred to as ICM-UNet.The simulation results obtained from ICM-UNet demonstrate their ability to represent the spatiotemporal variability of oceanic and atmospheric anomaly fields in the equatorial Pacific.In the ocean-only case study,theτUNet-derived wind stress anomaly fields are used to force the ocean component of the ICM,the results of which also indicate reasonable simulations of typical ENSO events.These results demonstrate the feasibility of integrating an AI-derived model with a physics-based dynamical model for ENSO modeling studies.Furthermore,the successful integration of the dynamical ocean models with the AI-based atmospheric wind model provides a novel approach to ocean-atmosphere interaction modeling studies.

Biochanin A attenuates spinal cord injury in rats during early stages by inhibiting oxidative stress and inflammasome activation

Previous studies have shown that Biochanin A,a flavonoid compound with estrogenic effects,can serve as a neuroprotective agent in the context of cerebral ischemia/reperfusion injury;howeve r,its effect on spinal cord injury is still unclea r. In this study,a rat model of spinal cord injury was established using the heavy o bject impact method,and the rats were then treated with Biochanin A(40 mg/kg) via intrape ritoneal injection for 14 consecutive days.The res ults showed that Biochanin A effectively alleviated spinal cord neuronal injury and spinal co rd tissue injury,reduced inflammation and oxidative stress in spinal cord neuro ns,and reduced apoptosis and pyroptosis.In addition,Biochanin A inhibited the expression of inflammasome-related proteins(ASC,NLRP3,and GSDMD)and the Toll-like receptor 4/nuclear factor-κB pathway,activated the Nrf2/heme oxygenase 1 signaling pathway,and increased the expression of the autophagy markers LC3 Ⅱ,Beclin-1,and P62.Moreove r,the therapeutic effects of Biochanin A on early post-s pinal cord injury were similar to those of methylprednisolone.These findings suggest that Biochanin A protected neurons in the injured spinal cord through the Toll-like receptor 4/nuclear factor κB and Nrf2/heme oxygenase 1 signaling pathways.These findings suggest that Biochanin A can alleviate post-spinal cord injury at an early stage.

Frequency Domain Fatigue Evaluation on SCR Girth-Weld Based on Structural Stress

The Steel Catenary Riser(SCR)is a vital component for transporting oil and gas from the seabed to the floating platform.The harsh environmental conditions and complex platform motion make the SCR’s girth-weld prone to fatigue failure.The structural stress fatigue theory and Master S-N curve method provide accurate predictions for the fatigue damage on the welded joints,which demonstrate significant potential and compatibility in multi-axial and random fatigue evaluation.Here,we propose a new frequency fatigue model subjected to welded joints of SCR under multiaxial stress,which fully integrates the mesh-insensitive structural stress and frequency domain random process and transforms the conventional welding fatigue technique of SCR into a spectrum analysis technique utilizing structural stress.Besides,a full-scale FE model of SCR with welds is established to obtain the modal structural stress of the girth weld and the frequency response function(FRF)of modal coordinate,and a biaxial fatigue evaluation about the girth weld of the SCR can be achieved by taking the effects of multi-load correlation and pipe-soil interaction into account.The research results indicate that the frequency-domain fatigue results are aligned with the time-domain results,meeting the fatigue evaluation requirements of the SCR.

Activation of the wnt/β-catenin/CYP1B1 pathway alleviates oxidative stress and protects the blood-brain barrier under cerebral ischemia/reperfusion conditions

Accumulating evidence suggests that oxidative stress and the Wnt/β-catenin pathway participate in stroke-induced disruption of the blood-brain barrier.However,the potential links between them following ischemic stroke remain largely unknown.The present study found that cerebral ischemia leads to oxidative stress and repression of the Wnt/β-catenin pathway.Meanwhile,Wnt/β-catenin pathway activation by the pharmacological inhibito r,TWS119,relieved oxidative stress,increased the levels of cytochrome P4501B1(CYP1B1)and tight junction-associated proteins(zonula occludens-1[ZO-1],occludin and claudin-5),as well as brain microvascular density in cerebral ischemia rats.Moreove r,rat brain microvascular endothelial cells that underwent oxygen glucose deprivation/reoxygenation displayed intense oxidative stress,suppression of the Wnt/β-catenin pathway,aggravated cell apoptosis,downregulated CYP1B1and tight junction protein levels,and inhibited cell prolife ration and migration.Overexpression ofβ-catenin or knockdown ofβ-catenin and CYP1B1 genes in rat brain mic rovascular endothelial cells at least partly ameliorated or exacerbated these effects,respectively.In addition,small interfering RNA-mediatedβ-catenin silencing decreased CYP1B1 expression,whereas CYP1B1 knoc kdown did not change the levels of glycogen synthase kinase 3β,Wnt-3a,andβ-catenin proteins in rat brain microvascular endothelial cells after oxygen glucose deprivatio n/reoxygenation.Thus,the data suggest that CYP1B1 can be regulated by Wnt/β-catenin signaling,and activation of the Wnt/β-catenin/CYP1B1 pathway contributes to alleviation of oxidative stress,increased tight junction levels,and protection of the blood-brain barrier against ischemia/hypoxia-induced injury.

Involvement of the ABA-and H_(2)O_(2)-Mediated Ascorbate-Glutathione Cycle in the Drought Stress Responses of Wheat Roots

Abscisic acid(ABA),hydrogen peroxide(H_(2)O_(2)) and ascorbate(AsA)–glutathione(GSH)cycle are widely known for their participation in various stresses.However,the relationship between ABA and H_(2)O_(2) levels and the AsA–GSH cycle under drought stress in wheat has not been studied.In this study,a hydroponic experiment was conducted in wheat seedlings subjected to 15%polyethylene glycol(PEG)6000–induced dehydration.Drought stress caused the rapid accumulation of endogenous ABA and H_(2)O_(2) and significantly decreased the number of root tips compared with the control.The application of ABA significantly increased the number of root tips,whereas the application of H_(2)O_(2) markedly reduced the number of root tips,compared with that under 15%PEG-6000.In addition,drought stress markedly increased the DHA,GSH and GSSG levels,but decreased the AsA levels,AsA/DHA and GSH/GSSG ratios compared with those in the control.The activities of the four enzymes in the AsA–GSH cycle were also markedly increased under drought stress,including glutathione reductase(GR),ascorbate peroxidase(APX),monodehydroascorbate reductase(MDHAR)and dehydroascorbate reductase(DHAR),compared with those in the control.However,the application of an ABA inhibitor significantly inhibited GR,DHAR and APX activities,whereas the application of an H_(2)O_(2) inhibitor significantly inhibited DHAR and MDHAR activities.Furthermore,the application of ABA inhibitor significantly promoted the increases of H_(2)O_(2) and the application of H_(2)O_(2) inhibitor significantly blocked the increases of ABA,compared with those under 15% PEG-6000.Taken together,the results indicated that ABA and H_(2)O_(2) probably interact under drought stress in wheat;and both of them can mediate drought stress by modulating the enzymes in AsA–GSH cycle,where ABA acts as the main regulator of GR,DHAR,and APX activities,and H_(2)O_(2) acts as the main regulator of DHAR and MDHAR activities.

Treatment with β-sitosterol ameliorates the effects of cerebral ischemia/reperfusion injury by suppressing cholesterol overload, endoplasmic reticulum stress, and apoptosis

β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unknown whetherβ-sitosterol treatment reduces the effects of ischemic stroke.Here we found that,in a mouse model of ischemic stroke induced by middle cerebral artery occlusion,β-sitosterol reduced the volume of cerebral infarction and brain edema,reduced neuronal apoptosis in brain tissue,and alleviated neurological dysfunction;moreover,β-sitosterol increased the activity of oxygen-and glucose-deprived cerebral cortex neurons and reduced apoptosis.Further investigation showed that the neuroprotective effects ofβ-sitosterol may be related to inhibition of endoplasmic reticulum stress caused by intracellular cholesterol accumulation after ischemic stroke.In addition,β-sitosterol showed high affinity for NPC1L1,a key transporter of cholesterol,and antagonized its activity.In conclusion,β-sitosterol may help treat ischemic stroke by inhibiting neuronal intracellular cholesterol overload/endoplasmic reticulum stress/apoptosis signaling pathways.

Heat stress induced hepatocyte apoptosis in largemouth bass Micropterus salmoides via IRE1α/TRAF2/ASK1/JNK pathway

Heat stress(HS)has been shown to adversely affect fish livers and can lead to extensive apoptosis.To investigate the relationship between endoplasmic reticulum(ER)stress and HS-induced apoptosis in fish livers,we isolated and cultured primary hepatocytes of largemouth bass,Micropterus salmoides by trypsin method,then established an in-vitro model of liver cells under HS(35℃).The contents of lactic dehydrogenase(LDH)and hydrogen peroxide(H2O2)were determined to evaluate the effects of HS on hepatocyte injury and oxidative stress.RT-qPCR was performed to discover the key genes in unfolded protein response(UPR)pathways involved at different HS duration.ERS inhibitor 4-PBA and IRE1αinhibitor 4μ8C were used to further investigate the effects of HS on IRE1αapoptosis pathway in hepatocytes.Results show that HS led to significant increases in the release of LDH,the content of H2O2,and the expressions of oxidative protein folding genes(ero1αand pdi)under HS,suggesting severe hepatocyte injury and oxidative stress happened in heat-stressed largemouth bass hepatocytes.The continuous activation of IRE1αpathway genes(grp78,grp94,atf6,perk,eif2a,atf4,chop,ire1α,traf2,ask1,jnk1,and jnk2)indicated that HS led significantly to ER stress.In particular,the mRNA expression levels of ER stress-related genes(grp78,grp94,atf6,perk,ire1α,chop,jnk1,and jnk2)in the high temperature(HT)+4-PBA group and the HT+4μ8C group were significantly down-regulated under HS.After 4μ8C treatment,the expression levels of apoptosis-related genes(caspase-2,caspase-3,caspase-6,caspase-7,caspase-8,caspase-9,and caspase-10)and LDH content were significantly decreased,whereas the cell survival rate was significantly increased when given 4-PBA or 4μ8C treatment.These findings demonstrate that HS could induce liver apoptosis of largemouth bass through the IRE1αpathway,which may act as a key switch mediating liver apoptosis of largemouth bass under HS.

Upregulation of the glycine-rich protein-encoding gene GhGRPL enhances plant tolerance to abiotic and biotic stressors by promoting secondary cell wall development

Abiotic and biotic stressors adversely affect plant survival,biomass generation,and crop yields.As the global availability of arable land declines and the impacts of global warming intensify,such stressors may have increasingly pronounced effects on agricultural productivity.Currently,researchers face the overarching challenge of comprehensively enhancing plant resilience to abiotic and biotic stressors.The secondary cell wall plays a crucial role in bolstering the stress resistance of plants.To increase plant resistance to stress through genetic manipulation of the secondary cell wall,we cloned a cell wall protein designated glycine-rich protein-like(GhGRPL)from cotton fibers,and found that it is specifically expressed during the period of secondary cell wall biosynthesis.Notably,this protein differs from its Arabidopsis homolog,AtGRP,since its glycine-rich domain is deficient in glycine residues.GhGRPL is involved in secondary cell wall deposition.Upregulation of GhGRPL enhances lignin accumulation and,consequently,the thickness of the secondary cell walls,thereby increasing the plant’s resistance to abiotic stressors,such as drought and salinity,and biotic threats,including Verticillium dahliae infection.Conversely,interference with GhGRPL expression in cotton reduces lignin accumulation and compromises that resistance.Taken together,our findings elucidate the role of GhGRPL in regulating secondary cell wall development through its influence on lignin deposition,which,in turn,reinforces cell wall robustness and impermeability.These findings highlight the promising near-future prospect of adopting GhGRPL as a viable,effective approach for enhancing plant resilience to abiotic and biotic stress factors.

Three-dimensional stress variation characteristics in deep hard rock of CJPL-Ⅱ project based on in-situ monitoring

In deep hard rock excavation, stress plays a pivotal role in inducing stress-controlled failure. While the impact of excavation-induced stress disturbance on rock failure and tunnel stability has undergone comprehensive examination through laboratory tests and numerical simulations, its validation through insitu stress tests remains unexplored. This study analyzes the three-dimensional stress changes in the surrounding rock at various depths, monitored during the excavation of B2 Lab in China Jinping Underground Laboratory Phase Ⅱ(CJPL-Ⅱ). The investigation delves into the three-dimensional stress variation characteristics in deep hard rock, encompassing stress components and principal stress. The results indicate changes in both the magnitude and direction of the principal stress during tunnel excavation. To quantitatively describe the degree of stress disturbance, a series of stress evaluation indexes are established based on the distances between stress tensors, including the stress disturbance index(SDI), the principal stress magnitude disturbance index(SDIm), and the principal stress direction disturbance index(SDId). The SDI indicates the greatest stress disturbance in the surrounding rock is 4.5 m from the tunnel wall in B2 Lab. SDIm shows that the principal stress magnitude disturbance peaks at2.5 m from the tunnel wall. SDId reveals that the largest change in principal stress direction does not necessarily occur near the tunnel wall but at a specific depth from it. The established relationship between SDI and the depth of the excavation damaged zone(EDZ) can serve as a criterion for determining the depth of the EDZ in deep hard rock engineering. Additionally, it provides a reference for future construction and support considerations.

Stress tensor determination by modified hydraulic tests on pre-existing fractures:Method and stress constraints

The hydraulic testing of pre-existing fractures(HTPF)is one of the most promising in situ stress measurement methods,particularly for three-dimensional stress tensor determination.However,the stress tensor determination based on the HTPF method requires at least six tests or a minimum of 14-15 tests(under different conditions)for reliable results.In this study,we modified the HTPF method by considering the shear stress on each pre-existing fracture,which increased the number of equations for the stress tensor determination and decreased the number of tests required.Different shear stresses were attributed to different fractures by random sampling;therefore,the stress tensors were obtained by searching for the optimal solution using the least squares criterion based on the Monte Carlo method.Thereafter,we constrained the stress tensor based on the tensile strength criterion,compressive strength criterion,and vertical stress constraints.The inverted stress tensors were presented and analyzed based on the tensorial nature of the stress using the Euclidean mean stress tensor.Two stress-measurement campaigns in Weifang(Shandong Province,China)and Mercantour road tunnel(France)were implemented to highlight the validity and efficiency of the modified HTPF(M-HTPF)method.The results showed that the M-HTPF method can be applied for stress tensor inversion using only three to four tests on pre-existing fractures,neglecting the stress gradient.The inversion results were confined to relatively small distribution dispersions and were significantly reliable and stable due to the shear stresses on the fractures and the stress constraints employed.The M-HTPF method is highly feasible and efficient for complete stress tensor determination in a single borehole.