POSS and PEG Contained Copolymer for Antibioadhesive Rigid Contact Lenses Materials Application.

Myopia is a global public health issue. Rigid contact lenses (RCLs) are an effective way to correct or control myopia. However, bioadhesion issues remain one of the significant obstacles limiting its clinical application. Although enhancing hydrophilicity through various surface treatments can mitigate this problem, the duration of effectiveness is short-lived and the processing involved is complex and costly. Herein, an antiadhesive RCLs material was designed via 8-armed methacrylate-POSS (8MA-POSS), and poly(ethylene glycol) methacrylate (PEGMA) copolymerization with 3-[tris(trimethylsiloxy)silyl] propyl methacrylate (TRIS). The POSS and PEG segments incorporated P(TRIS-co-PEGMA-co-8MA-POSS) (PTPM) material was obtained and their optical transparency, refractive index, resolution, hardness, surface charge, thermal features, and wettability were tested and optimized. The antibioadhesion activities, including protein, lipid, and bacteria, were evaluated as well. In vitro and in vivo results indicated that the optimized antibioadhesive PTPM materials present good biocompatibility and biosafety. Thus, such POSS and PEG segments containing material were a potential antibioadhesive RCL material option.

Free Radicals Generated in Perfluorocarbon-Water (Liquid-Liquid) Interfacial Contact Electrification and Their Application in Cancer Therapy.

Electron transfer during solid-liquid contact electrification has been demonstrated to produce reactive oxygen species (ROS) such as hydroxyl radicals (•OH) and superoxide anion radicals (•O2-). Here, we show that such a process also occurs in liquid-liquid contact electrification. By preparing perfluorocarbon nanoemulsions to construct a perfluorocarbon-water "liquid-liquid" interface, we confirmed that electrons were transferred from water to perfluorocarbon in ultrasonication-induced high-frequency liquid-liquid contact to produce •OH and •O2-. The produced ROS could be applied to ablate tumors by triggering large-scale immunogenic cell death in tumor cells, promoting dendritic cell maturation and macrophage polarization, ultimately activating T cell-mediated antitumor immune response. Importantly, the raw material for producing •OH is water, so the tumor therapy is not limited by the endogenous substances (O2, H2O2, etc.) in the tumor microenvironment. This work provides new perspectives for elucidating the mechanism of generation of free radicals in liquid-liquid contact and provides an excellent tumor therapeutic modality.

Melatonin alleviates chronic stress-induced hippocampal microglia pyroptosis and subsequent depression-like behaviors by inhibiting Cathepsin B/NLRP3 signaling pathway in rats.

Melatonin improves chronic stress-induced hippocampal damage and depression-like behaviors, but the mechanism needs further study. This study was to explore the mechanism of melatonin inhibiting microglia pyroptosis. In virtro experiments, melatonin improved corticosterone-induced the ultrastructure and microstructure damage of HAPI cells by inhibiting pyroptosis, thereby increasing cell survival rate. Protein-protein interaction network and molecular autodocking predicted that Cathespin B might be the target of melatonin inhibition of NLRP3-mediated pyroptosis. Melatonin inhibited corticosterone-induced Cathespin B expression. Both Cathepsin B inhibitor CA-074Me and NLRP3 knockout inhibited the HAPI cells pyroptosis. Similarly, melatonin inhibited Cathepsin B agonist Pazopanib-induced activation of Cathepsin B/NLRP3 signaling pathway and HAPI cells pyroptosis. In vivo studies, melatonin inhibited chronic restraint stress (CRS)-induced activation of Cathepsin B/NLRP3 signaling pathway and alleviated hippocampal microglia pyroptosis in rats. Inhibition of microglia pyroptosis improved CRS-induced depression-like behaviors of rats. In addition, inhibition of Cathepsin B and NLRP3 alleviated hippocampal pyroptosis. Melatonin inhibited Pazopanib-induced activation of Cathepsin B/NLRP3 signaling pathway and hippocampal pyroptosis. These results demonstrated that melatonin could alleviate CRS-induced hippocampal microglia pyroptosis by inhibiting Cathepsin B/NLRP3 signaling pathway, thereby improving depression-like behaviors in rats. This study reveals the molecular mechanism of melatonin in the prevention and treatment of chronic stress-related encephalopathy.

Recent Progress in Implantable Drug Delivery Systems.

In recent years, tremendous effort is devoted to developing platforms, such as implantable drug delivery systems (IDDSs), with temporally and spatially controlled drug release capabilities and improved adherence. IDDSs have multiple advantages: i) the timing and location of drug delivery can be controlled by patients using specific stimuli (light, sound, electricity, magnetism, etc.). Some intelligent "closed-loop" IDDS can even realize self-management without human participation. ii) IDDSs enable continuous and stable delivery of drugs over a long period (months to years) and iii) to administer drugs directly to the lesion, thereby helping reduce dosage and side effects. iv) IDDSs enable personalized drug delivery according to patient needs. The high demand for such systems has prompted scientists to make efforts to develop intelligent IDDS. In this review, several common stimulus-responsive mechanisms including endogenous (e.g., pH, reactive oxygen species, proteins, etc.) and exogenous stimuli (e.g., light, sound, electricity, magnetism, etc.), are given in detail. Besides, several types of IDDS reported in recent years are reviewed, including various stimulus-responsive systems based on the above mechanisms, radio frequency-controlled IDDS, "closed-loop" IDDS, self-powered IDDS, etc. Finally, the advantages and disadvantages of various IDDS, bottleneck problems, and possible solutions are analyzed to provide directions for subsequent research.

Effects of running time on biological activated carbon filters: water purification performance and microbial community evolution.

Ozone-biologically activated carbon (BAC) filtration is an advanced treatment process that can be applied to remove recalcitrant organic micro-pollutants in drinking water treatment plants (DWTPs). In this study, we continuously monitored a new and an old BAC filter in a DWTP for 1 year to compare their water purification performance and microbial community evolution. The results revealed that, compared with the new filter, the use of the old BAC filter facilitated a slightly lower rate of dissolved organic carbon (DOC) removal. In the case of the new BAC filter, we recorded general increases in the biomass and microbial diversity of the biofilm with a prolongation of operating time, with the biomass stabilizing after 7 months. For both new and old BAC filters, Proteobacteria and Acidobacteria were the dominant bacterial phyla. At the genus level, the microbial community gradually shifted over the course of operation from a predominance of Herminiimonas and Hydrogenophaga to one predominated by Bradyrhizbium, Bryobacter, Hyphomicrobium, and Pedomicrobium, with Bradyrhizobium being established as the most abundant genus in the old BAC filter. Regarding spatial distribution, we detected reductions in the biomass and number of operational taxonomic units with increasing biofilm depth, whereas there was a corresponding increase in microbial diversity. However, compared with the effects of time, the influence of depth on the composition of the biofilm microbial community was considerably smaller. Furthermore, co-occurrence network analysis revealed that the microbial community network of the new filter after 11 months of operation was the most tightly connected, although its modular coefficient was the lowest of those assessed. We speculate that the positive and negative interactions within the network may be attributable to symbiotic or competitive relationships among species. Moreover, there may have been a significant negative interaction between SWB02 and Acidovorax, plausibly associated with a competition for substrates.

Photo-controllable drug releasing bulk polyacrylic intraocular lens material for safer posterior capsular opacification prevention.

Posterior capsular opacification (PCO) is the most common complication that occurs after intraocular lens (IOL) implantation in cataract therapy. In recent years, IOLs have been developed as drug delivery platforms, but concerns over the safety of uncontrolled proliferative drug release have arisen. Therefore, a controlled drug release strategy is needed for safer PCO prevention. In this study, a new monomer contained coumarin group was introduced in material preparation, and poly(ethylene glycol phenyl ether methacrylate-co-2-(2-ethoxyethoxy) ethyl acrylate-co-7-(2-methacryloyloxyethoxy)-4-methylcoumarin) (PEEC) acrylic IOL materials were synthesized. The antiproliferative drug 5-fluorouracil (5-FU) could be chemically grafted to the PEEC IOL materials easily via a light induced [2 + 2] cycloaddition reaction with the coumarin group, getting drug-loaded IOL (PEEC@5-FU IOL). The PEEC@5-FU IOL exhibited excellent optical and mechanical properties and biocompatibility. More importantly, the loaded 5-FU could be easily controlled from release by light irradiation via photo-dissociation of the cyclobutane ring that was obtained by the [2 + 2] cycloaddition reaction of 5-FU and coumarin. The in vitro and in vivo experiments demonstrated that such photo-controllable drug release IOL could effectively prevent PCO after implantation in a safe way.

A review: Hydrochar as potential adsorbents for wastewater treatment and CO2 adsorption.

To valorize the biomass and organic waste, hydrothermal carbonization (HTC) stands out as a highly efficient and promising pathway given its intrinsic advantages over other thermochemical processes. Hydrochar, as the main product obtained from HTC, is widely applied as a fuel source and soil conditioner. Aside from these applications, hydrochar can be either directly used or modified as bio-adsorbents for environmental remediation. This potential arises from its tunable surface chemistry and its suitability to act as a precursor for activated or engineered carbon. In view of the importance of this topic, this review offers a thorough examination of the research progress for using hydrochar and its modified forms to remove organic dyes (cationic and anionic dyes), heavy metals, herbicides/pesticides, pharmaceuticals, and CO2. The review also sheds light on the fundamental chemistry involved in HTC of biomass and the major analytical techniques applied for understanding surface chemistry of hydrochar and modified hydrochar. The knowledge gaps and potential hurdles are identified to highlight the challenges and prospects of this research field with a summary of the key findings from this review. Overall, this article provides valuable insights and directives and pinpoints the areas meriting further investigation in the application potential of hydrochar in wastewater management and CO2 capture.

Effects of physical method and enzymatic hydrolysis on the properties of soybean fiber-rich stabilizer for oil in water emulsions.

BACKGROUND: Okara is a by-product from the soybean industry and an abundant resource of insoluble soybean fiber (ISF). ISF with various properties could be obtained by different extraction methods. It is an attractive option to utilize okara by taking advantage of ISF as an emulsifier or stabilizer. RESULTS: Compared with the untreated ISF (ISFUT ), superfine grinding reduced the particle size and viscosity of ISF (ISFSG ). Steam explosion increased the water solubility from 17.5% to 51.7% but decreased the water holding capacity and swelling capacity of ISF (ISFSE ) from 15.0 and 14.0 g/g to 4.2 and 3.3 g/g, respectively. Emulsions prepared by ISFUT and ISFSG before or after enzymatic hydrolysis presented large oil droplets and were unstable. Although emulsions prepared by ISFSE after enzymatic hydrolysis (ISFSE-E ) showed flocculation, the volume-weighted average diameter (19.7 µm) were the smallest while the viscosity and viscoelastic modulus were the highest, and exhibited excellent physical stability during storage. CONCLUSION: ISF obtained by physical and hydrolysis treatment displayed diverging physicochemical properties while ISF prepared by steam explosion-enzymatic hydrolysis presented the best potential to stabilize emulsions. The present study could provide novel information about the utilization of okara by the application of ISF as an emulsifier or stabilizer. © 2023 Society of Chemical Industry.

[Blood Lipid Indicators and Different Clinical Classifications of Dyslipidemia and Diabetic Kidney Disease: Correlation and Predictive Value].

Objective: To explore the relationship between blood lipid indicators and different clinical classifications of dyslipidemia and diabetic kidney disease (DKD) and to compare the value of different clinical classifications of dyslipidemia for predicting DKD. Methods: Continuously enrollment of subjects was conducted at the First Affiliated Hospital of Chongqing Medical University and the Yongchuan Hospital of Chongqing Medical University between October 2020 and October 2021. A total of 356 type 2 diabetes mellitus (T2DM) patients admitted to the two hospitals were enrolled. They were divided into DKD group ( n=126) and simple T2DM group ( n=230) according to whether their T2DM was combined with DKD. In addition, 250 healthy individuals undergoing physical examination during the same period were enrolled for the control group. The blood pressure, blood lipid, blood glucose, and the kidney function indicators of the three groups were measured. The effects of different classifications of dyslipidemia on DKD were analyzed with unconditional logistic regression models, the receiver operating characteristic (ROC) curve was constructed, the area under the curve ( AUC) of ROC was calculated, and the value of different classifications of dyslipidemia for predicting DKD was analyzed. Results: The diastolic blood pressure (DBP), systolic blood pressure (SBP), total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-C), serum creatinine (Scr), uric acid (UA), and glycosylated hemoglobin A1c (HbA1c) of the DKD group and the simple T2DM group were significantly higher than those of the control group, while the high-density lipoprotein cholesterol (HDL-C) levels of the DKD group and the simple T2DM group were lower than that of the control group (all P<0.05). The disease course of T2DM, DBP, SBP, TC, TG, Scr, UA and HbA1c of the DKD group were significantly higher than those of the T2DM group (all P<0.05). After adjusting for the effects of T2DM disease course, DBP, SBP, Scr, UA and HbA1c, the results showed that TC ( OR=1.426, 95% CI: 1.088-1.868) and TG ( OR=1.404, 95% CI: 1.075-1.833) were independent risk factors for DKD, and that hypercholesterolemia ( OR=1.817, 95% CI: 1.040-3.177) and mixed hyperlipidemia ( OR=2.148, 95% CI: 1.110-4.159) were independent risk factors for DKD (all P<0.05). The AUC (95% CI) of hypercholesterolemia was 0.789 (0.729-0.871). The AUC (95% CI) of mixed hyperlipidemia was 0.671 (0.579-0.760). Hypercholesterolemia showed better predictive value for the diagnosis and prediction of DKD. Conclusion: Among the blood lipid indicators, TC and TG are independent risk factors of DKD. In the clinical classifications of dyslipidemia, hypercholesterolemia and mixed hyperlipidemia are independent risk factors of DKD. Hypercholesterolemia can be used as a predictor to screen for DKD among T2DM patients and is well suited for extensive application in outpatient screening.

Recent advances in the synthesis and application of magnetic biochar for wastewater treatment.

Magnetic biochar (MBC) is a novel bio-carbon material with both desired properties as adsorbent and magnetic characteristics. This review provides an up-to-date summary and discussion on the latest development of MBC, which covers the progress on its synthesis, application, and techno-economic analysis. The review indicates that the direct hydrothermal synthesis has been catching more research attention to produce MBC due to its mild reaction conditions. Instead of the Fe-loaded MBC, there is a trend of using Mn for the magnetization. For the MBC application, how to improve its adsorption performance for water decontamination, ideally to match that of the biochar (BC) or activated carbon, is important. In addition, more studies on the environmental impacts of MBC and life-cycle assessment decoding the process optimization options are necessary. This review will provide valuable references for the development of MBC and MBC-based materials for wastewater treatment.

Cadmium-tolerant Bacillus cereus 2-7 alleviates the phytotoxicity of cadmium exposure in banana plantlets.

Bananas are the world's important fruit and staple crop in the developing countries. Cadmium (Cd) contamination in soils results in the decrease of crop yield and food safety. Bioremediation is an environmental-friendly and effective measure using Cd-tolerant plant growth promoting rhizobacteria (PGPR). In our study, a Cd-resistant PGPR Bacillus cereus 2-7 was isolated and identified from a discarded gold mine. It could produce multiple plant growth promoting biomolecules such as siderophores, indole-3-acetic acid (IAA), 1-aminocyclopropane-1-carboxylate (ACC)-deaminase and phosphatase. The extracellular accumulation was a main manner of Cd removal. Surplus Cd induced the expression of Cd resistance/transport genes of B. cereus 2-7 to maintain the intracellular Cd homeostasis. The pot experiment showed that Cd contents decreased by 50.31 % in soil, 45.43 % in roots, 56.42 % in stems and 79.69 % in leaves after the strain 2-7 inoculation for 40 d. Bacterial inoculation alleviated the Cd-induced oxidative stress to banana plantlets, supporting by the increase of chlorophyll contents, plant height and total protein contents. The Cd remediation mechanism revealed that B. cereus 2-7 could remodel the rhizosphere bacterial community structure and improve soil enzyme activities to enhance the immobilization of Cd. Our study provides a Cd-bioremediation strategy using Cd-resistant PGPR in tropical and subtropical area.

Effects of phosphate addition on the removal of disinfection by-product formation potentials by biological activated carbon filtration.

In drinking water treatment plants (DWTPs), the widely used biological activated carbon filters (BACFs), as the last barrier before disinfection, can remove dissolved organic matter (DOM) known as precursors of disinfection by-products (DBPs). Whether phosphate addition can improve water purification and DBP control of BACFs is still controversial. This study investigated short-term and long-term effects of phosphate addition on controlling DBP formation potentials (FPs) by BACFs via column and batch experiments. The BAC columns presented good water purification performance: they removed around 50 % DOM, nearly all fulvic acid-likes and humic acid-likes as well as 5 %-70 % chlor(am)innated THM4, HAA9 and HAN4 FPs (except chloraminated THM4 FPs)， which was mainly contributed by aerobic bacteria not anoxic bacteria. Phosphate addition within 7-14 days further improved removals of DOM, aromatic organics, fluorescence fractions in DOM as well as HAA9 and HAN4 FPs (especially TCAA FP and TCAN FP) to different extent. However, this improvement did not last longer, and removals of DOM, aromatic organics, two fluorescence fractions (soluble microbial byproduct-likes and humic acid-likes) and DBP FPs decreased despite long-term phosphate addition. Oxic and anoxic batch experiments showed that the positive response of water purification to short-term phosphate addition was also mainly attributed to aerobic bacteria and not to anoxic bacteria. For example, the former decreased DOM and DBP FPs, while the latter increased protein- and tryptophan-like substances as well as chloraminated THM4 FPs. Phosphate addition resulted in EPS increase in anoxic reactors and decrease in oxic reactors. These results indicated that a high dissolved oxygen in BACFs may be helpful for water purification and DBP control. Overall, short-term phosphate addition into phosphorus-limited water is beneficial for BACFs to control DBPs while long-term addition has no effect. Therefore, an intermittent phosphate addition into BACFs is suggested to control DBPs in DWTPs.

A High-Linearity Glucose Sensor Based on Silver-Doped Con A Hydrogel and Laser Direct Writing.

A continuous glucose monitoring (CGM) system is an ideal monitoring system for the blood glucose control of diabetic patients. The development of flexible glucose sensors with good glucose-responsive ability and high linearity within a large detection range is still challenging in the field of continuous glucose detection. A silver-doped Concanavalin A (Con A)-based hydrogel sensor is proposed to address the above issues. The proposed flexible enzyme-free glucose sensor was prepared by combining Con-A-based glucose-responsive hydrogels with green-synthetic silver particles on laser direct-writing graphene electrodes. The experimental results showed that in a glucose concentration range of 0-30 mM, the proposed sensor is capable of measuring the glucose level in a repeatable and reversible manner, showing a sensitivity of 150.12 Ω/mM with high linearity of R2 = 0.97. Due to its high performance and simple manufacturing process, the proposed glucose sensor is excellent among existing enzyme-free glucose sensors. It has good potential in the development of CGM devices.

Non-viral gene coating modified IOL delivering PDGFR-α shRNA interferes with the fibrogenic process to prevent posterior capsular opacification.

Posterior capsule opacification (PCO), the most common complication after cataract surgery, is caused by the proliferation, migration and epithelial-mesenchymal transition (EMT) of residual lens epithelial cells in the capsule bag. Although the surface modification and drug loading of intraocular lens (IOLs) have been effective in preventing PCO to some extent, the intraocular safety of anti-proliferative drug application is still a major limitation in clinical application. In this study, we used non-viral gene delivery systems in combination with layer-by-layer (LBL) self-assembly technology, and the modified IOL could effectively prevent the development of PCO by interfering with the EMT process mediated by the platelet-derived growth factor receptor-α (PDGFR-α). Herein, the gene fragments were wrapped by electrostatic conjugation using polyethyleneimine-graft-poly(ethylene glycol) to form gene complexes. Gene complexes were characterized by dynamic light scattering, transmission electron microscopy (TEM) and agarose gel electrophoresis, and evaluated for storage and serum stability. The layer assembly behavior of the IOL surface, changes in optical properties and the release behavior of the gene complexes were characterized using quartz crystal microbalance, UV-vis, contact angle and TEM. In vitro experiments showed that the IOL coating has good bio-compatibility and can achieve the corresponding transfection effect, and the released gene complexes exhibited excellent cell internalization and lysosomal escape behaviors, as well as effective inhibition of PDGFR-α expression and its mediated EMT process. The early PCO prevention effect and bio-compatibility evaluation of the modified IOL in vivo were evaluated by implantation into animal eyes. This study provides a new strategy for the development of surface modifications of small nucleic acid drugs and non-toxic EMT interference therapies for PCO.

Assessing Future Climate Change Impacts on Potato Yields - A Case Study for Prince Edward Island, Canada.

Crop yields are adversely affected by climate change; therefore, it is crucial to develop climate adaptation strategies to mitigate the impacts of increasing climate variability on the agriculture system to ensure food security. As one of the largest potato-producing provinces in Canada, Prince Edward Island (PEI) has recently experienced significant instability in potato production. PEI's local farmers and stakeholders are extremely concerned about the prospects for the future of potato farming industries in the context of climate change. This study aims to use the Decision Support System for Agrotechnology Transfer (DSSAT) potato model to simulate future potato yields under the Coupled Model Intercomparison Project Phase 6 (CMIP6) climate scenarios (including SSP1-1.9, SSP1-2.6, SSP2-4.5, SSP3-7.0, and SSP5-8.5). The study evaluates the combined effects of changing climatic conditions at local scales (i.e., warming temperature and changing precipitation patterns) and increasing carbon dioxide (CO2) concentration in the atmosphere. The results indicate future significant declines in potato yield in PEI under the current farming practices. In particular, under the high-emission scenarios (e.g., SSP3-7.0 and SSP5-8.5), the potato yield in PEI would decline by 48% and 60% in the 2070s and by 63% and 80% by 2090s; even under the low-emission scenarios (i.e., SSP1-1.9 and SSP1-2.6), the potato yield in PEI would still decline by 6-10%. This implies that it is important to develop effective climate adaptation measures (e.g., adjusting farming practices and introducing supplemental irrigation plans) to ensure the long-term sustainability of potato production in PEI.

Correction: Design of foldable, responsively drug-eluting polyacrylic intraocular lens bulk materials for prevention of postoperative complications.

Correction for 'Design of foldable, responsively drug-eluting polyacrylic intraocular lens bulk materials for prevention of postoperative complications' by Yueze Hong et al., J. Mater. Chem. B, 2022, DOI: https://doi.org/10.1039/d2tb01974d.

Design of foldable, responsively drug-eluting polyacrylic intraocular lens bulk materials for prevention of postoperative complications.

Posterior capsular opacification (PCO), resulting from undesired intracapsular cell proliferation, is the most common complication of intraocular lens (IOL) implantation after cataract surgery. In recent years, IOLs have been developed into a drug delivery platform. Compared with traditional eye drops, drug-loaded IOLs have the characteristics of independent patient compliance and no other operation except surgical implantation. In this work, a series of poly(glycidyl methacrylate-co-2-(2-ethoxyethoxy)ethyl acrylate) (PGE) acrylic intraocular lens materials were synthesized as drug delivery platforms. The PGE synthesized with 10% crosslinking agent has excellent optical, foldable, and thermomechanical properties. An aldehyde group was subsequently introduced into the PGE chains, and an antiproliferative drug (doxorubicin) was immobilized onto the PGE chains via an H+-sensitive imine bond. The IOL exhibits H+-dependent Dox release behavior in a simulated pathological environment. The in vitro and in vivo systematical evaluations indicate that such a responsively drug-eluting PGE IOL can effectively prevent PCO.

Overexpression of a ceramide synthase gene,GhCS1, inhibits fiber cell initiation and elongation by promoting the synthesis of ceramides containing dihydroxy LCB and VLCFA.

Cotton is an important natural fiber crop worldwide. Cotton fiber cell is regarded as an ideal material for studying the growth and development of plant cells. Sphingolipids are important components of biomembrane and bioactive molecules which participate in many processes such as plant growth, development regulation, stimulus sensing, and stress response. However, the functions of sphingolipids in the cotton fiber development are still unclear. In the present study, we identified a cotton ceramide synthase gene, GhCS1, which is predominantly expressed in fiber cell. The GhCS1 is located in the endoplasmic reticulum and has the conserved domains of ceramide synthase. Overexpression of GhCS1 gene inhibited both vegetative and reproductive growth in cotton. Importantly, the fiber cell initiation and elongation were severely inhibited when compared with control. Comparison of the sphingolipid profile in the 0-DPA (days past anthesis) ovule (with fiber cell) between control and transgenic cotton plants showed that the content of sphingosines (Sph) decreased significantly in transgenic ovules, whereas the content of phyto-sphingosines (Phyto-Sph) had no change. Meanwhile, the content of ceramide containing Sph and very-long-chain fatty acid (VLCFA) increased significantly in transgenic ovules, while ceramide containing Phyto-Sph and long-chain fatty acids (LCFA)/VLCFA significantly decreased. These results indicated that GhCS1 was a functional ceramide synthase, which preferentially used Sph and VLCFA as substrates and was different from the Arabidopsis ceramide synthase AtLOH1/AtLOH3, which preferentially used Phyto-Sph and VLCFA as substrates, and also different from AtLOH2, which preferentially used Sph and LCFA as substrates. It is suggested that GhCS1 might be a new ceramide synthase gene in the plant, play some roles in the development of fiber cells and cotton plants.

GhCYP710A1 Participates in Cotton Resistance to Verticillium Wilt by Regulating Stigmasterol Synthesis and Plasma Membrane Stability.

Cotton is an important economic crop. Cotton Verticillium wilt caused by Verticillium dahliae seriously damages production. Phytosterols play roles in plant-pathogen interaction. To explore the function and related mechanism of phytosterols in the interaction between Verticillium dahliae and cotton plants, and the resistance to Verticillium wilt, in this study, we analyzed the changes of sterol composition and content in cotton roots infected by Verticillium dahliae, and identified the sterol C22-desaturase gene GhCYP710A1 from upland cotton. Through overexpressing and silencing the gene in cotton plant, and ectopically expressing the gene in Arabidopsis, we characterized the changes of sterol composition and the resistance to Verticillium wilt in transgenic plants. The infection of Verticillium dahliae resulted in the content of total sterol and each sterol category decreasing in cotton root. The ratio of stigmasterol to sitosterol (St/Si) increased, indicating that the conversion of sitosterol to stigmasterol was activated. Consistently, the expression level of GhCYP710A1 was upregulated after infection. The GhCYP710A1 has the conservative domain that is essential for sterol C22-desaturase in plant and is highly expressed in root and stem, and its subcellular location is in the endoplasmic reticulum. The ectopic expression of GhCYP710A1 gene promoted the synthesis of stigmasterol in Arabidopsis. The St/Si value is dose-dependent with the expression level of GhCYP710A1 gene. Meanwhile, the resistance to Verticillium wilt of transgenic Arabidopsis increased and the permeability of cell membrane decreased, and the content of ROS decreased after V991 (a strain of Verticillium dahliae) infection. Consistently, the resistance to Verticillium wilt significantly increased in the transgenic cotton plants overexpressing GhCYP710A1. The membrane permeability and the colonization of V991 strain in transgenic roots were decreased. On the contrary, silencing GhCYP710A1 resulted in the resistance to Verticillium wilt being decreased. The membrane permeability and the colonization of V991 were increased in cotton roots. The expression change of GhCYP710A1 and the content alteration of stigmasterol lead to changes in JA signal transduction, hypersensitivity and ROS metabolism in cotton, which might be a cause for regulating the Verticillium wilt resistance of cotton plant. These results indicated that GhCYP710A1 might be a target gene in cotton resistance breeding.