Effect of Sugar on the Process of Cold-Acclimation-Induced Freezing Tolerance of Populus tomentosa

Populus tomentosa seedlings for cold-acclimating were pretreated wit h or without 20% saccharose. Changes in the concentrations of total soluble sugar , the survival rates, and freezing tolerance of seedlings during cold acclimatio n were investigated. The results showed that cold acclimation increased the conc entrations of total soluble sugar, the survival rates and freezing tolerance. Co ld acclimation, combined with the saccharose-pretreatment, enhanced the above- ment ioned effect of cold acclimation, and obviously increased the concentrations of total soluble sugar, the survival rates and freezing tolerance of seedlings. Fur ther analysis found that the concentrations of total soluble sugar in branches i ncreased greater than that in leaves during both cold acclimation with or withou t the pretreatment of saccharose. Moreover, an increase of the concentrations of total soluble sugar in branches and leaves was closely related to the freezing tolerance of seedlings. The results indicate the accumulation of soluble sugar i n seedlings induced by cold acclimation may be involved in the induction of free zing tolerance .

Effects of cold-hardening on compatible solutes and antioxidant enzyme activities related to freezing tolerance in Ammopiptanthus mongolicus seedlings

Cold acclimation is associated with many metabolic changes that lead to an increase of freezing tolerance. In order to investigate the biochemical process of cold acclimation in Ammopiptanthus mongolicus, seedlings were acclimated at 2℃ under 16-h photoperiod （150 μmol·m^-2·s^-1 photosynthetically active radiation） for 14 d. Freezing tolerance in seedlings increased after 14 d of cold-hardening. Contents of protein, proline and solute carbohydrate in cotyledon increased after cold acclimation. Patterns of isozymes of superoxide dismutase （SOD）, peroxidase, catalase and polyphenol oxidase （PPO） were investigated. The activities of SOD, peroxidase and PPO in cold acclimated plants were increased during cold-hardening. We deduced that compatible solutes and antioxidant enzymes play important roles in development of freezing tolerance during cold acclimation in this evergreen woody plant.

Anatomical Structure Comparison Between Leaves of Two Winter Wheat Cultivars with Different Cold/Freezing Tolerance Under Low Temperature Stress

Winter wheat （Triticum aestivum） cultivars Dongnongdongmai 1 with strong cold/freezing tolerance and Jimai 22 with weak cold/freezing tolerance were used for investigating the difference of microstructure and ultrastructure between leaves of two cultivars under low temperature stress （5℃ and -15 ℃） using optical and electron microscope. The results showed that there was no obvious difference between leaves of Dongnongdongmai 1 and Jimai 22 in microstructure. However, the difference between those leaves was distinct in ultrastructure. The grana lamella and stroma lamella were stacked regularly and arranged parallelly along the long axis of chloroplasts in cv. Dongnongdongmai 1, while the arrangement directions of thylakoids in Jimai 22＇s leaves were so irregular as to form various angles with the long axis of chloroplasts. At -15℃, the mitochondrias were swelled to be round and the structure of cristaes became blurry in both cultivars＇ leaves, while some cristaes of Jimai 22 disappeared. These results would provide theoretical evidence for selecting cold/freezing tolerant winter wheat germplasm resources

Overexpression of galactinol synthase 1 from Solanum commersonii(ScGolS1) confers freezing tolerance in transgenic potato

Potato(Solanum tuberosum L.) is the fourth largest food crop in the world. Low temperatures cause serious damage to potato plants every year, and freezing tolerance has become a hot spot in potato research. Galactinol synthase(GolS) is a key enzyme in the synthesis of raffinose family oligosaccharides(RFOs), and plays an important role in the response of plants to abiotic stress. In this study, the ScGolS1 gene from Solanum commersonii was cloned and introduced into the S. tuberosum cultivars 'Atlantic' and 'Desiree' via Agrobacterium-mediated transformation. Phenotyping assays showed that overexpression of ScGolS1 could significantly improve freezing tolerance in transgenic potato plants.Further physiological and biochemical experiments showed that the transgenic lines had lower relative conductivity, malondialdehyde content,and 3,3-diaminobenzidine staining and a higher plant survival rate compared with wild type(WT) under cold stress. Moreover, the C-repeat binding factors(CBF1, CBF2 and CBF3), the downstream cold-responsive genes COR413 and COR47, and the ethylene-responsive factor(ERF)transcription factor genes ERF3, ERF4 and ERF6, which function in the ethylene signaling pathway, were all induced by freezing treatment and expressed at higher levels in the ScGolS1 overexpression lines compared with WT. Besides, the expression of some genes such as MIPS, STS and RS from the RFO metabolic pathway was up-regulated under cold stress, resulting in changes in the content of some soluble sugars. This indicated that ScGolS1 overexpression altered the sugar composition and enhanced freezing tolerance in transgenic potato by inducing the ethylene and CBF signaling pathways. These results provided theoretical support and genetic resources for freezing tolerance breeding in potato.

Protease inhibitor ASP enhances freezing tolerance by inhibiting protein degradation in kumquat

Cold acclimation is a complex biological process leading to the development of freezing tolerance in plants.In this study,we demonstrated that cold-induced expression of protease inhibitor FmASP in a Citrus-relative species kumquat[Fortunella margarita(Lour.)Swingle]contributes to its freezing tolerance by minimizing protein degradation.Firstly,we found that only cold-acclimated kumquat plants,despite extensive leaf cellular damage during freezing,were able to resume their normal growth upon stress relief.To dissect the impact of cold acclimation on this anti-freezing performance,we conducted protein abundance assays and quantitative proteomic analysis of kumquat leaves subjected to cold acclimation(4◦C),freezing treatment(−10◦C)and post-freezing recovery(25◦C).FmASP(Against Serine Protease)and several non-specific proteases were identified as differentially expressed proteins induced by cold acclimation and associated with stable protein abundance throughout the course of low-temperature treatment.FmASP was further characterized as a robust inhibitor of multiple proteases.In addition,heterogeneous expression of FmASP in Arabidopsis confirmed its positive role in freezing tolerance.Finally,we proposed a working model of FmASP and illustrated how this extracellular-localized protease inhibitor protects proteins from degradation,therebymaintaining essential cellular function for post-freezing recovery.These findings revealed the important role of protease inhibition in freezing response and provide insights on how this role may help develop new strategies to enhance plant freezing tolerance.

The AaCBF4-AaBAM3.1 module enhances freezing tolerance of kiwifruit (Actinidia arguta)

Beta-amylase(BAM)plays an important role in plant resistance to cold stress.However,the specific role of the BAM gene in freezing tolerance is poorly understood.In this study,we demonstrated that a cold-responsive gene module was involved in the freezing tolerance of kiwifruit.In this module,the expression of AaBAM3.1,which encodes a functional protein,was induced by cold stress.AaBAM3.1-overexpressing kiwifruit lines showed increased freezing tolerance,and the heterologous overexpression of AaBAM3.1 in Arabidopsis thaliana resulted in a similar phenotype.The results of promoter GUS activity and cis-element analyses predicted AaCBF4 to be an upstream transcription factor that could regulate AaBAM3.1 expression.Further investigation of protein-DNA interactions by using yeast one-hybrid,GUS coexpression,and dual luciferase reporter assays confirmed that AaCBF4 directly regulated AaBAM3.1 expression.In addition,the expression of both AaBAM3.1 and AaCBF4 in kiwifruit responded positively to cold stress.Hence,we conclude that the AaCBF-AaBAM module is involved in the positive regulation of the freezing tolerance of kiwifruit.

Transcriptional regulation of bark freezing tolerance in apple(Malus domestica Borkh.)

Freezing tolerance is a significant trait in plants that grow in cold environments and survive through the winter.Apple(Malus domestica Borkh.)is a cold-tolerant fruit tree,and the cold tolerance of its bark is important for its survival at low temperatures.However,little is known about the gene activity related to its freezing tolerance.To better understand the gene expression and regulation properties of freezing tolerance in dormant apple trees,we analyzed the transcriptomic divergences in the bark from 1-year-old branches of two apple cultivars,“Golden Delicious”(G)and“Jinhong”(H),which have different levels of cold resistance,under chilling and freezing treatments.“H”can safely overwinter below−30℃in extremely low-temperature regions,whereas“G”experiences severe freezing damage and death in similar environments.Based on 28 bark transcriptomes(from the epidermis,phloem,and cambium)from 1-year-old branches under seven temperature treatments(from 4 to−29°C),we identified 4173 and 7734 differentially expressed genes(DEGs)in“G”and“H”,respectively,between the chilling and freezing treatments.A gene coexpression network was constructed according to this expression information using weighted gene correlation network analysis(WGCNA),and seven biologically meaningful coexpression modules were identified from the network.The expression profiles of the genes from these modules suggested the gene regulatory pathways that are responsible for the chilling and freezing stress responses of“G”and/or“H.”Module 7 was probably related to freezing acclimation and freezing damage in“H”at the lower temperatures.This module contained more interconnected hub transcription factors(TFs)and cold-responsive genes(CORs).Modules 6 and 7 contained C-repeat binding factor(CBF)TFs,and many CBF-dependent homologs were identified as hub genes.We also found that some hub TFs had higher intramodular connectivity(KME)and gene significance(GS)than CBFs.Specifically,most hub TFs in modules 6 and 7 were activated at the beginning of the early freezing stress phase and maintained upregulated expression during the whole freezing stress period in“G”and“H”.The upregulation of DEGs related to methionine and carbohydrate biosynthetic processes in“H”under more severe freezing stress supported the maintenance of homeostasis in the cellular membrane.This study improves our understanding of the transcriptional regulation patterns underlying freezing tolerance in the bark of apple branches.

Compositional and Hollow Engineering of Silicon Carbide/Carbon Microspheres as High-Performance Microwave Absorbing Materials with Good Environmental Tolerance

Microwave absorbing materials(MAMs)characterized by high absorption efficiency and good environmental tolerance are highly desirable in practical applications.Both silicon carbide and carbon are considered as stable MAMs under some rigorous conditions,while their composites still fail to produce satisfactory microwave absorption performance regardless of the improvements as compared with the individuals.Herein,we have successfully implemented compositional and structural engineering to fabricate hollow Si C/C microspheres with controllable composition.The simultaneous modulation on dielectric properties and impedance matching can be easily achieved as the change in the composition of these composites.The formation of hollow structure not only favors lightweight feature,but also generates considerable contribution to microwave attenuation capacity.With the synergistic effect of composition and structure,the optimized SiC/C composite exhibits excellent performance,whose the strongest reflection loss intensity and broadest effective absorption reach-60.8 dB and 5.1 GHz,respectively,and its microwave absorption properties are actually superior to those of most SiC/C composites in previous studies.In addition,the stability tests of microwave absorption capacity after exposure to harsh conditions and Radar Cross Section simulation data demonstrate that hollow SiC/C microspheres from compositional and structural optimization have a bright prospect in practical applications.

A dynamic soil freezing characteristic curve model for frozen soil

The soil freezing characteristic curve(SFCC)plays a fundamental role in comprehending thermohydraulic behavior and numerical simulation of frozen soil.This study proposes a dynamic model to uniformly express SFCCs amidst varying total water contents throughout the freezing-thawing process.Firstly,a general model is proposed,wherein the unfrozen water content at arbitrary temperature is determined as the lesser of the current total water content and the reference value derived from saturated SFCC.The dynamic performance of this model is verified through test data.Subsequently,in accordance with electric double layer(EDL)theory,the theoretical residual and minimum temperatures in SFCC are calculated to be-14.5℃to-20℃for clay particles and-260℃,respectively.To ensure that the SFCC curve ends at minimum temperature,a correction function is introduced into the general model.Furthermore,a simplified dynamic model is proposed and investigated,necessitating only three parameters inherited from the general model.Additionally,both general and simplified models are evaluated based on a test database and proven to fit the test data exactly across the entire temperature range.Typical recommended parameter values for various types of soils are summarized.Overall,this study provides not only a theoretical basis for most empirical equations but also proposes a new and more general equation to describe the SFCC.

Abiotic stress treatment reveals expansin like A gene OfEXLA1 improving salt and drought tolerance of Osmanthus fragrans by responding to abscisic acid

Sweet osmanthus(Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A(OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid(ABA). In yeast, we found OfEXLA1overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol(PEG)treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5(ABI5)-like genes(OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dualluciferase(dual-LUC) and yeast one hybrid(Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.

MIR1868 negatively regulates rice cold tolerance at both the seedling and booting stages

Low temperature causes rice yield losses of up to 30%–40%,therefore increasing its cold tolerance is a breeding target.Few genes in rice are reported to confer cold tolerance at both the vegetative and reproductive stages.This study revealed a rice-specific 24-nt miRNA,miR1868,whose accumulation was suppressed by cold stress.Knockdown of MIR1868 increased seedling survival,pollen fertility,seed setting,and grain yield under cold stress,whereas its overexpression conferred the opposite phenotype.Knockdown of MIR1868 increased reactive oxygen species(ROS)scavenging and soluble sugar content under cold stress by increasing the expression of peroxidase genes and sugar metabolism genes,and its overexpression produced the opposite effect.Thus,MIR1868 negatively regulated rice cold tolerance via ROS scavenging and sugar accumulation.

PHD17 acts as a target of miR1320 to negatively control cold tolerance via JA-activated signaling in rice

Plant Homeo Domain(PHD)proteins are involved in diverse biological processes during plant growth.However,the regulation of PHD genes on rice cold stress response remains largely unknown.Here,we reported that PHD17 negatively regulated cold tolerance in rice seedlings as a cleavage target of miR1320.PHD17 expression was greatly induced by cold stress,and was down-regulated by miR1320 overexpression and up-regulated by miR1320 knockdown.Through 5'RACE and dual luciferase assays,we found that miR1320 targeted and cleaved the 3'UTR region of PHD17.PHD17 was a nuclearlocalized protein and acted as a transcriptional activator in yeast.PHD17 overexpression reduced cold tolerance of rice seedlings,while knockout of PHD17 increased cold tolerance,partially via the CBF cold signaling.By combining transcriptomic and physiological analyses,we demonstrated that PHD17 modulated ROS homeostasis and flavonoid accumulation under cold stress.K-means clustering analysis revealed that differentially expressed genes in PHD17 transgenic lines were significantly enriched in the jasmonic acid(JA)biosynthesis pathway,and expression of JA biosynthesis and signaling genes was verified to be affected by PHD17.Cold stress tests applied with MeJA or IBU(JA synthesis inhibitor)further suggested the involvement of PHD17 in JA-mediated cold signaling.Taken together,our results suggest that PHD17 acts downstream of miR1320 and negatively regulates cold tolerance of rice seedlings through JA-mediated signaling pathway.

A Data Intrusion Tolerance Model Based on an Improved Evolutionary Game Theory for the Energy Internet

Malicious attacks against data are unavoidable in the interconnected,open and shared Energy Internet(EI),Intrusion tolerant techniques are critical to the data security of EI.Existing intrusion tolerant techniques suffered from problems such as low adaptability,policy lag,and difficulty in determining the degree of tolerance.To address these issues,we propose a novel adaptive intrusion tolerance model based on game theory that enjoys two-fold ideas:(1)it constructs an improved replica of the intrusion tolerance model of the dynamic equation evolution game to induce incentive weights;and (2)it combines a tournament competition model with incentive weights to obtain optimal strategies for each stage of the game process.Extensive experiments are conducted in the IEEE 39-bus system,whose results demonstrate the feasibility of the incentive weights,confirm the proposed strategy strengthens the system’s ability to tolerate aggression,and improves the dynamic adaptability and response efficiency of the aggression-tolerant system in the case of limited resources.

The ZOS7-MYB60 module confers drought-stress tolerance in rice

Shanlan upland rice is an important landrace resource with high drought stress(DS)tolerance.Despite its importance,genes responsible for yield in Shanlan upland rice have yet to be discovered.Our previous study identified a drought-responsive zinc finger protein,ZOS7,as highly expressed in Shanlandao upland rice.However,the function of this gene in controlling drought tolerance remains largely unexplored.In this study,we found that overexpressing ZOS7,a drought-responsive zinc finger protein,in rice increased biomass and yield under drought stress.Co-overexpressing ZOS7 and MYB60,encoding a protein with which ZOS7 interacted,intensified the yield increase.ZOS7 and MYB60 appear to form a module that confers drought tolerance by regulating stomatal density and wax biosynthesis.The ZOS7-MYB60module could be used in molecular breeding for drought tolerance in rice.

The UDP-glycosyltransferase OsUGT706D2 positively regulates cold and submergence stress tolerance in rice

In a genome-wide association study,we identified a rice UDP-glycosyltransferase gene,OsUGT706D2,whose transcription was activated in response to cold and submergence stress and to exogenous abscisic acid(ABA).OsUGT706D2 positively regulated the biosynthesis of tricin-4’-O-(syringyl alcohol)ether-7-O-glucoside at both the transcriptional and metabolic levels.OsUGT706D2 mediated cold and submergence tolerance by modulating the expression of stress-responsive genes as well as the abscisic acid(ABA)signaling pathway.Gain of function of OsUGT706D2 increased cold and submergence tolerance and loss of function of OsUGT706D2 reduced cold tolerance.ABA positively regulated OsUGT706D2-mediated cold tolerance but reduced submergence tolerance.These findings suggest the potential use of OsUGT706D2 for improving abiotic stress tolerance in rice.

Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp.PCC 6803 tolerant to both cadmium and high light

Photosynthetic cyanobacteria have shown great potential as“autotrophic cell factories”for the synthesis of fuels and chemicals.However,poor tolerance to various environmental stressors such as high light and heavy metals is an important factor limiting their economic viability.While numerous studies have focused on the tolerance mechanism of cyanobacteria to individual stressors,their response to simultaneous stresses remains to be recovered.To investigate the mechanism of cross tolerance to heavymetal Cd^(2+) and high light,the model cyanobacterium Synechocystis sp.PCC 6803 tolerant to both Cd^(2+) and high light was obtained via about 800 days’cross-adaptive laboratory evolution.Three evolutionary strains capable of tolerating both 5.5 μmol·L^(-1) Cd^(2+) and 600 μmol·m^(-2)·s^(-1) high light were successfully obtained,achieving about 83%enhancement of Cd^(2+) tolerance compared with the parent strain.The different response of parent and evolutionary strains to Cd^(2+) was elucidated via metabolomics.Furthermore,a total of 15 genes that were mutated during evolution were identified by whole-genome re-sequencing.Finally,by single-gene knockout and complementation analysis,four genes including ssl2615,sll1732,ssr1480,and sll1659 involved in the improvement of Cd^(2+) tolerance under high-light condition were successfully identified.This work explored the tolerance mechanism of Synechocystis sp.PCC 6803 to cadmium under high-light condition and provided valuable reference for deciphering multitolerance mechanism of cyanobacteria in the future.

Heterogeneous expression of stearoyl-acyl carrier protein desaturase genes SAD1 and SAD2 from Linum usitatissimum enhances seed oleic acid accumulation and seedling cold and drought tolerance in Brassica napus

Flax(Linum usitatissimum L.)is a versatile crop and its seeds are a major source of unsaturated fatty acids.Stearoyl-acyl carrier protein desaturase(SAD)is a dehydrogenase enzyme that plays a key role in oleic acid biosynthesis as well as responses to biotic and abiotic stresses.However,the function of SAD orthologs from L.usitatissimum has not been assessed.Here,we found that two LuSAD genes,LuSAD1 and LuSAD2,are present in the genome of L.usitatissimum cultivar‘Longya 10’.Heterogeneous expression of either LuSAD1 or LuSAD2 in Arabidopsis thaliana resulted in higher contents of total fatty acids and oleic acid in the seeds.Interestingly,ectopic expression of LuSAD2 in A.thaliana caused altered plant architecture.Similarly,the overexpression of either LuSAD1 or LuSAD2 in Brassica napus also resulted in increased contents of total fatty acids and oleic acid in the seeds.Furthermore,we demonstrated that either LuSAD1 or LuSAD2 enhances seedling resistance to cold and drought stresses by improving antioxidant enzyme activity and nonenzymatic antioxidant levels,as well as reducing membrane damage.These findings not only broaden our knowledge of the LuSAD functions in plants,but also offer promising targets for improving the quantity and quality of oil,and the abiotic stress tolerance of oil-producing crops,through molecular manipulation.

Thermal performance of cast-in-place piles with artificial ground freezing in permafrost regions

During the construction of cast-in-place piles in warm permafrost,the heat carried by concrete and the cement hydration reaction can cause strong thermal disturbance to the surrounding permafrost.Since the bearing capacity of the pile is quite small before the full freeze-back,the quick refreezing of the native soils surrounding the cast-in-place pile has become the focus of the infrastructure construction in permafrost.To solve this problem,this paper innovatively puts forward the application of the artificial ground freezing(AGF)method at the end of the curing period of cast-in-place piles in permafrost.A field test on the AGF was conducted at the Beiluhe Observation and Research Station of Frozen Soil Engineering and Environment(34°51.2'N,92°56.4'E)in the Qinghai Tibet Plateau(QTP),and then a 3-D numerical model was established to investigate the thermal performance of piles using AGF under different engineering conditions.Additionally,the long-term thermal performance of piles after the completion of AGF under different conditions was estimated.Field experiment results demonstrate that AGF is an effective method to reduce the refreezing time of the soil surrounding the piles constructed in permafrost terrain,with the ability to reduce the pile-soil interface temperatures to below the natural ground temperature within 3 days.Numerical results further prove that AGF still has a good cooling effect even under unfavorable engineering conditions such as high pouring temperature,large pile diameter,and large pile length.Consequently,the application of this method is meaningful to save the subsequent latency time and solve the problem of thermal disturbance in pile construction in permafrost.The research results are highly relevant for the spread of AGF technology and the rapid building of pile foundations in permafrost.

Cardiac Tolerance of Hydroalcoholic Extract of Bark of Terminalia mantaly H. Perrier (HAEBTM) in Wistar Rats

Terminalia mantaly H. Perrier is a plant used in traditional medicine for the treatment of various pathologies. However, Terminalia mantaly H. Perrier could present potential health effects on patients. In order to determine the possible cardiotoxic effects of the hydro-alcoholic extract of the bark of Terminalia mantaly H. Perrier, (HAEBTM) forty (40) rats distributed randomly into 4 groups, including 10 animals per group (5 males and 5 females) were used. Animals in group 1 received distilled water and were used as a control group. On the other hand, groups 2, 3, 4 received oral administration a volume of the hydroalcoholic extract of Terminalia mantaly H. Perrier corresponding to 1 mL/100g of body weight at 150 mg/kg, 300 mg/kg, 600 mg/kg, respectively. The extract was administered daily at the same time for 28 days and serum was collected once a week to evaluate cardiac biochemical markers using spectrophotometric methods using a Cobas C311 HITACHI biochemistry system. After one month of study, all rats were euthanized by overdose of ether, and the hearts of the rats were removed for gross morphological and histopathological analysis. Results were analysed using variance analysis (ANOVA) to compare outcomes as a function of doses administered and treatment times. The biochemical parameters ALT, LDH, CPK, CPKMB showed no significant change (p Terminalia mantaly showed no lesions, edema and necrosis. These results suggest that the hydroalcoholic extract of Terminalia mantaly did not interfere with the functioning or alter the integrity of the heart.

Artificial ground freezing of underground mines in cold regions using thermosyphons with air insulation

Current practice of underground artificial ground freezing(AGF)typically involves huge refrigeration systems of large economic and environmental costs.In this study,a novel AGF technique is proposed deploying available cold wind in cold regions.This is achieved by a static heat transfer device called thermosyphon equipped with an air insulation layer.A refrigeration unit can be optionally integrated to meet additional cooling requirements.The introduction of air insulation isolates the thermosyphon from ground zones where freezing is not needed,resulting in:(1)steering the cooling resources(cold wind or refrigeration)towards zones of interest;and(2)minimizing refrigeration load.This design is demonstrated using well-validated mathematical models from our previous work based on two-phase enthalpy method of the ground coupled with a thermal resistance network for the thermosyphon.Two Canadian mines are considered:the Cigar Lake Mine and the Giant Mine.The results show that our proposed design can speed the freezing time by 30%at the Giant Mine and by two months at the Cigar Lake Mine.Further,a cooling load of 2.4 GWh can be saved at the Cigar Lake Mine.Overall,this study provides mining practitioners with sustainable solutions of underground AGF.