Hypothermia Research Articles

Discovery and demonstration of the temperature stress response functions of Dermatophagoides farinae proteins 1 and 2

BackgroundDermatophagoides farinae proteins (DFPs) are abundantly expressed in D. farinae; however, their functions remain unknown. Our previous transcriptome sequencing analyses revealed that the basal expression of DFP1 and DFP2 in D. farinae was high and, more importantly, upregulated under temperature stress. Therefore, DFPs were speculated to exert a temperature stress response function.ResultsReal-time quantitative polymerase chain reaction detection revealed that both DFP1 and DFP2 were significantly upregulated under temperature stress. Particularly, DFP1 was upregulated under cold stress. Electrophoresis of D. farinae total proteins revealed an increased abundance of DFP1 and DFP2 (40–55 kDa bands) under temperature stress, which was corroborated by the mass spectrometry results. After silencing DFP1 and DFP2 further, temperature stress led to decreases in gene expression and survival rates. Moreover, DFP1 was identified as the upstream regulator of DFP2.ConclusionThis study highlights the temperature stress response functions of DFP1 and DFP2 at the mRNA and protein levels. These results provide important insights for applying DFP1 and DFP2 as potential target genes for the molecular prevention and control of D. farinae to prevent allergic diseases. The newly established methods provide methodological guidance for the study of genes with unknown functions in mites.

SlHB8 Is a Novel Factor in Enhancing Cold Resistance in Tomato Anthers by Modulating Tapetal Cell Death.

Tomato plants favor warmth, making them particularly susceptible to cold conditions, especially their reproductive development. Therefore, understanding how pollen reacts to cold stress is vital for selecting and improving cold-resistant tomato varieties. The programmed cell death (PCD) in the tapetum is particularly susceptible to cold temperatures which could hinder the degradation of the tapetal layer in the anthers, thus affecting pollen development. However, it is not clear yet how genes integral to tapetal degradation respond to cold stress. Here, we report that SlHB8, working upstream of the conserved genetic module DYT1-TDF1-AMS-MYB80, is crucial for regulating cold tolerance in tomato anthers. SlHB8 expression increases in the tapetum when exposed to low temperatures. CRISPR/Cas9-generated SlHB8-knockout mutants exhibit improved pollen cold tolerance due to the reduced temperature sensitivity of the tapetum. SlHB8 directly upregulates SlDYT1 and SlMYB80 by binding to their promoters. In normal anthers, cold treatment boosts SlHB8 levels, which then elevates the expression of genes like SlDYT1, SlTDF1, SlAMS, and SlMYB80; however, slhb8 mutants do not show this gene activation during cold stress, leading to a complete blockage of delayed tapetal programmed cell death (PCD). Furthermore, we found that SlHB8 can interact with both SlTDF1 and SlMYB80, suggesting the possibility that SlHB8 might regulate tapetal PCD at the protein level. This study sheds light on molecular mechanisms of anther adaptation to temperature fluctuations.

Age Specifics of Homeostasis Response to Cold Stress in Wistar Rats: Endurance Improvement or Disorganization?

Cold exposure, especially in combination with exercise, is a common procedure to fortify the body. However, its effects in old age are poorly understood. Using the homeostasis stability coefficient as an indicator, the body response to a 20-min swim in cold water was for the first time studied comprehensively in old and young Wistar rats. Effects on the hormonal, hematological, and morphofunctional systems of the liver and thymus were assessed. Unfavorable age-related changes in the stability of homeostasis were observed in old rats. The changes complicated the recovery after cold stress and required the involvement of a greater number of homeostatic mechanisms than in young rats. The liver was found to be the most vulnerable to cold stress. It was concluded that fortifying the body by cold exposure is possible to use in old age, but with due regard to age-related restrictions.

Electroacupuncture Reduced Fibromyalgia-Pain-like Behavior through Inactivating Transient Receptor Potential V1 and Interleukin-17 in Intermittent Cold Stress Mice Model.

Fibromyalgia (FM) is a widespread musculoskeletal pain associated with psychological disturbances, the etiopathogenesis of which is still not clear. One hypothesis implicates inflammatory cytokines in increasing central and peripheral sensitization along with neuroinflammation, leading to an elevation in pro-inflammatory cytokines, e.g., interleukin-17A (IL-17A), enhanced in FM patients and animal models. The intermittent cold stress (ICS)-induced FM-like model in C57BL/6 mice has been developed since 2008 and proved to have features which mimic the clinical pattern in FM patients such as mechanical allodynia, hyperalgesia, and female predominance of pain. Electroacupuncture (EA) is an effective treatment for relieving pain in FM patients, but its mechanism is not totally clear. It was reported as attenuating pain-like behaviors in the ICS mice model through the transient receptor potential vanilloid 1 (TRPV1) pathway. Limited information indicates that TRPV1-positive neurons trigger IL-17A-mediated inflammation. Therefore, we hypothesized that the IL-17A would be inactivated by EA and TRPV1 deletion in the ICS-induced FM-like model in mice. We distributed mice into a control (CON) group, ICS-induced FM model (FM) group, FM model with EA treatment (EA) group, FM model with sham EA treatment (Sham) group, and TRPV1 gene deletion (Trpv1-/-) group. In the result, ICS-induced mechanical and thermal hyperalgesia increased pro-inflammatory cytokines including IL-6, IL-17, TNFα, and IFNγ in the plasma, as well as TRPV1, IL-17RA, pPI3K, pAkt, pERK, pp38, pJNK, and NF-κB in the somatosensory cortex (SSC) and cerebellum (CB) lobes V, VI, and VII. Moreover, EA and Trpv1-/- but not sham EA countered these effects significantly. The molecular mechanism may involve the pro-inflammatory cytokines, including IL-6, IL-17, TNFα, and IFNγ. IL-17A-IL-17RA play a crucial role in peripheral and central sensitization as well as neuroinflammation and cannot be activated without TRPV1 in the ICS mice model. EA alleviated FM-pain-like behaviors, possibly by abolishing the TRPV1- and IL-17A-related pathways. It suggests that EA is an effective and potential therapeutic strategy in FM.

Integrating physiological and molecular insights in cotton under cold stress conditions

Integrating physiological and molecular insights in cotton under cold stress conditions

Cold mediates maize root hair developmental plasticity via epidermis-specific transcriptomic responses.

Cold stress during early development limits maize (Zea mays L.) production in temperate zones. Low temperatures restrict root growth and reprogram gene expression. Here, we provide a systematic transcriptomic landscape of maize primary roots, their tissues, and cell types in response to cold stress. The epidermis exhibited a unique transcriptomic cold response, and genes involved in root hair formation were dynamically regulated in this cell type by cold. Consequently, activation of genes involved in root hair tip growth contributed to root hair recovery under moderate cold conditions. The maize root hair defective mutants roothair defective 5 (rth5) and roothair defective 6 (rth6) displayed enhanced cold tolerance with respect to primary root elongation. Furthermore, dehydration response element-binding protein 2.1 (dreb2.1) was the only member of the dreb subfamily of AP2/EREB transcription factor genes upregulated in primary root tissues and cell types but exclusively downregulated in root hairs upon cold stress. Plants overexpressing dreb2.1 significantly suppressed root hair elongation after moderate cold stress. Finally, the expression of rth3 was regulated by dreb2.1 under cold conditions, while rth6 transcription was regulated by dreb2.1 irrespective of the temperature regime. We demonstrated that dreb2.1 negatively regulates root hair plasticity at low temperatures by coordinating the expression of root hair defective genes in maize.

Iodine-131 Metaiodobenzylguanidine (MIBG) and NP-59 uptakes in the Rat Normal Tissues: Increase of Cholesterol uptake in Cold-Activated Brown Adipose Tissue

Objective Aim: NP-59 (I-131-6ß-iodomethyl-19-norcholesterol) and MIBG (I-131-metaiodobenzylguanidine) both are useful to detect tumors. There is a great need to understand their distributions in normal organs and tissues as well as ways to reduce normal uptakes in tumor imaging and/or treatment. Activated Brown Adipose tissue (BAT) has been promising for its role in optimal treatment of obesity and hypercholesterolemia. The study of cholesterol uptake in BAT may have a significant impact in managing hyperlipidemia and protecting from atherosclerosis. Materials and Methods: Male Sprague-Dawley rats weighing 250—300 g were used in the study. All animals received 25 µCi of I-131-MIBG or NP-59 by the tail vein and were killed (N = 4 - 6 each) 3, 24 or 48 hours afterwards. For cold stressed animals were kept at 6 – 8 C in a cold room overnight prior to receiving labeled MIBG or NP-59. The animal continued to stay in cold room as compared to room temperature controls. To study the effect of norepinephrine-blocking agent, reserpine (4 mg/kg, IP) was given to rats 4 hours prior to I-131-MIBG infusion. Organs were immediately dissected. Radioactivity was counted and tissue concentrations were expressed as percent kilogram (body weight) dose per gram of tissue (% Kg dose/g). Findings: Higher uptakes of I-131-MIBG were observed in organs rich in sympathetic innervation [BAT (brown adipose tissue), heart and spleen] as compared to liver and muscle. Reserpine reduced uptakes in BAT, heart, adrenals and spleen but not in liver in room temperature controls. BAT, but not in other organs, showed a paradoxical increase in reserpine-treated animal kept in cold room as compared to room temperature controls. NP-59 showed a near 6 folds increase in adrenals from 30 minutes to 48 hours and unchanged in cold stressed animals. The only tissue showed significant increases in NP-59 uptake under cold stress was BAT from 1.84 folds at 30 minutes to 1.44 folds at 48 hours (p<0.05 as compared to room temperature controls). Conclusions: The uptake of MIBG is reduced in organs rich in sympathetic innervation under cold stress, but this reduction is partially blocked by reserpine. Findings may help to improve the efficacy of MIBG as a theragnostic agent. A noteworthy finding is the significant increase in NP-59, a cholesterol analog, in cold-activated BAT. This novel observation could serve as a valuable tool for studying the role of activated BAT in the overall cholesterol metabolism in humans. Furthermore, it presents a potent therapeutic avenue for mitigating hyperlipidemia and providing protection against atherosclerosis.

Enrichment of novel CD3+F4/80+ cells in brown adipose tissue following adrenergic stimulation.

Macrophages play a multifaceted role in maintaining tissue homeostasis, fighting infections, and regulating cold-induced thermogenesis. The brown adipose tissue (BAT) is crucial for maintaining body temperature during cold exposure. Cold stress triggers the sympathetic nervous system to release norepinephrine (NE), which activates BAT via β3-adrenergic receptors, initiating lipolysis and glycolysis. BAT-infiltrating macrophages can either hinder or enhance thermogenesis by controlling the interplay between BAT cells and sympathetic nerves. In this study we report on a unique population of CD3+F4/80+ dual lineage co-expressing (DE) cells within the interscapular BAT (iBAT), that increased following chronic adrenergic stimulation. In forward scatter/side scatter plots, they formed a cluster distinct from lymphocytes, appearing larger and more complex. These CD3+F4/80+ DE cells demonstrated the lack of T cell markers CD62L and TCRβ and expressed higher levels of Ly6C, F4/80, and CD11b markers compared to T cells and CD3- macrophages. Furthermore, analysis revealed two subpopulations within the CD3+F4/80+ DE population based on MHCII expression, with the proportion of MHCII-low subset increasing with adrenergic stimulation. This novel DE population within iBAT, unequivocally identified by the its unique surface marker profile, warrants further investigation into the intricate mechanisms governing adaptive thermogenesis regulation.

Integrated transcriptomic and epigenomic analyses to disclose the transcriptional regulatory mechanisms of lipid and energy metabolism under cold stress in grass carp

Cold-induced injuries significantly restrict the potential and efficacy of suivival and growth, necessitating a deeper understanding of cold response in ectothermic fish. Understanding the mechanisms by which fish respond to low-temperature stress has thus become a critical research focus in aquaculture. This study aims to investigate the effects of different temperature gradients (28 °C, 10 °C, and 2 °C) on global gene transcription in grass carp Ctenopharyngodon idellus. Additionally, the study examines cold-induced chromatin accessibility and transcriptional regulation through an integration of RNA sequencing (RNA-seq) and assay for transposase-accessible chromatin using sequencing (ATAC-seq) analyses. Transcriptome analysis revealed that mitochondrial damage and endoplasmic reticulum stress are major factors contributing to grass carp mortality under cold stress. The dataset also disclosed hepatocyte nuclear factor 1 beta (Hnf1b) and peroxisome proliferator-activated receptor alpha (PPARa) as key transcription factors for lipid and energy metabolism in grass carp under cold stress. Notably, Hnf1b and PPARa form a complex transcriptional regulatory network with other transcription factors, decreasd binding activity inactivates the expression of genes related to lipid and energy metabolism under cold stress. These results highlight the transcriptional regulation of lipid and energy metabolism in response to cold stress in grass carp. Overall, this study enhances our understanding of low-temperature response mechanisms in grass carp, providing valuable insights for further investigations.

Genome-wide identification and analysis of abiotic stress responsiveness of the mitogen-activated protein kinase gene family in Medicago sativa L.

BackgroundThe mitogen-activated protein kinase (MAPK) cascade is crucial cell signal transduction mechanism that plays an important role in plant growth and development, metabolism, and stress responses. The MAPK cascade includes three protein kinases, MAPK, MAPKK, and MAPKKK. The three protein kinases mediate signaling to downstream response molecules by sequential phosphorylation. The MAPK gene family has been identified and analyzed in many plants, however it has not been investigated in alfalfa.ResultsIn this study, Medicago sativa MAPK genes (referred to as MsMAPKs) were identified in the tetraploid alfalfa genome. Eighty MsMAPKs were divided into four groups, with eight in group A, 21 in group B, 21 in group C and 30 in group D. Analysis of the basic structures of the MsMAPKs revealed presence of a conserved TXY motif. Groups A, B and C contained a TEY motif, while group D contained a TDY motif. RNA-seq analysis revealed tissue-specificity of two MsMAPKs and tissue-wide expression of 35 MsMAPKs. Further analysis identified MsMAPK members responsive to drought, salt, and cold stress conditions. Two MsMAPKs (MsMAPK70 and MsMAPK75) responds to salt and cold stresses; two MsMAPKs (MsMAPK60 and MsMAPK73) responds to cold and drought stresses; four MsMAPKs (MsMAPK1, MsMAPK33, MsMAPK64 and MsMAPK71) responds to salt and drought stresses; and two MsMAPKs (MsMAPK5 and MsMAPK7) responded to all three stresses.ConclusionThis study comprehensively identified and analysed the alfalfa MAPK gene family. Candidate genes related to abiotic stresses were screened by analysing the RNA-seq data. The results provide key information for further analysis of alfalfa MAPK gene functions and improvement of stress tolerance.

Systematic Analysis of the BrHAT Gene Family and Physiological Characteristics of Brassica rapa L. Treated with Histone Acetylase and Deacetylase Inhibitors under Low Temperature.

Brassica rapa L. is an important overwintering oilseed crop in Northwest China. Histone acetyltransferases (HATs) play an important role in epigenetic regulation, as well as the regulation of plant growth, development, and responses to abiotic stresses. To clarify the role of histone acetylation in the low-temperature response of B. rapa L., we identified 29 HAT genes in B. rapa L. using bioinformatics tools. We also conducted a comprehensive analysis of the physicochemical properties, gene structure, chromosomal localization, conserved structural domains and motifs, cis-acting regulatory elements, and evolutionary relationships of these genes. Using transcriptome data, we analyzed the expression patterns of BrHAT family members and predicted interactions between proteins; the results indicated that BrHATs play an important role in the low-temperature response of B. rapa L. HAT inhibitor (curcumin; CUR) and histone deacetylase inhibitor (Trichostatin A; TSA) were applied to four B. rapa L. varieties varying in cold resistance under the same low-temperature conditions, and changes in the physiological indexes of these four varieties were analyzed. The inhibitor treatment attenuated the effect of low temperature on seed germination, and curcumin treatment was most effective, indicating that the germination period was primarily regulated by histone acetylase. Both inhibitor treatments increased the activity of protective enzymes and the content of osmoregulatory substances in plants, suggesting that histone acetylation and deacetylation play a significant role in the response of B. rapa L. to low-temperature stress. The qRT-PCR analyses showed that the expression patterns of BrHATs were altered under different inhibitor treatments and low-temperature stress; meanwhile, we found three significantly differentially expressed genes. In sum, the process of histone acetylation is involved in the cold response and the BrHATs gene plays a role in the cold stress response.

Hand cold stress testing among Arctic open-pit miners: a clinical study

Objectives. This study aimed to evaluate the influence of individual characteristics (sex, age, body mass index [BMI] and smoking habits) on the tolerance time, pain ratings and rewarming time of hand cold stress testing (CST). Methods. We included 153 subjects (63% men) working in a Swedish open-pit mine (participation rate 41%). The right hand was immersed in 3 °C circulating water for up to 45 s. Pain ratings were registered every fifth second using a visual analog scale. Results. The tolerance time (mean ± standard deviation) was 35 ± 12 s for men and 29 ± 14 s for women (p = 0.007). The youngest age group (18–29 years) had the longest tolerance time, while the oldest group (54–65 years) had the shortest (p = 0.005). Women had significantly higher pain ratings than men after 5, 10 and 25 s. The group with the highest BMI had the shortest rewarming time (p < 0.001). Conclusions. Age and sex influenced the tolerance time of hand CST, while only sex affected the pain ratings and BMI the rewarming time. When performing CST in future studies, these parameters should be considered.

Transcriptome Analysis Revealed ZmPTOX1 Is Required for Seedling Development and Stress Tolerance in Maize.

Plant seedling morphogenesis is considerably related to photosynthesis, pigment synthesis, and circadian periodicity during seedling development. We identified and cloned a maize zebra or crossbanding leaves mutant wk3735, which produces pale white kernels and was identified and plays a role in the equilibrium of the Redox state the in/out of ETC by active oxygen scavenging. Interestingly, it produces the zebra leaves during the production of the first seven leaves, which is apparently different from the mutation of homologs AtPTOX in Arabidopsis. It is intriguing to investigate how and why yellow crossbands (zebra leaf phenotype) emerge on leaves. As expected, chlorophyll concentration and photosynthetic efficiency both significantly declined in the yellow sector of wk3735 leaves. Meanwhile, we observed the circadian expression pattern of ZmPTOX1, which was further validated by protein interaction assays of the circadian clock protein TIM1 and ZmPTOX1. The transcriptome data of yellow (muW) and green (muG) sectors of knock-out lines and normal leaves of overexpression lines (OE) at the 5th-leaf seedling stage were analyzed. Zebra leaf etiolated sections exhibit a marked defect in the expression of genes involved in the circadian rhythm and rhythmic stress (light and cold stress) responses than green sections. According to the analysis of co-DEGs of muW vs. OE and muG vs. OE, terms linked to cell repair function were upregulated while those linked to environmental adaptability and stress response were downregulated due to the mutation of ZmPTOX1. Further gene expression level analyses of reactive oxygen species (ROS) scavenging enzymes and detection of ROS deposition indicated that ZmPTOX1 played an essential role in plant stress resistance and ROS homeostasis. The pleiotropic roles of ZmPTOX1 in plant ROS homeostasis maintenance, stress response, and circadian rhythm character may collectively explain the phenotype of zebra leaves during wk3735 seedling development.

Transcriptome, histology, and enzyme activities analysis of liver in Phoxinus lagowskii to the low temperature stress and recovery

Assessing the response and resilience of fish to low temperatures over different time scales can provide valuable insights into their mechanisms of adaptation to cold conditions. Farmed Amur minnows (Phoxinus lagowskii) frequently encounter low temperatures, especially during winter. However, the specific responses of P. lagowskii to low-temperature stress remain largely unexplored. In this study, we examined serum glucose and cortisol levels, histological changes, enzymes associated with phosphate and carbohydrate metabolism, triglyceride levels, and liver transcriptomics under various conditions: control (CK), short-term cold exposure (6 days, SC), prolonged cold exposure (14 days, PC), and recovery (RY) from cold exposure at 2 °C. Liver vacuolation was observed during short-term cold exposure. Additionally, we analyzed the enzymatic activity related to carbohydrate and lipid metabolism in serum and liver. Liver transcriptomic data revealed that the PPAR signaling pathway and autophagy-related genes were enriched during short-term cold exposure. Carbohydrate metabolism-related pathways, including the AMPK and MAPK signaling pathways, were significantly enriched after prolonged cold exposure. Metabolic pathways such as fat digestion and absorption, glycine, serine, and threonine metabolism, and arginine and proline metabolism were significantly enriched in the recovery group. Rapid warming after prolonged cold stress allowed P. lagowskii to recover quickly. These findings suggest that P. lagowskii has a strong adaptive capacity for energy metabolism during prolonged cold exposure and the ability to recover rapidly from cold stress. A comprehensive examination of the histological, physiological, biochemical, and molecular responses of P. lagowskii to low temperatures is crucial for developing effective strategies for cultivating this species in challenging environments.

Establishment and characterization of a gill cell line from Takifugu obscurus and transcriptome analysis of its gene expression profiles upon low temperature

Takifugu obscurus is a farmed fish of great economic importance in China. The rapid development of T. obscurus aquaculture industry has been accompanied by disease and low-temperature stress, resulting in huge economic losses. Cell lines are used extensively in teleost physiology and pathology as the most cost-effective platform for in vitro research. A novel gill cell line of T. obscurus (named TOG) was first successfully established, and passed through 52 generations. The optimal conditions for TOG growth were 20 % FBS concentration and 24 °C, TOG could be grown in both hypotonic (150 mOsmol-kg-1) and hypertonic (600 mOsmol-kg-1) environments. TOG was determined to be derived from T. obscurus by sequencing the mitochondrial COI gene. Karyotype analysis revealed that the chromosome number of TOG was 44 (2n = 44). Transfection experiment showed that TOG was able to express foreign genes. Furthermore, several immune-related genes were significantly up-regulated in TOG after LPS and poly (I:C) stimulation, including tlr3, isg15, il1β and il10. Additionally, transcriptome analysis of TOG under low-temperature stress (24 °C, 18 °C, 12 °C, 10 °C and 8 °C) found that differentially expressed genes (DEGs) were significantly clustered in several immunological and energy metabolic pathways, and cold stress could disrupt the immune barrier and reduce immunity by downregulating the immune-related pathways. Additionally, weighted gene co-expression network analysis (WGCNA) revealed that bule module and turquoise module, which were closely correlated with low temperature and the degree of fish damage, were both predominantly found in PPAR, NOD-like receptor and Toll-like receptor signaling pathway. Hub genes were identified in these two modules, including mre11, clpb, dhx15, ddx18 and utp15. TOG cell line will become an effective experimental platform for genetic and immunological research, and our results would help us gain a deeper insight into the molecular mechanism of cold tolerance in teleost.

Characterization and fine mapping of cold-inducible parthenocarpy in cucumber (Cucumis sativus L.)

Cold stress detrimentally influences fruit development, leading to a substantial yield reduction in many fruit-bearing vegetables. Cucumber, a vegetable of subtropical origin, is especially sensitive to cold. Cold-inducible parthenocarpy (CIP) promises fruit yield under cold conditions. Previously, we identified a CIP line EC5 in cucumber, which showed strong parthenocarpy and sustained fruit growth under cold conditions (16°C day/10°C night). However, the candidate gene and genetic mechanism underlying CIP in cucumber remain unknown. In this study, both BSA-seq and conventional QTL mapping strategies were employed on F2 populations to delve into the genetic control of CIP. A single QTL, CIP5.1, was consistently mapped across two winter seasons in 2021 and 2022. Fine mapping delimited the CIP locus into a 38.3 kb region on chromosome 5, harboring 8 candidate genes. Among these candidates, CsAGL11 (CsaV3_5G040370) was identified, exhibiting multiple deletions/insertions in the promoter and 5′UTR region. The CsAGL11 gene encodes a MADS-box transcription factor protein, which is homologous to the genes previously recognized as negative regulators in ovule and fruit development of Arabidopsis and tomato. Correspondingly, cold treatment resulted in decreased expression of CsAGL11 during the early developmental stage of the fruit in EC5. A promoter activity assay confirmed promoter polymorphisms leading to weak transcriptional activation of CsAGL11 under cold conditions. This study deepens our understanding of the genetic characteristics of CIP and elucidates the potential role of the CsAGL11 gene in developing cucumber cultivars with enhanced fruiting under cold conditions.

Physiological and transcriptomic profiles reveal key regulatory pathways involved in cold resistance in sunflower seedlings

During sunflower growth, cold waves often occur and impede plant growth. Therefore, it is crucial to study the underlying mechanism of cold resistance in sunflowers. In this study, physiological analysis revealed that as cold stress increased, the levels of ROS, malondialdehyde, ascorbic acid, and dehydroascorbic acid and the activities of antioxidant enzymes increased. Transcriptomics further identified 10,903 DEGs between any two treatments. Clustering analysis demonstrated that the expression of MYB44a, MYB44b, MYB12, bZIP2 and bZIP4 continuously upregulated under cold stress. Cold stress can induce ROS accumulation, which interacts with hormone signals to activate cold-responsive transcription factors regulating target genes involved in antioxidant defense, secondary metabolite biosynthesis, starch and sucrose metabolism enhancement for improved cold resistance in sunflowers. Additionally, the response of sunflowers to cold stress may be independent of the CBF pathway. These findings enhance our understanding of cold stress resistance in sunflowers and provide a foundation for genetic breeding.

An Innovative Heating Solution for Sustainable Agriculture: A Feasibility Study on the Integration of Phase Change Materials as Passive Heating Elements

In this study, we investigate an innovative option for the ecological management of agricultural land. The focus is on the use of phase change materials (PCMs) for passive temperature regulation in greenhouses and fruit crop fields in order to reduce yield losses due to unforeseen late frost events. The use of PCMs represents a novel approach to enhancing crop growth and extending growing seasons without relying on conventional energy-intensive methods, providing a stable microclimate that can protect plants from cold stress. This passive regulation of temperature helps to reduce the need for fossil fuel-based heating systems, thereby lowering greenhouse gas emissions and operational costs. The application of PCMs in agricultural settings is particularly innovative as it leverages naturally occurring temperature variations to create a self-sustaining, low-maintenance solution that aligns with the principles of sustainable farming. This approach not only improves energy efficiency but also contributes to the resilience of agricultural practices in the face of climate variability. This study focuses on the possible use of PCMs in passive heating modules for the protection of potted plants in greenhouses. Various PCMs such as paraffin, beeswax, and shea butter were tested. Experiments were then conducted, using one kind of paraffin-based PCM, in a specially designed module. In addition, an FEM simulation model (CFD) was built and tested. The model was used to perform detailed analyses of the heat transfer efficiency, fluid dynamics, and overall performance of the modules. The model can also be used for optimization purposes (e.g., efficiency improvements).

Two HbpA-like proteins HbpA1 and HbpA2 from Actinobacillus pleuropneumoniae protect bacteria from sulfur source limitation, oxidative and cold stresses, but not essential to virulence

Porcine pleuropneumonia is one of the respiratory diseases that pigs are susceptible to Actinobacillus pleuropneumoniae (A. pleuropneumoniae), poses a great threat to the global pig industry. Glutathione (GSH) is an important sulfur source, cellular antioxidant and virulence determinant of many pathogenic bacteria. In this study, roles of two HbpA-like proteins HbpA1 and HbpA2 of A. pleuropneumoniae were analyzed. A. pleuropneumoniae mutants without HbpA2 were basically unable to grow in chemically defined medium (CDM) with GSH as the sole sulfur source and had significantly reduced oxidative tolerance; whereas mutation in hbpA1 led to reduced survival under low-temperature environments. Neither HbpA1 nor HbpA2 affects utilization of heme. These two HbpA-like proteins are not associated with the virulence of A. pleuropneumoniae. Our results reveal the correlation of A. pleuropneumoniae HbpA1 and HbpA2 in GSH utilization, highlight the roles of HbpA1 in the cold stress resistance and HbpA2 in the anti-oxidative response. GSH limitation is not a way to attenuate colonization and pathogenicity of A. pleuropneumoniae.

Genome-Wide Identification of the DOF Gene Family in Kiwifruit (Actinidia chinensis) and Functional Validation of AcDOF22 in Response to Drought Stress.

DNA-binding one zinc finger (DOF) transcription factors are crucial plant-specific regulators involved in growth, development, signal transduction, and abiotic stress response generation. However, the genome-wide identification and characterization of AcDOF genes and their regulatory elements in kiwifruit (Actinidia chinensis) has not been thoroughly investigated. In this study, we screened the kiwifruit genome database and identified 42 AcDOF genes (AcDOF1 to AcDOF42). Phylogenetic analysis facilitated the categorization of these genes into five subfamilies (DOF-a, DOF-b, DOF-c, DOF-d, and DOF-e). We further analyzed the motifs, conserved domains, gene structures, and collinearity of the AcDOFgene family. Gene ontology (GO) enrichment analysis indicated significant enrichment in the "flower development" term and the "response to abiotic stress" category. Promoter prediction analysis revealed numerous cis-regulatory elements related to responses to light, hormones, and low-temperature and drought stress in AcDOF promoters. RNA-seq expression profiles demonstrated the tissue-specific expression of AcDOF genes. Quantitative real-time PCR results showed that six selected genes (AcDOF04, AcDOF09, AcDOF11, AcDOF13, AcDOF21, and AcDOF22) were differentially induced by abscisic acid (ABA), methyl jasmonate (MeJA), and cold, salt, and drought stresses, with AcDOF22 specifically expressed at high levels in drought-tolerant cultivars. Further experiments indicated that transient AcDOF22 overexpression in kiwifruit leaf disks reduced water loss and chlorophyll degradation. Additionally, AcDOF22 was localized to the nucleus and exhibited transcriptional activation, enhancing drought resistance by activating the downstream drought marker gene AcDREB2A. These findings lay the foundation for elucidating the molecular mechanisms of drought resistance in kiwifruit and offer new insights into drought-resistant breeding.