Search Results (939)

Background: Cymbidium goeringii, one of China’s traditional and valuable orchids, possesses significant ornamental and economic value. However, it is relatively sensitive to low temperature and other abiotic stresses, which severely restrict its application in landscaping and industrial development. WRKY transcription factors play important roles in plant responses to abiotic stresses, yet related research in C. goeringii remains limited. Methods: In this study, based on transcriptome data of C. goeringii under four different stresses, we identified and cloned the WRKY transcription factor gene CgWRKY53. Through bioinformatics analysis, quantitative real-time PCR, and heterologous transformation in Arabidopsis thaliana, we systematically investigated its structural characteristics, expression patterns, and function under cold stress. Results: The full-length CDS of CgWRKY53 is 1080 bp, encoding a protein of 359 amino acids with a molecular weight of 39.95 kDa. Group III subfamily of the WRKY family, possessing the conserved WRKYGQK domain and a C2HC-type zinc finger motif. CgWRKY53 is expressed in roots, pseudobulbs, leaves, and flowers of C. goeringii, with the highest expression observed in flowers. Under cold, heat, waterlogging, and ABA treatments, CgWRKY53 displayed significant changes in expression, with the most pronounced response occurring under cold stress, where its expression was significantly upregulated. Homozygous transgenic A. thaliana lines overexpressing CgWRKY53 exhibited dwarfed stature, with smaller and deformed leaves and notably shorter roots compared to wild-type plants. The overexpression lines also showed cold-sensitive phenotypes under low-temperature stress, and the expression of several cold-responsive genes was suppressed, suggesting that CgWRKY53 may act as a negative regulator in the response to cold stress. Conclusions: These results identify CgWRKY53 as a negative regulator of cold stress response in C. goeringii. This study provides important genetic resources and theoretical foundations for molecular breeding of stress-resistant orchids. Full article

Maize (Zea mays L.) originated in tropical and subtropical regions. During its growth and development, cold stress severely threatens seedling survival rates and final yield by inducing oxidative stress, compromising cell membrane integrity, and causing “physiological drought.” The Domain of Unknown Function 966 (DUF966) gene family comprises a class of regulatory factors containing conserved domains of undetermined function. Although they are considered to be extensively involved in plant growth, development, and stress response, their specific roles within the maize cold-tolerance regulatory network remain to be explored. In this study, 10 ZmDUF966 family members were identified via genome-wide analysis, and their phylogenetic relationships, gene structures, conserved motifs, chromosomal localizations, and cis-acting elements were systematically analyzed. The results indicate that the ZmDUF966 family is highly conserved among Poaceae species, and its promoters are enriched with stress-responsive elements such as LTR and ABRE. The core gene, ZmDUF966-10, was significantly up-regulated (approximately 35-fold at 48 h, p < 0.05) as validated by RT-qPCR under cold stress and is post-transcriptionally regulated by conserved miRNAs such as zma-miR159. Further yeast two-hybrid experiments revealed a preliminary physical interaction between the ZmDUF966-10 protein and an ABA/WDS-induced protein, suggesting its potential involvement in ABA-mediated stress signaling, though functional validation remains to be conducted. In conclusion, this study identifies ZmDUF966-10 as a promising candidate gene that responds to cold signals through multi-level regulatory networks, providing a valuable gene resource for further functional characterization and potential application in cold-tolerant maize improvement. Full article

PYL proteins are core components of the abscisic acid (ABA) signaling pathway and are involved in plant responses to biotic and abiotic stresses. In this study, a total of 19, four, and eight PYL genes were identified in Saccharum spontaneum, the Saccharum spp. hybrid R570, and Sorghum bicolor, respectively. Phylogenetic analysis classified these PYL genes into three distinct groups. Cis-acting element analysis, Gene Ontology annotation, and Kyoto Encyclopedia of Genes and Genomes pathway enrichment and gene expression profile indicated that members of the PYL gene family are mainly associated with hormone signaling and stress-related biological processes. The ScPYL8 gene (GenBank accession number: OR838856) was isolated from sugarcane cultivar QT3. Expression of the ScPYL8 gene was induced under stresses of cold, PEG, SA, MeJA, ABA, and brown stripe disease (Bipolaris setariae). The gene was expressed in roots, stems and leaves, with the highest expression level in leaves. Subcellular localization analysis showed that the ScPYL8 protein localized to the cytoplasm and nucleus. ScPYL8 overexpression in tobacco activated the reactive oxygen species defense system and regulated the ABA and jasmonic acid signaling pathways, enhancing its resistance against Fusarium solani var. coeruleum. These findings provide insights into the expression, function, and evolutionary characteristics of the PYL gene family in sugarcane, offering valuable genetic resources for future molecular breeding. Full article

Plant viruses can cause substantial yield losses, yet disease severity often varies between seasons because plants frequently experience heat or cold episodes before infection. In this study, we tested whether such temperature conditions affect the plant’s redox balance and alter its response to Tomato bushy stunt virus (TBSV) infection in Nicotiana benthamiana. Plants were exposed to short-term heat and cold stress, after which they recovered before virus inoculation. Following this, we assessed the reactive oxygen species (ROS) content, lipid peroxidation (LPO), oxidative DNA damage, stress-related proteins, redox-associated enzymes, and antioxidant metabolites. TBSV led to non-parallel ROS responses during infection, with consistently elevated hydrogen peroxide in infected plants but reduced superoxide relative to corresponding mock controls. Heat pre-exposure caused pronounced LPO that decreased further after infection, whereas cold pre-exposure stabilized malondialdehyde near levels observed at 25 °C. Both thermal stress and infection increased 8-oxo-dG and were associated with distinct changes in 8-oxoguanine glycosylase abundance. Infection strongly induced heat shock protein 90 (and moderately heat shock protein 70), while prior heat limited further chaperone induction by TBSV. These results indicate that viral infection develops within and is limited by the host’s oxidative state, where redox homeostasis may restrict infection-related processes, and infection leads to changes in this redox environment that are favorable for its development. Full article

Background: The DOF transcription factor family is involved in plant growth, development, and stress responses, but systematic comparative genomics studies across legume species are lacking. Methods: We identified the whole genome of the DOF gene family of five legume plants: Medicago truncatual (43), Cicer arietinum (43), Phaseolus vulgaris (44), Glycine max (79), and Lotus japonicus (32). Genome-wide identification of DOF genes was performed in five legume species, followed by phylogenetic analysis, gene structure characterization, duplication event identification, promoter element prediction, synteny analysis, and expression pattern profiling. Results: Phylogenetic comparison with Arabidopsis thaliana (47) and Oryza sativa (37) classified them into four subfamilies (Groups I–IV). The five legumes all had no more than 30% members of the subgroup. The same subfamily has similar protein structures and gene structures, and most of its members have motif1, with most plants having more than 30% of genes intronic. Gene duplication events were evenly distributed among the members of the DOF gene in all five legumes, and played an important role in its evolution. Moreover, the majority of the DOF genes showed tissue specificity in the five legumes, with most of these members being upregulated in flowers. Additionally, expression pattern analysis under abiotic stress in soybean revealed that members of different subfamilies exhibit divergent expression dynamics under salt, alkali, and cold stresses. The DOF gene family in legumes expanded primarily through segmental duplication and evolved under purifying selection. Conclusion: The subfamily-specific responses to abiotic stress and tissue-specific expression patterns provide candidate gene resources for functional studies aimed at improving stress tolerance and agronomic traits in legume crops. Full article

MLO (Mildew Resistance Locus O) genes encode seven-transmembrane proteins that function as critical regulators of powdery mildew resistance and abiotic stress responses. Despite their established importance, the MLO gene family in Gossypium hirsutum L. has not been systematically investigated under salt stress conditions. Here, we performed genome-wide identification of 46 GhMLO members using Hidden Markov Model and BLAST searches based on the latest cotton genome assembly. Phylogenetic analysis classified these genes into four distinct subfamilies. Transmembrane topology and conserved domain analyses revealed that all GhMLO proteins contain typical MLO domains and transmembrane structures, maintaining high structural similarity with dicotyledonous model plants. Synteny analysis demonstrated that the expansion of the GhMLO family was primarily driven by segmental and tandem duplications. Integration of transcriptomic data from the COTTONOMICS database revealed tissue-specific expression patterns, with higher transcript abundance in receptacles, stems, and roots, but lower levels in stamens and petals. Salt, drought, and cold stress treatments induced upregulation of GhMLO family members, with most genes showing increased expression over time. RT-qPCR analysis validated that five candidate GhMLO genes were significantly upregulated under salt stress. In summary, this study provides a comprehensive genome-wide characterization of the GhMLO gene family, elucidating their phylogenetic relationships and expression dynamics, which establishes a theoretical basis for identifying key regulatory genes involved in abiotic stress responses and offers novel genetic resources for improving stress tolerance in cotton molecular breeding. Full article

Sepsis is characterized by dysregulated immune responses induced by damage-associated molecular patterns, such as extracellular cold-inducible RNA-binding protein (eCIRP), that frequently lead to acute lung injury (ALI) and high mortality. Recently, a subset of CD4⁺ T cells possessing both T helper 1 (Th1) and regulatory T cell (Treg) phenotypes, termed Th1-Treg cells, has been identified; however, their function in sepsis remains unknown. In this study, we investigated the dynamics, induction mechanisms, and functional roles of Th1-Treg cells in the development of sepsis-induced ALI. Polymicrobial sepsis was induced in mice using cecal ligation and puncture. In vivo, Th1-Treg cell accumulation in the lungs was analyzed in WT and CIRP^−/− mice following sepsis. In vitro, isolated CD4⁺ T cells from WT and TLR4^−/− mice were treated with eCIRP to evaluate Th1-Treg cell differentiation and downstream signaling pathways. STAT1 and STAT5 activation were evaluated, and pharmacological inhibitors were used to assess their involvement. Adoptive transfer of Th1-Treg cells was conducted to determine their functional impact on ALI and mortality in septic mice. We observed a significant accumulation of Th1-Treg cells in the lungs of WT septic mice compared to sham mice. eCIRP drove the induction of Th1-Treg cells in vitro, and CIRP^−/− mice exhibited decreased Th1-Treg cell accumulation in the lungs compared to WT mice after sepsis. In parallel to Th1-Treg cell induction, eCIRP activated signal transducer and activator of transcription, STAT1 and STAT5. Both the induction of Th1-Treg cells and the activation of STAT1/5 proteins were significantly attenuated in TLR4^−/− mice. Furthermore, pharmacological inhibition of STAT1/5 signaling significantly reduced eCIRP-induced Th1-Treg cell differentiation. Intriguingly, adoptive transfer of Th1-Treg cells significantly exacerbated ALI, resulting in increased mortality in sepsis. Our findings indicate Th1-Treg cells induced by the eCIRP–TLR4–STAT1/5 axis aggravate ALI, worsening mortality in sepsis. Targeting these pathogenic cells potentially alleviates sepsis-induced ALI. Full article

Taxillus chinensis (DC.) Danser is an important hemiparasitic medicinal plant whose propagation is severely limited by the desiccation sensitivity of its recalcitrant seeds. Dehydrins (DHNs), which protect plants against dehydration-induced stresses such as salinity, drought, and low temperatures, may play a critical role in protecting recalcitrant seeds. However, the role of DHNs in the seeds of T. chinensis remains unclear. In this study, a differentially expressed gene was identified from the seed transcriptome of T. chinensis and designated TcDHN1. Sequence alignment and phylogenetic analyses revealed that TcDHN1 encodes a dehydrin protein. Heterologous overexpression of TcDHN1 in Arabidopsis did not affect growth under normal conditions. Under salt, drought, and cold stresses, transgenic lines exhibited higher seed germination rates, longer primary roots, and improved seedling growth compared with wild-type (WT) plants. The transgenic lines showed significantly increased activities of antioxidant enzymes, including superoxide dismutase, catalase, and peroxidase. In addition, ectopic overexpression of TcDHN1 in Arabidopsis conferred enhanced tolerance to abiotic stresses compared to WT plants, accompanied by increased expression of the stress-responsive genes Responsive to Desiccation 29A (AtRD29A) and Heat Shock Protein 70-1 (AtHSP70-1). The above results indicate that TcDHN1 confers enhanced tolerance to abiotic stresses. This study provides a functional characterization of an abiotic stress-responsive gene from recalcitrant seeds and identifies a potential genetic resource for molecular breeding. This could potentially improve abiotic stress resistance in T. chinensis and related medicinal plants. Full article

Low temperature and drought are among the most pervasive abiotic stresses limiting crop productivity worldwide, and their frequent co-occurrence or alternation imposes compounded constraints on agricultural sustainability. Increasing evidence supports cross-tolerance, whereby exposure to one stress enhances resistance to another, as an emergent property of shared signaling networks and integrative regulatory layers. In this review, we summarize recent advances in understanding cold–drought cross-talk, from early stress perception and secondary messengers to hormonal coordination via abscisic acid, transcriptional reprogramming centered on dehydration responsive element binding protein/C repeat binding factor (DREB/CBF) modules, and longer-term regulatory memory mediated by chromatin remodeling and biomolecular condensates. Importantly, we further discuss how these mechanistic insights can be translated into precision breeding strategies, including genome editing, allele mining, and backcross-assisted introgression, to accelerate the development of crop varieties with stable multi-stress tolerance. Finally, we highlight future directions for integrating multi-omics, high-throughput phenotyping, and data-driven approaches to enable efficient molecular design breeding for complex stress environments. Full article

Background/Objectives: Oncolytic viruses are an immunotherapeutic approach that can modulate the tumor microenvironment (TME), transforming immunologically ‘cold’ tumors into ‘hot’ ones. Insertion of genes encoding immunomodulatory proteins can further enhance antitumor immune responses. In this study, we compared the antitumor and immunomodulatory effects of the double recombinant vaccinia virus VV-GMCSF-Lact, which carries the human GM-CSF gene, with those of recombinant human GM-CSF (rhGM-CSF) in an immunocompetent murine GL261 glioma model. Methods: The study was conducted using a subcutaneous GL261 glioma model in immunocompetent C57BL/6 mice, comparing intratumoral VV-GMCSF-Lact and rhGM-CSF treatments with evaluation of immune cell populations by flow cytometry, tumor morphology by H&E staining, and tumor transcriptome profiles by RNA sequencing. Results: Flow cytometry showed that VV-GMCSF-Lact reduced the number of immunosuppressive cells in the TME of subcutaneously transplanted gliomas, targeting different components of the TME depending on animal sex. The immunotherapeutic effects of rhGM-CSF were less pronounced and primarily affected peripheral immune cells. Histological analysis revealed a decrease in mitotic figures in tumors from female mice after viral therapy. Transcriptome profiling of GL261 tumors demonstrated divergent gene expression patterns and cellular compositions between treatment groups. VV-GMCSF-Lact treatment was associated with a decreased proportion of malignant GL261 cells and CD8⁺ T lymphocytes, while rhGM-CSF treatment increased proportions of MDSCs, macrophages, NK cells, and tumor-associated neutrophils. Conclusions: Taken together, our data demonstrate that VV-GMCSF-Lact induces antitumor immune responses in murine GL261 glioma in vivo and modulates the tumor microenvironment more effectively than rhGM-CSF alone, supporting its potential for developing new strategies for glioma treatment. Full article

Isopentenyltransferase (IPT) is the rate-limiting enzyme in cytokinin biosynthesis and plays a critical role in plant acclimation to abiotic stress. To explore soybean IPT genes, we performed genome-wide identification, bioinformatics analysis, and molecular experimental validation to systematically characterize the features and functions of the soybean IPT (GmIPT) gene family. We identified 15 GmIPT genes in the soybean genome, which are unevenly distributed across 12 chromosomes; their evolutionary expansion is primarily driven by whole-genome duplication events. Phylogenetic analysis of soybean IPT proteins with those from Arabidopsis, rice and maize clustered them into four groups, exhibiting lineage-specific functional specialization. GmIPT genes exhibit significant variations in conserved motifs, gene structure, and cis-acting elements; their promoter regions are enriched in light-responsive, abiotic stress-responsive, and hormone-responsive elements, indicating their involvement in complex transcriptional regulatory networks. Tissue expression profiling revealed that GmIPT7 and GmIPT10 are highly expressed in various tissues, whereas GmIPT14 shows specific expression in flowers and the shoot apical meristem. Transcriptomic analysis and qRT-PCR validation demonstrated that GmIPT7, GmIPT10 and GmIPT15 respond differentially to drought, salt and low-temperature stress, with GmIPT15 exhibiting a transient upregulation at 3 h (p < 0.01) followed by a gradual decline to levels close to the pre-treatment control at 6–12 h under low-temperature stress. We further performed haplotype analysis of GmIPT15 and identified a putative elite haplotype (hap1) associated with cold tolerance based on low-temperature germination index assessment. This study provides useful insights for the future functional characterization of plant IPT genes and offers potential genetic resources and molecular markers that may support molecular-assisted breeding for soybean abiotic stress tolerance. Full article

Ram semen is highly susceptible to cold shock, which induces irreversible damage to the integrity and fluidity of membranes. Chilled semen is commonly used within 24 h of collection. However, while its storage at 5 °C extends semen lifespan, it is often accompanied by quality deterioration due to accumulation of reactive oxygen species (ROS). This study evaluated the potential of crocin, a carotenoid with antioxidant properties, to improve the quality of chilled ram semen stored at 5 °C for up to three days in a soybean lecithin–based extender supplemented with two crocin concentrations (0.5 and 1 mM). Sperm motility, viability, glutathione levels, the expression of proteins involved in the heat stress response (HSR), and apoptosis were assessed at 24 h intervals. Crocin preserved motility (up to Day 1), viability (up to Day 2,) and kinematic parameters (up to Day 3). In addition, crocin enhanced intracellular glutathione and Hsp70 levels and inhibited apoptotic levels dose-dependently, indicating the antioxidant and cytoprotective role of crocin. Despite 0.5 mM being effective up to Day 1, 1 mM crocin augmented antioxidant capacity, modulated stress response mechanisms, and preserved sperm quality during chilled storage up to Day 3, highlighting its potential as a valuable additive of ram semen extenders. Full article

The heterogeneity of colorectal cancer (CRC) represents a great challenge in therapy. We integrated multiomics and machine learning, interpreted by SHAP models to provide a clinical rationale, to identify Calcineurin B Homologous Protein 2 (CHP2) as a core candidate, which was further validated via in vitro and zebrafish models. The expression of CHP2 are decreased in CRC, which is associated with a poor prognosis and an immune suppressed “cold” TIME. Functionally, CHP2 overexpression inhibits cell growth and invasion by inducing PANoptosis. Clinically, specific CHP2 expression profiles discriminate patients at high risk that are resistant to standard chemotherapy (e.g., 5-FU) but sensitive to targeted inhibitors. CHP2 is a powerful dual-function biomarker—prognostic for survival and predictive for the response to therapy—that could lead to a personalized approach in treating drug-resistant CRC. Full article

Pitaya (Hylocereus undatus) is a typical Crassulacean Acid Metabolism (CAM) plant with strong drought tolerance but high sensitivity to low temperatures. In this study, the responses of pitaya cultivated in the karst areas of Guizhou Province in southwest China to drought and low temperature were examined in winter seasons. The stems of ‘Zihonglong’ pitaya were used as materials to investigate the physiological responses to cold temperatures of pitaya stems under different water conditions, so as to understand the effects of drought stress on the response to low temperatures. The results showed that the severity of chilling injury in pitaya stems was influenced by cold degree and duration and temperature variation. Under sustained low-temperature conditions, the lower the temperature and the longer the duration, the more severe the chilling injury, particularly at 4 °C and below. Drastic temperature rise after exposure to low temperature of 5 °C aggravated the damage, especially when the temperature rise exceeded 10 °C. Compared to normally irrigated plants, those subjected to drought pretreatment exhibited milder chilling injury and higher survival rates under a temperature shift from 5 to 20 °C. The drought-treated pitaya stems had significantly lower membrane leakage and malondialdehyde (MDA) and reactive oxygen species (ROS) contents compared with the well-watered control under different temperature increases starting from 5 °C. Drought significantly reduced soluble sugars and soluble proteins but increased proline under a temperature shift from 5 to 20 °C. It significantly enhanced the activities of catalase (CAT) and ascorbate peroxidase (APX) under temperature shifts from 5 to 10 or 20 °C, but had no significant effect on peroxidase (POD) and superoxide dismutase (SOD). Drought also significantly increased ascorbic acid (ASA) content but significantly reduced glutathione (GSH). It is concluded that a drastic post-cold temperature rise causes more severe damage than the cold temperature itself. Drought pretreatment increases the chilling tolerance of pitaya stems. This effect involves an enhanced ASA-GSH cycle, which strengthens ROS scavenging and prevents membrane damage. Full article

Longan (Dimocarpus longan Lour.) is highly sensitive to low temperature, which severely restricts its cultivation and industrial development. MYB transcription factors serve as key regulators in plant responses to cold stress. In this study, an R2R3-MYB gene DlMYB108 was cloned from ‘Shixia’ longan. Sequence analysis showed that DlMYB108 contains two typical MYB repeats and shares high homology with cold-responsive MYB108 proteins from other plants. Expression pattern analysis revealed that DlMYB108 is highly expressed in young leaves, which are more sensitive to cold stress, and is significantly induced by low-temperature treatment. Subcellular localization and transcriptional activation assays confirmed that DlMYB108 is a nuclear-localized transcriptional activator. Yeast one-hybrid and dual-luciferase assays demonstrated that DlMYB108 specifically binds to the promoters of DlCBF2 and DlCBF3 and activates their transcription. Heterologous expression of DlMYB108 in Arabidopsis significantly enhanced cold tolerance, accompanied by reduced ion leakage, malondialdehyde (MDA) content and reactive oxygen species (ROS) accumulation, as well as upregulated expression of CBF and cold-responsive genes. Collectively, DlMYB108 positively regulates longan cold tolerance through activating DlCBF2 and DlCBF3 expression, providing a valuable candidate gene for cold-tolerant longan breeding. Full article