Search Results (6,110)

Crocins are high-value apocarotenoid pigments with broad applications in pharmaceuticals, foods, and personal-care products, and they exhibit diverse bioactivities, including antioxidant, antidepressant, and antidementia effects. In this study, we achieved the heterologous biosynthesis of crocins in Solanum lycopersicum L. by introducing the GjCCD4a, GjALDH2C3, GjUGT74F8 and GjUGT94E13 of Gardenia jasminoides J.Ellis to the binary expression vector via in-fusion technology and self-cleaving 2A peptides. Following Agrobacterium-mediated transformation, the engineered tomato plants predominantly produced main active ingredient crocin I, which accounted for 97–99% of the total crocins. The transgenic fruits displayed mixed red-and-golden colouration. These results highlight S. lycopersicum as a promising chassis for crocin I biosynthesis, helping to address supply constraints and enabling colour-trait breeding through synthetic biology. Full article

Hemophilia, an X-linked recessive bleeding disorder, results from mutations in the F8 or F9 genes, leading to factor VIII (hemophilia A) or factor IX (hemophilia B) deficiency. While conventional treatment relies on regular factor replacement therapy, gene therapy has emerged as a promising alternative, offering the potential for sustained endogenous factor production after a single administration. This review provides an in-depth analysis of recent advances in gene therapy for both hemophilia A and B, with a focus on AAV-mediated liver-directed approaches and other approved modalities. Key limitations—such as vector immunogenicity, hepatic toxicity, waning transgene expression, and limited re-dosing capacity—are discussed. Additional gene delivery platforms, including lentiviral and retroviral vectors, genome editing techniques (e.g., CRISPR/Cas9), and non-viral systems like transposons and lipid nanoparticles, are also examined. Although gene therapy for hemophilia B demonstrates greater clinical durability, hemophilia A presents unique challenges due to factor VIII’s size, poor expression efficiency, and the need for higher vector doses. Future efforts will focus on overcoming immune barriers, improving delivery technologies, and developing approaches suitable for pediatric patients and individuals with pre-existing immunity. This review provides not only a descriptive overview but also a critical comparison of gene therapy approaches for hemophilia A and B. We emphasize that the durability of response is currently superior in hemophilia B, whereas hemophilia A still faces unique barriers, including declining FVIII expression and higher immunogenicity. By analyzing cross-platform challenges (AAV, lentiviral, CRISPR, and emerging LNPs), we highlight the most promising strategies for overcoming these limitations and provide a forward-looking perspective on the future of gene therapy. Full article

Broccoli (Brassica oleracea L. var. italica) is a biennial or annual herbaceous plant belonging to the species Brassica oleracea in the genus Brassica of the Cruciferae family. The green flower curd serves as the primary edible organ, with its development and preservation critically determining broccoli yield and quality. Given that these processes are regulated by flowering time, understanding the mechanisms underlying floral transition is essential for enhancing broccoli yield and quality. This study aimed to identify the MADS-box family in broccoli and to investigate the function of the BoAGL8 gene in floral induction. We identified a total of 176 MADS-box genes, of which 54 genes were up-regulated and 50 genes were down-regulated under low-temperature treatment. Notably, the expression of BoAGL8 was up-regulated by 6.70-fold under low-temperature induction, prompting us to select and clone this gene for further analysis. Tissue-specific expression profiling further revealed that BoAGL8 is expressed at relatively high level in both mature and young leaves. After 15 days of low-temperature treatment, BoAGL8 expression in shoot tip was significantly upregulated compared to untreated controls. Subcellular localization analysis showed that BoAGL8 protein was located to the nucleus. Ectopic over-expression of BoAGL8 in Arabidopsis exhibited accelerated bolting and flowering, reduced rosette leaf number, and increased seed yield per plant compared to wild-type plants. Furthermore, compared to wild-type controls, transgenic lines exhibited upregulated expression of AtFT, AtAP1 and AtSEP3, alongside downregulation of SVP expression. The above results indicate that BoAGL8 may play a key regulatory role in the process of floral organ development in broccoli, providing an important theoretical basis for future research on flowering time regulation and breeding in broccoli. Full article

Background: Periodontitis is a chronic disease triggered by disturbed oral microbiota. We have previously reported that hepatocyte growth factor (HGF) could mitigate early-stage experimental periodontitis but exacerbate the condition in its late stage. Here, we investigated the impact of HGF on the periodontal microbiome during periodontitis progression. Methods: We established ligation-induced periodontitis in wild-type (WT) mice and HGF high-expression transgenic (HGF-Tg) mice. We quantified the levels of IL-6 and TNF-α in periodontal tissues, as well as the serum concentrations of CTXI and PINP. Ligatures were collected on days 0, 7, and 28 after ligation for 16S rRNA sequencing and microbial analysis. Results: HGF significantly altered the diversity of ligatures during periodontitis. Interestingly, specific microbial genera, such as Lactobacillus, exhibited opposing trends between the two disease stages of HGF-Tg mice, aligning with the different effects of HGF on periodontitis progression. We also identified some taxa, such as Sphingomonas, associated with IL-6, TNF-α, CTXI, and PINP. The predicted inflammatory pathways (e.g., IL-17 signaling pathways) were enriched in HGF-Tg mice on day 28 but decreased on day 7. Conclusions: HGF exerted different influences on the microbiota of ligatures during early and late stages of periodontitis, which may account for the divergent effects of HGF on periodontitis progression. Full article

The cold-regulated (Cor413) gene family encodes plant-specific, multispanning transmembrane proteins that localize to the plasma and thylakoid membranes; these genes are regulated by environmental stimuli. In this study, the Cor413-1 gene, isolated from the drought and saline-tolerant wild species Saccharum spontaneum, was engineered into the elite sugarcane cultivar Co 86032 to produce a commercially superior cultivar with improved abiotic stress tolerance. Expression analysis of the Cor413-1 gene transgenic lines under drought and salinity stress exhibited distinct gene expression patterns. During stress conditions, transgenic events, such as Cor413-9 and Cor413-3, showed notable resilience to salt stress and had a high relative expression of the Cor413-1 gene and other stress-related genes. The evaluation of physiological parameters showed that under stress conditions, transgenic events experienced milder wilting and less cell membrane injury than the non-transgenic control. Transgenic lines also demonstrated elevated relative water content and better photosynthetic efficiency, with events like Cor413-10 and Cor413-12 showing exceptional performance. Biochemical analyses indicated elevated proline content, higher activity of enzymatic antioxidants such as sodium dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX), and a low level of malondialdehyde MDA production in the transgenic lines. Thus, demonstrating the potential of the Cor413-1 gene for developing multiple stress-tolerant cultivars. Full article

MicroRNA (miRNA), as a key post-transcriptional regulatory factor, plays a crucial role in embryonic development. The coordination of endoderm cell convergence and cardiac precursor cell (CPC) migration is critical for cardiac tube fusion. Defects in endoderm can impair the normal migration of CPCs towards the midline, leading to cardia bifida. Although the role of the microRNA-183 family (miR-183, miR-96 and miR-182) in cardiovascular diseases has been reported, the mechanism by which they regulate early heart development remains unclear. In this study, we used zebrafish as a model to elucidate the roles of the microRNA-183 family in early heart development. miRNA mimics were injected into Tg (cmlc2: eGFP) and Tg (sox17: eGFP) transgenic embryos to overexpress the miR-183 family. The results showed that, at 36 hpf, single or co-injection of miR-183/96/182 mimics caused defects in endoderm convergence, with a hole in the endoderm, and a significant down-regulation of the endoderm marker gene sox32. Additionally, embryos with single or co-injection of miR-183/96/182 mimics exhibited cardia bifida and tail blisters, with significantly down-regulated expression levels of genes related to heart development, including cmlc2, vmhc, amhc, nppa, gata4, gata5, nkx2.5, bmp2b, and bmp4. The phenotype caused by overexpression of the miR-183 family is highly consistent with loss of the sphingosine 1-phosphate receptor S1Pr2. Bioinformatics analysis result found that miR-183 can bind to 3′-UTR of the s1pr2 to regulate its expression; overexpression of miR-183 led to a significant decrease in the expression of the s1pr2 gene. Dual luciferase assay results suggest that s1pr2 is a bona fide target of miR-183. In summary, the miR-183 family regulates endoderm convergence and cardiac precursor cell migration via the S1Pr2 signaling pathway. This study reveals that the miR-183 family is a key regulatory factor in endoderm convergence and cardiac precursor cell migration during the early zebrafish development, elucidating the molecular mechanisms underlying early cardiac precursor cell and endoderm cell movement. Full article

Introduction/Background: Rift Valley fever virus (RVFV) and peste des petits ruminants virus (PPRV) are significant pathogens affecting small ruminants, causing substantial economic losses in the affected regions. The development of effective vaccines against both viruses is crucial for disease control. Recombinant viruses expressing heterologous antigens have shown promise as multivalent vaccine candidates. Unlike conventional PPRV vaccines, our recombinant RVFV-vectored vaccines offer a novel dual-protection strategy against RVF and PPR, combining safety, immunogenicity, and a DIVA strategy. Methods: Recombinant RVFVs (ZH548 strain) were generated to express either the hemagglutinin (H) or fusion (F) proteins from the PPRV strain Nigeria 75/1. The stability of these recombinant viruses was assessed through consecutive passages in cell culture. Immunogenicity studies were carried out in both mice and sheep to assess the induction of cellular and humoral immune responses capable of providing protection against RVFV and PPRV. These studies included intracellular cytokine staining (ICS), IFN-γ ELISAs, standard ELISAs for antibody detection, and virus neutralization assays. Results: The recombinant RVFVs expressing PPRV H or F proteins demonstrated stability in cell culture, maintaining high viral titers and consistent transgene expression over four passages. Immunization of mice resulted in the production of serum antibodies capable of neutralizing both RVFV and PPRV in vitro as well as cell-mediated immune responses specific to PPRV and RVFV antigens. In mice vaccinated with a high dose (10⁵ pfu), RVFV neutralizing titers reached ≥1:160 and PPRV neutralizing titers ranged from 1:40 to 1:80 by day 30 post-immunization. In sheep, neutralizing antibody titers against RVFV exceeded 1:160 as early as 2 days post-inoculation, while PPRV-specific neutralization titers reached up to 1:80 by day 21 in responsive individuals. In mice, administration of rZH548ΔNSs:F_PPRV elicited a detectable CD8+ IFNγ+ T-cell response against PPRV, with levels ranging from 1.29% to 1.56% for the low and high doses, respectively. In sheep, rZH548ΔNSs:F_PPRV also induced a robust IFNγ production against PPRV at 14 and 21 days post-infection (dpi). Conclusions: The successful generation and characterization of recombinant RVFVs expressing PPRV antigens demonstrate the potential of using rationally attenuated RVFV as a vector for multivalent vaccine development. Notably, the strategy proved more effective for the recombinant virus expressing the F protein, as it consistently induced more robust cellular and humoral immune responses. These results suggest that this approach could be a viable strategy for simultaneous immunization against Rift Valley fever and other prevalent ruminant diseases, such as peste des petits ruminants. Even though challenge studies were not performed in target species, the strong immune response observed supports including them in future studies. Full article

Miscanthus, a perennial grass, is renowned for its remarkable tolerance to abiotic stress. Excessive levels of drought severely impair plant growth and yield. Plant nuclear factor Y (NF-Y) transcription factors (TFs) play pivotal roles in regulating responses to drought stress in species such as Arabidopsis and maize. However, their functional roles in conferring drought tolerance in Miscanthus remain largely unexplored. This study’s genome-wide analysis and gene expression profiling of Miscanthus under dehydration/osmotic stress identified a transcription factors gene, MsNF-YA4, which was significantly upregulated under dehydration/osmotic stress. MsNF-YA4 overexpression in Arabidopsis significantly enhanced drought tolerance, leading to increased transcription of stress- and antioxidant enzyme-related genes. Compared with the wild type (WT), the transgenic lines exhibited markedly higher relative water content (RWC), chlorophyll content, proline level, and antioxidant enzyme activity. Furthermore, the MsNF-YA4/MsNF-YB3/MsNF-YC2 improved the transactivation of the Miscanthus P5CS1, SOD (Cu/Zn) and CAT1 promoters in the transient system. These results offer fresh perspectives on the role of Miscanthus NF-YAs in drought tolerance and offer promising genetic resources for developing drought-tolerant crops through breeding programs. Full article

The BRI1-EMS suppressor/Brassinazole-resistant (BES/BZR) transcription factors (TFs) act as regulators of the Brassinosteroid (BR) signaling pathway and play key roles in modulating plant growth, development, and abiotic stress tolerance. However, the function of BES/BZR TFs remains unknown in warm-season turfgrass species. In this study, ZjBZR2, a BES/BZR TF in Zoysia japonica was identified and shared the closest evolutionary relationship with OsBZR2 from Oryza sativa. ZjBZR2 was a nuclear-localized protein and had transcriptional activation activity. ZjBZR2 was predominantly expressed in roots, stems, and lamina joints, and could be significantly induced by BR treatment and osmotic stresses including PEG and salinity. ZjBZR2-overexpressing rice lines increased leaf angle compared with wild-type plants. Furthermore, overexpression of ZjBZR2 enhanced osmotic stress (PEG and salt) tolerance which is associated with the upregulation of stress-responsive and ROS-scavenging genes. These findings provide the first functional characterization of ZjBZR2 in rice and offer excellent genetic resources for the improvement of turfgrass cultivars. Full article

Bladder dysfunction, particularly overactive bladder (OAB), is increasingly recognized as a clinical concern in patients with sickle cell disease (SCD), yet its pathophysiological mechanisms remain poorly understood. This study investigated the relationship between oxidative stress, inflammation, and bladder dysfunction in the Townes transgenic SCD mouse model. Cystometric analysis revealed that SCD mice exhibit an OAB phenotype, characterized by increased frequencies of voiding and non-voiding contractions and reduced bladder compliance. In vitro functional assays demonstrated detrusor hypocontractility in SCD mice, associated with a significant reduction in carbachol- and EFS-induced contractions and downregulation of muscarinic M3 receptor expression. Purinergic signaling and calcium-dependent contractility remained preserved. Molecular analyses showed increased mRNA expression of NOX-2 and IL-1β, and elevated protein levels of 3-nitrotyrosine and myeloperoxidase (MPO) activity, indicating redox imbalance and chronic inflammation in bladder tissue. Together, these changes suggest that oxidative and nitrosative stress, combined with inflammation, contribute to bladder remodeling and dysfunction in SCD. This is the first study to characterize bladder alterations in Townes SCD mice, establishing this model as a valuable tool for investigating lower urinary tract complications in SCD. Our findings provide mechanistic insight into the genitourinary manifestations of SCD and identify redox and inflammatory pathways as potential therapeutic targets for bladder dysfunction in affected individuals. Full article

Salt stress is a major abiotic factor limiting crop productivity worldwide, as it disrupts plant growth, metabolism, and survival. In this study, we report that the genes PvPR10-2 and PvPR10-3 were significantly up-regulated in bean leaves and stems in response to combined salt and jasmonic acid (NaCl–JA) treatment. Foliar application of JA with salt induced physiological alterations, including stem growth inhibition, H₂O₂ accumulation, and activation of antioxidant enzymes. To investigate the role of PvPR10-3 in response to salt and phytohormones, we introduced this gene into Arabidopsis and found that its heterologous expression conferred salt tolerance to the transgenic lines. Interestingly, exogenous JA contributed to salt tolerance by reducing H₂O₂ levels, inducing ROS-scavenging enzymes, and promoting the accumulation of phenolic compounds and ABA. Furthermore, gene expression analysis of the transgenic lines revealed that PvPR10-3 expression under NaCl–JA stress is associated with the induction of JA-related genes like MYC2, JAZ2, JAZ11, and JAZ12, as well as SA-responsive genes, like ALD1 and TGA2, and two ABA-independent components DREB2A and ERD1, suggesting potential coordination between JA, ABA, and SA signaling in salt stress response. Additionally, key flowering regulators (FT, GI) were upregulated in transgenic lines under NaCl–JA treatment, suggesting a previously unexplored link between salt tolerance pathways and the regulation of flowering time. Taken together, our findings suggest a role of PvPR10-3 in enhancing salt stress tolerance and the involvement of exogenous JA in tolerance potentially by modulating ROS balance, hormone-associated gene expression, and protective secondary metabolites. Full article

Alfalfa (Medicago sativa L.) is a vital global forage crop. Transgenic technology promises enhanced yield and quality, but requires rigorous environmental risk assessment, particularly regarding pollen-mediated gene flow, for which standardized protocols are lacking. Based on an optimized in vitro culture medium, this study developed a method to assess alfalfa pollen viability. Using a single-factor experimental design, key assessment parameters were established at 1/4/8 h and 20/30/40 °C. A comparative analysis revealed no significant difference (p > 0.05) in pollen viability between the transgenic line SA6-8 and its non-transgenic parent “ZM-1” within this evaluation system. This result indicates that the genetic modification did not impact the pollen viability of SA6-8. By establishing this in vitro germination-based pollen viability assessment system and comparatively analyzing pollen viability between transgenic alfalfa and its non-transgenic parent under diverse environmental conditions, our approach provides crucial insights for optimizing transgenic alfalfa planting strategies and strengthening biosafety review protocols. Full article

Pre-harvest sprouting (PHS) reduces the quality and quantity of crop seeds. PHS can be imposed through the embryo or husk pathway of cereal crops. Most reported PHS seeds are imposed via the embryo pathway. Here, we generated transgenic rice plants overexpressing rice melatonin 2-hydroxylase (OsM2H), which catalyzes the hydroxylation of melatonin to 2-hydroxymelatonin (2-OHM). OsM2H overexpression (M2H-OE) showed PHS under paddy conditions. Germination assays revealed that intact seeds harvested at 26 and 36 days after heading (DAH) showed PHS, whereas dehusked seeds did not, indicating husk-imposed PHS. Overproduction of 2-OHM was observed in M2H-OE seeds compared to wild-type control. In addition, M2H-OE lines produced more hydrogen peroxide than the wild-type. 2-OHM-induced reactive oxygen species resulted in the induction of OsGA3ox2, a gibberellin (GA) biosynthesis gene, and repression of OsGA2ox3, a GA degradation gene, in caryopses at 2 DAH, but in the induction of the ABA degradation gene OsABA8ox3 in intact seeds at 26 DAH. In addition, M2H-OE seedlings were longer and showed increased levels of hydrogen peroxide and OsGA3ox2 expression versus the wild-type. This is the first report showing that 2-OHM can induce PHS via the husk pathway in rice seeds through the induction of GA biosynthetic and ABA degradation genes. Full article

Transgenic insect-resistant maize can effectively control insect pests, which is of great significance to improve maize yield and quality. Transgenic maize LD05 is an insect-resistant and herbicide-tolerant maize independently developed by Shandong Academy of Agricultural Sciences and highly resistant to major lepidopteran pests. In order to study the pest resistance of transgenic maize LD05 in different ecological areas of China, this study conducted a laboratory bioassay, and artificial inoculation test and natural pest investigation in field were carried out in one pilot of each of five maize ecological zones in China. The results of laboratory bioassay showed that transgenic maize LD05 had high resistance to Ostrinia furnacalis (Guenée), Mythimna separata (Walker), Helicoverpa armigera (Hübner) and Spodoptera frugiperda (J. E. Smith), the main lepidopteran pests threatening maize production in China. The results of artificial inoculation test and natural pest investigation in field showed that transgenic maize LD05 had high resistance to major lepidopteran pests in different ecological areas of China, which was consistent with the pest resistance management strategy, and can provide important theoretical basis and technical support for the industrialization of transgenic maize LD05 in the future. Full article

The cereal cyst nematode, Heterodera avena, is responsible for substantial economic losses in the global production of wheat, barley, and other cereal crops. Extracellular enzymes, particularly those from the glycoside hydrolase 18 (GH18) family, such as chitinases secreted by Trichoderma spp., play a crucial role in nematode control. However, the genome-wide analysis of Trichoderma longibrachiatum T6 (T6) GH18 family genes in controlling of H. avenae remains unexplored. Through phylogenetic analysis and bioinformatics tools, we identified and conducted a detailed analysis of 18 GH18 genes distributed across 13 chromosomes. The analysis encompassed gene structure, evolutionary development, protein characteristics, and gene expression profiles following T6 parasitism on H. avenae, as determined by RT-qPCR. Our results indicate that 18 GH18 members in T6 were clustered into three major groups (A, B, and C), which comprise seven subgroups. Each subgroup exhibits highly conserved catalytic domains, motifs, and gene structures, while the cis-acting elements demonstrate extensive responsiveness to hormones, stress-related signals, and light. These members are significantly enriched in the chitin catabolic process, extracellular region, and chitinase activity (GO functional enrichment), and they are involved in amino sugar and nucleotide sugar metabolism (KEGG pathway enrichment). Additionally, 13 members formed an interaction network, enhancing chitin degradation efficiency through synergistic effects. Interestingly, 18 members of the GH18 family genes were expressed after T6 parasitism on H. avenae cysts. Notably, GH18-3 (Group B) and GH18-16 (Group A) were significantly upregulated, with average increases of 3.21-fold and 3.10-fold, respectively, from 12 to 96 h after parasitism while compared to the control group. Meanwhile, we found that the GH18-3 and GH18-16 proteins exhibit the highest homology with key enzymes responsible for antifungal activity in T. harzianum, demonstrating dual biocontrol potential in both antifungal activity and nematode control. Overall, these results indicate that the GH18 family has undergone functional diversification during evolution, with each member assuming specific biological roles in T6 effect on nematodes. This study provides a theoretical foundation for identifying novel nematicidal genes from T6 and cultivating highly efficient biocontrol strains through transgenic engineering, which holds significant practical implications for advancing the biocontrol of plant-parasitic nematodes (PPNs). Full article