Search Results (394)

This study provides a comprehensive and refined framework for the micropropagation of the critically endangered Greek endemic Silene conglomeratica. Using a limited seed stock, a reliable in vitro propagation protocol was developed, supporting both ex situ conservation and potential commercial applications in floriculture and landscape architecture. Nodal explants excised from aseptic seedlings, established on half-strength Murashige and Skoog (MS) medium, were successfully used for culture initiation. Supplementation with 1.0 mg L⁻¹ meta-topolin (mT) and 0.2 mg L⁻¹ 1-naphthaleneacetic acid (NAA) promoted shoot proliferation. Subsequent subculturing on half-strength Rugini Olive Medium (OM/2) supplemented with 0.5 mg L⁻¹ 2-isopentenyladenine (2iP) resulted in high multiplication rates and a high frequency of spontaneous rooting. Rooting initiation was further optimized using OM/2 supplemented with 0.5 mg L⁻¹ indole-3-butyric acid (IBA). The high acclimatization percentage (80%) confirms the feasibility of this protocol for ex situ conservation and highlights its applicability for nursery production and specialized landscape use. Overall, this study contributes an efficient and scalable propagation strategy that supports both the conservation and sustainable utilization of this valuable endemic species. Future work should focus on refining these protocols through more targeted testing of concentrations and alternative combinations of growth regulators and nutrient compositions. Full article

The genus Isaria is a group of abundant and widely distributed entomopathogenic fungi that plays an important role in the history of traditional Chinese medicine. Entomopathogenic fungi with medicinal value were collected from the field, and optimal temperature and growth media compositions were investigated to establish a theoretical foundation for the future development of these strains. A strain of Isaria cateniobliqua, designated ICF, was isolated from soil in the Hualongshan National Nature Reserve in southern Shaanxi. The optimal cultivation temperature and nutrient solution were screened, and the effects of subcultivation on mycelium production, metabolite production, and hydroxyl radical scavenging activity of strain ICF were investigated. The optimal growth temperature for strain ICF was determined to be 21 °C, with the ideal culture medium consisting of glucose and tussah silkworm pupa powder supplemented with KH₂PO₄ and MgSO₄. Mycelium production and cordycepin content peaked in the fourth generation (G4), whereas peak metabolite production and cordycepic acid production occurred in the fifth generation (G5). Polysaccharide content was highest in the first generation (G1), and hydroxyl radical scavenging activity was optimal in G4. Exploring the optimal culture conditions of the strain provides a theoretical basis for its development, utilization, and industrial production for medicinal applications. Full article

The increased mobilization of Rare Earth Elements (REEs), due to emerging technologies, could impact human health. The study assessed the effects of CeO₂ nanopowder (100 μg/mL) in human intestinal cells (HT-29) following both acute (24 h) and, a novelty for in vitro study, sub-chronic exposure, treating subcultures of exposed cells to CeO₂ NP up to 35 days. Recovery was also examined in exposed cells’ progeny. CeO₂ NP internalization and acute cytotoxicity were dose and time dependent. A significant pro-oxidant effect was observed for up to 14 days. The highest mitochondrial impairment was detected after 7 days, but in post-exposure experiments the recovery was observed. Conversely, genotoxicity highlighted the saturation of the DNA repair mechanisms. The irreversible cell damage of sub-chronic exposure was highlighted by the percentage of death cells (p = 0.011) and by the weekly cell replication index (5.68 vs. 7.41). The homeostatic mitophagy pathway was able to counteract ROS-induced mitochondrial dysfunction, as shown by overexpression of ATG5, LC3, and BECN1 genes throughout the examined times. Instead, the overexpression of the pro-apoptotic gene Bax was very brief, highlighting that prolonged exposure might cause more widespread adverse effects, also involving cells that are not directly exposed to nanoceria. Full article

Metarhizium lepidiotae is an important entomopathogenic fungus with substantial agricultural value. However, prolonged subculturing often leads to phenotypic degeneration, including reduced conidiation and impaired metabolic activity, while the underlying molecular mechanisms remain poorly understood. Elucidating these mechanisms is essential for maintaining strain vitality and ensuring biocontrol efficacy. In this study, we found that M. lepidiotae exhibited a pronounced decline in conidiation during long-term serial subculturing. However, this degenerative phenotype could be effectively reversed by passage through insect hosts, leading to strain rejuvenation. Subsequently, comparative transcriptomic analyses were performed on the original strain (XMC-Y), the degenerated strain (XMC-T), and the rejuvenated strain (XMC-F) at 7 and 18 days of cultivation. Our results revealed that XMC-T initially compensates for defects in basal metabolism and signaling pathways by enhancing translational capacity, but progressively exhibits a profound collapse of RNA-processing systems and the translational machinery at later cultivation stages. Moreover, the significant downregulation of the peroxisome pathway indicates impaired peroxisome biogenesis and compromised reactive oxygen species (ROS) metabolic capacity, suggesting a weakened antioxidant defense and a potential increase in oxidative stress. Collectively, these findings indicate that disruptions in RNA regulatory networks and oxidative homeostasis are strongly associated with M. lepidiotae degeneration. This study provides important theoretical insights for maintaining strain stability during large-scale production and agricultural biocontrol applications. Full article

The rapid expansion of gaming and esports in Saudi Arabia has prompted new interest in understanding how social and regional factors shape digital play. One trend is the variation in gaming preferences and motivations across gender and geographic regions. Previous research on gaming behavior has often centered on Western contexts and cross-national comparisons, overlooking how subcultural variation may shape players’ motivations and genre choices. Through a nationwide survey of gamers in Saudi Arabia, distributed via gaming community platforms, we examined how gender and region relate to gaming motivations, preferred genres, and intention to continue playing. Quantitative data were analyzed using inferential statistical methods, while qualitative responses were examined through thematic analysis to identify recurring patterns and contextual insights. Findings reveal that men are more likely to report performance and competition-driven motivations, while women show stronger preferences for escapism and casual gameplay. These results indicate that regional subcultures within Saudi Arabia shape gaming behavior, challenging generalized assumptions about player preferences. By demonstrating systematic regional differences in gaming motivations and preferences, this study extends games user research and informs the development of culturally responsive game design strategies. Full article

While drill music is often talked about in relation to crime, it is often overlooked as an art form and cultural practice. Consequently, its artists are rarely heard from. To address this societal and academic gap, we have conducted in-depth interviews with ten UK artists from this genre and subculture. This article presents the shared meanings these UK drill artists attach to the motivation to make their own music, the music subgenre and its culture, and its ongoing criminalisation. We do so by conceptualising these meanings as counter-narratives. The article departs from the observation that these counter-narratives present themselves in drill, as a form of expression, on two dimensions: drill as the outcome of intra-group expression of emotions and social relations, and as the platform to engage with social injustice on the inter-group level. An interdisciplinary theoretical framework, combining psychological insights on needs, philosophical cues on (mis)recognition, and the lens intersectionality, allows us to study and bridge these two dimensions. We identify twelve counter-narratives that were validated by a majority of respondents. The study, besides analysing these in-depth counter-narratives, also foregrounds UK drill artists’ agency generally absent from both societal and academic discourse. Full article

The complex microbiota of cheese starters plays a key role in determining the structure and flavour of the final product, primarily through their acid-forming capacity, protease activity, and exopolysaccharide synthesis. However, the specific microbial communities underlying the unique qualities of artisanal cheeses remain poorly understood. This study presents the microbiological and molecular genetic characterisation of the microbiome isolated from an artisanal cheese starter in Kosh-Agach, Altai, Russia. Metagenomic analysis of this starter revealed the presence of three bacterial genomes corresponding to those of Lactococcus lactis. Pure cultures from this starter were obtained by sequential subculture, and seventeen colonies displaying distinct characteristics on differential media were selected. Genome sequencing was performed for each colony. Bioinformatic analysis based on the rpoB gene grouped the isolates into three clusters, each corresponding to a distinct strain of Lactococcus lactis subsp. diacetilactis. This classification was further confirmed by microbiological and microscopic analyses. A notable finding was that none of the strains produced the characteristic aroma compounds of L. l. subsp. diacetilactis, namely, diacetyl and CO₂. The functional properties and metabolic characteristics of this starter consortium are discussed. Full article

Background/Objectives: Lawsonia intracellularis (L. intracellularis) is an important intestinal pathogen that causes porcine proliferative enteropathy (PPE) in swine production worldwide. Currently, only a few commercially available vaccines are available for PPE prevention. Methods: In this study, an attenuated L. intracellularis variant of JS-G90 was obtained through subculturing of L. intracellularis JS isolates in McCoy cells for 90 generations, and its immune response was evaluated in pigs. Results: The results demonstrated that pigs who underwent intragastric administration of JS-G90 had lower fecal bacterial shedding and no histopathological lesions, indicating that it was safe in pigs. Therefore, JS-G90 was selected to develop the attenuated PPE vaccine. The immune response of JS-G90 in pigs was further evaluated based on fecal bacterial shedding, histopathological lesions, and humoral and cell-mediated immune responses following challenge with pathogenic L. intracellularis. The results revealed that JS-G90 significantly decreased the copies of L. intracellularis in rectal swabs containing feces and ileum infection (p < 0.001), reduced histopathological lesions in the ileum, and elicited non-specific humoral (IgG and sIgA) and cell-mediated immune responses (p < 0.001) compared with the challenge control and mock groups. Conclusions: In conclusion, the attenuated vaccine JS-G90 is safe and induced humoral and cell-mediated immune responses in pigs against pathogenic L. intracellularis infection. It may serve as an effective strategy for preventing and controlling PPE. Full article

In vitro-cultured plant calli are colonized by diverse endophytes. As these endophytes are inherited from the maternal plant and appear to be highly dependent on the eco-niche of the host cells, they have been termed host-dependent endophytes (HDEs). HDEs occupy the most intimate microbial environment of plant cells. Nevertheless, our understanding of HDEs and their microenvironmental effects on host plants remains limited due to their cultivation-recalcitrant nature. In this study, grapevine (Vitis vinifera L. × V. labrusca L.) callus was subjected to long-term cultivation in media containing different antibiotics (Q: penicillin; L: streptomycin; Z: nystatin) with the intention of creating grapevine calli with different HDEs. The treated calli were then transferred to an antibiotic-free medium for continuous cultivation. After three cycles of subculture over a total period of 45 days, the endophytic microbiota of the grapevine calli were profiled and their physiological parameters were analyzed. Our results revealed that antibiotic treatments can effectively shape HDEs and create distinct bacterial and fungal HDE microbiota in grapevine calli. Compared to treatment without antibiotics (CK), the Q-treated callus contained more Gram-positive bacterial HDEs but fewer Gram-negative and stress-resistant bacterial HDEs, whereas the Z-treated callus had fewer Gram-positive bacterial HDEs and more Gram-negative, stress-resistant and potentially pathogenic bacterial HDEs. More importantly, grapevine calli with different HDE communities showed varying physiological traits such as respiratory rate, peroxidase activity and total sugar content. Correlative analyses further revealed the functional associations between HDE taxa and callus traits. This work provides an example for studying and utilizing plant HDEs. Full article

Background/Objectives: We aimed to investigate the effects of tacrolimus on human Tenon’s fibroblast (HTF) migration, proliferation, and transdifferentiation in vitro. Methods: HTF cells were subcultured and serum-starved for 24 h before being treated with 10 ng/mL tacrolimus. After 1 h, 30 ng/mL platelet-derived growth factor (PDGF) or 10 ng/mL transforming growth factor beta-1 (TGF-β1) was administered to the HTFs. Migration, proliferation, and transdifferentiation were assessed using WST-1 assays, scratch-induced directional wounding, and western blot analysis. The involvement of the TGF-β signaling pathway was examined via western blotting to measure phosphorylated Smad2, Smad3, ERK, and Akt levels. Results: TGF-β1 and PDGF enhanced HTF migration, proliferation, and transdifferentiation, whereas tacrolimus inhibited these effects. Tacrolimus also inhibited the TGF-β1-induced upregulation of phosphorylated Smad2 and Smad3, suggesting its inhibitory effects occur through TGF-β1 signaling. Conclusions: Overall, tacrolimus can inhibit PDGF- and TGF-β1-induced HTF migration, proliferation, and transdifferentiation, primarily through the Smad-dependent TGF-β signaling pathway. To develop a new therapeutic modality, further longitudinal in vivo studies and human clinical trials are warranted. Full article

Chelerythrine (CHE) is a naturally occurring benzophenanthridine alkaloid obtained from plants such as Chelidonium majus L. It has received notable attention in pharmacology and microbial control because of its broad-spectrum activity and marked anti-inflammatory, apoptosis-inducing, and antibacterial effects. In this study, Bacillus tropicus, which frequently presents in the soil environment, was selected as the target microorganism to systematically examine the dose-dependent inhibitory influence of CHE on its growth curve, biofilm development, and survival rate. Furthermore, by simulating an antibiotic pressure environment in vitro, the original strain was subjected to continuous subculturing (30 times), and a highly drug-resistant B. tropicus strain capable of stable growth under high concentrations of CHE (300 mg/L) was successfully acclimated. After that, transcriptomics analysis was employed to compare the genetic differences between the wild-type bacterium and drug-resistant bacterium to determine how bacterial cells are able to resist CHE. A total of 868 genes in the CHE-resistant bacterium were revealed to be more active, while 539 genes were less active. These results indicate that the CHE resistance characteristics of the strain may be related to the adjustment of its sugar metabolism pathway and the biofilm formation pathway. As a widely used biological control bacterial strain, the successful acclimation of the B. tropicus strain with resistance to CHE has made it possible to use the combined formulation of these two agents for the prevention and control of plant diseases. Full article

White leaf spot disease [Neopseudocercosporella capsellae (Ellis & Everhart) S.I.R.Videira & P.W.Crous] poses a significant threat to rapeseed production globally. The herbicides atrazine and glyphosate are widely applied to herbicide-tolerant major crops, including rapeseed. Herbicides can affect disease levels directly and indirectly by stressing host plants, influencing pathogens, and altering abiotic and biotic stress levels in the environment. The specific effects of herbicides on the dimorphic pathogen N. capsellae regarding hyphal growth, conidial germination rate, and the morphological transformation from multi-celled hyphae or conidia into numerous single-celled blastospores remain unknown. Hence, studies were performed on two agar media [malt extract agar (MEA) and water agar (WA)] to determine how atrazine and glyphosate, each applied at 1 g a.i. L⁻¹ or the commercial recommended concentrations of 10 and 7.8 g a.i. L⁻¹, respectively, affect these characteristics in four highly pathogenic isolates of N. capsellae. Across a 32-day assessment period, the hyphal growth of all four isolates subcultured individually on MEA or WA was significantly restricted by both concentrations of atrazine and glyphosate. For both atrazine and glyphosate, restriction of hyphal growth was much greater at the higher commercial recommended concentration. Glyphosate restricted hyphal growth more than atrazine for each comparative concentration. Using a mixture of all four isolates, a similar trend of suppression by atrazine or glyphosate occurred in relation to conidial germination and the morphological transformation from multi-celled hyphae or conidia into numerous single-celled blastospores. These new insights into how herbicides constrain hyphal growth, conidial germination, and morphological transformation suggest their potential as a control measure in herbicide-tolerant crops to limit the epidemic spread and development of not only N. capsellae in rapeseed but other dimorphic fungal pathogens as well. Full article

Red Fruit Ginseng (Campanumoea lancifolia), widely cultivated in Yunnan, Guizhou, Sichuan, and Guangxi, in China, is valued for its sweet-tasting fruit and medicinal potential. In this study, leaves of the Yunnan Maguan variety were used as explants to establish an efficient tissue culture system for callus induction, adventitious bud regeneration, and rooting. Different combinations of cytokinins and auxins were tested to determine the optimal balance of plant growth regulators (PGRs). Our results indicated that when the combined concentration of kinetin (KT) and 6-benzylaminopurine (6-BA) in the primary culture medium was lower than 1.0 mg/L, callus induction was the most effective, with treatments of 0.5 mg/L KT + 0.2 mg/L 6-BA and 0.2 mg/L KT + 0.8 mg/L 6-BA showing higher induction rates compared to other groups. During the subculture, 0.5 mg/L 6-BA promoted vigorous adventitious bud growth, while higher levels inhibited development. For rooting, 0.5 mg/L indole-3-acetic acid (IAA) combined with 0.2 mg/L 1-naphthaleneacetic acid (NAA) induced the highest root number and healthiest plantlets, outperforming indole-3-butyric acid (IBA). The optimized protocol resulted in over 80% explant survival and produced vigorous seedlings suitable for acclimatization. This leaf-derived regeneration method provides a reliable approach for large-scale propagation, conservation, and potential pharmacological applications of Red Fruit Ginseng, contributing to the sustainable utilization and commercial development of this valuable medicinal plant. Full article

The aim of this study was to find a way to remove persistent bacteria inhabiting in vitro shoot cultures of raspberry. Often, decontamination treatments fail to reach bacteria residing in internal tissues, leading to contaminated cultures later. Three raspberry cultivars, each harboring a unique bacterial contaminant, were used in this study. Experiments were conducted to assess the potential for eliminating these bacteria using biocide infiltration at 30 mbar. The following biocides were used: mercuric chloride (HgCl₂ at 0.05 and 0.1%), Plant Preservative Mixture (PPM^TM 0.2–4%), rifampicin (50–200 mg L⁻¹), and sodium hypochlorite (NaOCl 0.1–60%). Only 0.05 or 0.1% HgCl₂ applied via infiltration successfully eliminated all of the bacteria from the shoots, which remained bacteria-free for several years, as confirmed by indexing explants on bacterial media at each subculture. While most treated shoots became necrotic and died due to infiltration, the surviving shoots remained vital and provided bacteria-free material for long-term propagation. Results from experiments comparing micropropagation potential in bacteria-contaminated and bacteria-free cultures showed that bacteria-free shoots produced longer shoots, and the total number of shoots did not differ, except for ‘Norna’/Curtobacbacteria-free cultures, which were more productive. Bacteria-contaminated shoots rooted at higher percentages, but roots were much shorter, and plantlets initiated growth during acclimatization later. Cultures that were contaminated did not survive storage at 4 °C in the dark for 4–6 months. Full article

The friability of callus is closely associated with its genetic transformation efficiency, and optimizing induction and transformation conditions is essential for establishing an efficient transformation system. In this study, we developed a high-efficiency friable callus induction and Agrobacterium-mediated transformation system for Chrysanthemum ‘Jimba’. Three plant growth regulator (PGR) combinations—6-Benzylaminopurine (6-BA) + Naphthaleneacetic Acid (NAA), 6-BA + 2,4-Dichlorophenoxyacetic Acid (2,4-D), and Thidiazuron (TDZ) + 2,4-D—were evaluated for their effects on callus morphology, proliferation, and transformation efficiency. The optimal PGR combination was identified as 1.0 mg/L 6-BA + 0.4 mg/L NAA, which produced highly friable calli with a loose structure, rapid proliferation, and the highest nuclear-to-cytoplasmic ratio. The optimal subculture time for maintaining friability and high proliferation was the 7th week, while the best Agrobacterium infection conditions were OD600 = 0.5 with a 10 min infection period, which achieved a transformation efficiency of 91%. This optimized protocol provides an efficient and rapid transformation method for future gene function studies using callus transformation. Full article