Search Results (2,446)

This study aimed to develop biofumigation strategies against chestnut fruit rot caused by Botryosphaeria dothidea. An endophytic strain, FPYF2509, was isolated from Castanea mollissima fruit and identified as Trichoderma nordicum using morphological and phylogenetic (tef1, rpb2) analyses. Antifungal volatile organic compounds (VOCs) were analyzed using headspace solid-phase microextraction and gas chromatography–mass spectrometry during dual-culture interactions with pathogens. The volatiles from the interaction exhibited to inhibit pathogen growth. Particularly an induced myrtenol, demonstrated strongly biofumigation activity in vitro, with a lowest observed effect concentration of 0.02 µL/mL, minimum inhibitory concentration and a minimum fungicidal concentration of 0.2 µL/mL against B. dothidea. In vivo, fumigation with 0.2 µL/mL myrtenol significantly reduced disease incidence from 83.3% to 17.39%, achieving a 79.1% control efficacy. This work presents endophytic T. nordicum FPYF2509 as a promising biocontrol agent and identifies myrtenol, of fungal interaction origin, as a novel and effective mycofumigant for postharvest disease management. Full article

Cryopreservation of preantral follicles (PAFs) is a promising tool for gene conservation and fertility preservation. However, standardized protocols for the cryopreservation and in vitro culture of isolated follicles—particularly in pigs—are still lacking. This study aimed to analyze the survival and developmental potential of porcine PAFs vitrified using two different methods: open pulled straw (OPS) and cryotube (CT). Ovaries of Hungarian Large White sows were collected from a local slaughterhouse and enzymatically digested to isolate preantral follicles. Morphologically normal follicles were assigned to three groups: fresh control, OPS-vitrified, and CT-vitrified. All follicles were cultured for 10 days in FSH-supplemented medium, with growth, survival, and estradiol (E2) production monitored. Survival rate was lower in the CT group (83.3%) than that of the control and OPS (97.4% and 94.4%, respectively). The follicular area was consistently larger in control than in CT and OPS, with no difference between vitrified groups. E2 production varied among treatments: OPS follicles showed lower E2 levels on Day 2, no differences were detected on Day 7, and CT follicles produced less E2 on Day 10. These results indicate that OPS is the more suitable vitrification method for porcine PAFs and that the culture system supports hormone production; however, it may require refinement to provide long-term follicle maintenance. Full article

Plant growth-promoting (PGP) bacteria are beneficial microbes that can help plants mitigate various biotic and abiotic stresses through different PGP functions. Flavins (FLs) are involved in flavoprotein-mediated reactions essential for plant metabolism and could act as PGP molecules. The aim of this study was to isolate and characterize potential FLs secreting bacteria from apple (Malus domestica [Suckow] Borkh) roots based on their fluorescence and to evaluate their PGP properties, including FLs secretion. A total of 26 bacteria with increased fluorescence in liquid culture were isolated from the apple roots. Based on 16S rRNA sequencing analysis, 11 genetically different strains mostly from Burkholderia and Rhizobia spp. were identified. All isolates secreted considerable amounts of riboflavin. In vitro plant assays showed that under nitrogen (N) limitation, inoculated alfalfa (Medicago sativa) plants yielded at least 25% more dry mass than non-inoculated plants, and inoculation with AK7 and FL112 enriched plant tissue N content compared to non-inoculated plants. This improved N acquisition was not linked to symbiotic N fixation. Additionally, the isolates exhibited some other PGP properties. However, no specific PGP functions were linked to improved plant N acquisition but could potentially be linked to the FLs secretion. For future investigation, the mechanisms underlying improved plant N uptake should be assessed to gain a more in-depth understanding. Full article

The increasing biomedical and conservation interest in porcine species has driven the development of advanced in vitro follicle culture systems designed to preserve genetic diversity and accurately model key stages of folliculogenesis. This study assessed a three-dimensional (3D) alginate-based system for the in vitro culture of porcine preantral follicles, aiming to overcome the structural limitations of conventional two-dimensional (2D) methods. A total of six experimental groups were established, consisting of group-cultured (four follicles/well) or individually cultured (one follicle/well) follicles maintained either without alginate (0%) or encapsulated in 0.5% or 1% alginate for 14 days in media supplemented with FSH, EGF, and IGF-I, with LH added from day 9. Follicular development was assessed by morphometric evaluation, image-based and histological analyses, and quantification of steroid hormones in media collected every 48 h. Group-cultured follicles encapsulated in 0.5% alginate most effectively maintained their 3D architecture, reached the largest diameters, and progressed more uniformly compared with other groups. In contrast, follicles cultured without alginate rapidly lost structural integrity, showed granulosa cell migration, and decreased in size, whereas those encapsulated in 1% alginate exhibited restricted growth. Estradiol and testosterone concentrations increased over time in the 0.5% alginate group, were lowest without alginate, and intermediate in 1% alginate. Individually cultured follicles exhibited reduced growth and lower total hormone production compared with group-cultured follicles; however, when normalized per-follicle, steroid secretion, particularly in the 0.5% alginate group, was enhanced, indicating increased steroidogenic efficiency on a per-follicle basis. These findings indicate that 0.5% alginate provides an optimal balance between structural support and physiological steroidogenesis during preantral follicle culture. This 3D system improves the biological relevance of porcine follicle culture and may support future applications in reproductive biology, conservation, and genetic resource preservation. Full article

Arbuscular mycorrhizal fungi (AMF) play a pivotal role in plant adaptation to arid ecosystems, yet their widespread agricultural use is constrained by the scarcity of high-quality, locally adapted inoculum. This study established a reliable monoxenic culture system for mass-producing an indigenous AMF isolate from the date palm (Phoenix dactylifera L.) rhizosphere in the Figuig oasis, southeastern Morocco. The isolate was identified as Rhizophagus irregularis based on spore morphology and Large Subunit ribosomal DNA (LSU rDNA) phylogeny. Two propagule types, surface-sterilized spores and mycorrhizal root fragments of Plantago lanceolata L., were compared for initiation of in vitro cultures on Ri T-DNA-transformed carrot (Daucus carota L.) hairy roots. By week 16, cultures initiated from mycorrhizal root fragments produced 1414 ± 65 spores per plate and showed significantly higher performance than spore-derived cultures in terms of propagule viability, root colonization, and hairy root growth. Propagule viability reached 84% and 68%, root colonization frequencies were 95% and 72%, and hairy root lengths averaged 81 and 63 cm in root fragment- and spore-derived cultures, respectively (p < 0.01). In a subsequent whole-plant assay using P. lanceolata, in vitro-produced spores induced markedly higher mycorrhizal colonization frequency (91.0 ± 1.6% compared with 74.8 ± 1.9%) and intensity (70.0 ± 1.6% compared with 55.0 ± 1.6%) than spores obtained from conventional trap cultures (p < 0.001). These results demonstrate that monoxenic root-organ culture using root fragments is a robust, reproducible method for generating abundant, contaminant-free, and functionally superior inoculum of native R. irregularis. This advance provides a solid platform for developing tailored bio-inoculants to enhance crop resilience and sustainability in arid and semi-arid agroecosystems. Full article

Adventitious shoot regeneration in woody species is regulated by interactions between plant growth regulators, endogenous hormone metabolism, and environmental cues such as light quality. Here, we investigated the effects of thidiazuron (TDZ) and the cytokinin oxidase/dehydrogenase (CKX) inhibitors INCYDE and phenyladenine (PA), in combination with different light spectra, on morphogenesis in Melia volkensii leaf explants. TDZ induced the highest frequencies of callus formation and adventitious shoot regeneration, particularly under white light. INCYDE promoted localized regeneration responses, including activation of dormant meristematic regions in secondary leaf axils, whereas PA showed limited regeneration efficiency. Light quality significantly influenced morphogenesis, with white and blue light favoring organized shoot development, while red and far-red light suppressed shoot regeneration and promoted callus formation. Cytokinin profiling revealed treatment-dependent shifts in endogenous cytokinin composition, most notably in isopentenyladenine (iP)-type cytokinins, which is consistent with altered cytokinin degradation dynamics. Cis-zeatin-type cytokinins were abundant across treatments, likely reflecting regulation associated with in vitro culture conditions. These findings indicate that cytokinin metabolism and light quality jointly influence organogenic competence in Melia volkensii Gürke, providing a physiological basis for optimizing regeneration strategies in woody plants. This study provides the first integrated analysis of cytokinin-modulating compounds and light spectra on adventitious shoot regeneration in Melia volkensii. The findings establish a physiological basis for improving regeneration protocols in recalcitrant woody species and support future biotechnological applications, including genetic improvement and advanced propagation strategies. Full article

Organoids are self-organizing multicellular structures generated in vitro that recapitulate the micro-architecture and function of an organ. They are commonly derived from stem cells but can also emerge from pieces of proliferative tissues. Organoid technology has opened novel ways to model development and disease, but it is not without challenges. Computational models of organoids have been established to elucidate organoid growth and facilitate the optimization of organoid cultures. This article is a systematic review of in silico organoid models constructed at single-cell or subcellular resolution. PubMed, Scopus, and Web of Science were searched for original papers published in peer-reviewed journals before 26 September 2025, yielding 439 records after deduplication. Two independent reviewers screened their titles and abstracts, retrieved 84 papers for full-text scrutiny, and identified 32 papers that met the inclusion criteria. They were grouped by organoid type: 12 intestinal, 1 airway, 2 pancreas, 3 neural, 1 kidney, 1 inner cell mass, 9 tumor, and 3 generic. The analysis of these works revealed that computer simulations guided experimental work. Parsimonious computational models provided insights into diverse organoid behaviors, such as the rotation of airway organoids, size oscillations of pancreatic organoids, epithelial patterning of neural tube organoids, or nephron segment formation in kidney organoids. Generally, a deep understanding was achieved through combined in silico and in vitro investigations (e.g., optic cup morphogenesis). Recent research trends suggest that next-generation computational models of organoids may emerge from a more detailed understanding of the complex regulatory circuits that govern stem cell fate, and machine-learning-based, high-throughput imaging of organoids. Full article

This research aimed to assess the effect of chitosan nanoparticles (ChNPs) during in vitro shoot proliferation of vanilla using temporary immersion bioreactors (TIB). TIB culture is a biotechnological process that uses semiautomated containers for the production of explants exposed in liquid culture medium. Concentrations of control, 25, 50, 100, 200, and 400 mg/L ChNPs were evaluated in Murashige and Skoog culture medium. Morphological characterization of ChNPs was performed using scanning electron microscopy. At 60 days of culture, survival (%), development variables, photosynthetic pigment content, lipid peroxidation expressed in malondialdehyde, total phenolic content (TPC), hydrogen peroxide (H₂O₂) content, and total antioxidant capacity (TAC) expressed in trolox equivalents were evaluated. The data were analyzed with analysis of variance, with a Tukey test (p ≤ 0.05) using SPSS statistics software, version 29. The results revealed that the greatest survival (%) was obtained at concentrations of control, 25, and 50 mg/L ChNPs, while the lowest survival (%) was observed at concentrations of 400 mg/L ChNPs. Growth stimulation was found, as well as an increase in chlorophyll and β-carotene at concentrations of 25 and 50 mg/L ChNPs. The level of H₂O₂ increased at 25 and 50 mg/L ChNPs. Lipid peroxidation showed no differences among treatments. TPC increased at 100 and 200 mg/L ChNPs, while TAC increased at 200 and 400 mg/L ChNPs. In conclusion, the administration of ChNPs at low concentrations can stimulate growth, while at high concentrations they can inhibit it, a response known as hormesis or hormetic effect. Full article

Inflammatory bowel disease (IBD) arises from chronic dysregulation at the epithelial–stromal interface, creating a need for in vitro systems that better capture these interactions. In this study, we developed a 3D co-culture platform in which HT-29 intestinal epithelial cells and IMR-90 fibroblasts are embedded within an alginate–gelatin hydrogel, alongside a complementary interface model using a plasma-treated electrospun mesh to spatially compartmentalize stromal and epithelial layers. We first assessed metabolic activity, viability, and proliferation across several epithelial-to-fibroblast ratios and identified 1:0.5 as the most supportive of epithelial expansion. The A1G7 hydrogel maintained high viability (>92%) and sustained growth in all mono- and co-cultures. To evaluate inflammatory competence, models were stimulated with lipopolysaccharide (LPS), administered either within the hydrogel or through the culture medium. LPS exposure increased TNF-α and IL-1β secretion in both configurations, with the magnitude of the response depending on the delivery route. Treatment with dexamethasone consistently reduced cytokine levels, confirming the model’s suitability for pharmacological testing. Together, these results demonstrate that the alginate–gelatin system provides a reproducible epithelial–stromal platform with quantifiable inflammatory readouts, offering a practical foundation for mechanistic studies and early-stage screening of anti-inflammatory therapeutics in IBD. Full article

Background: Non-absorbable polyvinyl alcohol sponges (Merocel) are widely used in otolaryngology for nasal and ear packing but are prone to bacterial colonization and biofilm formation, which may increase infection risk and drive frequent use of systemic antibiotics. Sustained-release drug delivery systems enable prolonged local antiseptic activity at the site of packing while minimizing systemic exposure. Methods: We developed a sustained-release varnish containing chlorhexidine (SRV-CHX) and coated sterile Merocel sponges. Antibacterial, in vitro, activity against Staphylococcus aureus and Pseudomonas aeruginosa was evaluated using kinetic diffusion assays on agar, optical density (OD₆₀₀) measurements of planktonic cultures, drop plate, ATP-based viability assays, biofilm analysis by MTT metabolic assay, crystal violet bio-mass staining, high-resolution scanning electron microscopy (HR-SEM), and spinning disk confocal microscopy. Results: SRV-CHX-coated sponges produced sustained zones of inhibition on agar plates for up to 37 days against S. aureus and 39 days against P. aeruginosa, far exceeding the usual 3–5 days of clinical sponge use. Planktonic growth was significantly reduced compared with SRV-placebo, and a bactericidal effect persisted for up to 16 days for S. aureus and 5 days for P. aeruginosa before becoming predominantly bacteriostatic. Biofilm formation was markedly inhibited, with suppression of metabolic activity and biomass for at least 33 days for S. aureus and up to 16 days for P. aeruginosa. HR-SEM and confocal imaging confirmed sparse, discontinuous biofilms and predominance of non-viable bacteria on SRV-CHX-coated sponges compared with dense, viable biofilms on the placebo controls. Conclusions: Coating Merocel sponges with SRV-CHX provides prolonged antibacterial and anti-biofilm activity against clinically relevant pathogens. This strategy may reduce dependence on systemic antibiotics and improve infection control in nasal and ear packing applications in otolaryngology. Full article

Salinity stress severely limits crop productivity worldwide. While plant growth-promoting rhizobacteria (PGPR) are known to alleviate abiotic stress, the specific mechanisms mediated by their volatile organic compounds (VOCs) remain largely elusive. In this study, an in vitro split-plate system was used to investigate the effects of VOCs emitted by Bacillus velezensis SQR9 on Arabidopsis thaliana seedlings under salt stress. Exposure to SQR9 VOCs significantly enhanced Arabidopsis salt tolerance, evidenced by increased biomass and root growth. Mechanistically, SQR9 VOCs mitigated salt-induced damage by increasing chlorophyll content, modulating osmolytes, and reducing malondialdehyde (MDA) levels. SQR9 VOCs alleviated oxidative stress by decreasing ROS (H₂O₂, O₂⁻) accumulation and enhancing antioxidant enzyme (SOD, CAT, POD) activities. Furthermore, SQR9 VOCs maintained ion homeostasis by significantly reducing leaf Na⁺ accumulation, maintaining a high K⁺/Na⁺ ratio, and upregulating key ion transporter genes. Analysis of the headspace from SQR9 cultured on MSgg medium identified 2,3-butanediol (2,3-BD) as a major active VOC. Exogenous application of 2,3-BD successfully mimicked the growth-promoting and salt-tolerance-enhancing effects of SQR9. Our findings demonstrate that SQR9 VOCs, particularly 2,3-BD, systemically prime Arabidopsis for salt tolerance by co-activating the antioxidant defense system and the SOS ion homeostasis pathway. Full article

Phosphorus (P) is an essential nutrient in plant development, but its availability in the soil is often limited due to chemical fixation and poor solubility. This study presents the development, characterization and evaluation of a novel granular bioinoculant formulated with Aspergillus tubingensis (P-solubilizing) and Trichoderma virens (P-mineralizing) using clinoptilolite (CZ) as a carrier to improve P bioavailability. The formulation process included the evaluation of the proposed components, the standardization of conidia production in different media cultures and conditions, the elaboration and characterization of the bioinoculant and its evaluation in plants. In this study, in vitro analysis demonstrated the synergistic effect of the components, showing that in all treatments with dual inoculation and CZ, the amount of soluble phosphorus (SP) was higher than in their counterparts (from 27.8 to 36.8 mg·L⁻¹). A concentration greater than 1 × 10⁹ CFU·mL⁻¹ was obtained by standardizing the production of conidia in different media (PDA, V8-Agar and Molasses Agar), which were then used to produce granular batches containing at least 2 × 10⁷ CFU·g⁻¹. Furthermore, the size (88% of the granules measured <4.5 mm), purity (<2 CFU·g⁻¹ in 10⁻⁴ dilution), and moisture content of the prototype granules (3.3–3.8%) were confirmed to be within established international quality parameters. Plant evaluations in chili and tomato demonstrated the formulation efficacy, showing an increase in both soluble and foliar P content (with at least 30% more than controls), alongside improvements in all parameters evaluated that are related to plant growth promotion (with at least 15% more growth than controls). The development of this formulation prototype represents a focused effort toward process standardization and optimization required to validate developed formulations, thus promoting the advancement of applied biotechnology. Full article

Metal and metal oxide nanoparticles (NPs) synthesized through biologically mediated reduction of metal ions using biomolecules derived from microorganisms, algae, or plants are attracting growing attention in plant biotechnology due to their multifunctional properties and environmental advantages compared with conventional physicochemical synthesis. This review provides a comprehensive analysis of biological approaches for NP production using bacteria, fungi, algae, cyanobacteria, whole plants, and in vitro plant cell cultures. The main biosynthetic mechanisms, types of reducing and capping metabolites, metal specificity, and typical NP characteristics are described for each system, with emphasis on their relative productivity, scalability, reproducibility, and biosafety. Special consideration is given to plant cell and tissue cultures as highly promising platforms that combine the metabolite diversity of whole plants with precise control over growth conditions and NP parameters. Recent advances highlight the significance of bioengineering of reductive capacity as a novel strategy to enhance the efficiency and controllability of NP biosynthesis. Since NP formation is driven by key biomolecules, targeted modification of biosynthetic pathways through metabolic and genetic engineering can substantially increase NP yield and allow fine-tuning of their structural and functional properties. The applications of biogenic NPs in plant biotechnology are systematically evaluated, including their use as environmentally safe disinfectants for explants and seed sterilization, modulators of callus induction and morphogenesis, and abiotic elicitors that enhance the accumulation of economically valuable secondary metabolites. Remaining challenges, such as variability in NP characteristics, limited scalability, and insufficient data on phytotoxicity and environmental safety, are discussed to outline future research priorities. The synthesis–function relationships highlighted here provide a foundation for developing sustainable NP-based technologies in modern agriculture. Full article

Net form net blotch disease, caused by Pyrenophora teres f. teres (Ptt), is one of the most destructive barley diseases, resulting in severe yield and grain quality losses worldwide. The increasing prevalence of fungicide-resistant Ptt strains, driven by the pathogen’s high genetic variability, highlights the urgent need for sustainable and eco-friendly disease management strategies. The present study provides novel insights into the use of native seed-borne endophytic bacteria naturally associated with barley as biological control agents against Ptt. Two endophytic bacterial strains isolated from healthy barley seeds were identified based on 16S rRNA gene sequencing as Bacillus subtilis PX491551 and Stenotrophomonas rhizophila PX494419. Their biocontrol potential against Ptt was evaluated through in vitro, greenhouse, and field experiments. In the dual-culture assay, B. subtilis and S. rhizophila inhibited the mycelial growth of Pyrenophora teres f. teres by 64.34% and 50.14%, respectively. Under greenhouse conditions, B. subtilis and S. rhizophila significantly reduced disease severity at the seedling stage, with scores of 2.00 and 4.00, respectively, compared to 9.33 in the untreated control. Beyond disease suppression, both endophytic bacteria markedly enhanced the host’s defense system. S. rhizophila induced the highest accumulation of total soluble phenolics, while B. subtilis significantly increased flavonoid content and boosted higher activities of superoxide dismutase and phenylalanine ammonia-lyase. In contrast, S. rhizophila showed the strongest induction of ascorbate peroxidase activity. Notably, field application of both bacteria consistently reduced net blotch severity over two consecutive growing seasons (2023–2024 and 2024–2025) and considerably improved chlorophyll content, 1000-grain weight, and grain yield. Overall, this study demonstrates that native seed-derived endophytic bacteria not only suppress barley net blotch but also enhance host antioxidant and defense responses, highlighting their potential as effective and sustainable biological control agents for barley disease management. Full article

White mold (Sclerotinia sclerotiorum) reduces potato yield and quality in Sinaloa, Mexico. This study first evaluated the in vitro efficacy of Trichoderma azevedoi, T. afroharzianum, T. asperellum and T. asperelloides in inhibiting S. sclerotiorum mycelial growth and sclerotia production. Field experiments then assessed a combination of these antagonists, their alternating application with synthetic fungicides, and a fungicide-alone treatment for disease control, sclerotia reduction and yield increase. In vitro, all four Trichoderma species significantly inhibited the pathogen, achieving 60.1–63.1% mycelial suppression in dual culture and 90.3–94.1% via volatile metabolites, with the latter also completely suppressing sclerotia formation. In the field, the Trichoderma combination significantly controlled white mold, reducing plant incidence and severity to 66.0 and 27.1% in 2021 and 55.6 and 18.8% in 2022, while lowering sclerotia production to 32.7 and 14.6 on ten plants, respectively. This control extended to tubers, where incidence and severity were reduced to 1.6% and 0.4% in 2021, and 1.3% and 0.3% in 2022. The alternating application of Trichoderma with synthetic fungicides proved statistically equivalent to the Trichoderma-alone treatment in disease control, while the fungicides-alone treatment was significantly less effective. Potato yield was highest in plots treated with the Trichoderma combination (46.0 and 52.9 t ha⁻¹ in 2021 and 2022, respectively). These results highlight the potential of using a mixture of these four Trichoderma species as a cornerstone of sustainable disease management in Sinaloa, offering effective control of potato white mold while significantly reducing dependence on synthetic fungicides. Full article