Search Results (1,620)

Solid-state fermentation (SSF) is a long-established biotechnological approach gaining renewed interest for its ability to enhance nutrient availability and improve the functional properties of agro-industrial by-products. This strategy is particularly relevant for early post-weaning piglets, which are highly susceptible to weaning stress due to an immature digestive system and a gut microbiota not yet adapted to solid feed. In this study, the fermentation parameters of flaxseed cake were optimized using a Plackett–Burman experimental design. Protease activity was selected as the response variable due to its relevance for improving protein degradation and potential digestibility in fermented feed ingredients. Accordingly, based on the statistical analysis, the conditions selected for the in vivo trial were 1% molasses, 0.5% yeast extract, 0.05% CaCl₂, 0.5% NaCl, 7.5% inoculum (4.12 × 10⁹ CFU/mL), 60% moisture, and 72 h fermentation. Fermentation time was identified as the main factor positively influencing protease production, while higher CaCl₂ concentrations and inoculum levels negatively affected enzyme activity. Optimization increased protease activity, microbial viability and free amino acid content. In addition, SSF reorganizes the carbohydrate profile by reducing structural fiber fractions, with neutral detergent fiber and acid detergent fiber decreasing by 27% and 29%, respectively, while simultaneously increasing soluble carbohydrates by 14.67%. Phytic acid content being also reduced by 23.81%. A pilot nutritional trial on post-weaned piglets (35 days old) showed that including 8% fermented flaxseed cakes (FFSC group) improved body weight, average daily gain, feed conversion ratio, and diarrhea score, without affecting average daily feed intake, compared with 8% unfermented flaxseed cakes (FSC group). These performance improvements were accompanied by changes in fermentation metabolites and gut microbial composition. Lower isovalerate concentrations suggested reduced proteolysis, while higher propionate levels may contribute to increased blood glucose availability in the FFSC group. These changes coincided with a shift in microbial composition, characterized by a reduced abundance of methanogenic archaea and increased abundances of taxa such as Lactobacillus, Enterococcus, and members of the Lachnospiraceae and Eubacteriaceae families. Full article

Yeast models are widely used to study molecular chaperones from diverse organisms, including plants, because of their well-characterized genetics and the conservation of the protein-folding machinery among eukaryotes. Cross-species complementation studies in yeast have yielded valuable insights into conserved biochemical activity and molecular functions that manage protein folding, assembly, and repair during stress. This study evaluated the functional capacity of three potato StBiP isoforms (StBiP1, StBiP2, and StBiP3) to complement the kar2 deletion (kar2Δ) strain under a range of environmental and ER stress conditions. All three StBiPs partially restored colony growth under normal conditions, demonstrating that they are functional orthologs of yeast KAR2 and can support core ER housekeeping functions. Under severe stress, however, the isoforms diverged: StBiP3 most effectively complemented the kar2Δ strain during heat- and chemically induced ER stress, whereas StBiP1 and StBiP2 provided weaker protection. Unfolded protein response (UPR) activation, monitored via HAC1 mRNA splicing, further highlighted isoform-specific differences in how the StBiPs support IRE1-HAC1 signaling under ER stress and oxidative stress. A conserved cysteine in the nucleotide-binding domain, previously implicated in Kar2 redox control, was also critical for StBiP3-mediated protection in yeast, although the same mutation led to different consequences in plant tissues. Together, these findings provide evidence of subfunctionalization among potato BiP isoforms, with StBiP3 emerging as a stress-specialized chaperone that is a promising target for improving ER stress resilience in solanaceous crops. Full article

RipP1 is a well-characterized avirulence effector that induces a hypersensitive response (HR) in three tobacco species. However, the molecular mechanisms by which host proteins recognize RipP1 to activate a defense response and modulate host–pathogen interactions remain largely unknown. In this study, we screened a Nicotiana benthamiana cDNA library via yeast two-hybrid assay and identified FRIGIDA-like protein 4a (FRL4a) as a host protein interacting with RipP1. Secondary structure analysis of FRL4a and construction of serial mutants revealed that the ClyA-like domain of FRL4a is the key region mediating its interaction with RipP1. Using virus-induced gene silencing (VIGS) and quantitative real-time PCR (qPCR) analysis, we found that the ability of RipP1 to induce HR was significantly attenuated in FRL4a-silenced plants, and RipP1 no longer suppressed the ethylene signaling pathway. Pathogenicity tests by inoculating R. solanacearum on N. benthamiana with different FRL4a expression levels showed enhanced bacterial wilt resistance in FRL4a-silenced plants but increased susceptibility in FRL4a-overexpressing plants. Collectively, these findings demonstrate that RipP1 suppresses the ethylene pathway through its interaction with FRL4a, and FRL4a acts as a negative regulator of tobacco resistance to bacterial wilt. Full article

Despite the importance of surface microbiota in postharvest quality, the effects of mechanical damage on microbial succession in Hypsizygus marmoreus during refrigerated storage remain insufficiently understood. In this study, 16S rRNA gene and ITS amplicon sequencing were used to characterize the bacterial and fungal communities on intact and mechanically damaged H. marmoreus during 15 days of storage at 4 °C. Storage time, rather than mechanical damage, was the main driver of whole-community variation, although mechanical damage accelerated visible spoilage assessed qualitatively. Bacterial communities showed pronounced temporal turnover, shifting from early Firmicutes-rich assemblages to late-stage Proteobacteria-dominated communities, especially Pseudomonas. In contrast, fungal communities remained largely dominated by Ascomycota throughout storage, although mechanically damaged mushrooms showed a greater late-stage occurrence of opportunistic yeasts such as Candida. Predicted functional and phenotypic analyses further suggested late-stage increases in Gram-negative, aerobic, biofilm-forming, stress-tolerant, and potentially pathogenic bacterial traits. Because these traits were inferred from 16S rRNA gene-based prediction rather than measured directly, they should be interpreted cautiously. Overall, the results suggest that maintaining the physical integrity of H. marmoreus during postharvest handling may help preserve quality and delay the emergence of spoilage-associated microbial traits during refrigerated storage. Full article

Soil salinization has become a major global constraint threatening ecosystem stability and agricultural production. As a prominent salt-tolerant turfgrass, Paspalum vaginatum (seashore paspalum) serves as an excellent material for exploring salt tolerance mechanisms. In this study, PvHAK12, a high-affinity K⁺ transporter (HAK) family gene isolated from seashore paspalum, was functionally characterized. PvHAK12 encodes a 788 amino acid protein with 13 transmembrane domains, belonging to the plasma membrane-localized ion transporters. It exhibits high sequence conservation with other HAK transporters and is predominantly expressed in roots and stems, with distinct tissue- and time-specific induction under salt stress. Yeast complementation assays revealed that PvHAK12 has no obvious K⁺ transport capacity but may mediate Na⁺ transport. Overexpression of PvHAK12 in Arabidopsis thaliana significantly reduced salt tolerance at germination, seedling and rosette stages, as reflected by lower germination rate, fresh weight, survival rate, the maximum quantum yield of photosystem II (Fv/Fm) value and chlorophyll content, accompanied by higher ion leakage. Under salt stress, transgenic plants accumulated more Na⁺ and less K⁺, leading to an elevated Na⁺/K⁺ ratio. Moreover, transgenic lines displayed weaker antioxidant enzyme activities and higher reactive oxygen species (ROS) accumulation. Transcript analysis further demonstrated that PvHAK12 overexpression suppressed the induction of multiple ion-transport and stress-responsive genes under salt conditions. These results indicate that PvHAK12 negatively regulates plant salt tolerance by disrupting ion homeostasis, antioxidant capacity and stress-related gene expression. Full article

Alchemilla alpina L. (Rosaceae), belongs to a genus well recognized in traditional medicine for treating gynecological disorders and hormonal imbalance; however, the specific bioactivity of A. alpina itself remains poorly characterized. This study aimed to elucidate the phenolic composition and the biological potential of the methanolic (MeOH) extract of A. alpina. LC–MS/MS analysis identified 39 phenolic compounds, with rutin, catechin, kaempferol-3-O-glucoside, and caffeic acid being the dominant constituents. The extract exhibited high total phenolic and flavonoid contents, consistent with strong antioxidant capacities. It demonstrated notable α-glucosidase and acetylcholinesterase inhibitory activities, indicating its potential relevance for metabolic and neurodegenerative disorders. The extract effectively reduced AAPH-induced ROS levels in MRC-5 fibroblasts, indicating cytoprotective and antioxidative effects. The cytotoxicity toward cervical cancer cells HeLa and ovarian cancer cells A2780 was moderate and concentration dependent. A yeast-based fluorescent screen revealed a strong and selective binding affinity toward estrogen receptor α (ERα) and selective inhibition of human recombinant AKR1C3 (59.5%), without affecting AKR1C4. Additionally, high COX-1/COX-2 inhibition (>70%) supported its anti-inflammatory potential. Collectively, these findings provide the first integrated evidence of A. alpina’s phenolic richness and multifunctional bioactivity, scientifically supporting its potential in managing hormone-dependent and oxidative stress-related disorders. Full article

Sigma factors (SIGs) are nuclear-encoded regulators of chloroplast gene transcription. We conducted a genome-wide analysis in Brassica napus, identifying 23 SIG genes that were phylogenetically classified into six distinct subfamilies. Characterization of gene structure, conserved motifs, and chromosomal locations indicated family expansion primarily through segmental duplication under purifying selection. Promoter analysis identified cold-responsive elements enriched in BnSIG5A. Expression profiling showed that BnSIG5 subfamily members, particularly BnSIG5A, are strongly induced by cold stress. Analysis of Arabidopsis SIG5 mutants confirmed previously reported roles of AtSIG5 in cold tolerance. Heterologous expression in yeast, and the strong cold induction of BnSIG5A together with its chloroplast localization, suggest that BnSIG5A may play a conserved role, providing a foundation for future functional studies in B. napus. This work establishes a genomic framework for the SIG family in rapeseed and identifies BnSIG5A as a high-priority candidate for further investigation. Subcellular localization confirmed chloroplast targeting of BnSIG5A. Heterologous expression in yeast and analysis of Arabidopsis SIG5 mutants suggest conserved functions in cold tolerance, providing a foundation for future functional studies in B. napus. This work establishes a genomic framework for understanding SIG-mediated stress responses in rapeseed and identifies BnSIG5A as a promising candidate for further investigation. 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

Whole-cell biocatalysis presents a sustainable and efficient approach for the selective reduction in α,β-unsaturated bonds in flavonoid derivatives. This study investigates the capability of yeast strains from the Yarrowia clade to catalyze the chemoselective reduction of 4′-methoxychalcone (1a) to its dihydro derivative. All tested strains exhibited similarly high hydrogenation activity, indicating a broadly conserved enoate reductase function within the clade. Among them, Yarrowia lipolytica KCh 71, previously reported and well characterized in the literature, was selected for preparative-scale transformation of a diverse series of synthetic methoxychalcones bearing additional methoxy groups in positions C-2, C-3, C-4, C-5, and C-6 of ring B. All derivatives were effectively converted into the corresponding dihydrochalcones, with yields ranging from 62% to 92%. Among the tested derivatives, the 2′,4′,6′-trimethoxy chalcone (7a) did not undergo biotransformation under our conditions, whereas mono- and di-methoxy derivatives (2a–6a) were efficiently reduced. These results confirm the broad substrate tolerance, high efficiency, and potential scalability of Y. lipolytica KCh 71, supporting its potential as a whole-cell biocatalyst for the sustainable synthesis of bioactive dihydrochalcones. The consistently high hydrogenation activity observed across 21 tested strains suggests the involvement of evolutionarily conserved enoate reductases. Bioinformatic analysis supports that the Yarrowia clade possesses a robust complement of Old Yellow Enzymes (OYE), providing a reliable enzymatic basis for the observed chemoselective reductions. All Yarrowia tested strains showed the same general transformation type, although the extent and rate of conversion differed among strains, and Y. lipolytica KCh 71 was one of the most tolerant. The broad reduction in α,β-unsaturated chalcones is consistent with the action of flavoenzymatic ene-reductases, particularly Old Yellow Enzyme (OYE)–like reductases. Bioinformatic analysis of Yarrowia genomes reveals putative OYE homologs, supporting this mechanistic interpretation, although the specific enzymes were not identified in this study. Full article

Kombucha fermentation is driven by a Symbiotic Culture of Bacteria and Yeast (SCOBY), a cellulose-rich biofilm that hosts a complex microbial consortium. While most kombucha studies focus on the liquid beverage, the SCOBY pellicle itself remains underexplored, particularly with respect to species-level microbial resolution and its intrinsic biological activities. In this study, a commercial kombucha SCOBY was characterized using full-length 16S rRNA gene and ITS amplicon sequencing based on Oxford Nanopore Technology, enabling species-level taxonomic resolution. In parallel, hydroalcoholic and aqueous extracts of dried SCOBY biomass were evaluated for in vitro antioxidant activity (DPPH and ABTS assays), antidiabetic-related enzyme inhibition (α-glucosidase and dipeptidyl peptidase-4, DPP4), and anti-aging-related enzyme inhibition (tyrosinase and elastase). The SCOBY bacterial community was strongly dominated by acetic acid bacteria, with Komagataeibacter saccharivorans and Acetobacter tropicalis accounting for more than 60% of total reads, reflecting a biofilm structure optimized for cellulose production and oxidative metabolism. The yeast community showed marked unevenness, with Brettanomyces bruxellensis representing over 80% of reads, consistent with its known role in ethanol production and stress tolerance within kombucha systems. In vitro assays revealed that hydroalcoholic SCOBY extracts consistently exhibited higher biological activity than aqueous extracts across all tested assays. However, both extracts showed substantially lower potency than purified reference compounds, indicating moderate but measurable bioactivity typical of complex fermented matrices. These findings support the potential valorization of SCOBY as a fermentation-derived biomaterial and functional ingredient while underscoring the need for further chemical characterization, mechanistic studies, and biological validation beyond enzyme-based assays. Full article

Background: The Calcineurin B-like (CBL) and CBL-interacting protein kinase (CIPK) system constitute critical signaling modules mediating plant responses to abiotic stress. Although these families have been studied across various species, their evolutionary dynamics across grasses and the functional plasticity of specific isoforms remain elusive. Methods: A genome-wide analysis of CBL and CIPK families was conducted across five major Poaceae species (Oryza sativa, Triticum aestivum, Zea mays, Sorghum bicolor, and Saccharum spontaneum). Phylogenetic and synteny analyses were analyzed to family expansion and evolution. Cis-regulatory elements analysis in gene promoter regions were examined to predict potential stress-responsive features. Expression profiles of OsCBL and OsCIPK gene families were examined by qRT-PCR under conditions involving PEG-induced osmotic stress, pathogen strain P6 inoculation, and exogenous application of the phytohormones abscisic acid (ABA) and methyl jasmonate (MeJA). Protein–protein interactions between selected CBL (OsCBL4) and CIPK pairs were assessed via Yeast Two-Hybrid (Y2H) and Luciferase Complementation Imaging assays (LCI). Results: Phylogenetic and synteny analyses indicated that segmental duplications have contributed substantially to the expansion of these gene families. Promoter analysis revealed that the majority of CBL and CIPK family members, exemplified by OsCBL4, traditionally characterized as a salt sensor, possesses a cis-element architecture (rich in ABREs and MBS) heavily biased towards dehydration responsiveness. Expression profiling showed that OsCBL4 is significantly hyper-induced by direct osmotic stress (PEG) but exhibits almost no response to exogenous ABA. A subset of kinases genes (e.g., OsCIPK2, 9, 18) displayed PEG-induced expression patterns resembling those of OsCBL4, whereas OsCIPK30 remained transcriptionally unresponsive under the same conditions. Protein interaction assays demonstrated that OsCBL4 physically interacts exclusively with PEG-responsive transcriptionally activated kinases such as OsCIPK9, but failed to interact with the non-responsive OsCIPK30. Conclusions: Our study provides a genomic characterization of CBL and CIPK families across five major Poaceae species. The combined expression and interaction data reveal that OsCBL4-assembles with specific CIPKs into signaling modules during osmotic stress responses in rice, pointing to roles that go beyond salt stress responses. The findings establish a foundation for further functional dissection of CBL-CIPK pathway diversification in abiotic stress adaptation. Full article

Cadmium (Cd) contamination severely threatens crop productivity and food safety, particularly in maize (Zea mays L.), which exhibits relatively high capacities for metal uptake and translocation. Metal tolerance proteins (MTPs) play essential roles in metal homeostasis and detoxification; however, the functions of maize MTP under Cd stress remain poorly understood. In this study, a comprehensive expression analysis of the maize MTP gene family revealed that two Zn-CDF members, ZmMTP1-1 and ZmMTP1-2, displayed the strongest and most consistent transcriptional induction in response to Cd stress, especially in roots. Phylogenetic and structural analyses confirmed that both genes are closely related to MTP1 homologs from other plant species, while exhibiting distinct gene structures and regulatory features. Functional characterization in transgenic Arabidopsis thaliana demonstrated that overexpression of ZmMTP1-1 or ZmMTP1-2 significantly enhanced tolerance to Cd and Zn stress, as reflected by improved seed germination, root growth, survival, and biomass accumulation. Enhanced metal tolerance was associated with elevated antioxidant enzyme activities, reduced oxidative damage, and coordinated upregulation of endogenous metal transporter genes. Moreover, heterologous expression of ZmMTP1-1 in yeast further supported its conserved role in Cd tolerance. Collectively, these findings indicate that ZmMTP1-1 and ZmMTP1-2 contribute to Cd detoxification through coordinated metal transport and stress-response pathways, providing potential genetic resources for improving heavy metal tolerance in maize. Full article

The WRKY group II subfamily, a major conserved and plant-specific WRKY transcription factor family, plays a central role in regulating plant responses to abiotic stresses. However, systematic characterization of WRKY group II genes and their involvement in saline–alkali stress responses in Hordeum brevisubulatum (Trin.) Link remains largely unexplored. In this study, 23 WRKY group II genes were identified at the genome-wide level in H. brevisubulatum. Phylogenetic analysis classified these genes into five subgroups, with members within each subgroup exhibiting highly conserved motif compositions and gene structures. Promoter analysis revealed abundant cis-acting elements related to stress and defense responses, phytohormone signaling, growth and development, and light responsiveness, suggesting diverse regulatory potential. Transcriptome sequencing and qRT-PCR analysis showed that several HbWRKY genes were responsive to NaHCO₃-induced saline-alkali stress. Notably, HbWRKY11 displayed sustained up-regulation and significantly enhanced yeast tolerance to NaHCO₃ stress. Overall, this study provides the first systematic analysis of WRKY group II genes transcription factors in H. brevisubulatum and identifies HbWRKY11 as a key candidate gene contributing to saline–alkali stress tolerance. Full article

This paper presents a mathematical modeling and advanced control strategy for the beer fermentation process, which is characterized by nonlinear biochemical kinetics and time-dependent dynamics. A biokinetic model was developed to describe the relationship between yeast growth, sugar consumption, and ethanol formation. The system was represented as a cascade of several continuous stirred-tank reactors (CSTRs), and experimental data confirmed a fermentation cycle of approximately 10 days. During this period, biomass concentration reached 6.8 g/L and ethanol levels exceeded 42 mmol/L. Substrate concentration (S) declined from 120 to 5 g/L, demonstrating effective conversion. The model was linearized around an operating point and reformulated into a 12-state-space system with input variables: temperature (set at 20–22 °C) and pH (maintained within 4.2–4.5). These inputs were controlled using fuzzy logic control (FLC) and model predictive control (MPC). Simulation results indicated that the FLC reduced temperature deviation to ±0.3 °C and minimized pH fluctuation below ±0.05. The MPC strategy improved substrate consumption efficiency by 8.5% and decreased fermentation time by 12 h under optimized input profiles. The combined FLC–MPC scheme demonstrated superior robustness, smooth trajectory tracking, and adaptability to biological variability compared to traditional methods. The developed framework supports intelligent brewery automation and provides a scalable foundation for further integration of digital fermentation technologies. Full article

Phenolic acids are the major bioactive compounds in Perilla frutescens (L.) Britt; however, the regulatory roles of NAC transcription factors (TFs) in their biosynthesis remain unclear. Here, we performed a genome-wide identification and characterization of the NAC family in P. frutescens and explored their involvement in phenolic acid production. A total of 108 PfNAC genes were identified and classified into 17 subfamilies. Expression and promoter analyses suggested potential roles in secondary metabolism. PfNAC29 is located in the plasma membrane and necleus, while PfNAC40 and PfNAC80 are located in the nucleus.Yeast one-hybrid and dual-luciferase assays demonstrated that these TFs bind to the CATGTG motif in the PfC4H promoter and activate its transcription. Overexpression in transgenic hairy roots significantly increased rosmarinic acid, caffeic acid, and ferulic acid accumulation, accompanied by upregulation of key biosynthetic genes. These results indicate that PfNAC29, PfNAC40, and PfNAC80 act as positive regulators of phenolic acid biosynthesis and provide promising targets for metabolic engineering in medicinal plants. Full article