When organisms encounter pathogens, they rapidly activate complex defense programs to ensure survival. While these immune responses are vital, they often also incur trade-offs, such as reduced growth and development and must therefore be tightly controlled. In this study, we reveal that the steroid hormones brassinosteroids (BRs) contribute to this control in Arabidopsis thaliana by repressing immunity-related genes. We provide evidence that the BR-regulated basic helix-loop-helix (bHLH) transcription factor CESTA (CES), along with its homologs BR ENHANCED EXPRESSION (BEE)1-3, mediate DNA methylation changes at transposable element (TE)-rich loci containing nucleotide-binding leucine-rich-repeat (NLR)-type receptor genes, including SUPPRESSOR OF NPR1-1 CONSTITUTIVE 1 (SNC1). These CES-induced methylation changes correlate with altered splicing of SNC1 pre-mRNA, a process that requires the BR receptor BRASSINOSTEROID INSENSITIVE 1 (BRI1). In support, we show that CES associates with components of the chromatin remodeling and splicing machinery. Together, our findings reveal a previously unrecognized BR-induced mechanism that modulates the epigenetic and post transcriptional regulation of immune genes, enabling plants to prioritize growth over defense.

This study investigated the quantitative changes in 33 stress- or resistance-related metabolites induced by Bipolaris sorokiniana in barley leaves of quantitatively resistant and susceptible barley lines of the multiparental nested association mapping (NAM) population HEB-25. The analyses were based on ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Twenty-nine infected and noninfected barley genotypes were analyzed at four different time points after inoculation. The method provided quantification of hordatines, phenolamides, hydroxycinnamic acids, flavone glucosides, hydroxynitrile glucosides, apocarotenoids, and indole derivatives. In leaves infected with B. sorokiniana, phenolamide levels were elevated compared to noninfected plants. A correlation between metabolite levels and the severity of infection showed that the more resistant barley lines contained higher amounts of hordatines and hordatine glucosides.

Little is known about regulatory mechanisms that crop plants use to respond to combinations of abiotic and biotic stress. We analysed four barley genotypes under simultaneous Fusarium culmorum infection and drought stress by phenotyping for Fusarium Head Blight (FHB) disease, drought stress responses, hormone profiling and transcriptome analysis. FHB severity was host genotype-dependent, with moderately resistant cultivars Avalon and Barke showing increased FHB under drought, while drought did not further increase FHB severity in highly susceptible Morex and Palmella Blue. Transcriptome analysis revealed largely additive effects of single stresses, with drought-dominated regulation with increasing drought severity. Co-expression analysis connected abscisic acid and auxin to gene expression modules functionally enriched with stress-specific physiological responses. Stress-response genes, uniformly expressed across genotypes, were linked to pathogen defence, detoxification and drought adaptation, whereas a cluster of hundreds of moderately Fusarium-responsive genes was limited in up-regulation under combined stress, possibly explaining enhanced FHB severity under drought. A multiple linear regression model accurately predicted combined stress expression from single stress responses, demonstrating that gene regulation under combined drought and Fusarium stress is largely driven by additive effects of individual stresses.

The sustainable production of microalgal biomass for food and feed applications requires efficient downstream processing methods, particularly for the disruption of recalcitrant microalgal cell walls. This study aimed to develop an enzymatic hydrolysis method using fungal enzyme mixtures for effective degradation of the cell wall of the green microalga Chlorella sorokiniana. Eight fungal strains were screened for their enzyme production and hydrolysis efficiency, with Aspergillus awamori, Aspergillus niger, and Ceratocystis paradoxa showing the highest performance and selected for detailed analysis. A Design of Experiment approach optimized hydrolysis parameters, revealing that all enzyme mixtures showed maximal sugar release at 60 °C and an enzyme-to-biomass ratio of 5

Plant growth regulation and responses to biotic and abiotic stress factors are mediated by phytohormones. Understanding the effects of the phytometabolome is essential for addressing future global agricultural and food sector challenges. Therefore, a liquid chromatography-mass spectrometry method was developed to improve the ability to determine the concentrations of various phytohormones. Using only 20 mg of plant material, a total of 27 hormones from different classes, including 18 gibberellins, abscisic acid, salicylic acid, auxin, and jasmonates, can be analyzed in 10 min via the stable isotope dilution assay approach. Using a carbodiimide-hydrazine one-pot multifunctional chemical derivatization approach, the analytes can be detected with quantitation limits ranging from 0.04 to 29.9 nM. Application of the method to tomato, maize, and cress verified its applicability, thus enabling quantitative multiclass phytohormone profiling.

Monosodium glutamate (MSG; L-glutamate monosodium salt) is among the most widely used flavor enhancers, yet its molecular effects on gut microbial physiology remain poorly understood. Here, we examined the strain-specific transcriptomic and metabolic responses of Clostridium butyricum and Bacteroides thetaiotaomicron grown in pure anaerobic culture with 0.1% (wt/vol) MSG. Although MSG exposure was not associated with major changes in total bacterial biomass dynamics, both species showed a temporal functional reprogramming. In C. butyricum, MSG rapidly intensified metabolic activity to capitalize on exogenous glutamate by activating central carbon metabolism, redox-balancing pathways, and the GABA shunt, thereby collectively enhancing butyrate synthesis, a metabolite associated with gut barrier integrity, energy regulation, and anti-inflammatory signaling. On the other hand, B. thetaiotaomicron adopted a conservative, homeostatic response, suppressing glycan utilization and central carbon and energy metabolism, and maintaining stable SCFA production through controlled regulation that buffered against abrupt dietary perturbations. Overall, MSG functioned as a potent metabolic signal, triggering distinct adaptive strategies in two dominant gut bacteria. IMPORTANCE:The impact of monosodium glutamate (MSG) as a highly consumed food additive on the gut microbiome is often overlooked, and community-level analyses reveal little change, masking distinct phenotypic responses of individual strains. By combining gene expression and metabolite profiling using two key human gut bacteria, we show that MSG is sensed as a metabolic signal. A butyrate-producing gut bacterium increases energy metabolism and butyrate production, and a fiber-degrading gut bacterium transiently moderates metabolism to maintain stable fermentation products. These differences suggest that MSG's physiological effects may depend on which bacterial groups dominate an individual's microbiome.

Bitter taste is a critical quality determinant in food systems, particularly those using sustainable protein hydrolysates, where the unpredictable formation of bitter peptides severely limits consumer acceptance. Achieving predictive control over flavor chemistry requires deciphering the complex sequence-activity relationship. To address this, we integrated the generative capacity of a protein language model with BitterPep-GCN, a Graph Convolutional Network (GCN) capable of robust in silico bitter/non-bitter classification, to target the de novo design of functional bitter and non-bitter sequences. We achieved this by generating two strategic peptide libraries: a targeted tripeptide library derived from known bitter and non-bitter peptide sequences, and a set of de novo designed sequences. For the de novo designed peptides, we fine-tuned the conditional language model ZymCTRL on our curated dataset of sensory-validated bitter peptides (BPS-1000). Both libraries were subjected to classification and rigorous filtering using BitterPep-GCN to select high-confidence candidates for validation. The selected peptides were purchased and rigorously assessed for high purity. Sensory tests were conducted by an expert human panel to determine intrinsic taste quality and taste recognition thresholds. The results validated the high predictive fidelity of our pipeline: out of the 31 tested peptides, 25 were correctly classified, including 15 confirmed bitter and 10 confirmed non-bitter sequences. This study successfully demonstrates the application of machine learning frameworks in the design of bioactive peptides. It provides a set of novel taste-active peptides that can be used to accelerate the rational mitigation of off-tastes in next-generation food products.

Fusarium Head Blight (FHB) is a devastating fungal disease of small grain cereals like wheat and barley, causing substantial yield and quality losses each year worldwide. FHB is caused by Fusarium species that produce mycotoxins such as deoxynivalenol (DON) that impairs protein biosynthesis. Although defense responses in barley to Fusarium infection have been described at the transcriptional level, it remains unclear to what extent these responses are translated into functional changes at the protein and metabolite levels. In this study, we employed comprehensive transcriptomics, proteomics, and metabolomics to dissect the defense responses of barley heads during infection with Fusarium culmorum. Our integrated analyses revealed a set of significantly regulated gene-protein pairs linked to biosynthetic pathways that consistently correspond to upregulated defense-related metabolites. These include tryptophan-derived stress metabolites such as tryptamine and serotonin, as well as barley-specific hydroxycinnamylamides, including conjugates from the trypthophan metabolism, hordatines, and their biosynthetic precursors. Integrating data across multiple omics layers identifies the upregulation of aromatic amino acid derived secondary metabolism as the most consistent barley response to FHB infection across diverse barley varieties that share barley-typical type II resistance to fungal spreading in the head rachis.

This study employed activity-guided fractionation to identify the compounds that are responsible for the bitter off-taste of fava bean protein isolates and concentrates. UHPLC-ToF-MS and 1D/2D NMR experiments led to the identification of three known bitter compounds, vicine, convicine, and 3'-O-β-d-glucopyranosyl-L-DOPA. In addition, eight previously unknown vicine and convicine derivatives were identified. The bitter thresholds of the analytes were determined and found to be in the range of 0.10 to 1.44 mmol/L. To assess the taste contribution, the corresponding dose-overthreshold (DoT) factors were calculated, and it was shown that convicine with a DoT > 230 is playing a central role for the bitter off-taste. Furthermore, the analysis of fatty acids and their oxidation products suggests that linolenic-, linoleic-, and oleic acid directly contribute to the off-taste of fava bean protein. In addition, cell-based studies showed activation of bitter receptors TAS2R16 and TAS2R43 by vicine and convicine, respectively.

The global demand for sustainable protein sources has led to a growing interest in plant-based alternatives, with sunflower products emerging as a promising yet underutilized option. This review provides a comprehensive overview and critical evaluation of current knowledge on the flavor and off-flavor profiles and codes of sunflower seeds and their by-products, with a focus on both volatile and non-volatile low-molecular-weight compounds. It can highlight the importance of the sensomics approach and the knowledge on key food odorants and tastants. Furthermore, this review underscores the importance of advanced analytical methodologies for linking chemical composition to sensory outcomes. While volatile compounds that activate human olfactory receptors, such as aldehydes, terpenes, and pyrazines, are well described in sunflower products, using the sensomics approach the key odor-active stimuli are just verified in sunflower oil. In addition, the roles of non-volatile components including lipids, proteins, carbohydrates, and secondary metabolites such as polyphenols require further investigation and experimental validation to confirm their role as key tastants and their effect on sensory perception. By compiling existing data, this review establishes a foundational database of known and potential flavor-relevant compounds in different sunflower products, providing a valuable resource to directly or indirectly guide sensory (sensomics) studies and promote sunflower-based product innovation. Identifying the key flavor contributors in the different sunflower-based products and raw materials would facilitate precise approaches in processing and product formulation to enhance sensory quality while mitigating off-flavors. Addressing these challenges will support the development of sunflower-based food products with optimized flavor and nutritional profiles, consistent with global sustainability goals and consumer acceptance.

This study explores the chemical space of bitter peptides through a curated data set, named Bitter Peptide Space (BPS)-1000, which includes experimentally validated bitter and nonbitter peptides. The data set integrates sensory data, bitter taste thresholds (BTTs), and bitter taste receptor (TAS2R) activity when available. The inclusion of modified peptides further expands the data set's diversity. The HELM (Hierarchical Editing Language for Macromolecules) and BILN (Boehringer Ingelheim Line Notation) notations have been generated to provide a unique representation for both canonical and modified peptides. Through sequence-based and structure-based analyses, the study highlights the role of hydrophobicity, molecular size, and specific amino acid composition in the bitter and nonbitter sets in canonical and modified peptides, suggesting differences that could contribute to bitterness and enhancing the understanding of bitter peptide characteristics.

Growing crops in controlled-environment indoor farming systems offers new ways of producing high-yield, pesticide-free, environmental-friendly food. However, it replaces soil with hydroponics and the sun with LED lights. Compared with the field, wheat grown indoors showed a much higher yield potential and bread-making quality parameters. Many mineral concentrations were higher due to the unrestricted water supply and nutrients in hydroponics. However, concentrations declined with increasing yields. The microbiome richness inside the grains of wheat grown without soil indoors was still within the range of wheat grown in the field. However, taxa were different among cultivars and treatments. There were differences in the presence of undefined secondary metabolites between indoor and outdoor wheat and across the indoor experiments. Regardless of the growing environment, immunoreactive proteins were present. Indoor-grown wheat had a higher share of ω5-gliadins but lower shares of γ-gliadins and low‐molecular‐weight glutenin subunits, which may affect the gluten protein immunoreactive potential for individuals with wheat-related disorders (allergy and celiac disease). Growing wheat without soil and sunlight indoors can produce high-yielding, high-quality grains. However, the food quality and health aspects associated with gluten proteins might deteriorate with a further, theoretically possible, yield increase in a controlled growing environment.

Arbuscular mycorrhiza (AM) improves mineral nutrient supply, stress tolerance, and growth of host plants through re-programing of plant physiology. We investigated the effect of AM on the root secondary metabolome of the model legume Lotus japonicus using untargeted metabolomics. Acetonitrile extracts of AM and control roots were analysed using ultra-high-performance liquid chromatography-electrospray ionization-ion mobility-time-of-flight-mass spectrometry (UPLC-ESI-IM-ToF-MS). We characterized AM-regulated metabolites using co-chromatography with authentic standards or isolation and structure identification from L. japonicus roots using preparative high-performance liquid chromatography and nuclear magnetic resonance spectroscopy. Arbuscular mycorrhiza triggered major changes in the root metabolome, with most features representing unknown compounds. We identified three novel polyphenols: 5,7-dihydroxy-4'-methoxycoumaronochromone (lotuschromone), 4-hydroxy-2-(2'-hydroxy-4'-methoxyphenyl)-6-methoxybenzofuran-3-carbaldehyde (lotusaldehyde), and 7-hydroxy-3,9-dimethoxypterocarp-6a-ene (lotuscarpene). Further AM-enhanced secondary metabolites included the previously known lupinalbin A and B, ayamenin D, biochanin A, vestitol, acacetin, coumestrol, and betulinic acid. Lupinalbin A, biochanin A, ayamenin D, liquiritigenin, isoliquiritigenin, lotuscarpene, medicarpin, daidzein, genistein, and 2'-hydroxygenistein inhibited Rhizophagus irregularis spore germination upon direct application. Our results show that AM enhances the production of polyphenols in L. japonicus roots and highlights a treasure trove of numerous unknown plant secondary metabolites awaiting structural identification and functional characterization.

The study focuses on the comprehensive analysis of glutamyl dipeptides in cheese, particularly their formation during the cheese ripening process and the influence of various factors, such as origin, the use of various mold cultures, and cheese types. For the first time, all three subgroups of glutamyl dipeptides, namely alpha-Glu-X, X-Glu, and gamma-Glu-X, are covered in a comprehensive analytical LC-MS/MS method offering robust quantitation of all 56 glutamyl dipeptides. The workflow includes a simplified extraction protocol and an optimized separation of the analytes on the stationary phase. Validation experiments demonstrate the method's reliability, including repeatability, detection limits, and recovery. The comprehensive analysis of all glutamyl dipeptides in 122 cheese samples with ripening times between 2 weeks and 15 years shows a strong increase in all peptide classes with prolonged ripening and particularly in the presence of mold.

A growing global population and climate change challenge conventional agriculture and global food safety. Microalgae are an emerging sustainable nutrient food resource, increasingly regarded as an important component for the human diet. The chlorophyte Chlorella sp. is regulated and commercialized for food applications. Commercial Chlorella preparations are either produced by heterotrophic fermentation or photoautotrophic cultivation in bioreactors or outdoor ponds. Products can differ significantly with regard to taxonomy of the production strain, changed genetic properties due to strain development, cultivation conditions, and downstream biomass processing methods. Notably, heterotrophic strains may be adapted to generate little or no photosynthetic pigments, resulting in yellow or white variant. In this study, eight different commercial Chlorella products from different EU suppliers were analyzed and compared regarding identity of the production strain, macro-and micronutrient profiles, sensory properties, as well as microbial load. The production processes featured different cultivation methods, production strains, genetic variants (yellow and white) and downstream processing methods. Data indicates significant variations between the Chlorella products. White and yellow Chlorella products showed different protein and taste profiles compared to green, photosynthetically competent wild type strains. We confirmed relatively high and stable protein concentrations but could detect variations in sugar and specifically lipid and vitamin profiles depending on strain, cultivation and downstream processing methods. The microbial load varied strongly between closed and open cultivation systems but was compliant with legislative (EFSA) specifications for all samples. The comparative nutrient- and sensory data set presented in this study will aid in selecting algae products for the development of innovative foods, thereby accelerating adoption and differentiation of algae food products.

Spot blotch of barley (Hordeum vulgare L.), caused by Bipolaris sorokiniana, is responsible for major losses in crop yield. Breeding-resistant barley varieties have proven to be an effective countermeasure for protecting agricultural production. Plants react to pathogen attacks by up-regulating secondary metabolites. Marker compounds for a B. sorokiniana infection are examined by untargeted UPLC-TOF-MS metabolomics and lipidomics techniques. Through the analysis of nine quantitatively resistant and susceptible barley genotypes, derived from the nested association mapping population HEB-25, followed by structure identification experiments and spore germination assays, 57 metabolites are identified. In addition to previously known metabolites, the unknown compounds 5-carboxydidehydroblumenol C-9-O-ß-d-glucoside (46) and grasshopper ketone 3-sulfate (47) were elucidated. 5-Carboxyblumenol C-9-O-ß-d-glucoside (45) was described for the first time in barley leaves. Pheophytin derivatives, oxylipins, linolenate-conjugated lipids, and flavone glycosides were described for the first time in connection with infections by phytopathogenic fungi or resistance in barley.