Journal of Agricultural and Food Chemistry

Sustainable agriculture has driven increased exploration of microbial inoculants as a promising strategy to boost plant growth for higher yield and enhance secondary metabolism to increase crop nutritional value. However, their influence on fruit-specific metabolites under field conditions remains understudied. This study investigated the impact of inoculating cranberry (Vaccinium macrocarpon) plants with Bacillus velezensis EB37 and Lachnum sp. EC5, applied individually and in combination, on cranberry fruit phytochemistry. Over two growing seasons (2019 and 2021), cranberries were collected from treated and control plots and analysed using untargeted and targeted LC-MS-based metabolomics. Multivariate analysis revealed no significant metabolomic differences due to treatments. However, samples clustered strongly by year of harvest, highlighting a pronounced environmental effect. Quantitative analysis of six representative phenolic compounds: chlorogenic acid, catechin, p-coumaric acid, phloridzin, myricetin, and quercetin, showed no statistically significant differences between treated and control cranberries. These findings indicate that microbial inoculation alone does not alter cranberry fruit metabolome, including phenolic levels, at field conditions. This study underscores how multiple factors, such as environmental conditions, can affect the outcome of microbial inoculation under field conditions and suggests that additional interventions may be required to achieve microbiome-based improvements in cranberry fruit quality.

Prenylated isoflavonoids are widely distributed specialized metabolites within the Fabaceae and contribute to various characteristic biological activities for both plants and humans. Several aromatic prenyltransferases (PTs) have been identified in Glycyrrhiza species, which are the most widely consumed crude drugs in traditional Chinese medicine. However, these enzymes do not sufficiently explain the structural diversity of prenylated flavonoids produced in the Glycyrrhiza genus. To identify additional novel PTs, we used elicited cultured Glycyrrhiza glabra roots as source material, in which elicitor treatment of cultured roots increased the accumulation of multiple prenylated flavonoids. To identify the responsible enzyme, PT candidates were screened using G. uralensis transcriptomes, currently the sole publicly available transcriptomic resource within the genus, and a homolog designated GgBSPT1 (BSPT; a broad-substrate prenyltransferase) was subsequently isolated from elicited cultured G. glabra roots. GgBSPT1 differed from previously identified Glycyrrhiza PTs in both amino acid sequence and enzymatic properties. GgBSPT1 catalyzed 3'-prenylation of isoliquiritigenin and 6-prenylation of five flavonoids, i.e., this PT displayed broad substrate acceptance across 20 distinct flavonoid structures. Overall, elicited cultured G. glabra roots enabled the identification of a previously unrecognized PT that is functionally distinct from earlier reported Glycyrrhiza PTs. This study provides a new insight into the metabolic plasticity of Glycyrrhiza species and expands the enzymatic toolkit for future metabolic engineering of prenylated phytochemicals by the unusually broad substrate specificity of GgBSPT1. Main conclusionUsing cultured Glycyrrhiza glabra roots, we identified a new prenyltransferase involved in the formation of a variety of flavonoids, thereby revealing novel prenylated isoflavonoid pathways in licorice.

In many tropical countries, fermentation of palm sap into palm wine is an important fermented beverage contributing to local economies, tradition, and culture. Traditionally made in villages and families, palm sap is not inoculated with starter cultures and fermentation commences spontaneously. It is therefore possible that fermentation is influenced by multiple ecological factors, which affect microbial dynamics and thus flavour outcomes. Here, we studied microbial communities during fermentation of palm sap from three different palm tree species (palmyra, coconut, and sugar palm) on the island of Bali, Indonesia in both the wet and dry seasons. Our results suggest that season of collection has a strong influence on microbial dynamics and succession, and these changes positively correlate to metabolite concentration. The change of the season from the dry to wet season led to the loss of microbial diversity with lower richness in the dry season. The dominance of Saccharomyces cerevisiae was not affected by season and fermentation time and was dominant in all samples. Potential spoilage species, such as Candida tropicalis were negatively correlated to ester production and more abundant in the dry season. As microbial species varied in incidence and thus biochemical activity, the chemical groups of esters from their metabolism related to the change of season and fermentation time, while volatile compounds and small molecules were highly discriminated by season in the resultant wines. Ethyl octanoate was consistently different across all variables through comparison by three-way ANOVA and is proposed as a biomarker of seasonal variation in palm sap fermentation. These findings improve our understanding of microbial dynamics in palm sap fermentation, revealing flavour differentiation within season and suggests that strategies for microbial management, product development and quality assurance will elevate this traditional product into the future.

Plants produce specialized metabolites that function in plant defense and as attractants to pollinators and symbionts. One class of specialized metabolites are terpenoids that are synthesized from universal C5 building blocks via activities including terpene synthases, cytochromes P450, and glycosyl transferases. Some terpenes are highly valued for their use as insect repellants, fragrances, antimicrobial compounds, low calorie sweeteners, flavors, and medicines. Low abundance in target tissues, present in complex mixtures, as well as challenging extraction logistics are barriers to economic sustainable production of these compounds from their native species. While heterologous expression of terpenoid biosynthetic genes is feasible, the potential derivation of the products into conjugates via endogenous cytochromes P450 and glycosyl transferases limits this approach. In this project, we used multiplex gene editing technologies to overcome these challenges by creating novel tomato chassis with altered terpenoid biosynthetic capacity in fruit. Excluding central metabolic genes to minimalize impacts on growth and development, we selected 23 known and potential terpene-related genes expressed specifically in the fruit for gene editing. Fruit production and metabolic profiles of three chassis lines with alterations in the major classes of fruit specialized metabolites indicate loss of these genes is tolerated for fruit production. These combinatorial knockouts also showed modulation of native carbon reallocation toward endogenous sinks beneficial for a biosynthetic chassis. Establishing metabolite-modified fruit chassis demonstrates efficient combinatorial editing of entire branches of plant specialized metabolism, facilitating engineering of heterologous terpenes of industrial interest in tomato fruit.

Understanding how plants respond to high temperature is critical under global warming. Metabolite markers can provide insights into stress-responsive mechanisms and help guide strategies to maintain crop quality. However, heat-associated metabolite markers in grape berries remain poorly defined, particularly at the green stage, a critical phase of berry development during which early metabolic perturbations can influence subsequent ripening and ultimately determine berry composition and quality. Here, we applied berry-scale heat treatments of eight durations of two major wine cultivars, Cabernet Sauvignon and Merlot. Untargeted LC-MS profiling revealed both conserved and cultivar-dependent responses to heat. Based on these patterns, three time points were selected for targeted GC-MS analysis, and subsequent statistical analyses identified robust "cultivar-common heat markers": glycine decreased, whereas galactinol increased consistently across time points and cultivars. "Cultivar-dependent heat markers" were identified: xylose, lyxose, citrulline, quinic acid, and glutamine, that consistently distinguished CS and Merlot fruits under heat stress. Notably, xylose, lyxose, citrulline, and quinic acid also differentiated the two cultivars under ambient conditions, underscoring their potential as stable cultivar-discriminating metabolites. Together, these results reveal dynamic metabolic remodeling in grape berries under heat stress, particularly in amino acid, nitrogen, central carbon metabolism, raffinose family oligosaccharides pathway and the glutathione-ascorbate cycle.

Chitooligomers can act as plant biostimulants or biopesticides, but todays chitosan-based agro-biologics often lack sufficient efficacy. This is due to a lack of scalable production processes for structurally well-controlled chitosans combined with a limited understanding of structure-function relationships. Chitosans differ in their degree of polymerization (DP), fraction and pattern of acetylation (FA and PA). While the influence of DP and FA on antimicrobial and phytostimulatory properties is at least partially known, this is not yet the case for PA. PA can be partially controlled by using enzymatic rather than acid hydrolysis for oligomer production. We have used recombinant chitinases and chitosanases to hydrolyse a well-characterised chitosan polymer, and purified oligomers with different DP. We have structurally characterised the products and tested their abilities to protect tobacco from viral disease. Chitinase products were dominated by GlcNAc units at their reducing and non-reducing ends, with GlcN units dominating their centers, and v.v. for chitosanase products. While the chitinase-derived hydrolysates were inactive, the chitosanase-derived oligomers possessed elicitor and priming activities and protected plants from disease, and their activity increased with increasing DP. Clearly, the Bacillus chitosanase used is well-suited to set up a scalable production process for chitosan oligomers with promising agro-biologic properties. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=95 SRC="FIGDIR/small/731087v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@c312e9org.highwire.dtl.DTLVardef@10eaf99org.highwire.dtl.DTLVardef@12a937corg.highwire.dtl.DTLVardef@38dc8c_HPS_FORMAT_FIGEXP M_FIG C_FIG

Mushroom production generates large amounts of by-products, particularly stipes, which can represent up to half of the fruiting body biomass. Due to their similar composition to mushroom caps, these residues represent a promising substrate for the development of value-added foods. In this study, oyster mushroom stipes were used as a substrate for solid-state fermentation (SSF) with a Neurospora crassa strain isolated in Albacete to produce a novel meat analogue inspired by the oncom. Fermentation generated a cohesive matrix bound by hyphae that adopted the shape of the mold and exhibited a meat-like color, although with a softer texture. Nutritional analysis revealed a product with relatively low protein content but a complete amino acid profile, enriched in dietary fiber and containing unsaturated fatty acids. These results demonstrate that SSF with N. crassa provides a strategy to upcycle oyster mushroom by-products into fiber-rich meat analogues with potential applications in sustainable food systems.

Fungal contamination of food with yeast and moulds is associated with major economic losses due to spoilage and also poses health risks in the form of mycotoxin production. The strain Pantoea agglomerans APC 4211 isolated from leaves of Ilex aquifolium (holly tree) has broad spectrum antifungal activity against a variety of food spoilage fungi. Genomic analysis of the strain confirmed the presence of biosynthetic gene clusters potentially encoding for the enzymatic machinery required for the production of the antifungal lipopeptide herbicolin A. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of the cell-free supernatant (CFS) confirmed the presence of molecular masses corresponding to herbicolin A (1300.8 Da), and herbicolin B (1138 Da). Purified herbicolin A has desirable properties for biotechnological applications, including potent antifungal activity against a range of spoilage fungi, thermal stability and resistance to proteases. Herbicolin A has low cytotoxicity against epithelial cell lines and has minimum inhibitory concentrations (MICs) lower than those of some commercial antifungal drugs (0.2 - 2.5 {micro}g/ml). In a model dairy system (10% skim milk), herbicolin A demonstrated excellent solubility and stability, effectively eliminating Aspergillus niger and Penicillium notatum at a concentration of 5 {micro}g/mL. In conclusion, herbicolin A is a potent, naturally occurring antifungal agent with the potential to be applied as a biopreservative in food systems, providing a safe, clean-label, and efficient compound for synthetic preservatives replacement. HighlightsO_LIHerbicolin A has a strong potential as a natural preservative for food protection C_LIO_LIHerbicolin A shows lower MICs than several antifungal agents C_LIO_LIHerbicolin A is stable under heat and resistant to proteolytic degradation C_LIO_LIHerbicolin A has strong solubility and stability in a model dairy system C_LIO_LIHerbicolin A indicates low cytotoxicity against epithelial cell lines C_LI Data summaryThe authors confirm all supporting data, code and protocols have been provided within the article or through supplementary data files.

Bread wheat (Triticum aestivum) is a staple food crucial for global caloric intake and food security. The current climate emergency demands the development of sustainable agricultural practices, particularly in the context of drought-induced yield reductions in bread wheat. Microalgae-based biostimulants have emerged as promising tools to enhance crop tolerance to drought stress while concurrently mitigating atmospheric CO2 accumulation. This study characterizes the transcriptomic responses to the foliar application of the microalgae-based biostimulant LRMTM in drought-stressed and fully irrigated wheat plants unveiling its mode of action. Drought stress at the tillering stage significantly altered gene expression activating key pathways related to phosphate starvation response (PSR), inositol phosphate signaling, and tocopherol biosynthesis. The application of the microalgae-based biostimulant LRMTM in drought-stressed plants further enhanced the expression of drought-responsive genes, particularly those involved in PSR and carbon fixation. Specific responses to LRMTM treatment in drought-stressed plants were also found related to abscisic acid (ABA) signaling activating genes involved in stomata closure, which plays a critical role in drought tolerance. In fully irrigated plants, LRMTM treatment was also beneficial modulating circadian rhythms, shade avoidance and attenuating stress responses. Phenotypic analysis showed that LRMTM-treated plants exhibited enhanced drought tolerance, increased height and spike length even under fully irrigated conditions. These results indicate that the microalgae-based biostimulant LRMTM not only enhances wheat response to drought but also promotes growth and productivity in both stressed and non-stressed conditions which could contribute to the development of sustainable agriculture in the face of the current climate challenges.

Lignocellulosic biomass represents a promising renewable feedstock for sustainable biorefinery applications, yet efficient enzymatic saccharification remains challenging due to the recalcitrant structure of plant cell walls. This study presents a comprehensive comparative analysis of enzymatic activities, saccharification performance, and secretome composition of six fungal species cultivated under solid-state fermentation (SSF) and submerged fermentation (SmF) conditions using untreated flax shives as substrate. While SmF yielded approximately 4-fold higher total protein concentrations (0.38 {+/-} 0.13 g.L-1 vs. 0.08 {+/-} 0.02 g.L-1), SSF-derived enzymes demonstrated superior specific enzymatic activities, particularly for endo-xylanase and endo-cellulase, resulting in more efficient biomass saccharification. Proteomics analysis revealed distinct secretome profiles between fermentation modes, with SSF showing higher proportions of polysaccharide metabolism proteins (71.0%) compared to SmF (49.3%), while SmF exhibited greater enzyme diversity including more lytic polysaccharide monooxygenases (LPMOs) and auxiliary activity enzymes. Trichoderma species consistently demonstrated the highest saccharification efficiency, with glucose yields reaching 2.37 mM under SSF conditions. A Scheffe simplex-lattice mixture design comprising 65 enzyme cocktail combinations revealed significant synergistic interactions between several cocktails, with the binary mixture of Trichoderma 2SA21 and P. chrysogenum achieving 54% synergy - in terms of higher sugar release above expectations - and the highest total monosaccharide release (1.80 mM). These findings provide practical guidance for developing cost-effective enzyme cocktails for lignocellulosic biorefinery applications, emphasizing the importance of fermentation mode selection and strategic strain combination over enzyme supplementation complexity. The methodology established here, combining systematic screening, comparative proteomics, and statistical mixture design, offers a robust framework for optimizing fungal enzyme systems across diverse biomass substrates. BULLET POINTSSuperior enzymatic activity (xylanase, cellulase) and saccharification in solid-state fermentation Superior total protein content and diversity in submerged fermentation Specific enzyme cocktails combination can lead to synergistic effects, justifying a combinatorial approach GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=107 SRC="FIGDIR/small/729743v1_ufig1.gif" ALT="Figure 1"> View larger version (25K): org.highwire.dtl.DTLVardef@1f3704dorg.highwire.dtl.DTLVardef@151e4bforg.highwire.dtl.DTLVardef@180ced7org.highwire.dtl.DTLVardef@18bb42f_HPS_FORMAT_FIGEXP M_FIG C_FIG

Although biostimulants have attracted attention for sustainable agricultural systems, their efficacy remains poorly understood. In this study, we evaluated the effects of fermented botanical product (FBP) produced by fermenting and aging 41 types of fruits, grains, seaweed, and root vegetables with brown sugar for more than three years. Three crops, tomato, rice, and komatsuna (Brassica rapa), were cultivated with the application of 5,000- or 10,000-fold diluted FBP in greenhouses or fields. Application of diluted FBP promoted plant growth, as indicated by increased fresh weights of shoots, leaves, and roots, fruit production in tomato, and rice husk yield. As diluted FBP contained low nutrient levels, an indirect mechanism of plant growth promotion was suggested. Bacterial community structure analysis indicated changes in alpha diversity, beta diversity, and the predominant phyla in FBP-applied soils without plants and in soils cultivated with tomato, rice, and komatsuna. In addition, the abundance of plant-growth-promoting bacteria, such as Arthrobacter, Pseudomonas, Paraburkholderia, and Planifilum, increased in soils treated with diluted FBP. Furthermore, ammonium formation activity was observed in komatsuna cultivation soils treated with diluted FBP, whereas phosphate-solubilizing activity was enhanced in soils from all three crop cultivation systems treated with diluted FBP. These results suggest that diluted FBP influences bacterial communities and promotes crop growth through indirect effects, including increases in plant-growth-promoting bacteria, ammonium production, and phosphate solubilization. Alternatively, FBP may directly stimulate plant growth. Therefore, FBP may be a useful biostimulant for sustainable agricultural systems. HighlightsO_LIDiluted FBP promoted the growth of tomato, rice, and komatsuna (Brassica rapa). C_LIO_LIDiluted FBP altered the bacterial community structure in cultivated soils. C_LIO_LIFBP increased the abundance of plant-growth-promoting bacteria in cultivated soils. C_LIO_LIFBP stimulated ammonium formation and phosphate solubilization in cultivated soils. C_LIO_LIFBP may be a useful biostimulant for sustainable agricultural systems. C_LI Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/728655v1_ufig1.gif" ALT="Figure 1"> View larger version (36K): org.highwire.dtl.DTLVardef@4772dorg.highwire.dtl.DTLVardef@12c40e8org.highwire.dtl.DTLVardef@594513org.highwire.dtl.DTLVardef@c5e5a6_HPS_FORMAT_FIGEXP M_FIG C_FIG

O_LIEuropean beech (Fagus sylvatica L.) is a widely distributed, ecologically and economically important deciduous tree species in European forests, but is increasingly threatened by drought stress. Volatile organic compounds (VOCs) are ubiquitous plant metabolites that may serve as non-invasive biomarkers of drought stress, yet they have rarely been studied in European beech. C_LIO_LIIn this study, we examined VOC responses of European beech to experimental drought across diverse genetic backgrounds in a common garden. The 72 four-year-old beech saplings represented three genetic clusters, seven provenances (geographic seed sources), and 12 maternal seed families. Half of the saplings were assigned to the drought treatment and received no water for 14 days, while the remaining saplings served as controls and were watered as required. VOC profiles, quantified as peak heights of mass spectral features, were measured for all individuals during pre-drought, drought, and rewatering periods. C_LIO_LIWe found that pre-drought VOC profiles, in particular monoterpenes, varied significantly among genetic backgrounds. Experimental drought significantly altered VOC profiles, characterized by increased green leaf volatiles and decreased monoterpenes, oxidized terpenoid derivatives, and other fatty acid derivatives. Reductions in monoterpenes persisted after rewatering, indicating a drought legacy effect. Drought responses were largely conserved across genetic backgrounds, with significant seed family-specific responses detected for only three VOC features. C_LIO_LIOur findings suggest that VOC profiles are genetically structured yet highly plastic under drought and highlight their potential as non-invasive biomarkers for monitoring drought stress in European beech under climate change. C_LI

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the most abundant protein on Earth, is an attractive and sustainable food ingredient owing to its favourable nutritional and techno-functional properties. Leafy vegetables are particularly rich sources of RuBisCO; however, large-scale vegetable production generates substantial quantities of residual biomass throughout agri-food supply chains. Drying is widely used to stabilise this biomass and facilitate storage, transport, and handling, yet most reported RuBisCO extraction methods have been developed for fresh material and are poorly suited to dried feedstocks. Here, we present a simple, scalable, and cost-effective process for the recovery of food-grade RuBisCO from dried leafy biomass. Using spinach, rocket, and kale as model systems, efficient protein extraction was achieved from both freshly dried leaves and commercially available leaf powders without the need for resource-intensive processing. Application of the method to spinach yielded approximately 75 mg of high-purity RuBisCO per 100 g fresh-leaf equivalent, corresponding to an extraction efficiency of [~]70%, which increased to [~]90% following supplementation with 20 mM CaCl2. The recovered protein fraction also exhibited favourable foaming capacity and foam stability, demonstrating its potential as a functional food ingredient. This work provides a practical route for the valorisation of dried vegetable residues and supports the development of circular, waste-to-value supply chains for sustainable plant protein production.

Green manure is a promising strategy for reducing dependence on chemical fertilizers in crop production. However, vigorous growth due to green manure often leads to high yield accompanied by lodging in rice, hindering its practical use in rice cultivation. Here, we show that rice cultivars carrying a semi-dwarfing sd1/ga20ox2 allele achieve high grain yield without lodging under hairy vetch-based green manure conditions. The leading Japanese cultivar Koshihikari exhibited enhanced vegetative growth, increased panicle number, and consequently higher grain yield and quality under green manure conditions in 2023 and 2024 compared with chemical fertilizer management, although this was accompanied by increased culm length and widespread lodging. Among the four GA20-oxidase genes, green manure significantly upregulated Sd1/GA20ox2 mRNA levels. A temperate japonica cultivar, Nijinokirameki, and an indica cultivar, Hokuriku-193, carrying a non-functional sd1/ga20ox2 allele exhibited no lodging under hairy vetch-based green manure management while achieving improved yield performance. Notably, yields obtained under our hairy vetch-based cultivation system were comparable to or exceeded a recently reported high-yield benchmark observed for Hokuriku-193 under chemical fertilizer management in the same region of Japan. These findings suggest that cultivars harboring non-functional sd1/ga20ox2 alleles enable the practical implementation of annual hairy-vetch-rice rotation for sustainable rice production.

BackgroundControlled-environment cultivation of medicinal cannabis (Cannabis sativa L.) typically optimizes light conditions to enhance the biosynthesis of pharmaceutically important metabolites like cannabinoids. Such experimental strategies may also influence other specialized metabolites like terpenoids, flavonoids, alkaloids, among others. Previous untargeted metabolomics studies testing short wavelength conditions like UV and blue light have shown that terpenoids and prenylated flavonoids in cannabis leaves respond differentially. However, since metabolomic studies in cannabis have so far mostly focused on floral cannabinoids, a comprehensive untargeted study into cannabis floral metabolome response to short wavelengths is currently lacking. ObjectivesOur study investigates the impact of short wavelength usage on cannabis specialized metabolism, and in particular the influence of UVB, UVA, and blue light on the cannabis floral flavonoid metabolome and associated glycosylation moieties. MethodsCannabis plants were grown under a white background light and exposed to supplemental UVB, UVA, or blue light during the generative phase of the cultivation cycle. Treatments were compared to a reference white background light without UV or blue light. Metabolites from floral tissue were extracted and analyzed via ultra-performance liquid chromatography-tandem mass spectrometry. A comparative metabolomics workflow was designed and used to characterize the floral flavonoid metabolome and associated glycosylation moieties. ResultsOur results demonstrate how short wavelengths differentially affect the metabolism of natural product compound classes including polyketides and phenylpropanoids/shikimates. Blue light induced flavonoids similarly to how UVB did, while both UVA and blue light specifically induced flavanones accumulation. UVB showed the strongest regulatory effect on flavonoids production and glycosylation patterns. ConclusionsUVB reshapes the cannabis floral flavonoid metabolome by selectively stimulating the accumulation and structural modification of flavonoids. Therefore, UVB application in cannabis cultivation represents a useful horticultural strategy to increase inflorescence medicinal quality without affecting cannabinoid levels.

Edible seaweed has the potential to become a valuable marine resource for food applications due to its potential health benefits and ecological sustainability. The brown seaweed Alaria esculenta is rich in essential minerals, vitamins, and dietary fibers, making it a nutritious food source. Fermentation, as a traditional preservation method, can enhance seaweed shelf-life and be useful for the development of new foods/ beverages. In this study, the effects of fermentation of A. esculenta, by the lactic acid bacterium (LAB) Lactiplantibacillus plantarum, on the nutritional profile, and the content of potentially toxic elements, was investigated. L. plantarum was successfully cultivated on A. esculenta using two modes of operation, submerged (SmF) and solid-state fermentation (SSF), resulting in production of cells and lactic acid, and reduction of the pH to below 4.3 within 3 days, which was not achieved in parallel spontaneous fermentations using indigenous seaweed microbiota. A. esculenta s macro-nutritional profile was altered, reducing mannitol but increasing fucose and glucose content (after acid hydrolysis) while also concentrating the protein content. LAB fermentation significantly increased the concentration of antioxidant phenolic compounds, such as phloroglucinol, syringic acid, and epicatechin, compared to untreated samples. However, lipophilic compounds like carotenoids decreased after both spontaneous and LAB-fermentation. A reduction in total mineral content was observed after LAB fermentation and water soaking, and SmF with L. plantarum effectively reduced arsenic and iodine levels. Overall, fermentation using L. plantarum showed potential as a bio-preservation method for the edible brown seaweed, A. esculenta, improving its nutritional profile and enhancing food safety.

{gamma}-Glutamyl cyclotransferases (GGCTs) belongs to class of cytosolic enzymes that are responsible for glutathione (GSH) degradation under stress conditions. They regulate GSH homeostasis through the {gamma}-glutamyl cycle which is responsible for maintaining the synthesis of GSH as well as its breakdown, enabling recycling of its constituent amino acids. Although GGCTs have been implicated in enhancing heavy metal (HMs) tolerance in plants, their role in biotic stress remains largely unexplored. Previously, OsGGCT1 was identified as a gene strongly upregulated in Fusarium stress. In this study, the GGCT1 homolog from Oryza sativa japonica was characterized for its role in conferring tolerance to Fusarium oxysporum (F.O.). Similar to abiotic factors, biotic stresses significantly impact crop yield and productivity. The rhizosphere harbors diverse microbial communities, including harmful pathogens such as F. oxysporum. Fusarium causes wilt disease in a variety of plant species, such as: tomato, legumes, rice, and Arabidopsis thaliana. Our results demonstrate that overexpression of OsGGCT1 enhanced tolerance to F. oxysporum in A. thaliana, primarily by reducing fungal spore accumulation. Transgenic plants showed elevated expression of OsGGCT1 along with AtGSH1 and AtGSH2, reduced levels of reactive oxygen species (ROS), improved growth and photosynthetic performance and enhanced activities of the antioxidant enzymes. OsGGCT1 serves as a key component in maintaining GSH homeostasis by supporting glutamate (Glu) regeneration necessary for sustained GSH biosynthesis. Overall, these findings identify OsGGCT1 as an important constituent of the GSH-mediated detoxification pathway against Fusarium oxysporum and provide valuable molecular insights for developing Fusarium-tolerant rice varieties with reduced fungal accumulation.

The prevalence of naturally occurring C13-acetoxy taxanes, together with the presence of a native C13-acetyltransferase in yew trees, suggests that the natural biosynthetic pathway for paclitaxel may involve a cryptic C13-O-deacetylation step. However, whether a putative taxane C13-O-deacetylase (T13dA) acts in the pathway of paclitaxel biosynthesis remains elusive. Here we functionally characterized two novel taxane C13-O-deacetylases (T13dA1 and T13dA2) from Taxus x media cell cultures, providing experimental evidence for the molecular and biochemical plausibility of C13-O-deacetylation in paclitaxel biosynthesis in Taxus species. Also, we identified a previously uncharacterized bifunctional taxane C7{beta}-O-, C9-O-deacetylase, designated T79dA, which demonstrates the functional promiscuity by enabling stepwise deacetylation at taxane C7{beta} and C9 positions in a single enzymatic reaction. Furthermore, T7dA1, a novel taxane C7{beta}-O-deacetylase with higher activity than the reported T7dA was discovered and characterized here. Moreover, we reconstituted two new pathways (an 18-gene and a 19-gene pathway) enabled by the integration of a C13-O-acetylation-deacetylation module for the de novo biosynthesis of baccatin III in Nicotiana benthamiana leaves. These pathways with the previously established 17-gene baccatin III pathway, further allow paclitaxel biosynthesis to be a network. Our reconstituted 19-gene pathway achieves a baccatin III yield of up to 23 g g-1 dried weight (DW) in N. benthamiana leaves, which is comparable to the yield reported for the 17-gene pathway. This work facilitates a better understanding, elucidation and reconstruction of metabolic network of paclitaxel biosynthetic pathway, and provides new enzymes and strategies for artificial pathway reconstruction and efficiently bio-chemical production of paclitaxel.

Rice (Oryza sativa) is highly sensitive to salinity, yet the metabolic mechanisms underlying salt tolerance remains incompletely understood. In this study, we performed leaf tissue-specific untargeted metabolomic profiling of the salt-tolerant introgression line JN100 (JN), its donor parent Nona Bokra (NB), and its recurrent parent Jupiter (JU) to characterize metabolic responses to salt stress. Comparative analysis identified differentially accumulated metabolites (DAMs) spanning diverse chemical classes, including amino acids, sugars and carbohydrates, lipids, organic acids, cofactors, electron carriers, and nucleotides. Under salt stress (SS), 201 DAMs (89 upregulated and 112 downregulated) were detected in JN relative to JU. Notably, metabolites such as allantoin, glycitin, nicotinamide ribotide, D-arabinono-1,4-lactone, violanthin, L-methionine S-oxide, ribitol, lysine, rutin, glutamine, pantothenic acid, and quinic acid, showed significant differential accumulation. Pathway enrichment analysis revealed significant enrichment of arginine biosynthesis, purine metabolism, and alanine, aspartate, and glutamate metabolism, indicating extensive reprogramming of nitrogen and energy-associated metabolic pathways under salinity stress. Integration of transcriptomic and metabolomic datasets from the SS experiments further identified ten differentially expressed genes (DEGs) associated with the metabolite network in the JN vs. JU comparison. Among these, OsDHQDT/SDH, OsFd-GOGAT, phenylalanyl-tRNA synthetase, OsP5CS1, OsP5CS2, and a pyridoxal phosphate-dependent transferase were linked to metabolites involved in shikimate, amino acid, and proline metabolism. Collectively, these results demonstrate that salinity tolerance in rice is associated with coordinated transcriptional and metabolic reprogramming that supports oxidative stress mitigation and adaptive stress responses.

In this study, the lipid content produced by Thraustochytrium sp. in a medium prepared from orange pulp residue was compared with that obtained in a conventional medium. The pulp residue was subjected to freezing, blending, and filtration in seawater to prepare three treatments: conventional medium (T1), filtrate (T2), and filtrate supplemented with KNO3 (T3). A growth curve was performed over six days, after which biomass and lipid content were analyzed. The results showed that T2 exhibited the highest cell growth and biomass yield (5.24 g/L). However, lipid content was higher in the conventional medium (38.75%), whereas the treatments containing orange extract showed lower values. These findings suggest that the medium prepared from orange pulp residue is feasible for the growth of Thraustochytrium sp., but requires optimization to enhance lipid accumulation and its potential use in sustainable bioprocesses.