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The occurrence of microplastics (MPs) and nanoplastics (NPs) in soils potentially induce morphological, physiological, and biochemical alterations in plants. The present study investigated the effects of MPs/NPs on lettuce (Lactuca sativa L. var. capitata) plants by focusing on (i) four different particle sizes of polyethylene micro- and nanoplastics, at (ii) four concentrations. Photosynthetic activity, morphological changes in plants, and metabolomic shifts in roots and leaves were investigated. Our findings revealed that particle size plays a pivotal role in influencing various growth traits of lettuce (biomass, color segmentation, greening index, leaf area, and photosynthetic activity), physiological parameters (including maximum quantum yield - Fv/Fmmax, or quantum yield in the steady-state Fv/FmLss, NPQLss, RfdLss, FtLss, FqLss), and metabolomic signatures. Smaller plastic sizes demonstrated a dose-dependent impact on aboveground plant structures, resulting in an overall elicitation of biosynthetic processes. Conversely, larger plastic size had a major impact on root metabolomics, leading to a negative modulation of biosynthetic processes. Specifically, the biosynthesis of secondary metabolites, phytohormone crosstalk, and the metabolism of lipids and fatty acids were among the most affected processes. In addition, nitrogen-containing compounds accumulated following plastic treatments. Our results highlighted a tight correlation between the qPCR analysis of genes associated with the soil nitrogen cycle (such as NifH, NirK, and NosZ), available nitrogen pools in soil (including NO3- and NH4), N-containing metabolites and morpho-physiological parameters of lettuce plants subjected to MPs/NPs. These findings underscore the intricate relationship between specific plastic contaminations, nitrogen dynamics, and plant performance.

Novel insights on the valorisation of onion (Allium cepa L.) are required to decipher its potential as nutritional and health-promoting agent. Wood vinegar (WV) was exogenously applied to onion plants to determine its effectiveness as an elicitor. An untargeted UHPLC-QTOF-MS metabolomics approach was combined with evaluating the bioactivities of onion extracts. WV promoted tissue and dose-specific effects on onion metabolome, involving both primary and secondary metabolites. Accordingly, the antioxidant and enzyme-inhibiting properties of onion were significantly enhanced after WV treatment. Moderate to high WV concentrations, ranging 250–500 µL L−1, were found optimal at conferring the highest rates of biological activities. Polyphenols were related to the bioactivities of onion extracts since phenolic acids, flavanols, and flavonols were suggested as the contributors to their health-enhancing properties. This approach provides insight into the characterisation of WV as a natural elicitor and opens wide perspectives on the research of onion tissues as biofactories of functional ingredients.

Elicited cell cultures of Taxus spp. are successfully used as sustainable biotechnological production systems of the anticancer drug paclitaxel, but the effect of the induced metabolomic changes on the synthesis of other bioactive compounds by elicitation has been scarcely studied. In this work, a powerful combinatorial approach based on elicitation and untargeted metabolomics was applied to unravel and characterize the effects of the elicitors 1 µM of coronatine (COR) or 150 µM of salicylic acid (SA) on phenolic biosynthesis in Taxus baccata cell suspensions. Differential effects on cell growth and the phenylpropanoid biosynthetic pathway were observed. Untargeted metabolomics analysis revealed a total of 83 phenolic compounds, mainly flavonoids, phenolic acids, lignans, and stilbenes. The application of multivariate statistics identified the metabolite markers attributed to elicitation over time: up to 34 compounds at 8 days, 41 for 16 days, and 36 after 24 days of culture. The most notable metabolic changes in phenolic metabolism occurred after 8 days of COR and 16 days of SA elicitation. Besides demonstrating the significant and differential impact of elicitation treatments on the metabolic fingerprint of T. baccata cell suspensions, the results indicate that Taxus ssp. biofactories may potentially supply not only taxanes but also valuable phenolic antioxidants, in an efficient optimization of resources.

Taxus cell cultures are a reliable biotechnological source of the anticancer drug paclitaxel. However, the interplay between taxane production and other metabolic pathways during elicitation remains poorly understood. In this study, we combined untargeted metabolomics and elicited Taxus baccata cell cultures to investigate variations in taxane-associated metabolism under the influence of 1 µM coronatine (COR) and 150 µM salicylic acid (SA). Our results demonstrated pleiotropic effects induced by both COR and SA elicitors, leading to differential changes in cell growth, taxane content, and secondary metabolism. Metabolite annotation revealed significant effects on N-containing compounds, phenylpropanoids, and terpenoids. Multivariate analysis showed that the metabolomic profiles of control and COR-treated samples are closer to each other than to SA-elicited samples at different time points (8, 16, and 24 days). The highest level of paclitaxel content was detected on day 8 under SA elicitation, exhibiting a negative correlation with the biomarkers kauralexin A2 and taxusin. Our study provides valuable insights into the intricate metabolic changes associated with paclitaxel production, aiding its potential optimization through untargeted metabolomics and an evaluation of COR/SA elicitor effects.

Individual health-related behavior is among the most influential yet modifiable factors affecting both climate change and chronic disease. To encourage behaviors bringing about environmental and health co-benefits, it is important to understand the underlying factors of behavior change for healthy and sustainable lifestyles. One area of potential overlap concerns people's health consciousness. The purpose of this study was to examine the relationship between health consciousness and pro-environmental behavior. We investigated whether health consciousness correlates with five clusters of pro-environmental behaviors: sustainable food consumption, recycling, green purchasing, sustainable mobility, and energy saving. Research data were collected via cross-sectional survey involving a representative sample of n = 1011 Italian citizens. Statistically significant differences emerged in the frequency of the different classes of pro-environmental behaviors: people living in Italy most frequently implement sustainable behaviors related to energy saving and recycling while sustainable mobility behaviors are the least implemented. Moreover, the stepwise linear regression model demonstrated the predictive role of citizens' health consciousness on the adoption of specific classes of pro-environmental behaviors showing how higher involvement in one's own health determines higher levels of pro-environmental behaviors. These results highlight the relevance of developing and testing complex programs featuring educational, sensitization, and structural strategies to increase citizens involvement in public health and pro-environmental behaviors.

The use of microbial biostimulants in agriculture is recognized as a sustainable approach to promoting crop productivity and quality due to improved nutrient uptake, enhanced stress tolerance, and improved ability to cope with non-optimal environments. The present paper aimed to comparatively investigate the effect of seven different commercial mycorrhizal-based treatments in terms of yield, phytochemical components, and technological traits of Malvasia di Candia Aromatica grape (Vitis vinifera L.) plants. Metabolomic analysis and photosynthetic performance were first investigated in leaves to point out biochemical differences related to plant growth. Higher photosynthetic efficiency and better PSII functioning were found in biostimulant-treated vines, reflecting an overall decrease in photoinhibition compared to untreated plants. Untargeted metabolomics followed by multivariate statistics highlighted a robust reprogramming of primary (lipids) and secondary (alkaloids and terpenoids) metabolites in treated plants. The analysis of berry yield and chemical components exhibited significant differences depending on the biostimulant product. Generally, berries obtained from treated plants displayed improved contents of polyphenols and sugars, while yield remained unchanged. These results elucidated the significant role of microbial biostimulants in determining the quality of grape berries and eliciting biochemical changes in vines.

Limited knowledge regarding the susceptibility of grape varieties to ochratoxin A (OTA)-producing fungi is available to date. This study aimed to investigate the susceptibility of different grape varieties to Aspergillus carbonarius concerning OTA contamination and modulation at the metabolome level. Six grape varieties were selected, sampled at early veraison and ripening, artificially inoculated with A. carbonarius, and incubated at two temperature regimes. Significant differences were observed across cultivars, with Barbera showing the highest incidence of moldy berries (around 30%), while Malvasia and Ortrugo showed the lowest incidence (about 2%). OTA contamination was the lowest in Ortrugo and Malvasia, and the highest in Croatina, although it was not significantly different from Barbera, Merlot, and Sauvignon Blanc. Fungal development and mycotoxin production changed with grape variety; the sugar content in berries could also have played a role. Unsupervised multivariate statistical analysis from metabolomic fingerprints highlighted cultivar-specific responses, although a more generalized response was observed by supervised OPLS-DA modeling. An accumulation of nitrogen-containing compounds (alkaloids and glucosinolates), phenylpropanoids, and terpenoids, in addition to phytoalexins, was observed in all samples. A broader modulation of the metabolome was observed in white grapes, which were less contaminated by OTA. Jasmonates and oxylipins were identified as critical upstream modulators in metabolomic profiles. A direct correlation between the plant defense machinery and OTA was not observed, but the information was acquired and can contribute to optimizing preventive actions.

Climate change and abiotic stress challenges in crops are threatening world food production. Among others, salinity affects the agricultural sector by significantly impacting yield losses. Plant biostimulants have received increasing attention in the agricultural industry due to their ability to improve health and resilience in crops. The main driving force of these products lies in their ability to modulate plant metabolic processes involved in the stress response. This study’s purpose was to investigate the effect of two biostimulant products, including a protein hydrolysate (Clever HX®) and a seaweed extract with high amino acids content (Ascovip®), and their combination, on the metabolomics profile of tomato crops grown under salt stress (150 mM NaCl). Several stress indicators (leaf relative water content, membrane stability index, and photosynthesis activity) and leaf mineral composition after salinity stress exposure were assessed to evaluate stress mitigation, together with growth parameters (shoot and root biomasses). After that, an untargeted metabolomics approach was used to investigate the mechanism of action of the biostimulants and their link with the increased resilience to stress. The application of the biostimulants used reduced the detrimental effect of salinity. In saline conditions, protein hydrolysate improved shoot dry weight while seaweed extracts improved root dry weight. Regarding stress indicators, the application of the protein hydrolysate was found to alleviate the membrane damage caused by salinity stress compared to untreated plants. Surprisingly, photosynthetic activity significantly improved after treatment with seaweed extracts, suggesting a close correlation between root development, root water assimilation capacity and photosynthetic activity. Considering the metabolic reprogramming after plant biostimulants application, protein hydrolysates and their combination with seaweed extracts reported a distinctive metabolic profile modula

Thymus comosus Heuff ex. Griseb. (Lamiaceae) is a wild thyme species endemic for Romanian Carpathian areas, frequently collected as substitute for collective herbal product Serpylli herba, cited as antibacterial and diuretic remedy in traditional medicine. The present study aimed to evaluate the in vivo diuretic effect and in vitro antimicrobial properties of three herbal preparations (infusion—TCI, tincture—TCT and an hydroethanolic extract prepared through an optimized ultrasound-assisted method—OpTC) obtained from the aerial parts of T. comosus Heuff ex. Griseb, also evaluating their comprehensive phenolic profile. In vivo diuretic effect was tested using Wistar rats treated orally with each herbal preparation (125 and 250 mg/kg dispersed in 25 ml/kg isotonic saline solution) and quantified based on cumulative urine output (ml), diuretic action and diuretic activity. Additionally, sodium and potassium excretion were monitored using a potentiometric method with selective electrodes. In vitro antibacterial and antifungal activities were assessed using p-iodonitrotetrazolium chloride assay against six bacterial strains and six fungal strains by monitoring minimum inhibitory concentration (MICs), minimum bactericidal concentrations (MBCs) and minimum fungicidal concentrations (MFCs). Finally, phenolic profile of the aforementioned herbal extracts was evaluated using an ultra-high-pressure liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) method to check the impact of the different preparations on the most abundant and significant compounds. All the extracts exerted a mild diuretic action, TCT and OpTC inducing the most intense diuretic effect. Both herbal preparations produced a statistically significant, dose-dependent and gradual increase of the urine output, the effect being more intense at 24 h (6.63–7.13 ml/24 h). Potentiometric evaluation of urine samples collected from treated rats revealed a clear and mild

Together with toxicity, beneficial effects on plant growth have been ascribed to nanoparticles (NPs). This study aimed to survey the growth performance and metabolome adjustment of beans grown in a growth medium containing ZnONPs at different concentrations and compared with bulk ZnSO4 as a positive control. Growth parameters showed a reduction in shoot height starting from the lowest (25 mg L−1) concentration of ZnONPs. In comparison, growth was inhibited from 50 mg L−1 ZnSO4, suggesting more toxic effects of nano forms of Zn. Untargeted metabolomics allowed us to unravel the biochemical processes involved in both promising and detrimental aspects. Multivariate statistics indicated that the tested Zn species substantially and distinctively altered the metabolic profile of both roots and leaves, with more metabolites altered in the former (435) compared with leaves (381). Despite having Zn forms in the growth medium, also leaf metabolome underwent a significant and extensive modulation. In general, the elicitation of secondary metabolism (N-containing compounds, phenylpropanoids, and phytoalexins) and the down-accumulation of fatty acid biosynthesis compounds were common responses to different Zn forms. However, an opposite trend could be observed for amino acids, fatty acids, carbohydrates, and cofactors being down-accumulated in ZnONPs treatment. Osmolytes, especially in ZnSO4 treatment, contributed to mitigating the effect of Zn toxicity and maintaining plant growth. Overall, the results indicated a complexity of tissue-specific and Zn-dependent response differences, resulting in distinctive metabolic perturbations.

Wheat is a prevalent food worldwide, although its production arises several tonnes of industrial by-products that merit valorisation, owing to sustainable demands. In this work, an untargeted metabolomics approach was applied to shed light on the profile of free and bound phenolic compounds in white and wholegrain flours, as well as the major wheat by-products, namely wheat bran, wheat shorts and wheat middlings. By-products and wholegrain flours were found to be rich sources of ferulic acid derivatives, lignans, and alkylresorcinols, mostly bound to wheat fibre components, whereas different flavonoids were found in their free forms. Afterwards, the in vitro gastrointestinal digestion of these matrices showed that flour phenolics, in particular lignans, were more bioaccessible than those from wheat by-products. These results support the carrier effect attributed to dietary fibre and open a wide perspective on the use of underexploited wheat by-products to formulate novel functional foods.

Wheat flour is one of the most prevalent foodstuffs for human consumption, and novel strategies are underway to enhance its nutritional properties. This work evaluated wholegrain flours from bread wheat lines with different amylose/amylopectin ratios through in vitro starch digestion and large intestine fermentation. High-amylose flours presented a higher resistant starch content and lower starch hydrolysis index. Moreover, UHPLC-HRMS metabolomics was carried out to determine the profile of the resulting in vitro fermentates. The multivariate analysis highlighted distinctive profiles between the flours derived from the different lines compared to the wild type. Peptides, glycerophospholipids, polyphenols, and terpenoids were identified as the main markers of the discrimination. The high-amylose flour fermentates showed the richest bioactive profile, containing stilbenes, carotenoids, and saponins. Present findings pave the way toward applying high-amylose flours to design novel functional foods.

The Camellia genus comprises a vast array of underexplored medicinal plants that merit a systematic valorization to exploit their potential as natural sources of phytochemicals with associated health-promoting properties. In this work, flower extracts from eight poorly characterized Camellia japonica L. cultivars were subjected to polyphenol profiling through untargeted metabolomics combined with in vitro functional analysis. Anthocyanins, mostly represented by cyanidin 3-O-glycosides, flavones, and flavonols, were found as the major constituents of C. japonica flowers, together with hydroxycinnamic acids, tyrosols, alkylphenols, and stilbenes, which were detected for the first time in this species. The application of multivariate statistics revealed a flower color-dependent fingerprint of C. japonica cultivars, featuring anthocyanins and other flavonoids as metabolite markers associated with color-flowered cultivars with respect to white-flowered ones. The accumulation of anthocyanins, especially reported in ‘Eugenia de Montijo’ flowers, was highly correlated with the cytotoxic and anti-inflammatory properties of the derived extracts, including AGS, Caco-2, and MCF7 cancer cell lines. Moreover, the flavones accumulation reported in ‘Carolyn Tuttle’ extracts was also associated with high rates of free-radical scavenging activity, as well as a potent cytotoxicity against the Caco-2 cell line. In general, C. japonica anthocyanin-enriched flower extracts were revealed as promising candidates for the industrial production of polyphenols with associated biological activities of high interest for critical sectors in the food, pharmaceutical, and cosmetic industries.

This work aimed to investigate the influence of high-power ultrasound (HPU) technology on the stability of bioactive compounds in strawberry juices obtained from fruits with different stages of ripeness (75% vs. 100%) and stored at 4 degrees C for 7 days. HPU parameters were amplitude (25, 50, 75, and 100%), pulses (50 vs. 100%) and treatment time (5 vs. 10 min). Amplitude and pulse had a significant effect (p <= 0.05) on all bioactive compounds except flavonols and hydroxycinnamic acids. The treatment duration of 5 min vs. 10 min had a significant positive impact on the content of anthocyanins, flavonols and condensed tannins, while the opposite was observed for total phenols, whereas no statistically significant effect was observed for hydroxycinnamic acids. The temperature changes during HPU treatment correlated positively with almost all HPU treatment parameters (amplitude, pulse, energy, power, frequency). Optimal parameters of HPU were obtained for temperature changes, where the highest content of a particular group of bioactive compounds was obtained. Results showed that by combining fruits with a certain ripeness and optimal HPU treatment, it would be possible to produce juices with highly preserved bioactive compounds, while HPU technology has prospects for application in functional food products.

The metabolic composition of thyme, one of the most used aromatic herbs, is influenced by environmental and post-harvest processing factors, presenting the possibility of exploiting thyme fingerprint to assess its authenticity. In this study, a comprehensive UHPLC-QTOF-HRMS fingerprinting approach was applied with a dual objective: (1) tracing thyme from three regions of production (Spain, Morocco, and Poland) and (2) evaluating the metabolic differences in response to processing, considering sterilized thyme samples. Multivariate statistics reveal 37 and 33 key origin and processing differentiation compounds, respectively. The findings highlighted the remarkable “terroir” influence on thyme fingerprint, noticing flavonoids, amino acids, and peptides among the most discriminant chemical classes. Thyme sterilization led to an overall metabolite enrichment, most likely due to the facilitated compound accessibility as a result of processing. The findings provide a comprehensive metabolomics insight into the origin and processing effect on thyme composition for product traceability and quality assessment.

The application of microbial biostimulants to plants has revealed positive effects related to nutrients uptake, stress tolerance, root development and phenological growth. However, little information is available exploiting the potential synergistic biostimulant action of microbes on the functional quality of the yields. The current research elucidated the effect of single or coupled action of biostimulants, associated with either optimal or reduced nitrogen application, on the functional quality of tomato fruits. Chemical assays and untargeted metabolomics were applied to investigate Rhizoglomus irregulare and Funneliformis mosseae administration (both being arbuscular mycorrhiza, AMF), under optimal or low N input conditions, alone or coupled to Trichoderma atroviride application. The coupling of AMF and Trichoderma fungal inoculations resulted in a synergistic biostimulant effect on tomato fruits under sub-optimal fertility, revealing improved concentrations of carotenoid compounds-B-carotene (0.647 +/- 0.243 mg/100 g), Z-carotene (0.021 +/- 0.021 mg/100 g), 13-z-lycopene (0.145 +/- 0.052 mg/100 g) and all-trans-lycopene (12.586 +/- 1.511 mg/100 g), and increased values for total phenolic content (12.9 +/- 2.9 mgGAE/g), total antioxidant activity (phosphomolybdenum, 0.9 +/- 0.2 mmolTE/g), radical scavenging activity (DPPH, 3.4 +/- 3.7 mgTE/g), reducing power (FRAP, 23.6 +/- 6.3 mgTE/g and CUPRAC, 37.4 +/- 7.6 mg TE/g), and enzyme inhibitory activity (AChE, 2.4 +/- 0.1 mg GALAE/g), when compared to control. However, evidence of carotenoid and bioactive compounds were exclusively observed under the sub-optimal fertility and no significant differences could be observed between the biostimulant treatment and control under optimal fertility.

A multi-omics approach was adopted to investigate the impact of lactic acid fermentation and seed germination on the composition and physicochemical properties of rye doughs. Doughs were prepared with either native or germinated rye flour and fermented with Saccharomyces cerevisiae, combined or not with a sourdough starter including Limosilactobacillus fermentum, Weissella confusa and Weissella cibaria. LAB fermentation significantly increased total titrable acidity and dough rise regardless of the flour used. Targeted metagenomics revealed a strong impact of germination on the bacterial community profile of sprouted rye flour. Doughs made with germinated rye displayed higher levels of Latilactobacillus curvatus, while native rye doughs were associated with higher proportions of Lactoplantibacillus plantarum. The oligosaccharide profile of rye doughs indicated a lower carbohydrate content in native doughs as compared to the sprouted counterparts. Mixed fermentation promoted a consistent decrease in both monosaccharides and low-polymerization degree (PD)-oligosaccharides, but not in high-PD carbohydrates. Untargeted metabolomic analysis showed that native and germinated rye doughs differed in the relative abundance of phenolic compounds, terpenoids, and phospholipids. Sourdough fermentation promoted the accumulation of terpenoids, phenolic compounds and proteinogenic and non-proteinogenic amino acids. Present findings offer an integrated perspective on rye dough as a multi-constituent system and on cereal-sourced bioactive compounds potentially affecting the functional properties of derived food products.

Plants have evolved diverse plant-species specific tolerance mechanisms to cope with salt stress. However, these adaptive strategies often inefficiently mitigate the stress related to increasing salinity. In this respect, plant-based biostimulants have gained increasing popularity since they can alleviate deleterious effects of salinity. Hence, this study aimed to evaluate the sensitivity of tomato and lettuce plants grown under high salinity and the possible protective effects of four biostimulants based on vegetal protein hydrolysates. Plants were set in a 2 × 5 factorial experimental design completely randomized with two salt conditions, no salt (0 mM) and high salt (120 mM for tomato or 80 mM for lettuce), and five biostimulant treatments (C: Malvaceae-derived, P: Poaceae-derived, D: Legume-derived commercial ‘Trainer®’, H: Legume-derived commercial ‘Vegamin®’, and Control: distilled water). Our results showed that both salinity and biostimulant treatments affected the biomass accumulation in the two plant species, albeit to different extents. The salinity stress induced a higher activity of antioxidant enzymes (e.g., catalase, ascorbate peroxidase, guaiacol peroxidase and superoxide dismutase) and the overaccumulation of osmolyte proline in both lettuce and tomato plants. Interestingly, salt-stressed lettuce plants showed a higher accumulation of proline as compared to tomato plants. On the other hand, the treatment with biostimulants in salt-stressed plants caused a differential induction of enzymatic activity depending on the plant and the biostimulant considered. Overall, our results suggest that tomato plants were constitutively more tolerant to salinity than lettuce plants. As a consequence, the effectiveness of biostimulants in alleviating high salt concentrations was more evident in lettuce. Among the four biostimulants tested, P and D showed to be the most promising for the amelioration of salt stress in both the plant species, thereby suggesting their

Extra-virgin olive oil (EVOO) is subjected to different frauds. This work aimed at integrating the untargeted phenolic and sterol signatures with supervised multivariate discriminant analysis (OPLS-DA) and Artificial Neural Networks (ANN) for tracing the authenticity (as a function of variety, origin, and the blending) of Taggiasca Ligure, a renowned Italian EVOO. Overall, 408 samples from three consecutive growing seasons were used. Despite the cultivar, season, growth altitude, and geographical origin were all contributing to phytochemical profile, OPLS-DA models allowed identifying specific markers of authenticity. Cholesterol-derivatives and phenolics (tyrosols and oleuropeins, stilbenes, lignans, phenolic acids, and flavonoids) were the best markers, based on statistics. Thereafter, ANN was used to discriminate authentic Taggiasca, and the sensitivity was 100% (32/32) thus indicating an excellent classification. Our results strengthen the concept of “terroir” for EVOO and indicate that profiling sterols and phenolics can support EVOO integrity if adequate data treatments are adopted.

The current consumers’ demand for food naturalness is urging the search for new functional foods of natural origin with enhanced health-promoting properties. In this sense, algae constitute an underexplored biological source of nutraceuticals that can be used to fortify food products. Both marine macroalgae (or seaweeds) and microalgae exhibit a myriad of chemical constituents with associated features as a result of their primary and secondary metabolism. Thus, primary metabolites, especially polysaccharides and phycobiliproteins, present interesting properties to improve the rheological and nutritional properties of food matrices, whereas secondary metabolites, such as polyphenols and xanthophylls, may provide interesting bioactivities, including antioxidant or cytotoxic effects. Due to the interest in algae as a source of nutraceuticals by the food and related industries, novel strategies should be undertaken to add value to their derived functional components. As a result, metabolomics is considered a high throughput technology to get insight into the full metabolic profile of biological samples, and it opens a wide perspective in the study of algae metabolism, whose knowledge is still little explored. This review focuses on algae metabolism and its applications in the food industry, paying attention to the promising metabolomic approaches to be developed aiming at the functional characterization of these organisms.

Untargeted metabolomics based on ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry was combined with sensory analysis to provide new insights into the impact of the feeding system from mountain regions (grassland deriving from permanent meadows) on the chemical fingerprint of Parmigiano Reggiano PDO hard cheese. In the framework of a representative investigation, two different ripening times (12 and 24 months) were also considered. Multivariate statistics allowed discriminating cheese samples from different feeding regimens according to their metabolomics signatures. Interestingly, mountain grassland-based cheese samples were characterized by a more favourable fatty acid profile, recording also feed-derived compounds (such as terpenoids and linoleic acid derivatives) potentially associated with both beneficial effects on human health and sensory properties. According to the sensory analysis, the impact of herbs and grass enhanced the colour and retro-olfactive complexity of Parmigiano Reggiano PDO cheese, with spicy, umami and intense vegetal aromatic notes representing distinctive features.

The modulation of plant secondary metabolism by the manipulation of nutrient solution (NS) electrical conductivity and biostimulant application has the potential to increase crop growth and the content of bioactive compounds, thus ameliorating the qualitative attributes of vegetables. A Graminaceae-derived protein hydrolysate (pH) and its molecular fractions PH1 (>10 kDa), PH2 (1 < x <10 kDa) and PH3 (<1 kDa) were applied on lettuce grown in a soilless system and irrigated with two levels of NaCl in the NS (0 and 30 mM). The different pH fractions provided distinct responses, with PH2 in saline conditions increasing lettuce fresh weight by 10.48%, and decreased Na, Cl, malate and citrate concentrations in the leaves by 37.24%, 15.45%,31.4% and 33.73% compared to control. Similar results were achieved by the fraction PH3. However, pH and its fractions PH2 and PH3 increased total flavonoids in salinity conditions, while pH and its fraction PH3 increased total phenolic acids in both saline and non-saline conditions. At the same time, all the 3 fractions of pH increased lutein in both conditions. Therefore, pH fractions should be casted, under tailored conditions, to achieve the desired growth and modulation of qualitative attributes. Nonetheless, omic approaches should be further recommended to support and clarify the mechanisms of these fractions.

Metallic nanoparticles (MNPs) are becoming widespread environmental contaminants. They are currently added to several food preparations and cause a fast-growing concern for human health. The present work aims to assess the impact of zinc oxide (ZnO), titanium dioxide (TiO2), and silver (Ag) nanoparticles (NPs) on the human gut metabolome and microbiome. Water samples spiked with two different concentrations of each MNPs were subjected to in-vitro gastrointestinal digestion and in-vitro large intestine fermentation. The effects of the treatments were determined through 16 S amplicon sequencing and untargeted metabolomics. Multi-omics data integration was then applied to correlate the two datasets. MNPs treatments modulated the microbial genera Bifidobacterium, Sutterella, Escherichia and Bacteroides. The treatments, especially the lower concentrations of Ag and ZnO, caused modulation of indole derivatives, peptides, and metabolites related to protein metabolism in the large intestine. Notably, these metabolites are implicated in ulcerative colitis and inflammatory bowel disease. TiO2 NPs treatment in all concentrations increased E.coli relative abundance and decreased the abundance of B. longum. Moreover, for TiO2, an enrichment in proinflammatory lipid mediators of arachidonic acid metabo-lites, such as prostaglandin E2 and leukotrienes B4, was detected. For all metals except TiO2, low NP concen-trations promoted differentiated profiles, thus suggesting that MNPs aggregation can limit adverse effects on living cells.

In the current climate change scenario, water stress is a serious threat to limit crop growth and yields. It is necessary to develop tolerant plants that cope with water stress and, for this purpose, tolerance mechanisms should be studied. NIBER® is a proven water stress- and salt-tolerant pepper hybrid rootstock (Gisbert-Mullor et al., 2020; López-Serrano et al., 2020), but tolerance mechanisms remain unclear. In this experiment, NIBER® and A10 (a sensitive pepper accession (Penella et al., 2014)) response to short-term water stress at 5 h and 24 h was studied in terms of gene expression and metabolites content in roots. GO terms and gene expression analyses evidenced constitutive differences in the transcriptomic profile of NIBER® and A10, associated with detoxification systems of reactive oxygen species (ROS). Upon water stress, transcription factors like DREBs and MYC are upregulated and the levels of auxins, abscisic acid and jasmonic acid are increased in NIBER®. NIBER® tolerance mechanisms involve an increase in osmoprotectant sugars (i.e., trehalose, raffinose) and in antioxidants (spermidine), but lower contents of oxidized glutathione compared to A10, which indicates less oxidative damage. Moreover, the gene expression for aquaporins and chaperones is enhanced. These results show the main NIBER® strategies to overcome water stress.

Lettuce (Lactuca sativa L., Asteraceae) is a popular vegetable leafy crop playing a relevant role in human nutrition. Nowadays, novel strategies are required to sustainably support plant growth and elicit the biosynthesis of bioactive molecules with functional roles in crops including lettuce. In this work, the polyphenolic profile of lettuce treated with glutamic acid (GA), humic acid (HA), and their combination (GA + HA) was investigated using an untargeted metabolomics phenolic profiling approach based on high-resolution mass spectrometry. Both aerial and root organ parts were considered, and a broad and diverse phenolic profile could be highlighted. The phenolic profile included flavonoids (anthocyanins, flavones, flavanols, and flavonols), phenolic acids (both hydroxycinnamics and hydroxybenzoics), low molecular weight phenolics (tyrosol equivalents), lignans and stilbenes. Overall, GA and HA treatments significantly modulated the biosynthesis of flavanols, lignans, low molecular weight phenolics, phenolic acids, and stilbene. Thereafter, antioxidant capacity was evaluated in vitro with 2,2-diphenyln-1-picrylhydrazyl (DPPH), 2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), and cupric ion reducing antioxidant capacity (CUPRAC) assays. In addition, this study examined the inhibitory properties of enzymes, including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), tyrosinase, alpha-amylase, and alpha-glucosidase. Compared to individual treatments, the combination of GA + HA showed stronger antioxidant abilities in free radical scavenging and reducing power assays in root samples. Moreover, this combination positively influenced the inhibitory effects of root samples on AChE and BChE and the tyrosinase inhibitory effect of leaf samples. Concerning Pearson’s correlations, antioxidant and enzyme inhibition activities were related to phenolic compounds, and lignans in particular correlated with radica

In the last years, animal proteins from meat and meat-derived products have represented one of the main concerns to achieve a sustainable food production. Within this perspective, the partial replacement of meat with nonmeat substances having a high-protein content, offers interesting opportunities to reformulate more sustainable meat products characterized also by potential health-promoting properties. Therefore, starting from these background conditions, this short review critically summarizes recent findings on extenders from different origins, namely pulses, plant-based ingredients, plant by-products, and unconventional sources, as valuable possibility to improve the technological profile and functional quality of meat, together with providing a focus on their ability to affect the sustainability of meat products.

In addition to be used as a plant protection agent, copper (Cu) is also an essential micronutrient for plant growth and development. The bioavailability of Cu in agricultural systems can be limited due to its specific physical–chemical characteristics, leading to imbalances in plant production. To address this issue, an experimental trial was conducted on Genovese basil (Ocimum basilicum L.) in protected conditions to comparatively evaluate the effects of a vegetable protein hydrolysate (VPH), free Cu and Cu complexed with peptides and amino acids of vegetal origin (Cu and Cu-VPH, respectively), and a combination of VPH and Cu-VPH (VPH+Cu-VPH). The study showed that the combined application of VPH+Cu-VPH led to a significant average increase of 16.3% in fresh yield compared to the untreated Control and Cu treatment. This finding was supported by an improved photosynthetic performance in ACO2 (+29%) and Fv/Fm (+7%). Furthermore, mineral analysis using ICP OES demonstrated that Cu and Cu-VPH treatments determined, on average, a 15.1-, 16.9-, and 1.9-fold increase in Cu in plant tissues compared to control, VPH, and VPH+Cu-VPH treatments, respectively. However, the VPH+Cu-VPH treatment induced the highest contents of the other analyzed ions, except for P. In particular, Mg, Mn, Ca, and Fe, which take part in the constitution of chlorophylls, water splitting system, and photosynthetic electron transport chain, increased by 23%, 21%, 25%, and 32% compared to respective controls. Indeed, this improved the photosynthetic efficiency and the carboxylation capacity of the plants, and consequently, the physiological and productive performance of Genovese basil, compared to all other treatments and control. Consistently, the untargeted metabolomics also pointed out a distinctive modulation of phytochemical signatures as a function of the treatment. An accumulation of alkaloids, terpenoids, and phenylpropanoids was observed following Cu treatment, suggesting an oxidative imbalan

Lentils (Lens culinaris spp.) are promising legumes whose seed coats, the by-products produced from dehulling, can represent a valuable source of functional components. This work aimed at valorizing lentils seed coat as functional ingredients, by investigating its profile across 11 genotypes grown in two different growing locations in Italy. The total phenolic, flavonoids, and condensed tannins contents (mg/g dry weight,) ranged from 30.60 to 70.37 gallic acid equivalents, 0.86 to 2.21 catechin equivalents, and 22.34 to 77.49 catechin equivalents, respectively. The untargeted phenolic profiling through UHPLC/QTOF-MS, followed by multivariate statistics, revealed differences in the quality and quantity of polyphenols. A broad and diverse profile could be highlighted, including 420 compounds, mainly ascribable to flavonoids, phenolic acids, and low-molecular-weight phenolic compounds. Unsupervised cluster analysis highlighted that origin is hierarchically more important than genotype in determining the phenolic profiles. Pearson’s correlation analysis showed that color was only partially related to phenolic content, while the geographic location and the genotype were more relevant. The significant accumulation of phenolics in some genotypes suggests that the genetic background should be taken into account, when lentils seed coat is to be included in food design as a sustainable source of phenolic compounds.

Root exudates play an essential role in helping plants to cope with abiotic stress. However, the modulation of root exudation profiles under multiple stress conditions is still poorly understood. Using targeted and untargeted metabolomics, here we tested the effect of drought, heat stress, and their combination on maize root exudates, also considering the differences that might exist between root types (seminal and primary) and root zones (apical and sub-apical). In addition, we built an analytical framework that relates the root exudation profile with the rhizosphere bacterial community, enabling us to dissect the interactions between specific root exudates and bacterial taxa. The composition of root exudates undergoes distinct modulation according to the single or combined stress and to the root zone, but not according to the root type. In addition, we found that stress-specific exudates can influence the relative abundance of specific bacterial taxa, some of which are known to be beneficial microorganisms. Our results contribute to the understanding of plant-soil interactions under the influence of abiotic stressors, which is key in paving the way towards an increased resilience to abiotic stresses, representing a powerful tool to craft the next generation of agricultural practices.

High nursery densities reduce the seedling quality due to the competition for light. High light intensity, shading, and blue light depletion activate morphophysiological and metabolomic responses in plants, resulting in size modification to gain an advantage over neighboring plants. Our research aimed to unravel the effects of light intensity and quality on nursery seedlings at the morphological and biochemical levels. To this aim, the effect of black shading and blue photoselective shading nets were investigated in terms of morphometric, ionomic, and untargeted metabolomics signatures in Cucurbita pepo L., Citrullus lanatus L., Solanum lycopersicum L., and Solanum melongena L. seedlings. Plant height, diameter, sturdiness index, leaf area, specific leaf area, shoot/root ratio, and mineral content (by ion chromatography-IC) were evaluated. In C. pepo L and C. lanatus L., the blue net reduced the shoot/root and chlorophyll a/b ratios and increased stem diameter and total chlorophyll content. The black net increased plant height, stem diameter, and sturdiness index in Solanum lycopersicum L. and Solanum melongena L. At the same time, unshading conditions reduced leaf area, specific leaf area, shoot/root ratio, and total chlorophyll content. The blue net improved the sturdiness index and quality of C. pepo L. and C. lanatus L. Such impact on morphological parameters induced by the different shading conditions was corroborated by a significant modulation at the metabolomics level. Untargeted metabolomic phytochemical signatures of the selected plants, and the subsequent multivariate analysis coupled to pathway analysis, allowed highlighting a broad and diverse biochemical modulation. Metabolomics revealed that both primary and secondary metabolism were largely affected by the different shading conditions, regardless of the species considered. A common pattern arose to point at the activation of plant energy metabolism and lipid biosynthesis, together with a generalized

Increased soil salinity is one of the main concerns in agriculture and food production, and it negatively affects plant growth and crop productivity. In order to mitigate the adverse effects of salinity stress, plant biostimulants (PBs) have been indicated as a promising approach. Indeed, these products have a beneficial effect on plants by acting on primary and secondary metabolism and by inducing the accumulation of protective molecules against oxidative stress. In this context, the present work is aimed at comparatively investigating the effects of microbial (i.e., Azospirillum brasilense) and plant-derived biostimulants in alleviating salt stress in tomato plants by adopting a multidisciplinary approach. To do so, the morphological and biochemical effects were assessed by analyzing the biomass accumulation and root characteristics, the activity of antioxidant enzymes and osmotic stress protection. Furthermore, modifications in the metabolomic profiles of both leaves and root exudates were also investigated by ultra-high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS). According to the results, biomass accumulation decreased under high salinity. However, the treatment with A. brasilense considerably improved root architecture and increased root biomass by 156% and 118% in non-saline and saline conditions, respectively. The antioxidant enzymes and proline production were enhanced in salinity stress at different levels according to the biostimulant applied. Moreover, the metabolomic analyses pointed out a wide set of processes being affected by salinity and biostimulant interactions. Crucial compounds belonging to secondary metabolism (phenylpropanoids, alkaloids and other N-containing metabolites, and membrane lipids) and phytohormones (brassinosteroids, cytokinins and methylsalicylate) showed the most pronounced modulation. Overall, our results suggest a better performance of A. brasilense in alleviating high salinity

Saffron production from Crocus sativus flowers produces large amounts of by-products that may represent an excellent source of polyphenols. The aim of this work was to evaluate infusions originating from different brewing processes and from different saffron flower portions, in terms of both functional and sensory traits. For this aim, total polyphenols and total flavonoids, in vitro antioxidant assays and an untargeted phenolic profiling were applied. In general, tepals showed higher polyphenol and flavonoid content than stamen infusions, and their bioactive content depended more on brewing temperature than brewing time. These findings were consistent with both antioxidant capacity and phenolic profiling. Multivariate statistics highlighted polyphenols discriminating ‘boiled’ vs. ‘cold’ infusions, being mainly flavonoids, phenolic acids and the alkylphenol 5-pentadecylresorcinol (showing a strong down-accumulation at the higher brewing temperatures). Positive correlations could be highlighted between anthocyanins, flavones, flavonols and lignans, and the in vitro antioxidant assays. In general, cold brewing was successful in extracting phenolic compounds and provided better sensory properties, thus indicating that this may represent a valuable strategy to develop saffron-based functional beverages with better consumers' acceptability.

Plant phenomics is becoming a common tool employed to characterize the mode of action of biostimulants. A combination of this technique with other omics such as metabolomics can offer a deeper understanding of a biostimulant effect in planta. However, the most challenging part then is the data analysis and the interpretation of the omics datasets. In this work, we present an example of how different tools, based on multivariate statistical analysis, can help to simplify the omics data and extract the relevant information. We demonstrate this by studying the effect of protein hydrolysate (PH)-based biostimulants derived from different natural sources in lettuce and tomato plants grown in controlled conditions and under salinity. The biostimulants induced different phenotypic and metabolomic responses in both crops. In general, they improved growth and photosynthesis performance under control and salt stress conditions, with better performance in lettuce. To identify the most significant traits for each treatment, a random forest classifier was used. Using this approach, we found out that, in lettuce, biomass-related parameters were the most relevant traits to evaluate the biostimulant mode of action, with a better response mainly connected to plant hormone regulation. However, in tomatoes, the relevant traits were related to chlorophyll fluorescence parameters in combination with certain antistress metabolites that benefit the electron transport chain, such as 4-hydroxycoumarin and vitamin K1 (phylloquinone). Altogether, we show that to go further in the understanding of the use of biostimulants as plant growth promotors and/or stress alleviators, it is highly beneficial to integrate more advanced statistical tools to deal with the huge datasets obtained from the -omics to extract the relevant information.

The use of plant biostimulants such as seaweed extracts (SWE) and protein hydrolysates (PH) has grown in the recent years due to their beneficial effects on yield under both optimal and sub-optimal conditions such as salt stress. The comprehension of the mode of action of these two important categories of biostimulants on plant performance will allow to use them more efficiently under different growth conditions. This study aimed to examine the efficacy of a seaweed and plant-based biostimulants on greenhouse lettuce (Lactuca sativa L.) grown under non-saline (0 mM NaCl) and saline conditions (40 mM NaCl) in terms of growth, yield, SPAD index, leaf mineral composition and metabolomic profiling. Shoot fresh weight of lettuce was reduced by 15.3% under 40 mM of NaCl. Nonetheless, it was boosted by both used biostimulants by 9 and 18%, on average, under 0 and 40 mM NaCl salinity, respectively. Na content under saline conditions was reduced in the presence of the biostimulants treatment, where PH reduced it significantly by 15.6% and SWE by 9.4%. On the other hand, Cl content was significantly reduced only under PH treatment. Both biostimulants elicited a broad metabolic reprogramming, involving the accumulation of stress-related compounds such as glucosinolates, terpenoid phytoalexins, and jasmonates. Interestingly, distinctive metabolomic signatures could be observed following the application of the different biostimulants under salinity conditions. In more detail, PH promoted the accumulation of glucosinolates and phytoalexins precursors, while SWE induced a down accumulation of secondary metabolites. Our findings indicate different processes being modulated by PH and SWE, with possible synergistic effects, thus paving the way towards integrated strategies to alleviate the detrimental effects of salinity in lettuce.

Microplastics (MP) are ubiquitous contaminants of great concern due to their high persistence and potential hazardous impact on the environment. Depending on their size and shape, as well as the chemical additives they can have in their polymeric structure, MP can be taken up by organisms, ultimately leading to direct and indirect toxic effects. In this review, we discuss the primary sources, fate, and impact of MP on the rhizosphere ecology, focusing in particular on how soil physical-chemical properties, plant physiology, and soil biodiversity are modulated by the interaction of MP in the plant-soil system. Current knowledge on soil ecotoxicology shows that MP directly affects soil quality and fertility via alteration of soil nutrient cycling and microbial communities and indirectly by changing soil bulk density, pH, porosity, electric conductivity, and nutrient bioavailability. MP is also known to affect soil animals by altering their feeding, mobility, and reproductive behavior. Toxic effects of MP on the multiple and co-occurring interactions among the soil, soil organisms and plants, particularly in the long-term, remain unstudied. Indeed, a better understanding of rhizosphere-MP interactions at a functional level (e.g., nutrient availability, pollutant immobilization, root exudates, etc.) is urgently needed to develop risk assessment frameworks of soil pollution by MP.

Several studies have demonstrated that stigmas of Crocus sativus contain several bioactive compounds with potential health-promoting properties. However, during the processing of stigmas, large amounts of floral bio-residues are normally discarded as by-products. In this study, using untargeted metabolomics, the comprehensive phytochemical composition of C. sativus stigma and tepals was investigated. Moreover, the antibacterial and anti-biofilm properties of the extracts of C. sativus stigmas and tepals were compared. The study was carried out using two methicillin-resistant staphylococcal reference strains (i.e., Staphylococcus aureus ATCC 43300 and Staphylococcus epidermidis ATCC 35984), representing important Gram-positive biofilm-forming human pathogens. The antibacterial properties were correlated with total polyphenol content, total terpenoid content, and in vitro antioxidant properties of tepals and stigmas. The results demonstrated that stigma and tepal extracts, at the sub-Toxic concentrations, were able to interfere with biofilm formation by ATCC 43300 and ATCC 35984. Besides, the higher antibacterial activity of tepals than stigmas was associated with higher levels of phycompounds. Therefore, our results demonstrated that C. sativus stigmas and bio-residues, such as tepals, are potential antioxidant sources and good candidates as antibacterial agents to prevent biofilm formation. Taken together, these findings showed that C. sativus could be used as functional ingredient by the food and pharmaceutical industries.

Tomato (Solanum lycopersicum L.) is globally recognised as a high-value crop both for commercial profit and nutritional benefits. In contrast to the extensive data regarding the changes in the metabolism of tomato fruit exposed to UV radiation, less is known about the foliar and root metabolome. Using an untargeted metabolomic approach through UHPLC-ESI-QTOF-MS analysis, we detected thousands of metabolites in the leaves (3000) and roots (2800) of Micro-Tom tomato plants exposed to 11 days of short daily UV radiation, applied only on the aboveground organs. Multivariate statistical analysis, such as OPLS-DA and volcano, were performed to allow a better understanding of the modifications caused by the treatment. Based on the unexpected modulation to the secondary metabolism, especially the phenylpropanoid pathway, of which compounds were down and up accumulated respectively in leaves and roots of treated plants, a phenolic profiling was carried out for both organs. The phenolic profile was associated with a gene expression analysis to check the transcription trend of genes involved in the UVR8 signalling pathway and the early steps of the phenolic biosynthesis. The retention of the modifications at metabolic and phenolic levels was also investigated 3 days after the UV treatment, showing a prolonged effect on the modulation once the UV treatment had ceased.

Thyme is one of Europe's most consumed aromatic herbs and represents a matrix susceptible to intentional mislabelling and food frauds. In this study, a phenolic profiling approach based on UHPLC-QTOF-HRMS untargeted metabolomics was used to trace its geographical origin, as well as to assess the effect of post-harvest processing by comparing sterilized vs non-sterilized thyme. Both unsupervised and supervised statistics led to reliable sample clustering, high-quality model parameters, as well as the identification of a total of 45 differential compounds (markers) for discrimination purposes. The phenolic signature was markedly affected by environmental conditions related to the region of production, leading to an overall higher abundance of flavonoids in Moroccan thyme (from Fez), flavanols in Polish one (from Lublin), and tyrosols and other phenolics in thyme cultivated in Spain (from Castilla-La Mancha). The processing was also shown to play an important role in phenolic profiling, noticing not only the decrease of thermolabile phenolics (such as flavonoids) but also the enhancement of other phenolic subfamilies in response to sterilization. This study opens the path to novel metabolomics-based strategies to support the integrity of thyme and possibly other spices, scarcely studied so far.

Biostimulants are gaining increasing interest because of their ability to provide a green and effective strategy towards sustainable crop production. Nonetheless, their mode of action remains often unknown. The object of this work was to unravel the mechanisms through which 4-Vita, a biostimulant plant extract, can mitigate drought stress in tomato. To this aim, tomato plants were treated with two foliar applications of 4-Vita and drought stress imposed to both treated and control plants. Phenomics investigations were coupled to mass spectrometric untargeted metabolomics, and raw data were elaborated by multivariate statistics and pathway analysis. The biostimulant elicited a broad reprogramming of the tomato’s secondary metabolism, including its phytohormones profile, corroborating an improved ability to cope with drought stress. A series of mechanisms could be identified in response to the biostimulant treatment under drought, pointing to the preservation of photosynthetic machinery functionality. The modulation of thylakoid membrane lipids, the increase in xanthins involved in ROS detoxification, and the modulation of chlorophylls synthesis could also be observed. Overall, a series of coordinated biochemical mechanisms were elicited by the biostimulant treatment, supporting the increased resilience to drought stress in tomato.

An optimized ultrasound-assisted extractive method was developed to obtain a polyphenol-enriched extract from the aerial parts of Thymus comosus Heuff. ex Griseb. et Schenk. Optimization process was conducted based on Design of Experiment (DoE) principles, determining the influence of three independent variables (time, ultrasound amplitude, ethanol concentration) on the total phenolic content of the extract (dependent variable). Additionally, the phenolic composition of the extract was characterized through UHPLC-HRMS, revealing beside the most abundant flavonoid-type compounds the presence of salvianolic acids C, D and L in high amounts. Phytochemical profile of the extract was correlated with its antioxidant activity (tested through five complementary assays) and enzyme-inhibitory potential, showing important antiglucosidase and anticholinesterase effects. Overall, it was concluded that the developed method is suitable for obtaining a good recovery of both phenolic and non-phenolic compounds from Thymus comosus aerial parts, and their presence in the optimized extract is responsible for its pharmacological potential.

Spices, widely used to improve the sensory characteristics of food, contain several bioactive compounds as well, including polyphenols, carotenoids, and glucosynolates. Acting through multiple pathways, these bioactive molecules affect a wide variety of cellular processes involved in molecular mechanisms important in the onset and progress of human diseases. Capparis spinosa L. is an aromatic plant characteristic of the Mediterranean diet. Previous studies have reported that different parts (aerial parts, roots, and seeds) of C. spinosa exert various pharmacological activities. Flower buds of C. spinosa contain several bioactive compounds, including polyphenols and glucosinolates. Two different subspecies of C. spinosa L., namely, C. spinosa L. subsp. spinosa, and C. spinosa L. subsp. rupestris, have been reported. Few studies have been carried out in C. spinosa L. subsp. rupestris. The aim of our study was to investigate the phytochemical profile of floral buds of the less investigated species C. spinosa subsp. rupestris. Moreover, we investigated the effect of the extract from buds of C. spinosa subsp. rupestris (CSE) on cell proliferation, intracellular ROS levels, and expression of the antioxidant and anti-apoptotic enzyme paraoxonase-2 (PON2) in normal and cancer cells. T24 cells and Caco-2 cells were selected as models of advanced-stage human bladder cancer and human colorectal adenocarcinoma, respectively. The immortalized human urothelial cell line (UROtsa) and human dermal fibroblast (HuDe) were chosen as normal cell models. Through an untargeted metabolomic approach based on ultra-high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF-MS), our results demonstrate that C. spinosa subsp. rupestris flower buds contain polyphenols and glucosinolates able to exert a higher cytotoxic effect and higher intracellular reactive oxygen species (ROS) production in cancer cells compared to normal cells. Moreover, upregulation of

Novel approaches to the characterization of medicinal plants as biofactories have lately increased in the field of biotechnology. In this work, a multifaceted approach based on plant tissue culture, metabolomics, and machine learning was applied to decipher and further characterize the biosynthesis of phenolic compounds by eliciting cell suspension cultures from medicinal plants belonging to the Bryophyllum subgenus. The application of untargeted metabolomics provided a total of 460 phenolic compounds. The biosynthesis of 164 of them was significantly modulated by elicitation. The application of neurofuzzy logic as a machine learning tool allowed for deciphering the critical factors involved in the response to elicitation, predicting their influence and interactions on plant cell growth and the biosynthesis of several polyphenols subfamilies. The results indicate that salicylic acid plays a definitive genotype-dependent role in the elicitation of Bryophyllum cell cultures, while methyl jasmonate was revealed as a secondary factor. The knowledge provided by this approach opens a wide perspective on the research of medicinal plants and facilitates their biotechnological exploitation as biofactories in the food, cosmetic and pharmaceutical fields.

Arbuscular mycorrhizal fungi (AMF) have gained remarkable importance, having been proved to alleviate drought stress-induced damage in wheat due to their ability to ameliorate plant water use efficiency and antioxidant enzyme activity. However, despite the current relevance of the topic, the molecular and physiological processes at the base of this symbiosis never consider the single cultivar affinity to mycorrhization as an influencing factor for the metabolic response in the AMF-colonized plant. In the present study, the mycorrhizal affinity of two durum wheat species (T. turgidum subsp. durum (Desf.)) varieties, Iride and Ramirez, were investigated. Successively, an untargeted metabolomics approach has been used to study the fungal contribution to mitigating water deficit in both varieties. Iride and Ramirez exhibited a high and low level of mycorrhizal symbiosis, respectively; resulting in a more remarkable alteration of metabolic pathways in the most colonised variety under water deficit conditions. However, the analysis highlighted the contribution of AMF to mitigating water deficiency in both varieties, resulting in the up- and down-regulation of many amino acids, alkaloids, phenylpropanoids, lipids, and hormones.

The untargeted metabolomic profile of five pigmented whole rice varieties, namely Ermes, Orange, Nerone, Venere, and Violet, was investigated. Samples were cooked to a comparable starch gelatinization level through an electric commercial rice cooker and in vitro digested to evaluate the fate of rice compounds during in vitro starch digestion. The untargeted metabolomics profiling depicted a broad phytochemical profile of rice grains, including phenolic compounds, terpenoids, and phytosterols, among others. Interestingly, the metabolomic profile showed distinctive shaping following cooking and in vitro digestion in a variety-dependent manner. Multivariate statistics revealed that glycosylated flavonoids and hydrolysis-products of tannins (such as 1,2-digalloyl-beta-D-glucopyranose) were the most discriminant compounds of the in vitro gastrointestinal digestion. The combination of cooking and simulated digestion was found to affect, in a matrix-driven manner, the profile of amino acids and lipids derivatives, together with sterols and terpenoids. Cooked Violet rice grains showed the lowest starch hydrolysis index (i.e., 58.2) and the highest resistant starch content (i.e., 4.8 g/100 g) compared to the other samples. Non-pigmented phenolic compounds could be included among discriminant compounds. After combining cooking and enzyme hydrolysis, Violet rice distinctively showed positive average LogFC values for flavonoids and phenolic acids. Overall, water-soluble anthocyanins were the most retained class of polyphenols following the cooking process, with peonidin 3-(6”-acetyl-glucoside) showing the highest discriminant potential (VIP score = 1.64).

Co-inoculation of arbuscular mycorrhizal fungi (AMF) and bacteria can synergically and potentially increase nitrogen use efficiency (NUE) in plants, thus, reducing nitrogen (N) fertilizers use and their environmental impact. However, limited research is available on AMF-bacteria interaction, and the definition of synergisms or antagonistic effects is unexplored. In this study, we adopted a response surface methodology (RSM) to assess the optimal combination of AMF (Rhizoglomus irregulare and Funneliformis mosseae) and Bacillus megaterium (a PGPR—plant growth promoting rhizobacteria) formulations to maximize agronomical and chemical parameters linked to N utilization in maize (Zea mays L.). The fitted mathematical models, and also 3D response surface and contour plots, allowed us to determine the optimal AMF and bacterial doses, which are approximately accorded to 2.1 kg ha–1 of both formulations. These levels provided the maximum values of SPAD, aspartate, and glutamate. On the contrary, agronomic parameters were not affected, except for the nitrogen harvest index (NHI), which was slightly affected (p-value of < 0.10) and indicated a higher N accumulation in grain following inoculation with 4.1 and 0.1 kg ha–1 of AMF and B. megaterium, respectively. Nonetheless, the identification of the saddle points for asparagine and the tendency to differently allocate N when AMF or PGPR were used alone, pointed out the complexity of microorganism interaction and suggests the need for further investigations aimed at unraveling the mechanisms underlying this symbiosis.

Traceability of food is considered as an important tool for ensuring food quality and safety, in turn increasing consumers' trust. Verification of food authenticity is currently carried out by officials and producers, typically through the use of paper documents that may not ensure a proper level of integrity. Therefore, independent analytical methods and forgery-proof labels are required by the agri-food sector, driving the development of "omics" traceability technologies for the authentication of foods. Despite the growing interest in these technologies from a technical-scientific point of view, no research has been carried out on consumers' perceptions. Given these premises, this study aimed at understanding consumers' orientations towards "omics" technologies applied to organic vegetables and unravelling the main psychological and socio-demographic variables that influence their adoption. Data were collected with a self-report questionnaire filled out by a sample of 807 Italians, representative of the population. We used descriptive statistics and multiple linear regression to analyze data. Results show that, despite the lack of subjective knowledge of "omics" technologies, Italians have positive attitudes and are interested in using and buying organic vegetables certified by this approach, even paying more. Furthermore, young people with positive attitudes and higher environmental concerns have a more positive attitude towards "omics" technologies. This research points out the need to educate consumers about what the "omics" technology is, enhancing consumers' attitudes and underlining its environmental contribution. This will increase consumers' acceptance and support the implementation of the technology.

Nutrigenomics is a multidisciplinary science focusing on the diet-genes interaction and exploiting different ‘omics’ technologies to comprehensively investigate the overall effects of different nutrients or diets on health. Recent scientific evidence highlighted the role of dietary polyphenols in modulating gene expression, transcription factors, and cellular processes, thus in potentially affecting health and diseases. Nutrigenomics contributes to the understanding of the molecular mechanisms underlying the direct beneficial effects of polyphenolic compounds in humans. This short review critically summarizes some recent findings on the nutrigenomics-related effects mediated by the major polyphenolic classes of nutritional interest.

An untargeted metabolomics approach combined with sensory analysis was used to depict the impact of different traditional Italian extraction methods (i.e., Espresso, Neapolitan, Moka) along with Filter, on Coffea arabica and Coffea canephora var. robusta beverages. To this aim, polyphenols, Maillard reaction products, and coffee metabolites were screened by high resolution mass spectrometry and elaborated through both unsupervised and supervised multivariate statistical approaches. Multivariate statistics showed a distinctive chemical profile for Espresso preparation, while Moka and Neapolitan were very similar. The orthogonal projection to latent structures and discriminant analysis allowed the identification of 86 compounds showing a high VIP discrimination score (i.e., > 0.8). The 2,5-dimethyl-3-(methyldithio)-furan was a marker for the Filter preparation, while 1,2-disinapoylgentiobiose characterized both Filter and Neapolitan extractions. Caffeine (known to be a bitter compound) accumulated highly in Filter vs. Espresso, although at the sensory profile, bitterness was more perceived in Espresso. Vegetal aroma carried by pyrazines, pyridines, and phenolic acids were markers of Espresso, with Robusta showing higher values than Arabica. Notwithstanding, our findings showed that the extraction process played a hierarchically higher role in driving the chemical composition of the beverages when compared to coffee species.

Ecosystems provide many services that are essential for human activities and for our well-being. Many regulation services are interconnected and are fundamental in mitigating and hindering the negative effects of several phenomena such as pollution. Pollution, in particular airborne particulate matter (PM), represents an important risk to human health. This perspective aims at providing a current framework that relates ecosystem services, regulating services, pollination, and human health, with particular regards to pollution and its impacts. A quantitative literature analysis on the topic has been adopted. The health repercussions of problems related to ecosystem services, with a focus on the effects of atmospheric particulate matter, have been highlighted in the work throughout a case study. In polluted environments, pollinators are severely exposed to airborne PM, which adheres to the insect body hairs and can be ingested through contaminated food resources, i.e., pollen and honey. This poses a serious risk for the health of pollinators with consequences on the pollination service and, ultimately, for human health.

The elicitation of plant secondary metabolism may offer interesting opportunities in the framework of sustainable approaches in plant science and in terms of their ability to prime resistance to biotic and abiotic stressors. The broad metabolic reprogramming triggered by different molecular elicitors, namely salicylate (SA), polyamines (PAs), and chitosan, was comprehensively investigated using a metabolomics approach and the tomato (Solanum lycopersicum L.) as the model crop. Six different treatments were compared: a negative control (no treatments), a second negative control treated with 1 M acetic acid (the reference for chitosan, since chitosan was solubilized in acetic acid), and four molecular elicitors, 1 mM 2,1,3-benzothiadiazole (BTH, a positive control), 10 mg/mL chitosan, 0.01 mM SA, and a 0.1 mM PA (putrescine, spermidine, and spermine). All treatments determined a slight increase in biomass, in particular following PA treatment. A broad reprogramming of secondary metabolism could be observed, including membrane lipid remodeling, phenylpropanoid antioxidants, and phytohormone crosstalk. Overall, our results suggest that PAs, SA, and BTH shared a systemic acquired resistance (SAR)-related response, whereas chitosan induced a more distinct induced systemic resistance (ISR)-like jasmonate-related response. These results pave the way towards the possible use of elicitors as a sustainable tool in plant science and agriculture by increasing crop resilience to biotic and abiotic stressors without detrimental effects on plant biomass.