Comprehensive lipid profile of ‘Maraština’ grape skins from Dalmatia

Lipids are a diverse group of organic compounds that serve essential roles due to their biological functions for all prokaryotic and eukaryotic organisms. Despite the fundamental role of lipid class in plants, there is still a relatively low level of knowledge regarding the composition of grape lipids. This research represents the first investigation into the lipids of the Croatian grape variety ‘Maraština’ (Vitis vinifera L.), employing a targeted approach and the UHPLC-MS/MS method to conduct a comprehensive analysis of the lipid profile of grape skins. The study investigated the profile of free fatty acids, glycerolipids, glycerophospholipids, triterpenoids, sphingolipids, and free fatty acid esters in eleven vineyards located along the cultivation area of ‘Maraština’ variety (Dalmatia, Croatia). Grape skins from vineyards that were exposed to more sunlight and higher daily temperature during the vegetation period mainly resulted in a richer profile of free fatty acids. Among lipids, ‘Maraština’ grape skins were characterized mainly by a triterpenoid compound; oleanolic acid (249.49 g/g of FW), and free fatty acids, especially palmitic acid C16:0 (26.09 g/g of FW), and lignoceric acid C24:0 (12.40 g/g of FW). Hierarchical clustering analysis represented valuable insights into the diversity of lipids in ‘Maraština’ grape skin

Grape lipidomics: an extensive profiling thorough UHPLC–MS/MS method

8openInternationalItalian coauthor/editorLipids play many essential roles in living organisms, which accounts for the great diversity of these amphiphilic molecules within the individual lipid classes, while their composition depends on intrinsic and extrinsic factors. Recent developments in mass spectrometric methods have significantly contributed to the widespread application of the liquid chromatography-mass spectrometry (LC–MS) approach to the analysis of plant lipids. However, only a few investigators have studied the extensive composition of grape lipids. The present work describes the development of an ultrahigh performance liquid chromatography tandem mass spectrometry (UHPLC–MS/MS) method that includes 8098 MRM; the method has been validated using a reference sample of grapes at maturity with a successful analysis and semi-quantification of 412 compounds. The aforementioned method was subsequently applied also to the analysis of the lipid profile variation during the Ribolla Gialla cv. grape maturation process. The partial least squares (PLS) regression model fitted to our experimental data showed that a higher proportion of certain glycerophospholipids (i.e., glycerophosphoethanolamines, PE and glycerophosphoglycerols, PG) and of some hydrolysates from those groups (i.e., lyso-glycerophosphocholines, LPC and lyso-glycerophosphoethanolamines, LPE) can be positively associated with the increasing °Brix rate, while a negative association was found for ceramides (CER) and galactolipids digalactosyldiacylglycerols (DGDG). The validated method has proven to be robust and informative for profiling grape lipids, with the possibility of application to other studies and matrices.openMasuero, Domenico; Škrab, Domen; Chitarrini, Giulia; Garcia-Aloy, Mar; Franceschi, Pietro; Sivilotti, Paolo; Guella, Graziano; Vrhovsek, UrskaMasuero, D.; Škrab, D.; Chitarrini, G.; Garcia-Aloy, M.; Franceschi, P.; Sivilotti, P.; Guella, G.; Vrhovsek, U

Exploratory analysis of commercial olive-based dietary supplements using untargeted and targeted metabolomics

8openInternationalItalian coauthor/editorThe market of olive-based dietary supplements (OBDS) is composed of a broad range of natural extracts claiming different health effects and often sold without a clear statement on their chemical composition. The aim of this survey was to characterize the chemical profiles of 14 commercially available OBDS. As many as 378 compounds were tentatively annotated in the analyzed samples. Although for most of metabolites the annotation at level I was prevented due to the lack of the analytical standard, the spectra obtained from high-resolution tandem mass spectrometry (MS/MS) measurements were very informative, allowing annotation of dozens of metabolites at level II or III. A targeted method allowed the quantification of 26 selected compounds. A large qualitative and quantitative variability was observed. The products obtained from buds by glyceric maceration were those with the lowest concentrations of all the quantified elements. The dose of 5 mg of hydroxytyrosol, corresponding to the European Food Safety Authority (EFSA) health claim, was only reached by four products, all of them originating from the olive fruit or the leaves. If we also take into consideration oleuropein, two additional products provide this daily amount. This work demonstrates the high complexity and diversity in the composition of OBDSopenGarcia Aloy, M.; Groff, N.; Masuero, D.; Nisi, M.; Franco, A.; Battelini, F.; Vrhovsek, U.; Mattivi, F.Garcia Aloy, M.; Groff, N.; Masuero, D.; Nisi, M.; Franco, A.; Battelini, F.; Vrhovsek, U.; Mattivi, F

Cluster Thinning and Vineyard Site Modulate the Metabolomic Profile of Ribolla Gialla Base and Sparkling Wines

Depending on the vineyard location, cluster thinning (CT) may represent an effective tool to obtain the desired grape composition and wine quality. The effect of 20% cluster thinning on Ribolla Gialla (Vitis vinifera L.) sparkling wine aroma, lipid compounds, and aromatic amino acid (AAA) metabolites composition was studied for three consecutive seasons in two vineyards located in the Friuli Venezia Giulia region, Italy. In the examined sparkling wines, the vintage meteorological conditions exhibited significant influences on the metabolic profile of the samples. Data were normalized by season, and the impact of the CT treatment was evaluated for each vineyard site separately. Crop removal showed a limited positive impact on aroma compounds in sparkling wines from vineyards located in the valley. Concerning the AAA compounds, their concentration was higher in the vineyard at the foot of the hills. Cluster thinning resulted in a drop in concentration, reducing the risk of atypical aging. Despite minor differences according to targeted metabolome profiling, the sensory analysis confirmed the effects of the CT treatment in the valley floor vineyard. Reducing crop in this site, where the yield was higher, promoted a moderate improvement of Ribolla Gialla sparkling wine. In contrast, for wine produced in the vineyard at the foot of the hills, the sensory analysis indicated a preference for wines from the unthinned control samples. Overall, the study indicates that cluster thinning is a viticultural technique that could potentially improve the quality of Ribolla Gialla sparkling wines, but only in situations of excessive grape production

Anti-inflammatory and anti-acne effects of Hamamelis virginiana Bark in human keratinocytes

12openInternationalItalian coauthor/editorCutibacterium acnes (C. acnes) is recognized as one of the main triggers of the cutaneous inflammatory response in acne vulgaris, a chronic skin disorder with a multifactorial origin. Witch hazel (Hamamelis virginiana L.) is a plant widely used for skin inflammatory conditions, with some preliminary anti-inflammatory evidence on the skin, but lacking data on acne conditions. This study aimed to evaluate the effect of a glycolic extract from Hamamelis virginiana bark (HVE) versus C. acnes-induced inflammation in human keratinocytes (HaCaT). Phytochemical investigations of HVE identified hamamelitannin (HT) and proanthocyanidins as the most abundant compounds (respectively, 0.29% and 0.30% w/wextract). HVE inhibited C. acnes-induced IL-6 release (IC50: 136.90 μg/mL), by partially impairing NF-κB activation; however, no antibacterial or antibiofilm activities were found. In addition, HVE showed greater anti-inflammatory activity when TNF-α was used as a proinflammatory stimulus (IC50 of 38.93 μg/mL for IL-8 release), partially acting by antioxidant mechanisms, as shown for VEGF inhibition. The effects of HVE are primarily based on the proanthocyanidin content, as HT was found inactive on all the parameters tested. These results suggest further investigations of HVE in other inflammatory-based skin diseasesopenPiazza, Stefano; Martinelli, Giulia; Vrhovsek, Urska; Masuero, Domenico; Fumagalli, Marco; Magnavacca, Andrea; Pozzoli, Carola; Canilli, Luisa; Terno, Massimo; Angarano, Marco; Dell’Agli, Mario; Sangiovanni, EnricoPiazza, S.; Martinelli, G.; Vrhovsek, U.; Masuero, D.; Fumagalli, M.; Magnavacca, A.; Pozzoli, C.; Canilli, L.; Terno, M.; Angarano, M.; Dell’Agli, M.; Sangiovanni, E

Assessment of the chemical and genetic variability among accessions of Cicerbita alpina (L.) Wallr., an alpine plant with anthelmintic properties

Cicerbita alpina (L.) Wallr, is a perennial alpine plant and a member of the Asteraceae family, typically found at altitudes above 1000 meters in the Italian Alps. Although previously utilized primarily as a local delicacy, recent studies have revealed strong antiparasitic activity through in vitro experiments. In Europe, numerous chemical drugs employed to combat nematodes — helminths that infest the digestive tract of livestock — are banned due to their environmental harm or show only reduced efficiency because of the development of resistance. Consequently, there is a growing demand for new alternative anthelmintic treatments in agricultural practices. Specialized metabolites found in the extracts of C. alpina could offer a sustainable and biological alternative to chemical drugs, specifically for nematode control. For this purpose, a unique germplasm collection originating from eight distinct natural populations in the Italian Alps was analyzed for its chemical diversity using state-of-the-art targeted LC-MS/MS spectrometry, including quantification based on multiple reaction monitoring. The predominant metabolites identified from the species were the caffeic acid derivatives chicoric acid, chlorogenic acid, and 3. 5-dicaffeoylquinic acid, the sesquiterpene lactone derivative 8-O-acetyl-15-ß-D-glucopyranosyl lactucin and the flavone glycosides, apigenin-7-O-glucoside and luteolin-7-Oglucoside, alongside their precursors apigenin and luteolin, respectively. Additionally, the genetic diversity of eighty individual plants within the germplasm collection was evaluated using ten DNA molecular markers (Simple Sequence Repeats), successfully transferred from two closely related species (Cichorium intybus and Tanacetum parthenium). This investigation unveiled a significant range of genetic diversity within the examined populations, resulting in the establishment of three distinct genetic groups. The findings were further correlated with the original ecological environment and local climate conditions spanning a biennial period, indicating substantial variations among the different accessions and the intricate interplay between genetic background and environmental factors. These results could serve as a basis for future domestication of the species through plant breeding programs ensuring product quality, but also facilitating the cultivation of C. alpina in more diverse geographic region

Valorization of traditional Italian walnut (Juglans regia L.) production: genetic, nutritional and sensory characterization of locally grown varieties in the Trentino region

15openYesJuglans regia (L.) is cultivated worldwide for its nutrient-rich nuts. In Italy, despite the growing demand, walnut cultivation has gone through a strong decline in recent decades, which led to Italy being among the top five net importing countries. To promote the development of local high-quality Italian walnut production, we devised a multidisciplinary project to highlight the distinctive traits of three varieties grown in the mountainous region Trentino (northeast of Italy): the heirloom ‘Bleggiana’, a second local accession called local Franquette and the French cultivar ‘Lara’, recently introduced in the local production to increase yield. The genetic characterization confirmed the uniqueness of ‘Bleggiana’ and revealed local Franquette as a newly described autochthonous variety, thus named ‘Blegette’. The metabolic profiles highlighted a valuable nutritional composition of the local varieties, richer in polyphenols and with a lower ω-6/ω-3 ratio than the commercial ‘Lara’. ‘Blegette’ obtained the highest preference scores from consumers for both the visual aspect and tasting; however, the volatile organic compound profiles did not discriminate among the characterized cultivars. The described local varieties represent an interesting reservoir of walnut genetic diversity and quality properties, which deserve future investigation on agronomically useful traits (e.g., local adaptation and water usage) for a high-quality and sustainable production.Di Pierro, Erica A.; Franceschi, Pietro; Endrizzi, Isabella; Farneti, Brian; Poles, Lara; Masuero, Domenico; Khomenko, Iuliia; Trenti, Francesco; Marrano, Annarita; Vrhovsek, Urska; Gasperi, Flavia; Biasioli, Franco; Guella, Graziano; Bianco, Luca; Troggio, MichelaDi Pierro, E.A.; Franceschi, P.; Endrizzi, I.; Farneti, B.; Poles, L.; Masuero, D.; Khomenko, I.; Trenti, F.; Marrano, A.; Vrhovsek, U.; Gasperi, F.; Biasioli, F.; Guella, G.; Bianco, L.; Troggio, M

Simultaneous editing of two DMR6 genes in grapevine results in reduced susceptibility to downy mildew

The reduction of pesticides’ treatments is of paramount importance for the sustainability of viticulture, and it can be achieved by a combination of strategies including the cultivation of vines (Vitis vinifera) that are resistant or tolerant to diseases such as downy mildew (DM). In many crops, the knock-out of Downy Mildew Resistant 6 (DMR6) proved successful in controlling DM-resistance, but the effect of mutations in DMR6 genes in not yet known in grapevine.Today, gene editing serves crop improvement with small and specific mutations while maintaining the genetic background of commercially important clones. Moreover, recent technological advances allowed to produce non-transgenic grapevine clones by regeneration of protoplasts edited with the CRISPR/Cas9 ribonucleoprotein. This approach may revolutionize the production of new grapevine varieties and clones, but it requires knowledge on the targets, and on the impact of editing on plant phenotype and fitness in different cultivars. In this work we generated single and double knock-out mutants by editing DMR6 susceptibility (S) genes using CRISPR/Cas9, and showed that only the combined mutations in VviDMR6-1 and VviDMR6-2 are effective in reducing susceptibility to DM in two table-grape cultivars by increasing the levels of endogenous salicylic acid. Therefore, editing both genes may be necessary for effective DM control in real-world agricultural settings, which could potentially lead to unwanted phenotypes. Additional research, including trials conducted in experimental vineyards, is required to gain a deeper understanding of DMR6-based resistance

Comparative transcriptome and metabolite survey reveal key pathways involved in the control of the chilling injury disorder superficial scald in two apple cultivars, ‘Granny Smith’ and ‘Ladina’

The low temperature normally applied to prevent fruit decay during the storage of apples, can also triggers the onset of a chilling injury disorder known as superficial scald. In this work, the etiology of this disorder and the mechanism of action of two preventing strategies, such as the application of 1-MCP (1-methylcyclopropene) and storage at low oxygen concentration in ‘Granny Smith’ and ‘Ladina’ apple cultivars were investigated. The metabolite assessment highlighted a reorganization of specific metabolites, in particular flavan-3-ols and unsaturated fatty acids, while the genome-wide transcriptomic analysis grouped the DEGs into four functional clusters. The KEGG pathway and GO enrichment analysis, together with the gene-metabolite interactome, showed that the treatment with 1-MCP prevented the development of superficial scald by actively promoting the production of unsaturated fatty acids, especially in ‘Granny Smith’. ‘Ladina’, more susceptible to superficial scald and less responsive to the preventing strategies, was instead characterized by a higher accumulation of very long chain fatty acids. Storage at low oxygen concentration stimulated a higher accumulation of ethanol and acetaldehyde together with the expression of genes involved in anaerobic respiration, such as malate, alcohol dehydrogenase and pyruvate decarboxylase in both cultivars. Low oxygen concentration, likewise 1-MCP, through a direct control on ethylene prevented the onset of superficial scald repressing the expression of PPO, a gene encoding for the polyphenol oxidase enzyme responsible of the oxidation of chlorogenic acid. Moreover, in ‘Granny Smith’ apple, the expression of three members of the VII subgroups of ERF genes, encoding for elements coordinating the acclimation process to hypoxia in plants was observed. The global RNA-Seq pattern also elucidated a specific transcriptomic signature between the two cultivars, disclosing the effect of the different genetic background in the control of this disorder

A Metabolomic Approach to the Study of Wine Micro-Oxygenation

Wine micro-oxygenation is a globally used treatment and its effects were studied here by analysing by untargeted LC-MS the wine metabolomic fingerprint. Eight different procedural variations, marked by the addition of oxygen (four levels) and iron (two levels) were applied to Sangiovese wine, before and after malolactic fermentation