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Background: Protein hydrolysates (PHs) can enhance plant nitrogen nutrition and improve the quality of vegetables, depending on their bioactive compounds. A tomato greenhouse experiment was conducted under both optimal (14 mM) and suboptimal (2 mM) nitrogen (N-NO 3 ) conditions. Tomatoes were treated with a new Malvaceae-derived PH (MDPH) and its molecular fractions (MDPH1, >10 kDa; MDPH2, 1-10 kDa and MDPH3, <1 kDa)., Results: Under optimal N conditions, the plants increased biomass and fruit yield, and showed a higher photosynthetic pigment content in leaves in comparison with suboptimal N, whereas under N-limiting conditions, an increase in dry matter, soluble solid content (SSC) and lycopene, a reduction in firmness, and changes in organic acid and phenolic compounds were observed. With 14 mM N-NO 3 , MDPH3 stimulated an increase in dry weight and increased yield components and lycopene in the fruit. The MDPH2 fraction also resulted in increased lycopene accumulation in fruit under 14 mM N-NO 3 . At a low N level, the PH fractions showed distinct effects compared with the whole MDPH and the control, with an increase in biomass for MDPH1 and MDPH2 and a higher pigment content for MDPH3. Regardless of N availability, all the fractions affected fruit quality by increasing SSC, whereas MDPH2 and MDPH3 modified organic acid content and showed a higher concentration of flavonols, lignans, and stilbenes., Conclusion: The molecular weight of the peptides modifies the effect of PHs on plant performance, with different behavior depending on the level of N fertilization, confirming the effectiveness of fractioning processes. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Background: Microgreens constitute dietary sources of bioactive compounds imparting numerous health benefits and enhancing sensory experience. They can be successfully cultivated in soilless systems where biostimulants can be easily integrated as seed-priming and post-germination agents improving the sustainability of a crop's final production. Compared to an untreated control, three priming agents (a commercial legume-derived protein hydrolysate (A250), a novel protein hydrolysate derived from peanut biomass (H250) and hydropriming (H 2 O)) were applied to Komatsuna and Mibuna seeds grown as microgreens and compared for their effects on yield parameters, mineral composition, ABTS and FRAP antioxidant capacity, carotenoid concentration and phenolic compounds., Results: Significant effects of the main experimental factors and their interactions were identified on antioxidant capacity. Compared to the control and hydropriming, the highest ABTS and FRAP values were observed in Mibuna with the A250 and H250 treatments, respectively. Additionally, the H250 treatment increased the total concentrations of phenolic acid derivatives and flavonoid derivatives in Mibuna and Komatsuna, in tune with the levels of total flavonoids. Concerning mineral composition, the highest concentrations in both species were those of phosphorus and nitrate., Conclusion: These results highlight the potential of select plant-based biostimulants as priming agents to enhance the antioxidant capacity, nutrient content and bioactive compound content, thus further increasing their functional and nutritive quality. In the light of this, the possibility of reducing the application of fertilizers by promoting a green transition for the intensive production of microgreens could subsequently be evaluated. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The need to increase agricultural production to feed a steadily growing population may clash with the more environmentally friendly but less efficient production methods required. Therefore, it is important to try to reduce the use of chemical inputs without compromising production. In this scenario, natural biostimulants have become one of the most sought-after and researched technologies. In the present study, the results of a greenhouse experiment on hydroponic tomatoes ( Solanum lycopersicum L.) are presented, which involved comparing the use of ordinary NPK fertilizer (Cerbero ® ) with the use of NPK fertilizers enriched with 0.5% protein hydrolysate of plant origin (Cerbero Green ® ) at both standard (100%) and reduced (70%) fertilization rates. The results highlight how the use of Cerbero Green ® fertilizers improves the production performance of tomatoes. More specifically, they show that the use of Cerbero Green ® leads to higher marketable yields, especially under reducing fertilizer use, ensuring a positive net change in profit for the grower. In addition, carbon footprint analysis has revealed that the use of Cerbero Green ® reduces the environmental impact of hydroponic tomato growing practices by up to 8%. The observed higher yield of hydroponically grown tomatoes even with reduced fertilization rates underlines once again the key role of natural biostimulants in increasing both the economic and environmental sustainability of horticultural production.

Molybdenum (Mo) is a crucial microelement for both, humans and plants. The use of agronomic biofortification techniques can be an alternative method to enhance Mo content in vegetables. Concomitantly, arbuscular mycorrhizal fungi (AMF) application is a valuable strategy to enhance plant performances and overcome plant abiotic distresses such as microelement overdose. The aim of this research was to estimate the direct and/or indirect effects of Mo supply at four doses [0.0, 0.5 (standard dose), 2.0 or 4.0 μmol L -1 ], alone or combined with AMF inoculation, on plant performances. In particular, plant height and first flower truss emission, productive features (total yield, marketable yield and average marketable fruit weight) and fruit qualitative characteristics (fruit dry matter, soluble solids content, titratable acidity, ascorbic acid, lycopene, polyphenol, nitrogen, copper, iron and molybdenum) of an established cherry tomato genotype cultivated in soilless conditions were investigated. Moreover, proline and malondialdehyde concentrations, as well as Mo hazard quotient (HQ) in response to experimental treatments were determined. A split-plot randomized experimental block design with Mo dosages as plots and +AMF or -AMF as sub-plots was adopted. Data revealed that AMF inoculation enhanced marketable yield (+50.0 %), as well as some qualitative traits, such as fruit soluble solids content (SSC) (+9.9 %), ascorbic acid (+7.3 %), polyphenols (+2.3 %), and lycopene (+2.5 %). Molybdenum application significantly increased SSC, polyphenols, fruit Mo concentration (+29.0 % and +100.0 % in plants biofortified with 2.0 and 4.0 μmol Mo L -1 compared to those fertigated with the standard dose, respectively) and proline, whereas it decreased N (-25.0 % and -41.6 % in plants biofortified with 2.0 and 4.0 μmol Mo L -1 compared to those fertigated with the standard dose, respectively). Interestingly, the application of AMF mitigated the detrimental effect of high Mo dosages (2.0 or 4.0 μmol L -1 ). A pronounced advance in terms of plant height 45 DAT, fruit lycopene concentration and fruit Fe, Cu and Mo concentrations was observed when AMF treatment and Mo dosages (2.0 or 4.0 μmol Mo L -1 ) were combined. Plants inoculated or not with AMF showed an improvement in the hazard quotient (HQ) in reaction to Mo application. However, the HQ - for a consumption of 200 g day -1 of biofortified cherry tomato - remained within the safety level for human consumption. This study suggests that Mo-implementation (at 2.0 or 4.0 μmol L -1 ) combined with AMF inoculation could represent a viable cultivation protocol to enhance yield, produce premium quality tomato fruits and, concomitantly, improve Mo dose in human diet. In the light of our findings, further studies on the interaction between AMF and microelements in other vegetable crops are recommended., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Modern agriculture urgently requires viable alternatives to synthetic chemical substances, such as pesticides and fertilizers, to comply with new and stringent international regulations and meet the growing demands of consumers who prefer chemical-free food. Consequently, organic agriculture has garnered increasing interest over time. To compensate for yield reduction resulting from opting out of the use mineral fertilizers, research has focused on the use of biostimulants to sustain the productivity of horticultural crops. To this end, a greenhouse experiment was conducted to assess the effects of three nonmicrobial biostimulants (a plant extract, vegetable protein hydrolysate, and a seaweed extract) and an untreated control on the production and mineral content of wild rocket ( Diplotaxis tenuifolia (L.) DC.) cultivated under organic conditions and harvested three times during the growth cycle. In general, the nitrate content, which defines the commercial quality of wild rocket, was not influenced by the application of biostimulants. At each harvest, the application of biostimulants resulted in improved production performance, although this was not always accompanied by an increase in mineral content. Specifically, the best results were obtained with the use of plant-derived protein hydrolysate and plant extract, which led to an improvement in total yield of 32.1% and 27.2%, respectively compared to that of control plants. These results reconfirm that biostimulants represent a valid and indispensable tool for organic growers.

Plant biostimulants are widely applied in agriculture for their ability to improve plant fitness. In the present work, the impact of Graminaceae-derived protein hydrolysate (P) and its lighter molecular fraction F3 (< 1 kDa) on lettuce plants, subjected to either no salt or high salt conditions, was investigated through the combination of metabolomics and transcriptomics. The results showed that both treatments significantly modulated the transcriptome and metabolome of plants under salinity stress, highlighting an induction of the hormonal response. Nevertheless, P and F3 also displayed several peculiarities. F3 specifically modulated the response to ethylene and MAPK signaling pathway, whereas P treatment induced a down-accumulation of secondary metabolites, albeit genes controlling the biosynthesis of osmoprotectants and antioxidants were up-regulated. Moreover, according with the auxin response modulation, P promoted cell wall biogenesis and plasticity in salt-stressed plants. Notably, our data also outlined an epigenetic control of gene expression induced by P treatment. Contrarily, experimental data are just partially in agreement when not stressed plants, treated with P or F3, were considered. Indeed, the reduced accumulation of secondary metabolites and the analyses of hormone pathways modulation would suggest a preferential allocation of resources towards growth, that is not coherent with the down-regulation of the photosynthetic machinery, the CO 2 assimilation rate and leaves biomass. In conclusion, our data demonstrate that, although they might activate different mechanisms, both the P and F3 can result in similar benefits, as far as the accumulation of protective osmolytes and the enhanced tolerance to oxidative stress are concerned. Notably, the F3 fraction exhibits slightly greater growth promotion effects under high salt conditions. Most importantly, this research further corroborates that biostimulants' mode of action is dependent on plants' physiological status and their composition, underscoring the importance of investigating the bioactivity of the different molecular components to design tailored applications for the agricultural practice., (© 2024. The Author(s).)

The application of protein hydrolysates (PH) biostimulants is considered a promising approach to promote crop growth and resilience against abiotic stresses. Nevertheless, PHs bioactivity depends on both the raw material used for their preparation and the molecular fraction applied. The present research aimed at investigating the molecular mechanisms triggered by applying a PH and its fractions on plants subjected to nitrogen limitations. To this objective, an integrated transcriptomic-metabolomic approach was used to assess lettuce plants grown under different nitrogen levels and treated with either the commercial PH Vegamin® or its molecular fractions PH1(>10 kDa), PH2 (1-10 kDa) and PH3 (<1 kDa). Regardless of nitrogen provision, biostimulant application enhanced lettuce biomass, likely through a hormone-like activity. This was confirmed by the modulation of genes involved in auxin and cytokinin synthesis, mirrored by an increase in the metabolic levels of these hormones. Consistently, PH and PH3 upregulated genes involved in cell wall growth and plasticity. Furthermore, the accumulation of specific metabolites suggested the activation of a multifaceted antioxidant machinery. Notwithstanding, the modulation of stress-response transcription factors and genes involved in detoxification processes was observed. The coordinated action of these molecular entities might underpin the increased resilience of lettuce plants against nitrogen-limiting conditions. In conclusion, integrating omics techniques allowed the elucidation of mechanistic aspects underlying PH bioactivity in crops. Most importantly, the comparison of PH with its fraction PH3 showed that, except for a few peculiarities, the effects induced were equivalent, suggesting that the highest bioactivity was ascribable to the lightest molecular fraction., (© 2024 Scandinavian Plant Physiology Society.)

Introduction: High-throughput phenotyping technologies together with metabolomics analysis can speed up the development of highly efficient and effective biostimulants for enhancing crop tolerance to drought stress. The aim of this study was to examine the morphophysiological and metabolic changes in tomato plants foliarly treated with two protein hydrolysates obtained by enzymatic hydrolysis of vegetal proteins from Malvaceae (PH1) or Fabaceae (PH2) in comparison with a control treatment, as well as to investigate the mechanisms involved in the enhancement of plant resistance to repeated drought stress cycles., Methods: A phenotyping device was used for daily monitoring morphophysiological traits while untargeted metabolomics analysis was carried out in leaves of the best performing treatment based on phenotypic results. Results: PH1 treatment was the most effective in enhancing plant resistance to water stress due to the better recovery of digital biomass and 3D leaf area after each water stress event while PH2 was effective in mitigating water stress only during the recovery period after the first drought stress event. Metabolomics data indicated that PH1 modified primary metabolism by increasing the concentration of dipeptides and fatty acids in comparison with untreated control, as well as secondary metabolism by regulating several compounds like phenols. In contrast, hormones and compounds involved in detoxification or signal molecules against reactive oxygen species were downregulated in comparison with untreated control., Conclusion: The above findings demonstrated the advantages of a combined phenomics-metabolomics approach for elucidating the relationship between metabolic and morphophysiological changes associated with a biostimulant-mediated increase of crop resistance to repeated water stress events., Competing Interests: PB is Founder & CEO at oloBion SL and Member of Arcadia s.r.l. while GC is Founder & Member of Arcadia s.r.l. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Leporino, Rouphael, Bonini, Colla and Cardarelli.)

Essential oils are of great interest due to their potent pharmaceutical and biological activities. In this study, essential oils extracted from Origanum compactum and Thymus zygis originating from the Middle Atlas of Morocco were investigated. Their chemical compositions were analyzed using gas chromatography and mass spectrometry, while the assessment of the trapping power of the radical (DPPH: 1,1-diphenyl-2-picrylhydrazyl) and the reducing antioxidant potential of ferric ions (FRAP: Ferric Reducing Antioxidant Power) were performed in order to evaluate the antioxidant activity. Their antibacterial potency was tested against six bacterial strains through the disk diffusion method. The chromatography analyses of the extracted essential oils highlighted the presence of two main components, namely carvacrol at 75.70% in O. compactum and thymol at 40.67% in T. zygis . The antioxidant activity tests showed that both essential oils demonstrated a significant antioxidant activity comparable to the positive control (e.g., ascorbic acid). The antibacterial activity results showed a strong antimicrobial effect for both essential oils, compared to synthetic antibiotics. This study affirms the presence of bioactive components with interesting antioxidant and antibacterial activities in the essential oils extracted from Origanum compactum and Thymus zygis , which could find several applications in the food and pharmaceutical industries through the substitution of synthetic antioxidants and antibiotics.

The demand for high-quality strawberries continues to grow, emphasizing the need for innovative agricultural practices to enhance both yield and fruit quality. In this context, the utilization of natural products, such as biostimulants, has emerged as a promising avenue for improving strawberry production while aligning with sustainable and eco-friendly agricultural approaches. This study explores the influence of a bacterial filtrate (BF), a vegetal-derived protein hydrolysate (PH), and a standard synthetic auxin (SA) on strawberry, investigating their effects on yield, fruit quality, mineral composition and metabolomics of leaves and fruits. Agronomic trial revealed that SA and BF significantly enhanced early fruit yield due to their positive influence on flowering and fruit set, while PH treatment favored a gradual and prolonged fruit set, associated with an increased shoot biomass and sustained production. Fruit quality analysis showed that PH-treated fruits exhibited an increase of firmness and soluble solids content, whereas SA-treated fruits displayed lower firmness and soluble solids content. The ionomic analysis of leaves and fruits indicated that all treatments provided sufficient nutrients, with heavy metals within regulatory limits. Metabolomics indicated that PH stimulated primary metabolites, while SA and BF directly affected flavonoid and anthocyanin biosynthesis, and PH increased fruit quality through enhanced production of beneficial metabolites. This research offers valuable insights for optimizing strawberry production and fruit quality by harnessing the potential of natural biostimulants as viable alternative to synthetic compounds., Competing Interests: Two authors are from Atens Company Spain who manufactured a tested product. However, they were not involved in the trial management at Tuscia University (Italy), measurements, and analysis, interpretation of the results. They role was mainly in the manuscript writing. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Cardarelli, El Chami, Rouphael, Ciriello, Bonini, Erice, Cirino, Basile, Corrado, Choi, Kim and Colla.)