Foliar applications of a Malvaceae-derived protein hydrolysate and its fractions differentially modulate yield and functional traits of tomato under optimal and suboptimal nitrogen application.

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-NO3) 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-NO3, 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-NO3. 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.

Integrated multi-omic approach reveals the effect of a Graminaceae-derived biostimulant and its lighter fraction on salt-stressed lettuce plants.

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 CO2 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.

Unravelling the biostimulant activity of a protein hydrolysate in lettuce plants under optimal and low N availability: a multi-omics approach.

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.

Plant-derived biostimulant as priming agents enhanced antioxidant and nutritive properties in brassicaceous microgreens.

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 (H2O)) 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.

Copper boosts the biostimulant activity of a vegetal-derived protein hydrolysate in basil: morpho-physiological and metabolomics insights.

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 imbalance upon metal exposure. In contrast, a mitigation of oxidative stress was highlighted in Cu-VPH and VPH+Cu-VPH, where the treatments reduced stress-related metabolites. Overall, these results highlight an interaction between Cu and VPH, hence paving the way towards the combined use of Cu and biostimulants to optimize agronomic interventions.

Applying productivity and phytonutrient profile criteria in modelling species selection of microgreens as Space crops for astronaut consumption.

Introduction: Long-duration missions in outer Space will require technologies to regenerate environmental resources such as air and water and to produce food while recycling consumables and waste. Plants are considered the most promising biological regenerators to accomplish these functions, due to their complementary relationship with humans. Plant cultivation for Space starts with small plant growth units to produce fresh food to supplement stowed food for astronauts' onboard spacecrafts and orbital platforms. The choice of crops must be based on limiting factors such as time, energy, and volume. Consequently, small, fast-growing crops are needed to grow in microgravity and to provide astronauts with fresh food rich in functional compounds. Microgreens are functional food crops recently valued for their color and flavor enhancing properties, their rich phytonutrient content and short production cycle. Candidate species of microgreens to be harvested and eaten fresh by crew members, belong to the families Brassicaceae, Asteraceae, Chenopodiaceae, Lamiaceae, Apiaceae, Amarillydaceae, Amaranthaceae, and Cucurbitaceae. Methods: In this study we developed and applied an algorithm to objectively compare numerous genotypes of microgreens intending to select those with the best productivity and phytonutrient profile for cultivation in Space. The selection process consisted of two subsequent phases. The first selection was based on literature data including 39 genotypes and 25 parameters related to growth, phytonutrients (e.g., tocopherol, phylloquinone, ascorbic acid, polyphenols, lutein, carotenoids, violaxanthin), and mineral elements. Parameters were implemented in a mathematical model with prioritization criteria to generate a ranking list of microgreens. The second phase was based on germination and cultivation tests specifically designed for this study and performed on the six top species resulting from the first ranking list. For the second selection, experimental data on phytonutrients were expressed as metabolite production per day per square meter. Results and discussion: In the final ranking list radish and savoy cabbage resulted with the highest scores based on their productivity and phytonutrient profile. Overall, the algorithm with prioritization criteria allowed us to objectively compare candidate species and obtain a ranking list based on the combination of numerous parameters measured in the different species. This method can be also adapted to new species, parameters, or re-prioritizing the parameters for specific selection purposes.

Iodine-Biofortified Microgreens as High Nutraceutical Value Component of Space Mission Crew Diets and Candidate for Extraterrestrial Cultivation.

The success of Space missions and the efficacy of colonizing extraterrestrial environments depends on ensuring adequate nutrition for astronauts and autonomy from terrestrial resources. A balanced diet incorporating premium quality fresh foods, such as microgreens, is essential to the mental and physical well-being of mission crews. To improve the nutritional intake of astronaut meals, two levels of potassium iodide (KI; 4 µM and 8 µM) and an untreated control were assessed for iodine (I) biofortification, and overall nutraceutical profile of four microgreens: tatsoi (Brassica rapa L. subsp. narinosa), coriander (Coriandrum sativum L.), green basil, and purple basil (Ocimum basilicum L.). A dose-dependent increase in I was observed at 8 µM for all species, reaching concentrations of 200.73, 118.17, 93.97, and 82.70 mg kg-1 of dry weight, in tatsoi, coriander, purple basil, and green basil, respectively. Across species, I biofortification slightly reduced fresh yield (-7.98%) while increasing the antioxidant activity (ABTS, FRAP, and DPPH). LC-MS/MS Q extractive orbitrap analysis detected 10 phenolic acids and 23 flavonoids among microgreen species. The total concentration of phenolic acids increased (+28.5%) in purple basil at 8 µM KI, while total flavonoids in coriander increased by 23.22% and 34.46% in response to 4 and 8 µM KI, respectively. Both doses of KI increased the concentration of total polyphenols in all species by an average of 17.45%, compared to the control.

Modulation of Morpho-Physiological and Metabolic Profiles of Lettuce Subjected to Salt Stress and Treated with Two Vegetal-Derived Biostimulants.

Salinity in water and soil is a critical issue for food production. Using biostimulants provides an effective strategy to protect crops from salinity-derived yield losses. The research supports the effectiveness of protein hydrolysate (PH) biostimulants based on their source material. A greenhouse experiment was performed on lettuce plants under control (0 mM NaCl) and high salinity conditions (30 mM NaCl) using the Trainer (T) and Vegamin (V) PH biostimulants. The recorded data included yield parameters, mineral contents, auxiliary pigments, and polyphenolics. The plant sample material was further analyzed to uncover the unique metabolomic trace of the two biostimulants. The results showed an increased yield (8.9/4.6%, T/V) and higher photosynthetic performance (14%) compared to control and salinity treatments. Increased yield in salinity condition by T compared to V was deemed significant due to the positive modulation in stress-protecting molecules having an oxidative stress relief effect such as lutein (39.9% 0 × T vs. 30 × V), ß-carotene (23.4% vs. V overall), and flavonoids (27.7% vs. V). The effects of PH biostimulants on the physio-chemical and metabolic performance of lettuce plants are formulation dependent. However, they increased plant growth under stress conditions, which can prove profitable.

Productive, Morpho-Physiological, and Postharvest Performance of Six Basil Types Grown in a Floating Raft System: A Comparative Study.

Basil (Ocimum sp.) is one of the world's most famous culinary fresh herbs, characterized by rapid growth that makes it particularly suitable for hydroponic cultivation. This study aimed to evaluate the adaptability of six types of basil to a closed-loop hydroponic system (floating raft system) and their post-harvest performance. Twenty-three days after transplantation, productivity, morpho-physiological performance, and mineral profile (by ion chromatography) were evaluated. At 3, 6, and 9 days after harvest, the loss of water from the from leaves stored at 10 °C in the dark was evaluated. Although the total fresh production of Thai, Mexican, and Genovese did not differ significantly, the latter provided a higher fresh leaf weight (16.52 g of plant-1) despite a lower leaf number (30.06 n. of plant-1). Nine days after harvest, Thai and Mexican showed the lowest water loss. Although Mexican Purple had the lowest net CO2 assimilation, it accumulated the highest concentration of ascorbic acid (909.41 mg 100 g fw-1).

Nutritive and Phytochemical Composition of Aromatic Microgreen Herbs and Spices Belonging to the Apiaceae Family.

Microgreens represent a new generation of food products, commonly used to garnish and embellish culinary dishes, and recently associated with an increasing interest in their nutraceutical and phytochemical profiles. Four Apiaceae species: Pimpinella anisum L. (anise), Anthriscus cerefolium L. (chervil), Carum carvi L. (caraway), and Anethum graveolens L. (dill) were assessed for fresh yield, macro- and microminerals, total chlorophylls, total ascorbic acid, carotenoids, polyphenols, and their antioxidant activity. Anise was the species yielding the most (2.53 kg m-2) and having the highest lutein content (18.4 µg g-1 dry weight (DW)). Chervil and dill were characterized by the highest total ascorbic acid content (~151 mg AA g-1 fresh weight (FW)). The phenolic profile highlighted the presence of five flavonoid derivatives and 12 phenolic acid derivatives, with quinic acid derivatives being the most abundant phenols in the species tested. In addition, anise, caraway, and dill proved to be considerably rich in total polyphenols (~11056 µg g-1 DW). Caraway and dill were characterized by the highest antioxidant activity measured by the DPPH and ABTS methods, whereas the FRAP method revealed caraway as having the highest antioxidant activity. Such results highlight the potential of Apiaceae species as an alternative to other families which are commonly used for microgreens production.

Hemp microgreens as an innovative functional food: Variation in the organic acids, amino acids, polyphenols, and cannabinoids composition of six hemp cultivars.

Hemp (Cannabis sativa L.) is a multi-functional crop cultivated for fiber, seeds, or phytochemical extraction. Once a major industrial crop in several agro-environments, its cultivation strongly declined in developed countries since World War II. Exploiting hemp vegetative tissue as innovative food has remained largely unexplored. The current work examined the potential production of hemp microgreens. Six cultivars were assessed for yield and composition of organic acids, amino acids, polyphenols and phytocannabinoids, through IC, FLD-HPLC and UHPLC-HRMS, respectively. Bioactive composition was strongly related to the hemp variety. 'Silvana' demonstrated the highest total content of amino acids and essential amino acids, high concentrations of cannflavin A and B, and moderate levels of cannabidiol and cannabigerol. 'Finola' distinguished by the highest concentration of cannflavins and total polyphenols, and the lowest levels of Δ9-THC. Regardless of varietal differences, hemp microgreens proved widely safe in terms of Δ9-THC content.

Morpho-Anatomical, Physiological, and Mineral Composition Responses Induced by a Vegetal-Based Biostimulant at Three Rates of Foliar Application in Greenhouse Lettuce.

A promising strategy for sustainably increasing the quality and yield of horticultural products is the use of natural plant biostimulants. In this work, through a greenhouse experiment, we evaluated the effect of a legume-derived biostimulant at three dose treatments (0.0 control, 2.5 mL L-1, and 5.0 mL L-1) on the yield performance, nutrients traits, leaf anatomical traits, gas exchanges, and carbon photosynthetic assimilation of greenhouse lettuce. The lettuce plants were foliar sprayed every 7 days for 5 weeks. The application of plant biostimulant, at both lower and higher dosages, increased the nutrient use efficiency, root dry weight, and leaf area. However, it is noteworthy that the 5.0 mL L-1 dose enhanced photosynthetic activity in the early phase of growth (15 DAT), thus supplying carbon skeletons useful for increasing the number of leaves and their efficiency (higher SPAD), and for boosting nutrient uptake (P, S, and K) and transport to leaves, while the 2.5 mL L-1 dose exerted specific effects on roots, increasing their dimension and enabling them to better use nitrate and Ca. A higher dose of biostimulant application might find its way in shorter growing cycle, thus presenting new horizons for new lines of research in baby leaves production.

Tracking the Biostimulatory Effect of Fractions from a Commercial Plant Protein Hydrolysate in Greenhouse-Grown Lettuce.

Protein hydrolysate biostimulants are environmentally friendly options for the reduction of nitrogen input, but their plant growth-promoting mechanisms are still not completely unveiled. Here, to put the "signaling peptide theory" to the test, a greenhouse experiment was undertaken using low (1 mM) and optimal (8 mM) NO3-treated butterhead lettuce and three molecular fractions (PH1 (>10 kDa), PH2 (1−10 kDa) and PH3 (<10 kDa) fractions), in addition to the whole product Vegamin®: PH, in a randomized block design. PH1 and PH3 significantly increased fresh yield (+8%) under optimal (lighter leaves), but not under low (darker leaves) NO3 conditions. Total ascorbic acid, lutein and ß-carotene increased with PH3, and disinapoylgentobiose and kaempferol-3-hydroxyferuloyl-sophorosie-7-glucoside content increased with PH (whole/fractions) treatments, particularly under low NO3 conditions. The complete hydrolysate and analyzed peptide fractions have differential biostimulatory effects, enhancing the growth and nutritional quality of lettuce.

Pearl Grey Shading Net Boosts the Accumulation of Total Carotenoids and Phenolic Compounds That Accentuate the Antioxidant Activity of Processing Tomato.

Tomato (Solanum lycopersicum L.) is one of the most consumed vegetables worldwide due to its low caloric intake and high fiber, minerals, and phenolic compounds, making it a high-quality functional food. However, fruit quality attributes can be affected by pre-harvest factors, especially environmental stresses. This research aimed to evaluate the influence of two shading nets (white net -30% and pearl grey net -40% shading degree) on the yield and phytochemical profile of tomato fruits grown in summer under the Mediterranean climate. Mineral and organic acid content (by ion chromatography-IC), phenolic profile (by ultra-high performance liquid chromatography-UHPLC coupled with an Orbitrap high-resolution mass spectrometry-HRMS), carotenoid content (by high-performance liquid chromatography with diode array detection-HPLC-DAD), and antioxidant activities DPPH, ABTS, and FRAP (by UV-VIS spectrophotometry) were determined. Tomato fruits grown under the pearl grey net recorded the highest values of total phenolic compounds (14,997 µg 100 g-1 of fresh weight) and antioxidant activities DPPH, ABTS, and FRAP, without affecting either fruit color or marketable yield. The reduction of solar radiation through pearl grey nets proved to be an excellent tool to increase the phytochemical quality of tomato fruits during summer cultivation in a Mediterranean environment.

Biostimulant Substances for Sustainable Agriculture: Origin, Operating Mechanisms and Effects on Cucurbits, Leafy Greens, and Nightshade Vegetables Species.

Climate change is a pressing matter of anthropogenic nature to which agriculture contributes by abusing production inputs such as inorganic fertilizers and fertigation water, thus degrading land and water sources. Moreover, as the increase in the demand of food in 2050 is estimated to be 25 to 70% more than what is currently produced today, a sustainable intensification of agriculture is needed. Biostimulant substances are products that the EU states work by promoting growth, resistance to plant abiotic stress, and increasing produce quality, and may be a valid strategy to enhance sustainable agricultural practice. Presented in this review is a comprehensive look at the scientific literature regarding the widely used and EU-sanctioned biostimulant substances categories of silicon, seaweed extracts, protein hydrolysates, and humic substances. Starting from their origin, the modulation of plants' hormonal networks, physiology, and stress defense systems, their in vivo effects are discussed on some of the most prominent vegetable species of the popular plant groupings of cucurbits, leafy greens, and nightshades. The review concludes by identifying several research areas relevant to biostimulant substances to exploit and enhance the biostimulant action of these substances and signaling molecules in horticulture.

Bioformulations with Beneficial Microbial Consortia, a Bioactive Compound and Plant Biopolymers Modulate Sweet Basil Productivity, Photosynthetic Activity and Metabolites.

Increasing attention is being given to the development of innovative formulations to substitute the use of synthetic chemicals to improve agricultural production and resource use efficiency. Alternatives can include biological products containing beneficial microorganisms and bioactive metabolites able to inhibit plant pathogens, induce systemic resistance and promote plant growth. The efficacy of such bioformulations can be increased by the addition of polymers as adjuvants or carriers. Trichoderma afroharzianum T22, Azotobacter chroococcum 76A and 6-pentyl-α-pyrone (6PP; a Trichoderma secondary metabolite) were administrated singularly or in a consortium, with or without a carboxymethyl cellulose-based biopolymer (BP), and tested on sweet basil (Ocimum basilicum L.) grown in a protected greenhouse. The effect of the treatments on basil yield, photosynthetic activity and secondary metabolites production was assessed. Photosynthetic efficiency was augmented by the applications of the bioformulations. The applications to the rhizosphere with BP + 6PP and BP + T22 + 76A increased the total fresh weight of basil by 26.3% and 23.6%, respectively. Untargeted LC-MS qTOF analysis demonstrated that the plant metabolome was significantly modified by the treatments. Quantification of the profiles for the major phenolic acids indicated that the treatment with the T22 + 76A consortium increased rosmarinic acid content by 110%. The use of innovative bioformulations containing microbes, their metabolites and a biopolymer was found to modulate the cultivation of fresh basil by improving yield and quality, thus providing the opportunity to develop farming systems with minimal impact on the environmental footprint from the agricultural production process.

Root-Associated Bacterial Community Shifts in Hydroponic Lettuce Cultured with Urine-Derived Fertilizer.

Recovery of nutrients from source-separated urine can truncate our dependency on synthetic fertilizers, contributing to more sustainable food production. Urine-derived fertilizers have been successfully applied in soilless cultures. However, little is known about the adaptation of the plant to the nutrient environment. This study investigated the impact of urine-derived fertilizers on plant performance and the root-associated bacterial community of hydroponically grown lettuce (Lactuca sativa L.). Shoot biomass, chlorophyll, phenolic, antioxidant, and mineral content were associated with shifts in the root-associated bacterial community structures. K-struvite, a high-performing urine-derived fertilizer, supported root-associated bacterial communities that overlapped most strongly with control NPK fertilizer. Contrarily, lettuce performed poorly with electrodialysis (ED) concentrate and hydrolyzed urine and hosted distinct root-associated bacterial communities. Comparing the identified operational taxonomic units (OTU) across the fertilizer conditions revealed strong correlations between specific bacterial genera and the plant physiological characteristics, salinity, and NO3-/NH4+ ratio. The root-associated bacterial community networks of K-struvite and NPK control fertilized plants displayed fewer nodes and node edges, suggesting that good plant growth performance does not require highly complex ecological interactions in hydroponic growth conditions.

Preharvest Nutrient Deprivation Reconfigures Nitrate, Mineral, and Phytochemical Content of Microgreens.

While imparting gastronomic novelty and sensory delight, microgreens also constitute rudimentary leafy greens packed with nutrients and phytochemicals. As such, they comprise an upcoming class of functional foods. However, apart from bioactive secondary metabolites, microgreens also accumulate antinutritive agents such as nitrate, especially under conducive protected cultivation conditions. The current work examined nutrient deprivation before harvest (DBH), applied by replacing nutrient solution with osmotic water for six and twelve days, as a strategy for reducing microgreen nitrate levels in different species (lettuce, mustard, and rocket). The three species were sown on a peat-based substrate, cultivated in a controlled climate chamber, and harvested 18 days after sowing, when the first two true leaves emerged. DBH impact on major constituents of the secondary metabolome, mineral content, colorimetric, and yield traits was appraised. Nitrate and mineral content were determined through ion chromatography, phenolic composition through UHPLC-Q-Orbitrap HRMS, and carotenoid composition through HPLC-DAD. Nutrient deprivation was effective in reducing nitrate content; however, effective treatment duration differed between species and decline was more precipitous in nitrate hyperaccumulating species such as rocket. Quercetin and kaempferol glycosides were the flavonol glycosides most abundant in brassicaceous microgreens, whereas lettuce microgreens were steeped in caffeoyl quinic acid. DBH interacted with species as it increased the total phenolic content of lettuce, decreased that of rocket, but did not affect mustard. Further research to link changes in phenolic composition to the sensory and in vivo bioactive profile of microgreens is warranted. Notably, brief (≤6 days) DBH can be applied across species with moderate or no impact on the phenolic, carotenoid, and mineral composition of microgreens. Brief DBH applications also have limited impact on microgreens' yield and colorimetric traits hence on the commercial value of the product. They can therefore be applied for reducing microgreen nitrate levels without significantly impacting key secondary metabolic constituents and their potential bioactive role.

Divergent Leaf Morpho-Physiological and Anatomical Adaptations of Four Lettuce Cultivars in Response to Different Greenhouse Irradiance Levels in Early Summer Season.

Lettuce (Lactuca sativa L.) is a winter-spring leafy vegetable, but the high demand for fresh products available year-round requires off-season production. However, the warm climate of the Mediterranean areas can impair the summer production of lettuce, thus requiring the adoption of genotypes tolerant to high irradiance as well as useful agronomic strategies like shading net installations. The aim of our research was to assess the leaf morpho-physiological and anatomical changes, in addition to productive responses, of four lettuce cultivars ('Ballerina', 'Maravilla De Verano Canasta', 'Opalix', and 'Integral') grown under shading and non-shading conditions to unveil the adaptive mechanisms of this crop in response to sub-optimal microclimate (high irradiance and temperature) in a protected environment. Growth and yield parameters, leaf gas exchanges, chlorophyll fluorescence and morpho-anatomical leaf traits (i.e., leaf mass area, stomatal density and epidermal cell density) were determined. Under shading conditions, the fresh yields of the cultivars 'Ballerina', 'Opalix' ('Oak leaf') and 'Integral' ('Romaine') increased by 16.0%, 26.9% and 13.2% respectively, compared to non-shading conditions while both abaxial and adaxial stomatal density decreased. In contrast, 'Canasta' under non-shading conditions increased fresh yield, dry biomass and instantaneous water use efficiency by 9.6%, 18.0% and 15.7%, respectively, while reduced abaxial stomatal density by 30.4%, compared to shading conditions. Regardless of cultivar, the unshaded treatment increased the leaf mass area by 19.5%. Even though high light intensity and high temperature are critical limiting factors for summer lettuce cultivation in a protected environment, 'Canasta' showed the most effective adaptive mechanisms and had the best production performance under sub-optimal microclimatic conditions. However, greenhouse coverage with a white shading net (49% screening) proved to be a suitable agricultural practice that ensured an adequate microclimate for the off-season growth of more sensitive cultivars 'Ballerina', 'Oak leaf' and 'Romaine'.

Isosmotic Macrocation Variation Modulates Mineral Efficiency, Morpho-Physiological Traits, and Functional Properties in Hydroponically Grown Lettuce Varieties (Lactuca sativa L.).

The management of mineral elements in agriculture is important for their nutritional role for plants and dietary value for humans, sparking interest in strategies that can increase mineral use efficiency and accumulation in plant food. In this work, we evaluated the effects of the isosmotic variations of the concentration on three macrocations (K, Ca, and Mg) in lettuce (Lactuca sativa L.). Our aim was to improve the nutritional components of this valuable dietary source of minerals. Using a full factorial design, we analyzed mineral utilization efficiency (UtE), leaf morphology, gas exchange parameters, phenolic profiles (through ultra-high performance liquid chromatography coupled to a quadrupole-time-of-flight (UHPLC-QTOF) mass spectrometry), and enzymatic activities in two phytochemically diverse butterhead lettuce varieties (red or green). Plants were fed in hydroponics with three nutrient solutions (NSs) with different ratios of K, Ca, and Mg. The variation of these minerals in the edible product was associated with alterations of the morphology and physiology of the leaves, and of the quality and functional properties of lettuce, with a trade-off between total accumulation and mineral UtE. Moreover, in non-limiting conditions of nutrient availability, significant mineral interactions were also present. The flexibility of the plant response to the different ratios of macrocations, and the observed large intraspecific variation, were adequate to provide mineral-specific phytochemical profiles to the edible product. Specifically, the full-red lettuce provided more interesting results in regard to the compositional and functional attributes of the leaves.