Plant-derived protein hydrolysates (PHs) are gaining prominence as biostimulants due to their potential to improve yield and nutritional quality even under suboptimal nutrient regimens. In this study, we investigated the effects of foliar application of a legume-derived PH (0 or 4 mL L⁻¹) on greenhouse baby spinach (Spinacia oleracea L.) under four nitrogen (N) fertilization levels (0, 15, 30, or 45 kg ha⁻¹) by evaluating morphological and colorimetric parameters, mineral composition, carbohydrates, proteins, and amino acids. The fresh yield in untreated and biostimulant-treated spinach plants increased in response to an increase in N fertilization from 1 up to 30 kg ha⁻¹, reaching a plateau thereafter indicating the luxury consumption of N at 45 kg ha⁻¹. Increasing N fertilization rate, independently of PH, lead to a significant increase of all amino acids with the exception of alanine, GABA, leucine, lysine, methionine, and ornithine but decreased the polyphenols content. Interestingly, the fresh yield at 0 and 15 kg ha⁻¹ was clearly greater in PH-treated plants compared to untreated plants by 33.3% and 24.9%, respectively. This was associated with the presence in of amino acids and small peptides PH ‘Trainer^®’, which act as signaling molecules eliciting auxin- and/or gibberellin-like activities on both leaves and roots and thus inducing a “nutrient acquisition response” that enhances nutrients acquisition and assimilation (high P, Ca, and Mg accumulation) as well as an increase in the photochemical efficiency and activity of photosystem II (higher SPAD index). Foliar applications of the commercial PH decreased the polyphenols content, but on the other hand strongly increased total amino acid content (+45%, +82%, and +59% at 0, 15, and 30 kg ha⁻¹, respectively) but not at a 45-kg ha⁻¹-rate. Overall, the use of PH could represent a sustainable tool for boosting yield and nitrogen use efficiency and coping with soil fertility problems under low input regimens.

Selenium (Se) is considered essential for human nutrition as it is involved in the metabolic pathway of selenoproteins and relevant biological functions. Microgreens, defined as tender immature greens, constitute an emerging functional food characterized by overall higher levels of phytonutrients than their mature counterparts. The nutraceutical value of microgreens can be further improved through Se biofortification, delivering Se-enriched foods and potentially an enhanced content of bioactive compounds. The current study defined the effect of sodium selenate applications at three concentrations (0, 8, and 16 μM Se) on the bioactive compounds and mineral content of coriander, green basil, purple basil, and tatsoi microgreens grown in soilless cultivation. Analytical emphasis was dedicated to the identification and quantification of polyphenols by UHPLC-Q-Orbitrap-HRMS, major carotenoids by HPLC-DAD, and macro micro-minerals by ICP-OES. Twenty-seven phenolic compounds were quantified, of which the most abundant were: Chlorogenic acid and rutin in coriander, caffeic acid hexoside and kaempferol-3-O(caffeoyl) sophoroside-7-O-glucoside in tatsoi, and cichoric acid and rosmarinic acid in both green and purple basil. In coriander and tatsoi microgreens, the application of 16 μM Se increased the total phenols content by 21% and 95%, respectively; moreover, it improved the yield by 44% and 18%, respectively. At the same Se dose, the bioactive value of coriander and tatsoi was enhanced by a significant increase in rutin (33%) and kaempferol-3-O(feruloyl)sophoroside-7-O-glucoside (157%), respectively, compared to the control. In green and purple basil microgreens, the 8 μM Se application enhanced the lutein concentration by 7% and 19%, respectively. The same application rate also increased the overall macroelements content by 35% and total polyphenols concentration by 32% but only in the green cultivar. The latter actually had a tripled chicoric acid content compared to the untreated control. All microgreen genotypes exhibited an increase in the Se content in response to the biofortification treatments, thereby satisfying the recommended daily allowance for Se (RDA-Se) from 20% to 133%. The optimal Se dose that guarantees the effectiveness of Se biofortification and improves the content of bioactive compounds was 16 μM in coriander and tatsoi, and 8 μM in green and purple basil.

Research has been increasingly focusing on the environmentally friendly biostimulation of vegetable crop performances under sustainable farming management. An experiment was carried out in southern Italy on Diplotaxis tenuifolia to assess the effects of two plant biostimulants (Legume-derived protein hydrolysate, Trainer^®; Tropical plant extract, Auxym^®) and a non-treated control, in factorial combination with three crop cycles (autumn–winter; winter; and winter–spring) on leaf yield, photosynthetic and colour status, quality, elemental composition, antioxidant content and activity. Both biostimulants prevalently contain amino acids and soluble peptides, showing the major effects on crop performances, though Auxym also has a small percentage of phytohormones and vitamins. The biostimulants enhanced plant growth and the productivity of perennial wall rocket. The winter–spring cycle led to higher leaf yield than the winter one. The two plant biostimulants enhanced leaf dry matter, oxalic and citric acids, Ca and P concentrations, phenols and ascorbic acid content as well as antioxidant activity, but did not increase nitrate content. A presumed mechanism involved in the enhancement of crop production could be attributed to the improvement of mineral nutrient availability and uptake. The winter–spring cycle elicited higher antioxidant content and activity than winter crops. Our current study shows that both the legume-derived protein hydrolysate and tropical plant extract represent an effective tool for boosting the yield, nutritional and functional quality of vegetable produce in the view of sustainable crop systems.

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.

The present study examined the modulatory effects of natural fiber substrates (agave fiber, coconut fiber and peat moss) and synthetic alternatives (capillary mat and cellulose sponge) on the nutritive and phytochemical composition of select microgreens species (coriander, kohlrabi and pak choi) grown in a controlled environment. Polyphenols were analyzed by UHPLC-Q-Orbitrap-HRMS, major carotenoids by HPLC-DAD, and macro-minerals by ion chromatography. Microgreens grown on peat moss had outstanding fresh and dry yield but low dry matter content. Natural fiber substrates increased nitrate and overall macro-mineral concentrations in microgreens compared to synthetic substrates. The concentrations of chlorophylls, carotenoids and ascorbate were influenced primarily by species. On the contrary, variability in polyphenols content was wider between substrates than species. Out of twenty phenolic compounds identified, chlorogenic acid and quercetin-3-O-rutinoside were most abundant. Hydroxycinnamic acids and their derivatives accounted for 49.8% of mean phenolic content across species, flavonol glycosides for 48.4% and flavone glycosides for 1.8%. Peat moss provided optimal physicochemical conditions that enhanced microgreens growth rate and biomass production at the expense of phenolic content. In this respect, the application of controlled stress (eustress) on microgreens growing on peat moss warrants investigation as a means of enhancing phytochemical composition without substantial compromise in crop performance and production turnover. Finally, nitrate deprivation practices should be considered for microgreens grown on natural fiber substrates in order to minimize consumer exposure to nitrate.

An optimized nitrogen (N) fertilization may have a positive effect on leafy vegetables by increasing growth, yield and nutrient content of plants. Nevertheless, crop performance must be coupled with an increase in Nitrogen Use Efficiency (NUE) in order to limit external N inputs and to avoid N surpluses associated with environmental and health problems. The aim of the current study was to assess the effects of a legume-derived plant hydrolysates (LDPH; Trainer^®) and N fertilization levels (0, 2.25 and 4.5 g N m⁻² for spinach and 0, 2.5 and 5.0 g N m⁻² for lamb’s lettuce; N0%, N50%, N100%, respectively) on agronomical, biochemical, qualitative responses and NUE of these two important greenhouse leafy vegetables. Spinach and lamb’s lettuce were sprayed four times during the growing period (at a concentration of 4 mL L⁻¹ of LDPH). In baby spinach, the LDPH application elicited a significant increase at the three levels of N fertilization: +16.8%, +14.2%, and 39.4% at 0, 2.25 and 4.5 g N m⁻², respectively. Interestingly, in lamb’s lettuce, the N50% plants treated with LDPH reached similar values of marketable yield in comparison to treated and non-treated plants under N100% conditions. The presumed mechanism involved in the enhancement of yield response in the two leafy greens could be associated to a better activity of the photosystem II (higher SPAD index), biochemical (higher content of chlorophyll a, b and total) and leaf nitrate status. The foliar application of LDPH produced a major fortification in lipophilic and hydrophilic antioxidant activities (+11.6 and 6.3% for spinach and lamb’s lettuce, respectively). The biostimulant application also improved N-use efficiency and N-uptake efficiency compared to untreated plants: +17.8% and +18.8%, and +50% and +73.3%, for spinach and lamb’s lettuce, respectively.