Plant biostimulants (PBs) such as protein hydrolysates and seaweed extracts are attracting the increasing interest of scientists and vegetable growers for their potential toenhance yield and nutritional quality. The current study assessed crop productivity, leaf colorimetry, mineral profile and bioactive compounds of greenhouse spinach in response to the foliar application of three PBs: legume-derived protein hydrolysate [PH], extract of seaweed Ecklonia maxima or mixture of vegetal oils, herbal and seaweed Ascophyllum nodosum extracts. Plants were PB-treated at a rate of 3 mL L⁻¹ four times during their growth cycle at weekly intervals. Foliar PB applications enhanced fresh yield, dry biomass and leaf area of spinach in comparison with untreated plants. Improved yield performance with PB applications was associated with improved chlorophyll biosynthesis (higher SPAD index). The three PB treatments elicited an increase in bioactive compounds (total phenols and ascorbic acid), thus raised the functional quality of spinach. The application of PH enhanced K and Mg concentrations and did not result in increased nitrate accumulation as observed with the other two PB treatments. Our findings can assist vegetable farmers and the agro-food industry in adopting innovative and sustainable tools such as PB for complementing a high yield with premium quality.

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.

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.

Nitrogen (N) fertilizers play a crucial role in agriculture, representing a powerful tool for farmers for increasing yields throughout the seasons under both optimal and suboptimal conditions. At the same time, their synthetic/chemical nature could have several influences on ecosystems and human health. For this reason, there is an urgent need to find new and more sustainable means of production to increase plant productivity and optimize nitrogen use. An experiment was conducted in a plastic tunnel to assess the response of baby lettuce crop to the foliar application of three plant biostimulants (PBs): Legume-derived protein hydrolysate (LDPH) ‘Trainer^®’, tropical plant extract (TPE) ‘Auxym^®’ and seaweed extract (SwE) from Ecklonia maxima ‘Kelpak^®’ under different N rates of 0, 10, 20 and 30 kg N·ha⁻¹. The responses of baby lettuce plants were assessed in terms of yield, growth parameters and physicochemical composition of the leaves. The fresh yield of baby lettuce in both biostimulant-treated and untreated plants was positively affected by increasing N rates from 0 to 20 kg N·ha⁻¹, reaching a plateau thereafter indicating luxury N conditions at 30 kg N·ha⁻¹. However, high N fertilizer application (20 and especially 30 kg N·ha⁻¹) resulted in undesirable decreases in antioxidant activities and total ascorbic acid (TAA). Under non-fertilized regimens, foliar PBs application boost growth and yield of baby lettuce in comparison to non-treated plants. Foliar spray with LDPH and especially SwE elicited significant increases in marketable fresh yield (averaging 14%, 6% and 7% at 10, 20 and 30 kg N·ha⁻¹, respectively) compared to TPE and untreated plants. Improved agronomical performance of baby lettuce under optimal (10 kg N ha⁻¹) and especially suboptimal N regimens (0 kg N ha⁻¹) was associated with increasing photochemical efficiency and a better activity of photosystem II (higher Soil Plant Analysis Development-SPAD index and chlorophyllous pigments biosynthesis). The application of LDPH enhanced antioxidant capacity and TAA in baby lettuce leaf and did not increased nitrate content as recorded in SwE and TPE treatments. Overall, plant biostimulants may be considered as a sustainable tool of production to increase leafy vegetable productivity in low fertility soils.

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.

Nitrogen is the primary technical means responsible for food production increase, but on the other hand, wise management is needed because its excessive use can have a negative impact on the environment and on green leafy vegetable quality, such as that rocket. Rocket has the characteristics of accumulating nitrate in leaves with possible impacts on human health. In order to overcome this issue, researchers are focusing their attention on the use of alternative means, such as plant biostimulant application. The scope of this study was to assess the effect of legume-derived protein hydrolysate(LDPH) and tropical plant extract(TPE), combined with various doses of nitrogen (0 kg ha⁻¹ non-fertilized; N0); 60 kg ha⁻¹ (sub-optimal; N1); 80 kg ha⁻¹ (optimal; N2); and 100 kg ha⁻¹ (supra-optimal; N3)), in order to reduce nitrogen use, boost yield, and enhance the chemical and nutritional value of leaves without significantly accumulating nitrate. Both vegetal-based plant biostimulants enhanced plant growth, boosted the marketable yield (especially at N0 and N1 levels, by 38.2% and 28.2%, respectively, compared to the non-treated control), and increased the SPAD (Soil Plant Analysis Development) index and leaf pigments content, such as chlorophyll and carotenoids, especially in treated-LDPH rocket. The plant-based biostimulants also produced a major amplification in lipophilic antioxidant activity (+ 48%) and total ascorbic acid content (average + 95.6%), especially at low nitrogen fertilization levels, and maintained nitrate content under the legal European Comission limits.