From urine to food and oxygen: effects of high and low NH4+:NO3- ratio on lettuce cultivated in a gas-tight hydroponic facility.

In situ production of food, water and oxygen is essential for long-duration human space missions. Higher plants represent a key element in Bioregenerative Life Support Systems (BLSS), where crop cultivation can be based on water and nutrients recovered from waste and wastewater. Human urine exemplifies an important waste stream with potential to provide crops with nitrogen (N) and other nutrients. Dynamic waste composition and treatment processes may result in mineralized fractions with varying ammonium (NH4 +) to nitrate (NO3 -) ratios. In this study, lettuce was cultivated in the unique ESA MELiSSA Plant Characterization Unit, an advanced, gas-tight hydroponic research facility offering controlled environment and continuous monitoring of atmospheric gas composition. To evaluate biological and system effects of nutrient solution NH4 +:NO3 - ratio, two crop tests were run with different NH4 + to total N ratio (NH4 +:N) and elevated concentrations of Na+ and Cl- in line with a urine recycling scenario. Plants cultivated at 0.5 mol·mol-1 NH4 +:N (HiNH4 +) achieved 50% lower shoot biomass compared to those cultivated at 0.1 mol·mol-1 NH4 +:N (LoNH4 +), accompanied by higher shoot dry weight content and lower harvest index. Analyses of projected leaf area over time indicated that the reduced biomass observed at harvest could be attributed to a lower specific growth rate during the close-to-exponential growth phase. The HiNH4 + crop produced 40% less O2 over the full cultivation period. However, normalization of the results indicated a marginal increase in O2 production per time and per projected leaf area for the HiNH4 + crop during the exponential growth phase, in line with a higher shoot chlorophyll content. Mineral analysis demonstrated that the biomass content of NH4 + and NO3 - varied in line with the nutrient solution composition. The ratio of consumed NH4 + to consumed N was higher than the NH4 +:N ratio of the nutrient solution for both crop tests, resulting in decreasing NH4 +:N ratios in the nutrient solution over time. The results provide enhanced insight for design of waste processes and crop cultivation to optimize overall BLSS efficiency and hold valuable potential for improved resource utilization also in terrestrial food production systems.

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.

Foliar Nutrition Influences Yield, Nut Quality and Kernel Composition in Hazelnut cv Mortarella.

In hazelnut, foliar nutrition is utilized globally to integrate microelement deficiencies and optimize their assimilation and effects on yield performances. Nevertheless, nut quality and kernel composition can be positively affected by foliar nutrition. Recently, several studies pointed out the need for increasing the sustainability of orchard nutrition by proposing the management of not only micronutrients, but also main components, such as nitrogen, through foliar spraying. In our study, different foliar fertilizers were used to understand the effectiveness of supporting hazelnut productivity and nut and kernel quality. Water was used as a control. Foliar fertilizations affected tree annual vegetative growth, improved kernel weight and decreased the incidence of blanks compared to the control. Differences in fat, protein, and carbohydrate concentration were also found among treatments, with increased fat concentrations and total polyphenols content in fertilized treatments. Foliar fertilization improved the oil composition of the kernels, though fatty acid composition responded differently to nutrients spray. Oleic acid concentration was promoted, while palmitic acid concentration was reduced in fertilized plants compared to control trees. Furthermore, CD and B trees were characterized by an increase in the ratio of unsaturated/saturated fatty acids compared to untreated trees. Finally, foliar spraying improved lipid stability compared to the control due to higher total polyphenol concentration.

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).

Can Lunar and Martian Soils Support Food Plant Production? Effects of Horse/Swine Monogastric Manure Fertilisation on Regolith Simulants Enzymatic Activity, Nutrient Bioavailability, and Lettuce Growth.

To make feasible the crewed missions to the Moon or Mars, space research is focusing on the development of bioregenerative life support systems (BLSS) designed to produce food crops based on in situ resource utilisation (ISRU), allowing to reduce terrestrial input and to recycle organic wastes. In this regard, a major question concerns the suitability of native regoliths for plant growth and how their agronomic performance is affected by additions of organic matter from crew waste. We tested plant growth substrates consisting of MMS-1 (Mars) or LHS-1 (Lunar) simulants mixed with a commercial horse/swine monogastric manure (i.e., an analogue of crew excreta and crop residues) at varying rates (100:0, 90:10, 70:30, 50:50, w/w). Specifically, we measured: (i) lettuce (Lactuca sativa L. cultivar 'Grand Rapids') growth (at 30 days in open gas exchange climate chamber with no fertilisation), plant physiology, and nutrient uptake; as well as (ii) microbial biomass C and N, enzymatic activity, and nutrient bioavailability in the simulant/manure mixtures after plant growth. We discussed mechanisms of different plant yield, architecture, and physiology as a function of chemical, physico-hydraulic, and biological properties of different substrates. A better agronomic performance, in terms of plant growth and optically measured chlorophyll content, nutrient availability, and enzymatic activity, was provided by substrates containing MMS-1, in comparison to LHS-1-based ones, despite a lower volume of readily available water (likely due to the high-frequency low-volume irrigation strategy applied in our experiment and foreseen in space settings). Other physical and chemical properties, along with a different bioavailability of essential nutrients for plants and rhizosphere biota, alkalinity, and release of promptly bioavailable Na from substrates, were identified as the factors leading to the better ranking of MMS-1 in plant above and below-ground mass and physiology. Pure Mars (MMS-1) and Lunar (LHS-1) simulants were able to sustain plant growth even in absence of fertilisation, but the amendment with the monogastric manure significantly improved above- and below-ground plant biomass; moreover, the maximum lettuce leaf production, across combinations of simulants and amendment rates, was obtained in treatments resulting in a finer root system. Increasing rates of monogastric manure stimulated the growth of microbial biomass and enzymatic activities, such as dehydrogenase and alkaline phosphomonoesterase, which, in turn, fostered nutrient bioavailability. Consequently, nutrient uptake and translocation into lettuce leaves were enhanced with manure supply, with positive outcomes in the nutritional value of edible biomass for space crews. The best crop growth response was achieved with the 70:30 simulant/manure mixture due to good availability of nutrients and water compared to low amendment rates, and better-saturated hydraulic conductivity compared to high organic matter application. A 70:30 simulant/manure mixture is also a more sustainable option than a 50:50 mixture for a BLSS developed on ISRU strategy. Matching crop growth performance and (bio)chemical, mineralogical, and physico-hydraulic characteristics of possible plant growth media for space farming allows a better understanding of the processes and dynamics occurring in the experimental substrate/plant system, potentially suitable for an extra-terrestrial BLSS.

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.

Protein Hydrolysates from Animal or Vegetal Sources Affect Morpho-Physiological Traits, Ornamental Quality, Mineral Composition, and Shelf-Life of Chrysanthemum in a Distinctive Manner.

Protein hydrolysates (PHs) are a prominent category of plant biostimulants, mainly constituted of amino acids, oligopeptides and polypeptides, obtained by partial hydrolysis of animal or plant protein sources. Despite scientific evidence supporting the biostimulant action of PHs on vegetables, the morphological, physiological, and shelf-life performances underlying the PH action on cut flowers are still poorly explored. Accordingly, the aim of this research is to assess the effects of three commercial biostimulants, one animal PH (PH A, Hicure®) and two plant PHs (PH V1, Trainer® and PH V2, Vegamin©), on two chrysanthemum (Chrysanthemum morifolium) cultivars (Pinacolada and Radost). In both cultivars, only the plant-derived PH (V1 and V2) treatments recorded significantly higher fresh plant biomass than the control (on average +18%, in both cultivars). The foliar application of the vegetal-derived PHs but not the animal one, particularly in Pinacolada, improved the status of plants, stimulating stem elongation and the apical flower diameter. In Pinacolada, applications with PH V1 resulted in a significant increase in nitrate and P concentration in leaves and Ca content in flowers compared with the control (+43%, +27%, and +28% for nitrate, P, and Ca, respectively). In Radost, PH A and PH V2 applications caused a significant reduction in nitrate concentration in both leaves and flowers compared with the control. One week after harvest, in both cultivars, PH A applications caused flower stems to wilt faster than the control. In contrast, plants treated with PH V1 revealed significantly slower flower stem senescence compared to the control. Flower wilting during vase life was correlated to a decrease in the K-to-Na ratio in flowers due to an inability to transport K to the flowers from the leaves rather than an increase in Na in the flowers themselves.

Bioactive Compounds and Antioxidant Activity of Lettuce Grown in Different Mixtures of Monogastric-Based Manure With Lunar and Martian Soils.

The supplementation of bioactive compounds in astronaut's diets is undeniable, especially in the extreme and inhospitable habitat of future space settlements. This study aims to enhance the Martian and Lunar regolith fertility (testing two commercial simulants) through the provision of organic matter (manure) as established by in situ resource utilization (ISRU) approach. In this perspective, we obtained 8 different substrates after mixing Mojave Mars Simulant (MMS-1) or Lunar Highlands Simulant (LHS-1), with four different rates of manure (0, 10, 30, and 50%, w/w) from monogastric animals. Then, we assessed how these substrates can modulate fresh yield, organic acid, carotenoid content, antioxidant activity, and phenolic profile of lettuce plants (Lactuca sativa L.). Regarding fresh biomass production, MMS-1-amended substrates recorded higher yields than LHS-1-ones; plants grown on a 70:30 MMS-1/manure mixture produced the highest foliar biomass. Moreover, we found an increase in lutein and ß-carotene content by + 181 and + 263%, respectively, when applying the highest percentage of manure (50%) compared with pure simulants or less-amended mixtures. The 50:50 MMS-1/manure treatment also contained the highest amounts of individual and total organic acids, especially malate content. The highest antioxidant activity for the ABTS assay was recorded when no manure was added. The highest content of total hydroxycinnamic acids was observed when no manure was added, whereas ferulic acid content (most abundant compound) was the highest in 70:30 simulant/manure treatment, as well as in pure LHS-1 simulant. The flavonoid content was the highest in pure-simulant treatment (for most of the compounds), resulting in the highest total flavonoid and total phenol content. Our findings indicate that the addition of manure at specific rates (30%) may increase the biomass production of lettuce plants cultivated in MMS-1 simulant, while the phytochemical composition is variably affected by manure addition, depending on the stimulant. Therefore, the agronomic practice of manure amendment showed promising results; however, it must be tested with other species or in combination with other factors, such as fertilization rates and biostimulants application, to verify its applicability in space colonies for food production purposes.

Untargeted Phenolic Profiling and Functional Insights of the Aerial Parts and Bulbs of Drimia maritima (L.) Stearn.

Drimia maritima (L.) Stearn (squill), belonging to the Asparagaceae family, is acknowledged as a medicinally valuable species from the Drimia genera. In this study, water, methanol, and ethyl acetate extracts of D. maritima aerial parts and bulbs were investigated for their polyphenols profile and evaluated for their antioxidant and enzyme inhibition properties. Phenolics were profiled through an untargeted metabolomics approach using an ultra-high pressure liquid chromatograph coupled to quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF-MS). This analysis revealed an enrichment of low molecular weight phenolics and flavonoids in the aerial parts of D. maritima, while lignans mainly characterized bulb extracts. Antioxidant capacity was investigated by different assays, including phosphomolybdenum assays, radical scavenging (DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)), as well as reducing ability (CUPRAC: cupric reducing antioxidant capacity; FRAP: ferric reducing antioxidant power), and metal chelating. In radical scavenging and reducing power assays, the water extract of aerial parts exhibited the strongest ability (DPPH: 36.99 mg trolox equivalent (TE)/g; ABTS: 85.96 mg TE/g; CUPRAC: 87.37 mg TE/g; FRAP: 55.43 mg TE/g). In general, the ethyl acetate extracts from aerial parts and bulbs provided the weakest antioxidant capacity. Concerning enzyme inhibitory activities, the water extracts of the bulb were poorly active, while the ethyl acetate extracts from both plant portions displayed the best α-amylase inhibitory abilities. The best acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) abilities were recorded by ethyl acetate extract of aerial parts (2.36 mg galantamine equivalent (GALAE)/g) and bulbs (5.10 mg GALAE/g), respectively. Overall, these results support the medicinal aptitude of D. maritima and its possible use as a natural source of antioxidants and enzyme inhibitors with functional potential.

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'.

Ontogenetic Variation in the Mineral, Phytochemical and Yield Attributes of Brassicaceous Microgreens.

Microgreens constitute novel gastronomic ingredients that combine visual, kinesthetic and bioactive qualities. The definition of the optimal developmental stage for harvesting microgreens remains fluid. Their superior phytochemical content against mature leaves underpins the current hypothesis of significant changes in compositional profile during the brief interval of ontogeny from the appearance of the first (S1) to the second true leaf (S2). Microgreens of four brassicaceous genotypes (Komatsuna, Mibuna, Mizuna and Pak Choi) grown under controlled conditions and harvested at S1 and S2 were appraised for fresh and dry yield traits. They were further analyzed for macro- and micromineral content using inductively coupled plasma optical emission spectrometry (ICP-OES), carotenoid content using high-performance liquid chromatography with a diode-array detector (HPLC-DAD), volatile organic compounds using solid-phase microextraction followed by gas chromatography-mass spectrometry (SPME-GC/MS), anthocyanins and polyphenols using liquid chromatography-high resolution-tandem mass spectrometry (LC-MS/MS) with Orbitrap technology and for chlorophyll and ascorbate concentrations, well as antioxidant capacity by spectrophotometry. Analysis of compositional profiles revealed genotype as the principal source of variation for all constituents. The response of mineral and phytochemical composition and of antioxidant capacity to the growth stage was limited and largely genotype-dependent. It is, therefore, questionable whether delaying harvest from S1 to S2 would significantly improve the bioactive value of microgreens while the cost-benefit analysis for this decision must be genotype-specific. Finally, the lower-yielding genotypes (Mizuna and Pak Choi) registered higher relative increase in fresh yield between S1 and S2, compared to the faster-growing and higher-yielding genotypes. Although the optimal harvest stage for specific genotypes must be determined considering the increase in yield against reduction in crop turnover, harvesting at S2 seems advisable for the lower-yielding genotypes.

Morpho-Physiological Responses and Secondary Metabolites Modulation by Preharvest Factors of Three Hydroponically Grown Genovese Basil Cultivars.

Sweet basil (Ocimum basilicum L.) is an economically important leafy vegetable especially in Mediterranean countries. In Italian gastronomy, the large elliptical leaves of the Genovese type are mostly used for the well-known pesto sauce, and almost all (>90%) professional production is for the food industry. The growing demand for fresh leaves with standardized technological and sensory characteristics has prompted basil producers to adopt advanced cultivation methods such as the floating raft system (FRS). The aim of this study was to evaluate the productive, qualitative, and physiological performance of three Genovese basil cultivars ("Aroma 2," "Eleonora," and "Italiano Classico") in two successive harvests and at two densities (159 and 317 plants m-2). Caffeic, chicoric, rosmarinic, and ferulic acid were determined through the high-performance liquid chromatography (HPLC) system, whereas the extraction and quantification of the volatile organic compounds (VOCs) were performed by solid-phase microextraction (SPME) and gas chromatography coupled to a mass spectrometer (GC/MS). "Aroma 2" showed the highest fresh yield and photosynthetic rate together with the lowest nitrate content. For all the tested cultivars, the higher density, while reducing the number of leaves per plant, resulted in higher fresh and dry production per unit area, without altering the aroma profile. Successive harvests resulted in a significant increase in both the yield (37.5%) and the total phenolic acids (75.1%) and favored Eucalyptol and 1-octen-3-ol accumulation (+25.9 and +15.1%, respectively). The here presented comprehensive and multifactorial assessment of the productive and qualitative response of basil provides evidence of the positive effects (from biomass to specialized metabolites) that can be obtained from the management of the pre-harvest factors in soilless cultivation. In addition, it also highlights the role and constraints of the genetic factor in the observed response. We also discuss the implications of our work considering the impact for the food processing industry. Future research may explore the phenolic acids accumulation as a possible fortification means to extend the pesto sauce shelf life, reducing the need of added antioxidants and thermal processing.

Sweet Basil Functional Quality as Shaped by Genotype and Macronutrient Concentration Reciprocal Action.

Basil (Ocimum basilicum L.) is among the most widespread aromatic plants due to its versatility of use and its beneficial health properties. This aromatic plant thrives in hydroponics, which is a valid tool to improve the production and functional quality of crops, but nevertheless, it offers the possibility to de-seasonalize production. A floating raft system was adopted to test the production and quality potential during autumn season of three different genotypes of Genovese basil (Aroma 2, Eleonora and Italiano Classico) grown in three nutrient solutions with crescent electrical conductivity (EC: 1, 2 and 3 dS m-1). The aromatic and phenolic profiles were determined by GC/MS and HPLC analysis, respectively. The combination Aroma 2 and the EC 2 dS m-1 resulted in the highest production, both in terms of fresh weight and dry biomass. The 2 dS m-1 treatment determined the major phenolic content, 44%, compared to the other two EC. Italiano Classico showed a higher total polyphenolic content in addition to a different aromatic profile compared to the other cultivars, characterized by a higher percentage of Eucalyptol (+37%) and Eugenol (+107%) and a lower percentage of linalool (-44%). Correct management of the nutritional solution combined with adequate genetic material managed an improvement in the production and the obtainment of the desired aromatic and phenolic profiles.

Improved Porosity of Insect Proof Screens Enhances Quality Aspects of Zucchini Squash without Compromising the Yield.

In a global climate change environment, assuring optimal growing conditions is a difficult challenge, compromising the food supply for a rapidly rising population. The climatic conditions in the protected environment lead to high temperatures and fast insect development, impacting productivity and vegetables qualitative attributes. Consumers' interest in healthy food requires sustainable tools to manage biotic and abiotic factors and, from this perspective, anti-insect nets represent an excellent "green" solution. For this purpose, our goal was to compare two different anti-insect nets on microclimate, production, and qualitative traits of Cucurbita pepo L. fresh fruits. The experiment was conducted in three separate polyethylene high tunnels, with 50 mesh anti-insect nets of different porosities being installed on the openings of two tunnels, while the third tunnel was a control without nets. Microclimate measurements, as well as yield, physiological, and phytochemicals variables, were assessed. The 50 mesh net led to a decrease in marketable yield (22.5%), fruit number (18.0%), CO2 net assimilation rate (6.0%), and transpiration rate (29.5%). Total soluble solids, antioxidant activities and total ascorbic acid concentration had an opposite trend. The 50 mesh AirPlus net improved quality aspects of zucchini fruits by increasing total ascorbic acid, total phenols, and antioxidant compounds, with no negative impact on yield.

The Metabolic Reprogramming Induced by Sub-Optimal Nutritional and Light Inputs in Soilless Cultivated Green and Red Butterhead Lettuce.

Sub-optimal growing conditions have a major effect on plants; therefore, large efforts are devoted to maximizing the availability of agricultural inputs to crops. To increase the sustainable use of non-renewable inputs, attention is currently given to the study of plants under non-optimal conditions. In this work, we investigated the impact of sub-optimal macrocations availability and light intensity in two lettuce varieties that differ for the accumulation of secondary metabolites (i.e., 'Red Salanova' and 'Green Salanova'). Photosynthesis-related measurements and untargeted metabolomics were used to identify responses and pathways involved in stress resilience. The pigmented ('Red') and the non-pigmented ('Green Salanova') lettuce exhibited distinctive responses to sub-optimal conditions. The cultivar specific metabolomic signatures comprised a broad modulation of metabolism, including secondary metabolites, phytohormones, and membrane lipids signaling cascade. Several stress-related metabolites were altered by either treatment, including polyamines (and other nitrogen-containing compounds), phenylpropanoids, and lipids. The metabolomics and physiological response to macrocations availability and light intensity also implies that the effects of low-input sustainable farming systems should be evaluated considering a range of cultivar-specific positive and disadvantageous metabolic effects in addition to yield and other socio-economic parameters.

Understanding the Morpho-Anatomical, Physiological, and Functional Response of Sweet Basil to Isosmotic Nitrate to Chloride Ratios.

Sweet basil (Ocimum basilicum L.) is a leafy green with a short-production cycle that is emerging as a model species among aromatic plants. Modulating the mineral composition of the nutrient solution has proved to be a valuable tool to uncover the mechanisms and responses that higher plants adopt in relation to the availability of mineral nutrients. The aim of this work was to examine the effects on basil of four isosmotic nutrient solutions with different nitrate to chloride ratios. These two anions share uptake and transport mechanisms in plants and are often considered antagonist. To this goal, we analyzed morpho-anatomical and physiological parameters as well as quality-related traits, such as the antioxidant capacity, the leaf color, the mineral composition, and the aromatic profile in relation to the nutrient ratios. Moreover, using a full factorial design, we analyzed leaves in two consecutive harvests. The data indicated a broad, multifaceted plant response to the different nutritional ratios, with almost all the recorded parameters involved. Overall, the effects on basil can be explained by considering an interdependent combination of the nitrate and chloride roles in plant nutrition and physiology. Our work revealed the extent of the modification that can be achieved in basil through the modification of the nutrient solution. It also provided indications for more nutrient efficient growing conditions, because a moderate increase in chloride limits the expected negative impact of a sub-optimal nitrate fertilization.

Shading Affects Yield, Elemental Composition and Antioxidants of Perennial Wall Rocket Crops Grown from Spring to Summer in Southern Italy.

Shading nets have been increasingly drawing research interest, as they allow us to improve the environmental conditions for greenhouse-grown crops. The effects of two shading nets (50% and 79% shading degree), plus an unshaded control, on yield, mineral composition and antioxidants of perennial wall rocket (Diplotaxis tenuifolia L.-D.C.) grown under tunnels in southern Italy were determined. The shading application resulted in a yield decrease, compared to the unshaded control, except for the highest production under 50% shading in July. The highest yield was recorded in the April-May and May-June and the lowest in July. Similar trends were recorded for plant dry weight, leaf number per rosette and mean weight, but the latter showed the highest value under 79% light extinction in July. The rocket leaves were brighter in the summer cycles than in the spring ones. Leaf nitrate was highest in spring and under 79% shading. Potassium, phosphorus, calcium and magnesium showed the highest values in spring and in the unshaded control. The lipophilic antioxidant activity showed the highest values under the 79% shading net in the spring cropping seasons, whereas in July it did not significantly differ from 50% light extinction. The hydrophilic antioxidant activity always attained the highest values in the unshaded control. The unshaded leaves had the highest total phenol accumulation when grown in April-May and the lowest in July. The total ascorbic acid content was always highest in the unshaded control leaves compared to the shading treatments. Fifty percent crop shading is, therefore, an effective sustainable tool for increasing the yield of perennial wall rocket leaves in July, when the light intensity under the plastic tunnel exceeds the plant requirements, also resulting in a mineral composition that is not significantly different from that of the unshaded crops.

Design of a Module for Cultivation of Tuberous Plants in Microgravity: The ESA Project "Precursor of Food Production Unit" (PFPU).

Plant cultivation systems for Bioregenerative Life-Support Systems in Space developed on Earth need to be tested in space, where reduced gravity alters the liquid and gas behavior both within the plant and between the plant and its surrounding environment, making the distribution of water and nutrients a critical issue. The ESA project "Precursor of Food Production Unit" (PFPU) aims to design a modular cultivation system for edible tuberous plants (such as potato and sweet potato) in microgravity, to be preliminary tested in ground conditions in the view of successive space application. Among the different modules of the PFPU demonstrator, the Root Module (RM) is the component physically hosting the plant and accommodating tubers and roots. This paper describes the step-by-step procedure adopted to realize the RM, including the design, the building, and the ground testing of its prototype. Specifically, the hydrological characterization of possible cultivation substrates, the set-up of the water distribution system, and the validation test of the assembled prototype in a tuber-to-tuber growing cycle of potato plants are described. Among six substrates tested, including three organic materials and three synthetic materials, cellulosic sponge was selected as the best one, based on the hydrological behavior in terms of air and water transport and water retention capacity. The water sensor WaterScout was successfully calibrated to monitor the water status in cellulosic sponge and to drive irrigation and fertigation management. The designed porous tubes-based distribution system, integrated with water sensors, was able to provide water or nutrient solution in a timely and uniform way in cellulosic sponge.

Air Distribution in a Fully-Closed Higher Plant Growth Chamber Impacts Crop Performance of Hydroponically-Grown Lettuce.

The MELiSSA Pilot Plant (MPP) is testing in terrestrial conditions regenerative life support technologies for human exploration in Space. One of its components is a controlled Higher Plant Chamber (HPC) accommodating hydroponic plant cultures. It consists of a 9 m3 single closed growth chamber providing adequate environmental conditions for growing plants, enabling the production of food, water and oxygen for the crew. A critical aspect for a reliable HPC performance is to achieve homogeneous air distribution. The initial experiment carried out in the MPP with lettuce as salad crop, showed uneven plant growth throughout the HPC, which was attributed to inadequate air distribution due to non-homogeneous air velocity profile along the inlet-vents. After a detailed computational fluid dynamics (CFD) analysis, the heating, ventilation, and air conditioning subsystem of the HPC was upgraded and a new experiment was carried out in optimized air flow conditions. Nine-day seedlings of lettuce cultivar "Grand Rapids" were transplanted into the HPC and harvested at the end of the growing cycle, where shoot fresh weight, dry biomass, and shoot mineral composition were analyzed. During the experiment, the environmental control system performed remarkably well based on the biometric measurements as well as the mineral composition leading to a vast homogeneous growth. Overall, the results demonstrated the beneficial effect of an adequate air distribution system in HPCs and the effectiveness of CFD-analysis to design properly the gas distribution. The obtained results are of high relevance for life support systems in space involving plants growth.

Mars Regolith Simulant Ameliorated by Compost as in situ Cultivation Substrate Improves Lettuce Growth and Nutritional Aspects.

Heavy payloads in future shuttle journeys to Mars present limiting factors, making self-sustenance essential for future colonies. Therefore, in situ resources utilization (ISRU) is the path to successful and feasible space voyages. This research frames the concept of planting leafy vegetables on Mars regolith simulant, ameliorating this substrate's fertility by the addition of organic residues produced in situ. For this purpose, two butterhead lettuce (Lactuca sativa L. var. capitata) cultivars (green and red Salanova®) were chosen to be cultivated in four different mixtures of MMS-1 Mojave Mars simulant:compost (0:100, 30:70, 70:30 and 100:0; v:v) in a phytotron open gas exchange growth chamber. The impact of compost rate on both crop performance and the nutritive value of green- and red-pigmented cultivars was assessed. The 30:70 mixture proved to be optimal in terms of crop performance, photosynthetic activity, intrinsic water use efficiency and quality traits of lettuce. In particular, red Salanova® showed the best performance in terms of these quality traits, registering 32% more phenolic content in comparison to 100% simulant. Nonetheless, the 70:30 mixture represents a more realistic scenario when taking into consideration the sustainable use of compost as a limited resource in space farming, while still accepting a slight significant decline in yield and quality in comparison to the 30:70 mixture.