The role of light quality of photoperiodic lighting on photosynthesis, flowering and metabolic profiling in Ranunculus asiaticus L.

Photoperiodic light quality affects flowering of long day plants, by influencing the phytochrome photoequilibria (PPE) at plant level; however, the most effective light spectrum to promote flowering is still unknown for most of the flower crops. We evaluated the influence of light spectrum of three light sources, with different induced PPE, on photosynthesis, metabolic profiling, plant growth and flowering in two hybrids of Ranunculus asiaticus L., MBO (early flowering) and MDR (medium earliness). Three photoperiodic treatments were compared to natural day length (NL): white fluorescent light (PPE 0.84), light emitting diodes (LEDs) with red:far red (R:FR) light at 3:1 ratio (PPE, 0.84) and LEDs with R:FR light at 1:3 ratio (PPE 0.63). Under natural light, net photosynthesis was higher in MDR than in MBO, while photochemistry was similar in the hybrids. Compared to NL, photoperiodic treatments did not affect net photosynthesis, while they promoted the quantum yield of PSII and reduced the non-photochemical quenching. Under NL, plant growth was greater in MBO, while flowering started earlier in MDR and flowers characteristics were similar in the hybrids. Despite the greater sensitivity of MDR plants in terms of metabolism, photoperiodic lighting improved plant growth and reduced the flowering time only in MBO, with a stronger effect under R:FR 3:1 light. MDR plants were characterized by higher soluble sugars, polyphenols, photosynthetic pigments and proteins, while MBO plants by higher starch and amino acid content. The morphological effects of photoperiodic light quality and the hybrid-specific response should be taken into account to optimize lighting protocols in commercial farms.

Effects of plant size, temperature, and light intensity on flowering of Phalaenopsis hybrids in Mediterranean greenhouses.

Mediterranean greenhouses for cultivation of Phalaenopsis orchids reproduce the warm, humid, and shaded environment of tropical underbrush. Heating represents the highest production cost, due to the high thermal requirements and the long unproductive phase of juvenility, in which plants attain the critical size for flowering. Our researches aimed to investigate the effect of plant size, temperature, and light intensity, during the phase of flower induction, on flowering of modern genotypes selected for Mediterranean greenhouses. Three experiments were carried out to compare (i) plant size: reduced size versus size considered optimal for flowering (hybrids "Sogo Yukidian," "Chain Xen Diamond," and "Pinlong"); (ii) temperature: moderate reduction of temperature versus standard thermal regime (hybrid "Premium"); (iii) light intensity: supplemental lighting versus reference light intensity (hybrid "Premium"). The premature exposure of plants to the inductive treatment delayed the beginning of flowering and reduced the flower stem quality, in all the tested hybrids. In "Premium," the lower temperature did not affect flowering earliness and commercial quality of flower stems compared to the standard regime, whereas it promoted stem branching. In the same hybrid, supplemental lighting anticipated flowering and promoted the emission of the second stem and the stem branching, compared to the reference light regime.

Integrating Smart Greenhouse Cover, Reduced Nitrogen Dose and Biostimulant Application as a Strategy for Sustainable Cultivation of Cherry Tomato.

Fruit yield and quality of greenhouse tomatoes are strongly influenced by light conditions and nitrogen (N) availability, however, the interaction between these factors is still unclear. We evaluated the effects on cherry tomatoes of two tunnel plastic covers with different optical properties and three N doses, also in combination with a biostimulant treatment. We compared a diffuse light film (Film1) and a conventional clear film (Film2), and three N levels, corresponding to 50% (N50), 75% (N75) and 100% (N100) of the optimal dose, with and without a microbial plus a protein hydrolysed biostimulant, compared to a non-treated control. The three experimental treatments significantly interacted on several yield and quality parameters. In control plants (untreated with biostimulants), the early yield was higher at reduced N doses compared to N100, with greater increments under the diffusive Film1 compared to the clear Film2 (+57.7% and +37.0% vs. +31.7% and +16.0%, in N50 and N75 respectively). Film1 boosted the total fruit production at all the N rates and with or without biostimulants, compared to Film2, with stronger effects under sub-optimal N (+29.4% in N50, +21.2% in N75, and +7.8% in N100, in plants untreated with biostimulant). Total yield decreased with decreasing N levels, while it always increased with the application of biostimulants, which counterbalanced the detrimental effects of N shortage. Quality traits were mainly affected by the cover film and the biostimulant treatment. The diffusive film increased the content of carotenoids, lycopene and total phenols compared to the clear one, and the biostimulants increased texture, soluble solids, phenols and ascorbic acid compared to the untreated control. It is worth noting that in plants fertilized at 75% of the reference N dose, the biostimulants determined higher yield than the N100 untreated control, under both the covers (+48% in Film1 and +20% in Film2). In conclusion, the diffusive film improved the fruit yield and quality of greenhouse tomatoes in the spring-summer period, presumably avoiding plant stress due to high-intensity direct light. Reduced N rates limited the plant productivity, however, the biostimulant application was effective in compensating for the detrimental effects of sub-optimal supply of N synthetic fertilizers.

Assessment of Fertility Dynamics and Nutritional Quality of Potato Tubers in a Compost-Amended Mars Regolith Simulant.

Mars exploration will foresee the design of bioregenerative life support systems (BLSSs), in which the use/recycle of in situ resources might allow the production of food crops. However, cultivation on the poorly-fertile Mars regolith will be very challenging. To pursue this goal, we grew potato (Solanum tuberosum L.) plants on the MMS-1 Mojave Mars regolith simulant, pure (R100) and mixed with green compost at 30% (R70C30), in a pot in a cold glasshouse with fertigation. For comparison purposes, we also grew plants on a fluvial sand, pure (S100) and amended with 30% of compost (S70C30), a volcanic soil (VS) and a red soil (RS). We studied the fertility dynamics in the substrates over time and the tuber nutritional quality. We investigated nutrient bioavailability and fertility indicators in the substrates and the quality of potato tubers. Plants completed the life cycle on R100 and produced scarce but nutritious tubers, despite many critical simulant properties. The compost supply enhanced the MMS-1 chemical/physical fertility and determined a higher tuber yield of better nutritional quality. This study demonstrated that a compost-amended Mars simulant could be a proper substrate to produce food crops in BLSSs, enabling it to provide similar ecosystem services of the studied terrestrial soils.

Light Intensity Affects the Assimilation Rate and Carbohydrates Partitioning in Spinach Grown in a Controlled Environment.

The cultivation of spinach (Spinacia oleracea L.) has been increasing during the last years in controlled environment agriculture, where light represents a key factor for controlling plant growth and development and the highest energetic costs. The aim of the experiment was to evaluate the plant's response to two light intensities, corresponding to an optimal and a reduced level, in terms of the photosynthetic process, photoassimilates partitioning, and the biosynthesis of sucrose and starch. Plants of spinach cv. 'Gigante d'Inverno' were grown in a phytotron under controlled conditions, comparing two values of photosynthetic photon flux density (PPFD), 800 µmol m-2 s-1 (800 PPFD) and 200 µmol m-2 s-1 (200 PPFD), at a 10 h light/14 h dark regime. Compared to 800 PPFD, under 200 PPFD, plants showed a reduction in biomass accumulation and a redirection of photoassimilates to leaves, determining a leaf expansion to optimize the light interception, without changes in the photosynthetic process. A shift in carbon partitioning favouring the synthesis of starch, causing an increase in the starch/sucrose ratio at the end of light period, occurred in low-light leaves. The activity of enzymes cFBAse, SPS, and AGPase, involved in the synthesis of sucrose and starch in leaves, decreased under lower light intensity, explaining the rate of accumulation of photoassimilates.

Metabolic Profiling in Tuberous Roots of Ranunculus asiaticus L. as Influenced by Vernalization Procedure.

Ranunculus asiaticus L. is an ornamental geophyte. In commercial practice, it is mainly propagated by rehydrated tuberous roots. Vernalization before planting is a common practice to overcome the natural dormancy of tuberous roots; however, little is known about the mechanisms underlying the plant's response to low temperatures. We investigated the influence of three preparation procedures of tuberous roots, only rehydration (control, C), and rehydration plus vernalization at 3.5 °C for 2 weeks (V2) and for 4 weeks (V4), on plant growth, leaf photosynthesis, flowering, and metabolism in plants of two hybrids, MBO (early flowering, pale orange flower) and MDR (medium earliness, bright orange flower), grown in pots in an unheated greenhouse. We reported the responses observed in the aerial part in a previous article in this journal. In this paper, we show changes in the underground organs in carbohydrate, amino acids, polyphenols, and protein levels throughout the growing cycle in the different plant stages: pre-planting, vegetative growth, and flowering. The metabolic profile revealed that the two hybrids had different responses to the root preparation procedure. In particular, MBO synthesized GABA and alanine after 2 weeks and sucrose after 4 weeks of vernalization. In contrast, MDR was more sensitive to vernalization; in fact, a higher synthesis of polyphenols was observed. However, both hybrids synthesized metabolites that could withstand exposure to low temperatures.

Vernalization Procedure of Tuberous Roots Affects Growth, Photosynthesis and Metabolic Profile of Ranunculus asiaticus L.

In Ranunculus asiaticus L., vernalization of propagation material is a common practice for the production scheduling of cut flowers, however little is known about the plant physiology and metabolism of this species as affected by cold treatments. We investigated the influence of two hybrids, MBO and MDR, and three preparation procedures of tuberous roots, only rehydration (control, C), and rehydration plus vernalization at 3.5 °C for 2 weeks (V2) and for 4 weeks (V4), on plant growth and flowering, leaf photosynthesis, and leaf metabolic profile in plants grown in pot in a cold greenhouse. Net photosynthesis (NP) was higher in MDR than in MBO. In the two genotypes, the NP did not change in V2 and increased in V4 compared to C in MBO, while was unaffected by vernalization in MDR. Quantum yield of PSII electron transport (ΦPSII), linear electron transport rate (ETR) and non-photochemical quenching (NPQ) did not differ in the two hybrids, whereas maximal PSII photochemical efficiency (Fv/Fm) was higher in MBO than in MDR. Fluorescence indexes were unaffected by the preparation procedure, except for ETR, which decreased in V2 compared to C and V4 in MDR. A significant interaction between genotype and preparation procedure was found in plant leaf area, which was reduced only in V4 in MBO, while decreased in both the vernalization procedures in MDR. In Control plants, flowering started in 65 days in MBO and 69 days in MDR. Compared to controls, both the vernalization treatments anticipated flowering in MDR, while they were detrimental or only slightly efficient in promoting flowering in MBO. Vernalization always reduced the quality of flower stems in both the hybrids.

Modulation of Antioxidant Compounds in Fruits of Citrus reticulata Blanco Using Postharvest LED Irradiation.

Phlegrean mandarin fruits are already known for health-promoting properties due to the high concentration of phytochemicals in peel, pulp, and seed. Biotic and abiotic factors, including light, may modulate their biosynthesis, metabolism, and accumulation. In this context, light-emitting diodes (LED) have recently been applied to control nutritional traits, ripening process, senescence, fruit shelf-life, and pathogenic microbial spoilage of fruits. This study investigated the effect of the seven-day exposure of Phlegrean mandarin fruits to two LED regimes, white (W) and red-blue (RB), to test the possibility that the storage under specific light wavelengths may be used as green preservation technology that enhances fruit phytochemical properties. To pursue this aim, the antioxidant activity and polyphenolic profile of the pulp and peel of mandarins under W and RB light regimes were evaluated and compared with Control fruits not exposed to LED treatment. Our results indicated that storage under W and RB treatments modulates the antioxidant content in pulp and peel differently. Compared to W, the RB regime increases the ascorbic acid, flavonoid, anthocyanin, and carotenoid concentrations, while the polyphenol profile analysis reveals that the number of important phytochemicals, i.e., quercetin rutinoside, chlorogenic acid, sinensetin, and rutin, are higher under W. The overall data demonstrated that postharvest LED irradiation is a valid tool for modifying fruit phytochemical properties, which also boosts specific bioactive compounds.

Defining growth requirements of microgreens in space cultivation via biomass production, morpho-anatomical and nutritional traits analysis.

During long-term manned missions to the Moon or Mars, the integration of astronauts' diet with fresh food rich in functional compounds, like microgreens, could strengthen their physiological defenses against the oxidative stress induced by the exposure to space factors. Therefore, the development of targeted cultivation practices for microgreens in space is mandatory, since the cultivation in small, closed facilities may alter plant anatomy, physiology, and resource utilization with species-specific responses. Here, the combined effect of two vapor pressure deficit levels (VPD: 0.14 and 1.71 kPa) and two light intensities (150 and 300 µmol photons m-2 s-1 PPFD) on two species for microgreen production (Brassica oleracea var. capitata f. sabauda 'Vertus' and Raphanus raphanistrum subsp. sativus 'Saxa'), was tested on biomass production per square meter, morpho-anatomical development, nutritional and nutraceutical properties. Microgreens were grown in fully controlled conditions under air temperature of 18/24°C, on coconut fiber mats, RGB light spectrum and 12 h photoperiod, till they reached the stage of first true leaves. At this stage microgreens were samples, for growth and morpho-anatomical analyses, and to investigate the biochemical composition in terms of ascorbic acid, phenols, anthocyanin, carotenoids, carbohydrates, as well as of anti-nutritional compounds, such as nitrate, sulfate, and phosphate. Major differences in growth were mostly driven by the species with 'Saxa' always presenting the highest fresh and dry weight as well as the highest elongation; however light intensity and VPDs influenced the anatomical development of microgreens, and the accumulation of ascorbic acid, carbohydrates, nitrate, and phosphate. Both 'Saxa' and 'Vertus' at low VPD (LV) and 150 PPFD increased the tissue thickness and synthetized high ß-carotene and photosynthetic pigments. Moreover, 'Vertus' LV 150, produced the highest content of ascorbate, fundamental for nutritional properties in space environment. The differences among the treatments and their interaction suggested a relevant difference in resource use efficiency. In the light of the above, microgreens can be considered suitable for cultivation in limited-volume growth modules directly onboard, provided that all the environmental factors are combined and modulated according to the species requirements to enhance their growth and biomass production, and to achieve specific nutritional traits.

Flowering Mechanisms and Environmental Stimuli for Flower Transition: Bases for Production Scheduling in Greenhouse Floriculture.

The scheduling of plant production is a critical aspect in modern floriculture since nowadays, sales are not oriented toward the recurring holidays as in the past, but always more toward impulse buying, implying a more diverse and constant demand on the market. This requires continuous production, often regulated by precise commercial agreements between growers and buyers, and between buyers and dealers, particularly in large-scale retail trade. In this scenario, diverse techniques to modulate the duration of the growing cycle, by hastening or slowing down plant growth and development, have been developed to match plant flowering to the market demand. Among the numerous approaches, the manipulation of climatic parameters in the growth environment is one of the most common in greenhouse floriculture. In this review, we summarize the physiological and biochemical bases underlying the main mechanisms of flowering, depending on the plant reaction to endogenous signals or environmental stimuli. In addition, the strategies based on the control of temperature (before or after planting) and light environment (as light intensity and spectrum, and the photoperiod) in the scheduling of flower and ornamental crop production are briefly described.

Manipulation of light quality is an effective tool to regulate photosynthetic capacity and fruit antioxidant properties of Solanum lycopersicum L. cv. 'Microtom' in a controlled environment.

Light quality plays an essential role in setting plant structural and functional traits, including antioxidant compounds. This paper aimed to assess how manipulating the light spectrum during growth may regulate the photosynthetic activity and fruit bioactive compound synthesis in Solanum lycopersicum L. cv. 'Microtom' to improve plant physiological performance and fruit nutritional value. Plants were cultivated under three light quality regimes: red-green-blue LEDs (RGB), red-blue LEDs (RB) and white fluorescent lamps (FL), from sowing to fruit ripening. Leaf functional traits, photosynthetic efficiency, Rubisco and D1 protein expression, and antioxidant production in fruits were analyzed. Compared to FL, RGB and RB regimes reduced height and increased leaf number and specific leaf area, enhancing plant dwarf growth. The RGB regime improved photosynthesis and stomatal conductance despite lower biomass, favoring Rubisco synthesis and carboxylation rate than RB and FL regimes. The RB light produced plants with fewer flowers and fruits with a lower ascorbic acid amount but the highest polyphenol content, antioxidant capacity and SOD and CAT activities. Our data indicate that the high percentage of the green wavelength in the RGB regime promoted photosynthesis and reduced plant reproductive capacity compared to FL and RB. Conversely, the RB regime was the best in favoring the production of health-promoting compounds in tomato berries.

Can Peat Amendment of Mars Regolith Simulant Allow Soybean Cultivation in Mars Bioregenerative Life Support Systems?

Higher plants will play a key role in human survival in Space, being able to regenerate resources and produce fresh food. However, the creation of a fertile substrate based on extra-terrestrial soils is still a challenge for space cultivation. We evaluated the adaptability of soybean (Glycine max (L.) Merr.) cultivar 'Pr91M10' to three substrates, the Mojave Mars regolith Simulant MMS-1, alone (R100), and in a mixture with blond sphagnum peat at two different volumes, 85:15 (R85P15) and 70:30 (R70P30), in plants directly sown on the substrates or transplanted after sowing on peat. The low pH of peat (4.34) allowed the mitigation of the alkalinity of the Mars regolith simulant (pH 8.86), lowering the initial pH to neutral (6.98, R85P15), or subacid to neutral (6.33, R70P30) values. Seed germination reached the highest percentage in the shortest time in the mixture of regolith simulant with 15% of peat. The cultivation substrate did not affect the plant growth and nutritional status. However, a significant interaction between the substrate and planting method was found in several growth parameters, with the highest positive effects observed in plants resulting from direct sowing on the regolith mixture with peat.

Light response of photosynthesis and stomatal conductance of rose leaves in the canopy profile: the effect of lighting on the adaxial and the abaxial sides.

We investigated the light response of leaf photosynthesis, stomatal conductance and optical properties in rose plants grown in a glasshouse with bending technique. Leaves were lighted from the adaxial or the abaxial side during measurements, performed in four positions in the upright and bent shoots: top leaves, middle leaves, bottom leaves, and bent shoot leaves. Moreover, the effect of the irradiation on the adaxial or abaxial leaf side on whole canopy photosynthesis was estimated through model simulation. No significant differences were found in light transmission, reflection and absorption of leaves and in photosynthesis light response curves among the four positions. In all the leaf positions, light absorption, stomatal conductance and photosynthesis were higher when leaves were lighted from the adaxial compared with the abaxial side. The model showed that a substantial part of the light absorbed by the crop originated from light reflected from the greenhouse floor, and thus the abaxial leaf properties have impact on whole crop light absorbance and photosynthesis. Simulations were performed for crops with leaf area index (LAI) 1, 2 and 3. Simulation at LAI 1 showed the highest reduction of simulated crop photosynthesis considering abaxial properties; however, to a lesser extent photosynthesis was also reduced at LAI 2 and 3. The overall results showed that the model may be helpful in designing crop systems for improved light utilisation by changing lamp position or level of leaf bending and pruning.

Metabolic Profile and Performance Responses of Ranunculus asiaticus L. Hybrids as Affected by Light Quality of Photoperiodic Lighting.

Ranunculus asiaticus is a quantitative long day plant grown for cut flowers and flowering potted plants production. We evaluated the influence of light spectrum of three light sources for end-of-day photoperiodic treatments, with different phytochrome photoequilibria (PPE) induced at plant level, on the metabolic profiling of two hybrids of R. asiaticus L., MBO and MDR, in plants from vernalized tuberous roots. The following treatments were compared with natural day length (NL): white fluorescence lamp (FL, PPE 0.84), light emitting diodes (LEDs) Red:Far Red light at 3:1 ratio (R:FR 3:1, PPE 0.84), and LEDs Red:Far Red light at 1:3 ratio (R:FR 1:3, PPE 0.63). Measurements were carried out to evaluate the time course of carbohydrate, amino acid, and protein levels throughout the growing cycle in tuberous roots and leaves, in relation to the different plant stages (pre-planting, vegetative phase, and flowering). The study of metabolic profiling suggested that the differences between the tuberous root reserves of the two R. asiaticus hybrids could be responsible for the capacity of MBO to exert an early flowering. In particular, the proton-consuming synthesis during the pre-planting of two amino acids, alanine and γ-aminobutyric acid (GABA), is able to buffer the cytoplasmic acidosis and pH altered by the vernalization process, and GABA itself can efficiently scavenge reactive oxygen species. This fast response to the stress caused by vernalization allows MBO plants to accelerate the process of vegetative development and flowering. Some other changes in metabolites profile were certainly related to the different responses to day length and photoperiodic light quality in the two hybrids, such as dose exerted by low R:FR lighting in both MBO and MDR. However, most of the responses are under a strict genetic control.

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.

Photosynthesis in Ranunculus asiaticus L.: The Influence of the Hybrid and the Preparation Procedure of Tuberous Roots.

Ranunculus asiaticus L. is a quantitative long-day geophyte, grown in a cold greenhouse for cut flowers and potted plants. Flowering in ranunculus is a complex process, strongly steered by temperature and photoperiodism. Vernalization of rehydrated tuberous roots anticipate sprouting and leaf rosette formation and flowering. It is known that the time for flowering and the sensitivity to cold treatment, in terms of flowering anticipation, varies in numerous hybrids, while no information seems to be available on the influence of hybrids and on the vernalization on the photosynthetic process and primary metabolite profiling. We investigated the influence of two ranunculus hybrids, MDR and MBO, and two preparation procedures of tuberous roots, only rehydration (Control, C) and rehydration followed by vernalization (V), on the photosynthesis and photochemistry of plants grown in a climatic chamber, under a controlled environment. In addition, in MBO plants, in which the vernalization showed the main effects, carbohydrate, amino acid and protein levels were also investigated. In control plants, the response of leaf photosynthesis, to increasing white light, revealed higher photosynthetic activity in MDR than in MBO. The quantum yield of PSII (ϕPSII), electron transport rate (ETR) and non-photochemical quenching (NPQ) did not differ between the two hybrids. The maximal photochemical efficiency (Fv/Fm) was higher in MBO than in MDR and showed a decrease in both hybrids after vernalization. The preparation treatment of propagation material affected the light response of photosynthesis in the two hybrids differently, which increased in plants from vernalized tuberous roots, compared to those from only rehydrated in MBO and decreased in MDR, in accordance to the effects of vernalization observed in leaf photosynthetic pigments. In MBO vernalized tuberous roots, starch was rapidly degraded, and the carbon skeletons used to synthesize amino acids. Control plants of MBO, developed more leaves than those of MDR and a consequent larger plant leaf area. Compared to only rehydration, vernalization of rehydrated tuberous roots increased the plant leaf area in both the hybrids. Compared to the control, vernalized tuberous roots of MBO showed higher concentrations of sucrose and free amino acids, which could act as a long-distance signal promoting floral transition in young leaf primordia.

Changes in Leaf Anatomical Traits Enhanced Photosynthetic Activity of Soybean Grown in Hydroponics with Plant Growth-Promoting Microorganisms.

The use of hydroponic systems for cultivation in controlled climatic conditions and the selection of suitable genotypes for the specific environment help improving crop growth and yield. We hypothesized that plant performance in hydroponics could be further maximized by exploiting the action of plant growth-promoting organisms (PGPMs). However, the effects of PGPMs on plant physiology have been scarcely investigated in hydroponics. Within a series of experiments aimed to identify the best protocol for hydroponic cultivation of soybean [Glycine max (L.) Merr.], we evaluated the effects of a PGPMs mix, containing bacteria, yeasts, mycorrhiza and trichoderma beneficial species on leaf anatomy, photosynthetic activity and plant growth of soybean cv. 'Pr91m10' in closed nutrient film technique (NFT). Plants were grown in a growth chamber under semi-aseptic conditions and inoculated at seed, seedling and plant stages, and compared to non-inoculated (control) plants. Light and epi-fluorescence microscopy analyses showed that leaves of inoculated plants had higher density of smaller stomata (297 vs. 247 n/mm2), thicker palisade parenchyma (95.0 vs. 85.8 µm), and larger intercellular spaces in the mesophyll (57.5% vs. 52.2%), compared to non-inoculated plants. The modifications in leaf functional anatomical traits affected gas exchanges; in fact starting from the reproductive phase, the rate of leaf net photosynthesis (NP) was higher in inoculated compared to control plants (8.69 vs. 6.13 µmol CO2 m-2 s-1 at the beginning of flowering). These data are consistent with the better maximal PSII photochemical efficiency observed in inoculated plants (0.807 vs. 0.784 in control); conversely no difference in leaf chlorophyll content was found. The PGPM-induced changes in leaf structure and photosynthesis lead to an improvement of plant growth (+29.9% in plant leaf area) and seed yield (+36.9%) compared to control. Our results confirm that PGPMs may confer benefits in photosynthetic traits of soybean plants even in hydroponics (i.e., NFT), with positive effects on growth and seed production, prefiguring potential application of beneficial microorganisms in plant cultivation in hydroponics.

Effect of bacterial root symbiosis and urea as source of nitrogen on performance of soybean plants grown hydroponically for Bioregenerative Life Support Systems (BLSSs).

Soybean is traditionally grown in soil, where root symbiosis with Bradyrhizobium japonicum can supply nitrogen (N), by means of bacterial fixation of atmospheric N2. Nitrogen fertilizers inhibit N-fixing bacteria. However, urea is profitably used in soybean cultivation in soil, where urease enzymes of telluric microbes catalyze the hydrolysis to ammonium, which has a lighter inhibitory effect compared to nitrate. Previous researches demonstrated that soybean can be grown hydroponically with recirculating complete nitrate-based nutrient solutions. In Space, urea derived from crew urine could be used as N source, with positive effects in resource procurement and waste recycling. However, whether the plants are able to use urea as the sole source of N and its effect on root symbiosis with B. japonicum is still unclear in hydroponics. We compared the effect of two N sources, nitrate and urea, on plant growth and physiology, and seed yield and quality of soybean grown in closed-loop Nutrient Film Technique (NFT) in growth chamber, with or without inoculation with B. japonicum. Urea limited plant growth and seed yield compared to nitrate by determining nutrient deficiency, due to its low utilization efficiency in the early developmental stages, and reduced nutrients uptake (K, Ca, and Mg) throughout the whole growing cycle. Root inoculation with B. japonicum did not improve plant performance, regardless of the N source. Specifically, nodulation increased under fertigation with urea compared to nitrate, but this effect did not result in higher leaf N content and better biomass and seed production. Urea was not suitable as sole N source for soybean in closed-loop NFT. However, the ability to use urea increased from young to adult plants, suggesting the possibility to apply it during reproductive phase or in combination with nitrate in earlier developmental stages. Root symbiosis did not contribute significantly to N nutrition and did not enhance the plant ability to use urea, possibly because of ineffective infection process and nodule functioning in hydroponics.