Soybean cultivation for Bioregenerative Life Support Systems (BLSSs): The effect of hydroponic system and nitrogen source

Soybean [Glycine max (L.) Merr.] is one of the plant species selected within the European Space Agency (ESA) Micro-Ecological Life Support System Alternative (MELiSSA) project for hydroponic cultivation in Biological Life Support Systems (BLSSs), because of the high nutritional value of seeds. Root symbiosis of soybean with Bradirhizobium japonicum contributes to plant nutrition in soil, providing ammonium through the bacterial fixation of atmospheric nitrogen. The aim of this study was to evaluate the effects of two hydroponic systems, Nutrient Film Technique (NFT) and cultivation on rockwool, and two nitrogen sources in the nutrient solution, nitrate (as Ca(NO₃)₂ and KNO₃) and urea (CO(NH₂)₂), on root symbiosis, plant growth and seeds production of soybean. Plants of cultivar ‘OT8914’, inoculated with B. japonicum strain BUS-2, were grown in a growth chamber, under controlled environmental conditions.     

Pressure,O2, and CO2, in aquatic Closed Ecological Systems

Pressure increased during net photosynthetic O₂ production in the light and decreased during respiratory O₂ uptake during the dark in aquatic Closed Ecological Systems (CESs) with small head gas volumes. Because most CO₂ will be in the liquid phase as bicarbonate and carbonate anions, and CO₂ is more soluble than O₂, volumes of gaseous CO₂ and gaseous O₂ will not change in a compensatory manner, leading to the development of pressure. Pressure increases were greatest with nutrient rich medium with NaHCO₃ as the carbon source. With more dilute media, pressure was greatest with NaHCO₃, and less with cellulose or no-added carbon. Without adequate turbulence, pressure measurements lagged dissolved O₂ concentrations by several hours and dark respiration would have been especially underestimated in our systems (250–1000 ml). With adequate turbulence (rotary shaker), pressure measurements and dissolved O₂ concentrations generally agreed during lights on/off cycles, but O₂ measurements provided more detail. At 20 °C, 29.9 times as much O₂ will distribute into the gas phase as in the liquid, per unit volume, as a result of the limited solubility of O₂ in water and according to Henry’s Law. Thus even a small head gas volume can contain more O₂ than a larger volume of water. When both dissolved and gaseous O₂ and CO₂ are summed, the changes in Total O₂ and CO₂ are in relatively close agreement when NaHCO₃ is the carbon source. These findings disprove an assumption made in some of Taub’s earlier research that aquatic CESs would remain at approximately atmospheric pressure because approximately equal molar quantities of O₂ and CO₂ would exchange during photosynthesis and respiration; this assumption neglected the distribution of O₂ between water and gas phases. High pressures can occur when NaHCO₃ is the carbon source in nutrient rich media and if head-gas volumes are small relative to the liquid volume; e.g., one “worse case” condition developed 800 mm Hg above atmospheric pressure and broke the glass container. Plastic screw cap closures are likely to leak at high pressures and should not be assumed to seal unless tested at appropriate pressures. Pressure can be reduced by having larger head-gas volumes and using less concentrated nutrient solutions. It is important that pressure changes be considered for both safety and closure, and if total O₂ is used as the measure of net photosynthesis and respiration, the O₂ in the gas phase must be added to the dissolved O₂.     

Possible nutrient limiting factor in long term operation of closed aquatic ecosystem

To investigate nutrient limitation effect on the community metabolism of closed aquatic ecosystem and possible nutrient limiting factors in the experimental food chains, depletion of inorganic chemicals including carbon, nitrogen and phosphorous was tested. A closed aquatic ecosystem lab module consisting of Chlorella pyrenoidosa and Chlamydomonas reinhardtii, Daphnia magna and associated unidentified microbes was established. Closed ecological systems receive no carbon dioxide; therefore, we presumed carbon as a first limiting factor. The results showed that the algae population in the nutrient saturated group was statistically higher than that in the nutrient limited groups, and that the chlorophyll a content of algae in the phosphorus limited group was the highest among the limited groups. However, the nitrogen limited group supported the most Daphnia, followed by the carbon limited group, the nutrient saturated group and the phosphorus limited group. Redundancy analysis showed that the total phosphorus contents were correlated significantly with the population of algae, and that the amount of soluble carbohydrate as feedback of nutrient depletion was correlated with the number of Daphnia. Thus, these findings suggest that phosphorus is the limiting factor in the operation of closed aquatic ecosystem. The results presented herein have important indications for the future construction of long term closed ecological system.     

Physiologic and metabolic responses of wheat seedlings to elevated and super-elevated carbon dioxide

The metabolic consequence of suboptimal (400 μmol mol⁻¹ or ppm), near-optimal (1500 ppm) and supra-optimal (10,000 ppm) atmospheric carbon dioxide concentrations [CO₂] was investigated in an attempt to reveal plausible underlying mechanisms for the differential physiological and developmental responses to increasing [CO₂]. Both non-targeted and targeted metabolite profiling by GC–MS and LC–MS were employed to examine primary and secondary metabolites in wheat (Triticum aestivum, cv Yocoro rojo) continuously exposed to these [CO₂] levels for 14, 21 and 28 days. Metabolite profile was altered by both [CO₂] and physiological age. In general, plants grown under high [CO₂] exhibited a metabolite profile characteristic of older plants under ambient CO₂. Elevated [CO₂] resulted in higher levels of phosphorylated sugar intermediates, though no clear trend in the content of reducing sugars was observed. Transient starch content was enhanced by increasing [CO₂] to a much greater extent at 10,000 ppm CO₂ than at 1500 ppm CO₂. The percentage increase of starch content resulting from CO₂ enrichment declined as plants develope. In contrast, elevated [CO₂] promoted the accumulation of secondary metabolites (flavonoids) progressively to a greater extent as plants became mature. Elevated [CO₂] to 1500 ppm induced a higher initial growth rate, while super-elevated [CO₂] appeared to negate such initial growth promotion. However, after 4 weeks, there was no difference in vegetative growth between 1500 and 10,000 ppm CO₂-grown plants, both elevated CO₂ levels resulted in an overall 25% increase in biomass over the control plants. More interestingly, elevated atmospheric [CO₂] reduced evapotranspiration rate (ET), but further increase to the supra-optimal level resulted in increased ET (a reversed trend), i.e. ET at 1500 ppm < ET at 10,000 ppm < ET at 400 ppm. The differential effect of elevated and super-elevated CO₂ on plants was further reflected in the nitrogen dynamics. These results provide the potential metabolic basis for the differential productivity and stomatal function of plants grown under elevated and super-elevated CO₂ levels.     

Influence of the interaction between light intensity and CO2 concentration on productivity and quality of spinach (Spinacia oleracea L.) grown in fully controlled environment

The effects of the factorial combination of two light intensities (200 and 800 μmol m⁻² s⁻¹) and two CO₂ concentrations (360 and 800 ppm) were studied on the productivity and nutritional quality of spinach (Spinacia oleracea L.) grown under controlled environment. After 6 weeks within a growth chamber, spinach plants were sampled and analyzed for productivity and quality. There were no statistically significant interactions between the effects of light and CO₂ for all of the variables studied, except for the nitrate and oxalic acid content of the leaves. High light and high CO₂ independently one from the other, promoted spinach productivity, and the accumulation of ascorbic acid, while their interactive effect limited the accumulation of nitrate and oxalic acid in the spinach leaves. The results highlight the importance of considering the effects of the interaction among environmental variables on maximizing production and the nutritional quality of the food when cultivating and modeling the plant response in controlled environment systems such as for bioregenerative life support.     

Efficacy of oxygen-supplying capacity of Azolla in a controlled life support system

Azolla shows high growth and propagation rates, strong photosynthetic O₂-releasing ability and high nutritional value. It is suitable as a salad vegetable and can be cultured on a multi-layered wet bed. Hence, it possesses potential as a fresh vegetable, and to release O₂ and absorb CO₂ in a Controlled Ecological Life Support System in space. In this study, we investigated the O₂-providing characteristics of Azolla in a closed chamber under manned, controlled conditions to lay a foundation for use of Azolla as a biological component in ground simulation experiments for space applications. A closed test chamber, representing a Controlled Ecological Life Support System including an Azolla wet-culture device, was built to measure the changes in atmospheric O₂ and CO₂ concentrations inside the chamber in the presence of coexisting Azolla, fish and men. The amount of O₂ consumed by fish was 0.0805–0.0831 L kg⁻¹ h⁻¹ and the level of CO₂ emission was 0.0705–0.0736 L kg⁻¹ h⁻¹; O₂ consumption by the two trial volunteers was 19.71 L h⁻¹ and the volume of respiration-released CO₂ was 18.90 L h⁻¹. Under 7000–8000 Lx artificial light and Azolla wet-culture conditions, human and fish respiration and Azolla photosynthesis were complementary, thus the atmospheric O₂ and CO₂ concentrations inside chamber were maintained in equilibrium. The increase in atmospheric CO₂ concentration in the closed chamber enhanced the net photosynthesis efficiency of the Azolla colony. This study showed that Azolla has strong photosynthetic O₂-releasing ability, which equilibrates the O₂ and CO₂ concentrations inside the chamber in favor of human survival and verifies the potential of Azolla for space applications.     

Development of a ground-based space micro-algae photo-bioreactor

The purpose of the research is to develop a photo-bioreactor which may produce algae protein and oxygen for future astronauts in comparatively long-term exploration, and remove carbon dioxide in a controlled ecological life support system. Based on technical parameters and performance requirements, the project planning, design drafting, and manufacture were conducted. Finally, a demonstration test for producing algae was done. Its productivity for micro-algae and performance of the photo-bioreactor were evaluated. The facility has nine subsystems, including the reactor, the illuminating unit, the carbon dioxide (CO₂) production unit and oxygen (O₂) generation unit, etc. The demonstration results showed that the facility worked well, and the parameters, such as energy consumption, volume, and productivity for algae, met with the design requirement. The density of algae in the photo-bioreactor increased from 0.174 g (dry weight) L⁻¹ to 4.064 g (dry weight) L⁻¹ after 7 days growth. The principle of providing CO₂ in the photo-bioreactor for algae and removing O₂ from the culture medium was suitable for the demand of space conditions. The facility has reasonable technical indices, and smooth and dependable performances.     

Effects of different carbon dioxide and LED lighting levels on the anti-oxidative capabilities of Gynura bicolor DC

Gynura bicolor DC is not only an edible plant but also a kind of traditional Chinese herbal medicine. G. bicolor DC grown in controlled environmental chambers under 3 CO₂ concentrations [450 (ambient), 1500 (elevated), 8000 (super-elevated) μmol mol⁻¹] and 3 LED lighting conditions [white (WL), 85% red + 15% blue (RB15), 70% red + 30% blue (RB30) ] were investigated to reveal plausible antioxidant anabolic responses to CO₂ enrichment and LED light quality. Under ambient and elevated CO₂ levels, blue light increasing from 15% to 30% was conducive to the accumulation of anthocyanins and total flavonoids, and the antioxidant activity of extract was also increased, but plant biomass was decreased. These results demonstrated that the reinforcement of blue light could induce more antioxidant of secondary metabolites, but depress the effective growth of G. bicolor DC under ambient and elevated CO₂ levels. In addition, compared with the ambient and elevated CO₂ levels, the increased anthocyanins, total flavonoids contents and antioxidant enzyme activities of G. bicolor DC under super-elevated CO₂ level could serve as important components of antioxidative defense mechanism against CO₂ stress. Hence, G. bicolor DC might have higher tolerance to CO₂ stress.     

Effects of mineral nutrition conditions on heat tolerance of chufa (Сyperus esculentus L.) plant communities to super optimal air temperatures in the BTLSS

The use of mineralized human wastes as a basis for nutrient solutions will increase the degree of material closure of bio-technical human life support systems. As stress tolerance of plants is determined, among other factors, by the conditions under which they have been grown before exposure to a stressor, the purpose of the study is to investigate the level of tolerance of chufa (Cyperus esculentus L.) plant communities grown in solutions based on mineralized human wastes to a damaging air temperature, 45 °C. Experiments were performed with 30-day-old chufa plant communities grown hydroponically, on expanded clay aggregate, under artificial light, at 690 μmol m⁻² s⁻¹ PAR and at a temperature of 25 °C. Plants were grown in Knop’s solution and solutions based on human wastes mineralized according to Yu.A. Kudenko’s method, which contained nitrogen either as ammonium and urea or as nitrates. The heat shock treatment lasted 20 h at 690 and 1150 μmol m⁻² s⁻¹ PAR. Chufa heat tolerance was evaluated based on parameters of CO₂ gas exchange, the state of its photosynthetic apparatus (PSA), and intensity of peroxidation of leaf lipids. Chufa plants grown in the solutions based on mineralized human wastes that contained ammonium and urea had lower heat tolerance than plants grown in standard mineral solutions. Heat tolerance of the plants grown in the solutions based on mineralized human wastes that mainly contained nitrate nitrogen was insignificantly different from the heat tolerance of the plants grown in standard mineral solutions. A PAR intensity increase from 690 μmol m⁻² s⁻¹ to 1150 μmol m⁻² s⁻¹ enhanced heat tolerance of chufa plant communities, irrespective of the conditions of mineral nutrition under which they had been grown.     

Antioxidant capacity reduced in scallions grown under elevated CO2 independent of assayed light intensity

Long-duration manned space missions mandate the development of a sustainable life support system and effective countermeasures against damaging space radiation. To mitigate the risk of inevitable exposure to space radiation, cultivation of fresh fruits and vegetables rich in antioxidants is an attractive alternative to pharmacological agents. However it has yet to be established whether antioxidant properties of crops can be preserved or enhanced in a space environment where environmental conditions differ from that which plants have acclimated to on earth. Scallion (Allium fistulosum) rich in antioxidant vitamins C and A, and flavonoids was used as a model plant to study the impact of a range of CO₂ concentrations and light intensities that are likely encountered in a space habitat on food quality traits. Scallions were hydroponically grown in controlled environmental chambers under a combination of 3 CO₂ concentrations of 400, 1200 and 4000 μmol mol⁻¹ and 3 light intensity levels of 150, 300, 450 μmol m⁻² s⁻¹. Total antioxidant activity (TAA) of scallion extracts was determined using a radical cation scavenging assay. Both elevated CO₂ and increasing light intensity enhanced biomass accumulation, but effects on TAA (based on dry weight) differed. TAA was reduced for plants grown under elevated CO₂, but remained unchanged with increases in light intensity. Elevated CO₂ stimulated greater biomass production than antioxidants, while an increase in photosynthetic photo flux promoted the synthesis of antioxidant compounds at a rate similar to that of biomass. Consequently light is a more effective stimulus than CO₂ for antioxidant production.     

Origin and stability of lunar polar volatiles

Temperature regime at the LCROSS impact site is studied. All detected species in the Cabeus crater as well as CH₄ and CO clathrate hydrates except H₂, CO, and CH₄ are stable against evaporation at the LCROSS impact site. CO and CH₄ can be chemisorbed at the surface of the regolith particles and exist in the form of clathrate hydrates in the lunar cold traps. Flux rates of delivery of volatile species by asteroids, micrometeoroids, O-rich, C-rich, and low-speed comets into the permanently shadowed regions are estimated. Significant amounts of H₂O, CO, H₂, H₂S, SO₂, and CO₂ can be impact-produced during collisions between asteroids and O-rich comets with the Moon while CH₃OH, NH₃ and complex organic species survive during low-speed comet impacts as products of disequilibrium processes. C-rich comets are main sources of CH₄, and C₂H₄.     

Physical implications of discrepancy between summer and winter PC indices observed in the course of magnetospheric substorms

The PC index based on a statistically justified relationship between the polar cap magnetic activity and the interplanetary electric field E_KL has been derived as a value standardized for the E_KL intensity regardless of season, UT and hemisphere. As a result, the summer and winter PC indices are consistent with one another under ordinary conditions. Discrepancies between the summer and winter PC indices arising in the course of magnetospheric substorms are analyzed in this paper. It is argued that the channel of enhanced conductivity, formed in the auroral oval owing to intense auroral particle precipitation, strongly improves the conditions for closure of the Region 1 field-aligned currents in the winter dark polar region but only trivially affects the conditions of the Region 1 FAC closure in the summer sunlit ionosphere. Since the coefficients describing the relationship between E_KL and the polar cap magnetic activity were derived for statistically justified (i.e., mean) conditions, their application to such abnormal situation, as intense field-aligned currents in the winter dark polar region, leads to overestimation of the winter PC index. The summer and winter PC indices level off as soon as the intense auroral particle precipitation terminates and the auroral ionosphere in the winter and summer polar caps returns to the ordinary (statistically justified) state.     

Nostoc sphaeroides Kützing,an excellent candidate producer for CELSS

Some phytoplankton can be regarded as possible candidates in the establishment of Controlled Ecological Life Support System (CELSS) for some intrinsic characteristics, the first characteristic is that they should grow rapidly, secondly, they should be able to endure some stress factors and develop some corresponding adaptive strategies; also it is very important that they could provide food rich in nutritious protein and vitamins for the crew; the last but not the least is they can also fulfill the other main functions of CELSS, including supplying oxygen, removing carbon dioxide and recycling the metabolic waste. According to these characteristics, Nostoc sphaeroides, a potential healthy food in China, was selected as the potential producer in CELSS. It was found that the oxygen average evolution rate of this algae is about 150 μmol O₂ mg⁻¹ h⁻¹, and the size of them are ranged from 2 to 20 mm. Also it can be cultured with high population density, which indicated that the potential productivity of Nostoc sphaeroides is higher than other algae in limited volume. We measured the nutrient contents of the cyanobacterium and concluded it was a good food for the crew. Based on above advantages, Nostoc sphaeroides was assumed to a suitable phytoplankton for the establishment of Controlled Ecological Life Support System. We plan to develop suitable bioreactor with the cyanobacterium for supplying oxygen and food in future space missions.     

Seasonal patterns of condensation and sublimation cycles in the cryptic and non-cryptic regions of the South Pole

The South Pole of Mars is characterized by an asymmetric residual ice cap composed of water ice and CO₂ ice. On the opposite side of the residual cap, there exists an area called cryptic region which is relatively free of ice during summer time. Many fan-shaped km-scale structures apparently caused by a wind-blown system of dust-laden gas jets occurred dozens degrees of Ls before the complete sublimation of the CO₂ frost layer. We have examined the seasonal cycles of condensation and sublimation in the cryptic and non-cryptic regions by using the topographic data from the MOLA/MGS measurements. Using the MOLA topography data collected over one Martian year (1999–2001), we have studied the temporal elevation change and the seasonal cycle of the carbon dioxide frost on the southern polar caps. We have produced mapping of the seasonal CO₂ frost thickness variation for seven Ls (30°, 60°, 90°, 120°, 150°, 180°, 210°, 240°, 270° and 330°). It is found that the time variations of the CO₂ frost thickness in these two regions are quite similar. The greatest thickness of the CO₂ frost layer is about 0.76–0.78 m in both places occurs at Ls = 150°.     

Cowpeas and pinto beans: Performance and yields of candidate space crops in the laboratory biosphere closed ecological system

An experiment utilizing cowpeas (Vigna unguiculata L.), pinto beans (Phaseolus vulgaris L.) and Apogee ultra-dwarf wheat (Triticum sativa L.) was conducted in the soil-based closed ecological facility, Laboratory Biosphere, from February to May 2005. The lighting regime was 13 h light/11 h dark at a light intensity of 960 μmol m⁻² s⁻¹, 45 mol m⁻² day⁻¹ supplied by high-pressure sodium lamps. The pinto beans and cowpeas were grown at two different planting densities. Pinto bean production was 341.5 g dry seed m⁻² (5.42 g m⁻² day⁻¹) and 579.5 dry seed m⁻² (9.20 g m⁻² day⁻¹) at planted densities of 32.5 plants m⁻² and 37.5 plants m⁻², respectively. Cowpea yielded 187.9 g dry seed m⁻² (2.21 g m⁻² day⁻¹) and 348.8 dry seed m⁻² (4.10 g m⁻² day⁻¹) at planted densities of 20.8 plants m⁻² and 27.7 plants m⁻², respectively. The crop was grown at elevated atmospheric carbon dioxide levels, with levels ranging from 300–3000 ppm daily during the majority of the crop cycle. During early stages (first 10 days) of the crop, CO₂ was allowed to rise to 7860 ppm while soil respiration dominated, and then was brought down by plant photosynthesis. CO₂ was injected 27 times during days 29–71 to replenish CO₂ used by the crop during photosynthesis. Temperature regime was 24–28 °C day/deg 20–24 °C night. Pinto bean matured and was harvested 20 days earlier than is typical for this variety, while the cowpea, which had trouble establishing, took 25 days more for harvest than typical for this variety. Productivity and atmospheric dynamic results of these studies contribute toward the design of an envisioned ground-based test bed prototype Mars base.     

Is the North Atlantic Oscillation modulated by solar and lunar cycles? Some evidences from Hurst autocorrelation analysis

Several studies have suggested that the Sun and Moon cycles affect the Earth climatic dynamics. Nevertheless, there is a long-standing controversy whether solar variability and tides can significantly generate climate change, and how this may occur. Spectral analysis of climatic indices has provided only indirect evidences of the effects of solar–tidal periodicities in the Earth climate. This work addresses the issue by considering the dynamics of the daily North Atlantic Oscillation index over the period from 1950 to 2009. In contrast to previous studies, this work proposes that external cycles can be detected in the autocorrelation dynamics rather than in the raw North Atlantic Oscillation index series. Here, the R/S-scaling analysis is used to quantify, via the so-called Hurst exponent, the presence of autocorrelations along the studied years. Fourier analysis scan of the autocorrelation series thus show two prominent spectral components near (±3%) the lunar tidal 4.425 and the solar 11 years cycles. Intermediate spectral components near 6.4, 7.75 and 8.9 years are proposed to be, at least partially, a result of energy capture from internal mechanisms into cycles resulting from the nonlinear resonance of the fundamental solar–tidal cycles. The dominant effect of the solar variability is clarified by showing that in about 70% of the studied period the sunspot number and the Hurst exponent phases are synchronized, indicating that a higher solar activity enhances the North Atlantic Oscillation index predictability.     

Chlorella vulgaris culture as a regulator of CO2 in a bioregenerative life support system

It is the primary task for a bioregenerative life support system (BLSS) to maintain the stable concentrations of CO₂ and O₂. However, these concentrations could fluctuate based on various factors, such as the imbalance between respiration/assimilation quotients of the heterotrophic and autotrophic components. They can even be out of balance through catastrophic failure of higher plants in the emergency conditions. In this study, the feasibility of using unicellular Chlorella vulgaris of typically rapid growth as both “compensatory system” and “regulator” to control the balance of CO₂ and O₂ was analyzed in a closed ecosystem. For this purpose, a small closed ecosystem called integrative experimental system (IES) was established in our laboratory where we have been conducting multi-biological life support system experiments (MLSSE). The IES consists of a closed integrative cultivating system (CICS) and a plate photo-bioreactor. Four volunteers participated in the study for gas exchange by periodical breathing through a tube connected with the CICS. The plate photo-bioreactor was used to cultivate C. vulgaris. Results showed that the culture of C. vulgaris could be used in a situation of catastrophic failure of higher plant under the emergencies. And the productivity could recover itself to the original state in 3 to 5 days to protect the system till the higher plant was renewed. Besides, C. vulgaris could grow well and the productivity could be affected by the light intensity which could help to keep the balance of CO₂ and O₂ in the IES efficiently. Thus, C. vulgaris could be included in the design of a BLSS as a “compensatory system” in the emergency contingency and a “regulator” during the normal maintenance.