受控生态生保系统内红萍供氧特性研究

研究红萍载人供O₂特征, 为红萍生物部件进行系统总体地面模拟试验及空间应用奠定基础,构建了受控生态生保系统密闭试验舱和红萍栽培装置, 在“红萍-鱼-人”共存情况下, 测定密闭舱内O₂, CO₂浓度的变化. 试验结果显示, 单位重量的鱼耗O₂量. 0805～0.0831 L·kg^-1·h^-1, 排放CO₂量为0.0705～0.0736 L·kg^-1·h^-1; 试验志愿者耗O₂量19.71 L·h^-1, 呼吸释放CO₂量18.90 L·h^-1. 人工光照保持7000～8000 lx条件下, 红萍的光合作用与人和鱼的呼吸作用相辅相成, 舱内O₂, CO₂浓度趋于平衡. 密闭舱内CO₂浓度升高对促进红萍群体净光合效率有明显效果, 红萍光合放O₂能力很强, 能有效促使密闭舱内O₂, CO₂浓度朝着有利于人生存的环境方向平衡, 进而验证了红萍的空间应用前景.     

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.     

Community metabolism of aquatic Closed Ecological Systems: Effects of nitrogen sources

To investigate the effect of nitrogen sources on Closed Ecological Systems (CESs), three nitrogen sources (NaNO₃, sodium nitrate; NH₄Cl, ammonium chloride; and NH₄NO₃, ammonium nitrate) were each tested in freshwater CESs consisting of a chemically defined medium, three species of green algae (Ankistrodesmus, Scenedesmus, and Selenastrum), the grazer Daphnia magna, and associated microbes, under 12 h light/12 h dark cycles. It had been hypothesized that the development of high pH in earlier CESs was the result of nitrate utilization, and that ammonium might result in acid conditions, while ammonium nitrate might result in more moderate pH. The three nitrogen sources supported similar densities of algae (estimated by in vivo fluorescence) and similar Daphnia populations. The experiments showed that pH levels rapidly increased when grazers were absent or at low abundances irrespective of the nitrogen source. Consequently, it is hypothesized that carbon cycles, rather than nitrogen sources, are responsible for the pH dynamics. Oxygen diurnal (light:dark) cycles tended to come into balance more quickly than pH. It may be more feasible to convert O₂ data to energy units (using “oxycalorific” values) than CO₂ data since CO₂ dynamics may include other chemical reactions than just photosynthesis and respiration. The feasibility of sustaining grazer populations for at least several weeks in small, simple CESs was demonstrated, along with the ability to monitor algae-grazer dynamics, and the recording of O₂ and pH measurements.     

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.     

Potential of salt-accumulating and salt-secreting halophytic plants for recycling sodium chloride in human urine in bioregenerative life support systems

This study addresses the possibility of growing different halophytic plants on mineralized human urine as a way to recycle NaCl from human wastes in a bioregenerative life support system (BLSS). Two halophytic plant species were studied: the salt-accumulating Salicornia europaea and the salt-secreting Limonium gmelinii. During the first two weeks, plants were grown on Knop’s solution, then an average daily amount of urine produced by one human, which had been preliminarily mineralized, was gradually added to the experimental solutions. Nutrient solutions simulating urine mineral composition were gradually added to control solutions. NaCl concentrations in the stock solutions added to the experimental and control solutions were 9 g/L in the first treatment and 20 g/L in the second treatment. The mineralized human urine showed some inhibitory effects on S. europaea and L. gmelinii. The biomass yield of experimental plants was lower than that of control ones. If calculated for the same time period (120 d) and area (1 m²), the amount of sodium chloride taken up by S. europaea plants would be 11.7 times larger than the amount taken up by L. gmelinii plants (486 g/m² vs. 41 g/m²). Thus, S. europaea is the better choice of halophyte for recycling sodium chloride from human wastes in BLSS.     

Testing anti-fungal activity of a soil-like substrate for growing plants in bioregenerative life support systems

The object of this research is to study a soil-like substrate (SLS) to grow plants in a Bioregenerative Life Support System (BLSS). Wheat and rice straw were used as raw materials to prepare SLS. Anti-fungal activity of SLS using test cultures of Bipolaris sorokiniana, a plant-pathogenic fungus which causes wheat root rot was studied. Experiments were conducted with SLS samples, using natural soil and sand as controls. Infecting the substrates, was performed at two levels: the first level was done with wheat seeds carrying B. sorokiniana and the second level with seeds and additional conidia of B. sorokiniana from an outside source. We measured wheat disease incidence and severity in two crop plantings. Lowest disease incidence values were obtained from the second planting, SLS: 26% and 41% at the first and the second infection levels, respectively. For soil the values were 60% and 82%, respectively, and for sand they were 67% and 74%, respectively. Wheat root rot in the second crop planting on SLS, at both infection levels was considerably less severe (9% and 13%, respectively) than on natural soil (20% and 33%) and sand (22% and 32%). SLS significantly suppressed the germination of B. sorokiniana conidia. Conidia germination was 5% in aqueous SLS suspension, and 18% in clean water. No significant differences were found regarding the impact on conidia germination between the SLS samples obtained from wheat and rice straw. The anti-fungal activity in SLS increased because of the presence of worms. SLS also contained bacteria stimulating and inhibiting B. sorokiniana growth.     

Sweetpotato vine management for confined food production in a space life-support system

Sweetpotato (Ipomea batatas L.) ‘Whatley–Loretan’ was developed for space life support by researchers at Tuskegee University for its highly productive, nutritious storage roots. This promising candidate space life-support crop has a sprawling habit and aggressive growth rate in favorable environments that demands substantial growing area. Shoot pruning is not a viable option for vine control because removal of the main shoot apex drastically inhibits storage-root initiation and development, and chemical growth retardants typically are not cleared for use with food crops. As part of a large effort by the NASA Specialized Center of Research and Training in Advanced Life Support to reduce equivalent system mass (ESM) for food production in space, the dilemma of vine management for sweetpotato was addressed in effort to conserve growth area without compromising root yield. Root yields from unbranched vines trained spirally around wire frames configured either in the shapes of cones or cylinders were similar to those from vines trained horizontally along the bench, but occupying only a small fraction of the bench area. This finding indicates that sweetpotato is highly adaptable to a variety of vine-training architectures. Planting a second plant in the growth container and training the two vines in opposite directions around frames enhanced root yield and number, but had little effect on average length of each vine or bench area occupied. Once again, root yields were similar for both configurations of wire support frames. The 3–4-month crop-production cycles for sweetpotato in the greenhouse spanned all seasons of multiple years during the course of the study, and although electric lighting was used for photoperiod control and to supplement photosynthetic light during low-light seasons, there still were differences in total light available across seasons. Light variations and other environmental differences among experiments in the greenhouse had more effects on vine length than on root yield. Average vine length correlated positively with total hours of daylight received across seasons, and responses for one plant per container were higher above a threshold duration of solar exposure, suggesting that the vines of two plants per container compete for available light. In addition to the adaptability of sweetpotato to various vine-training architectures and across seasons in terms of maintaining root productivity, the open, interior volumes of the support frames tested in this study will provide future opportunity to enhance sweetpotato root yield in space by adding novel interior lighting, such as from intracanopy arrays of light-emitting diodes. This work was sponsored by NASA grant NAG 5 1286.     

Blue light-dependent phosphorylations of cryptochromes are affected by magnetic fields in Arabidopsis

The blue light receptor cryptochrome that could form radical pairs after exposure to blue light was suggested to be a magnetoreceptor based on the proposition that radical pairs were involved in the magnetoreception. But the effects of magnetic fields on the function of cryptochrome are poorly understood. Phosphorylation of cryptochrome in Arabidopsis was closely associated with the function of this photoreceptor. Here, we grew Arabidopsis seedlings in a 500 μT magnetic field and a near-null magnetic field and found that the 500 μT magnetic field enhanced the blue light-dependent phosphorylations of CRY1 and CRY2, and the near-null magnetic field weakened the blue light-dependent phosphorylation of CRY2 but not CRY1. Dephosphorylations of CRY1 and CRY2 in the darkness were slowed down in the 500 μT magnetic field, whereas dephosphorylations of CRY1 and CRY2 were accelerated in the near-null magnetic field. These results suggest that magnetic field with strength higher or weaker than the local geomagnetic field affects the activated states of cryptochromes, which thus modifies the functions of cryptochromes.     

Low light intensity effects on the growth,photosynthetic characteristics,antioxidant capacity,yield and quality of wheat (Triticum aestivum L.) at different growth stages in BLSS

Minimizing energy consumption and maximizing crop productivity are major challenges to growing plants in Bioregenerative Life Support System (BLSS) for future long-term space mission. As a primary source of energy, light is one of the most important environmental factors for plant growth. The purpose of this study is to investigate the effects of low light intensity at different stages on growth, pigment composition, photosynthetic efficiency, biological production and antioxidant defence systems of wheat (Triticum aestivum L.) cultivars during ontogenesis. Experiments were divided into 3 intensity-controlled stages according to growth period (a total of 65 days): seedling stage (first 20 days), heading and flowering stage (middle 30 days) and grain filling stage (last 15 days). Initial light condition of the control was 420 μmol m⁻² s⁻¹ and the light intensity increased with the growth of wheat plants. The light intensities of group I and II at the first stage and the last stage were adjusted to the half level of the control respectively. For group III, the first and the last stage were both adjusted to half level of the control. During the middle 30 days, all treatments were kept the same intensity. The results indicated that low-light treatment at seedling stage, biomass, nutritional contents, components of inedible biomass and healthy index (including peroxidase (POD) activity, malondialdehyde (MDA) and proline content) of wheat plants have no significant difference to the control. Furthermore, unit kilojoule yield of group I reached 0.591 × 10⁻³ g/kJ and induced the highest energy efficiency. However, low-light treatment at grain filling stage affected the final production significantly.     

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.     

Survival and death of the haloarchaeon Natronorubrum strain HG-1 in a simulated martian environment

Halophilic archaea are of interest to astrobiology due to their survival capabilities in desiccated and high salt environments. The detection of remnants of salty pools on Mars stimulated investigations into the response of haloarchaea to martian conditions. Natronorubrum sp. strain HG-1 is an extremely halophilic archaeon with unusual metabolic pathways, growing on acetate and stimulated by tetrathionate. We exposed Natronorubrum strain HG-1 to ultraviolet (UV) radiation, similar to levels currently prevalent on Mars. In addition, the effects of low temperature (4, −20, and −80 °C), desiccation, and exposure to a Mars soil analogue from the Atacama desert on the viability of Natronorubrum strain HG-1 cultures were investigated. The results show that Natronorubrum strain HG-1 cannot survive for more than several hours when exposed to UV radiation equivalent to that at the martian equator. Even when protected from UV radiation, viability is impaired by a combination of desiccation and low temperature. Desiccating Natronorubrum strain HG-1 cells when mixed with a Mars soil analogue impaired growth of the culture to below the detection limit. Overall, we conclude that Natronorubrum strain HG-1 cannot survive the environment currently present on Mars. Since other halophilic microorganisms were reported to survive simulated martian conditions, our results imply that survival capabilities are not necessarily shared between phylogenetically related species.     

Tolerance of chufa (Cyperus esculentus L.) plants,representing the higher plant compartment in bioregenerative life support systems,to super-optimal air temperatures

Plants intended to be included in the photosynthesizing compartment of the bioregenerative life support system (BLSS) need to be studied in terms of both their production parameters under optimal conditions and their tolerance to stress factors that might be caused by emergency situations. The purpose of this study was to investigate tolerance of chufa (Cyperus esculentus L.) plants to the super-optimal air temperature of 45 ± 1 °C as dependent upon PAR (photosynthetically active radiation) intensity and the duration of the exposure to the stress factor. Chufa plants were grown hydroponically, on expanded clay, under artificial light. The nutrient solution was Knop’s mineral medium. Until the plants were 30 days old, they had been grown at 690 μmol m⁻² s⁻¹ PAR and air temperature 25 °C. Thirty-day-old plants were exposed to the temperature 45 °C for 6 h, 20 h, and 44 h at PAR intensities 690 μmol m⁻² s⁻¹ and 1150 μmol m⁻² s⁻¹. The exposure to the damaging air temperature for 44 h at 690 μmol m⁻² s⁻¹ PAR caused irreversible damage to PSA, resulting in leaf mortality. In chufa plants exposed to heat shock treatment at 690 μmol m⁻² s⁻¹ PAR for 6 h and 20 h, respiration exceeded photosynthesis, and CO₂ release in the light was recorded. Functional activity of photosynthetic apparatus, estimated from parameters of pulse-modulated chlorophyll fluorescence in Photosystem 2 (PS 2), decreased 40% to 50%. After the exposure to the stress factor was finished, functional activity of PSA recovered its initial values, and apparent photosynthesis (P_apparent) rate after a 20-h exposure to the stress factor was 2.6 times lower than before the elevation of the temperature. During the first hours of plant exposure to the temperature 45 °C at 1150 μmol m⁻² s⁻¹ PAR, respiration rate was higher than photosynthesis rate, but after 3–4 h of the exposure, photosynthetic processes exceeded oxidative ones and CO₂ absorption in the light was recorded. At the end of the 6-h exposure, P_apparent rate was close to that recorded prior to the exposure, and no significant changes were observed in the functional activity of PSA. At the end of the 20-h exposure, P_apparent rate was close to its initial value, but certain parameters of the functional activity of PSA decreased 25% vs. their initial values. During the repair period, the parameters of external gas exchange recovered their initial values, and parameters of pulse-modulated chlorophyll fluorescence were 20–30% higher than their initial values. Thus, exposure of chufa plants to the damaging temperature of the air for 20 h did not cause any irreversible damage to the photosynthetic apparatus of plants at either 690 μmol m⁻² s⁻¹ or 1150 μmol m⁻² s⁻¹ PAR, and higher PAR intensity during the heat shock treatment enhanced heat tolerance of the plants.     

Molecular analysis of the graviperception signal transduction in the flagellate Euglena gracilis: Involvement of a transient receptor potential-like channel and a calmodulin

Euglena gracilis, a unicellular, photosynthetic flagellate is a model system for environmentally controlled behavior responses. The organism shows pronounced negative gravitaxis. This movement is based on physiological mechanisms, which in the past had been only indirectly assessed. It was shown that mechano-sensitive calcium channels are involved in the gravitaxis response. Recent studies have demonstrated that members of the transient receptor potential (TRP) family function as mechano-sensitive channels in several different cell types. We have sequenced part of a TRP gene in Euglena and applied RNA interference (RNAi) to confirm that these channels are involved in graviperception. It was found that RNAi against the putative TRP channel abolished gravitaxis. The genes of three calmodulins were sequences in Euglena, one of which was previously known in its protein structure (cal 1). The other two were unknown (cal 2 and cal 3). Cal 2 has been analyzed in detail. The biosynthesis of the corresponding proteins of cal 1 and cal 2 was inhibited by means of RNA interference to see whether this blockage impairs gravitaxis. RNAi of cal 1 leads to a long-term loss of free swimming in the cells (while euglenoid movement persists). It induced pronounced cell form aberrations and the division of cells was hampered. After recovery from RNAi the cell showed precise negative gravitaxis again. Thus cal 1 does not seem to be involved in gravitaxis. In contrast, the blockage of cal 2 has no pronounced influence on motility and cell form but leads to a complete loss of gravitactic orientation for more than 30 days showing that this calmodulin is an element in the signal transduction chain. The data are discussed in the context of the current model of the gravitaxis signal transduction chain in Euglena gracilis.     

Spaceflight results in increase of thick filament but not thin filament proteins in the paramyosin mutant of Caenorhabditis elegans

We have investigated the effect of microgravity during spaceflight on body-wall muscle fiber size and muscle proteins in the paramyosin mutant of Caenorhabditis elegans. Both mutant and wild-type strains were subjected to 10 days of microgravity during spaceflight and compared to ground control groups. No significant change in muscle fiber size or quantity of the protein was observed in wild-type worms; where as atrophy of body-wall muscle and an increase in thick filament proteins were observed in the paramyosin mutant unc-15(e73) animals after spaceflight. We conclude that the mutant with abnormal muscle responded to microgravity by increasing the total amount of muscle protein in order to compensate for the loss of muscle function.     

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.     

Involvement of nitric oxide in the mechanism of biochemical alterations induced by simulated microgravity in Microcystis aeruginosa

Simulated microgravity (SMG) can inhibit proliferation and enhance microcystin production of Microcystis aeruginosa. We investigated the role of nitric oxide (NO) in regulating the SMG induced changes of proliferation, photochemical system II photochemical activity, pigment, soluble protein and microcystin production in M. aeruginosa. M. aeruginosa was exposed to 0.1 mM sodium nitroprusside (SNP, NO donor) or 0.02 mM 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO, NO scavenger) alone or in combination with SMG for 48 h. SMG and SNP inhibited the growth of M. aeruginosa while c-PTIO had no effect on cell number. As to yield, the negative effect of SMG was augmented by SNP and suppressed by c-PTIO. The intracellular concentrations of chlorophyll a, carotenoid, phycocyanin, soluble protein and microcystin were increased by SMG after 48 h. The effects of SMG on these metabolic processes could be enhanced by SNP and be partly eliminated by c-PTIO. Moreover, SNP and c-PTIO only functioned in these biochemical processes under SMG, unlike in the regulation of cell proliferation and yield. These results showed that the effects of SMG could be enhanced by adding exogenous NO and be mitigated by scavenging endogenous NO, revealing the involvement of NO in the changes in biochemistry processes induced by SMG in M. aeruginosa.