Ethylene production by plants in a closed environment.

Ethylene production by 20-m2 stands of wheat, soybean, lettuce and potato was monitored throughout growth and development in NASA's Controlled Ecological Life Support System (CELSS) Biomass Production Chamber. Chamber ethylene concentrations rose during periods of rapid growth for all four species, reaching 120 parts per billion (ppb) for wheat, 60 ppb for soybean, and 40 to 50 ppb for lettuce and potato. Following this, ethylene concentrations declined during seed fill and maturation (wheat and soybean), or remained relatively constant (potato). Lettuce plants were harvested during rapid growth and peak ethylene production. The highest ethylene production rates (unadjusted for chamber leakage) ranged from 0.04 to 0.06 ml m-2 day-1 during rapid growth of lettuce and wheat stands, or approximately 0.8 to 1.1 nl g-1 fresh weight h-1. Results suggest that ethylene production by plants is a normal event coupled to periods of rapid metabolic activity, and that ethylene removal or control measures should be considered for growing crops in a tightly closed CELSS.     

Direct utilization of human liquid wastes by plants in a closed ecosystem.

Model experiments in phytotrons have shown that urea is able to cover 70% of the demand in nitrogen of the conveyer cultivated wheat. At the same time wheat plants can directly utilize human liquid wastes. In this article by human liquid wastes the authors mean human urine only. In a long-term experiment on "man-higher plants" system with two crewmen, plants covered 63 m2, with wheat planted to--39.6 m2. For 103 days, complete human urine (total amount--210.7 l) was supplied into the nutrient solution for wheat. In a month and a half NaCl supply into the nutrient solution stabilized at 0.9-1.65 g/l. This salination had no marked effect on wheat production. The experiment revealed the realistic feasibility to directly involve liquid wastes into the biological turnover of the life support system. The closure of the system, in terms of water, increased by 15.7% and the supply of nutrients for wheat plants into the system was decreased.     

The importance of restoration of the atmospheric electrical environment in closed Bioregenerative Life Supporting Systems.

Earth based Bioregenerative Life Supporting Systems (BLSS) are subject to 4 main physical factors: gravity, light, temperature and electrical environment. The first 3 are obvious to everyone, the Electrical Environment (EE) is not under the majority of prevailing conditions perceived directly by our senses. The EE is one of the important physical factors directly influencing some plants and in a less obvious way also the majority of plants. There are only two long range forces in nature: the electromagnetic and the gravitational forces. Gravity is very much weaker than the electromagnetic forces FG/FEL=10(-38), where FG is the gravitational Force and FEL are the electromagnetic Forces. The atmospheric electric field prevails all the time over the entire Earth with a mean intensity of 130 V/m. It is therefore a potent factor which may be used by some plants exposed throughout their entire life time to the atmospheric electric field. What effect should the normal atmospheric electric field have on plants? All living plants are good electrical conductors for electrostatic fields. The plants distort the normally vertical field lines, which have to be perpendicular to the plant tissue everywhere in order to avoid the extraction of energy from the field. The meristems concentrate the field lines, thus the electrically charged nutrients are supplied to the growing parts of the plant exposed to the field. This results in electrotropism in some plants. It is very well known that plants do have adaptive capabilities as compared to animals, it is important for their survival, because they cannot run away from trouble. It is found by careful observations of the behaviour of different plants that some plants do respond to the presence of the atmospheric electric field while other plants exposed to the same environment are indifferent to the atmospheric electric field. The plants growing in the Biosphere II were shielded by the metal structure of the Biosphere II. Because these plants which do make use of the atmospheric electric field are also to be found in the Biosphere II and must be deprived of this natural atmospheric electric field and in consequence of some of their natural nutrients. I have experimental evidence that at least some plants do respond to the atmospheric electric field. This effect is the most likely cause of the oxygen depletion and of the carbon dioxide accumulation in Biosphere II. Under the micro gravity encountered in space habitats the restoration of electric fields is even more important for plant growth than it is on the Earth.     

Effects of lighting and air movement on temperatures in reproductive organs of plants in a closed plant growth facility

Temperature increases in plant reproductive organs such as anthers and stigmas could cause fertility impediments and thus produce sterile seeds under artificial lighting conditions without adequately controlled environments in closed plant growth facilities. There is a possibility such a situation could occur in Bioregenerative Life Support Systems under microgravity conditions in space because there will be little natural convective or thermal mixing. This study was conducted to determine the temperature of the plant reproductive organs as affected by illumination and air movement under normal gravitational forces on the earth and to make an estimation of the temperature increase in reproductive organs in closed plant growth facilities under microgravity in space. Thermal images of reproductive organs of rice and strawberry were captured using infrared thermography at air temperatures of 10–11 °C. Compared to the air temperature, temperatures of petals, stigmas and anthers of strawberry increased by 24, 22 and 14 °C, respectively, after 5 min of lighting at an irradiance of 160 W m⁻² from incandescent lamps. Temperatures of reproductive organs and leaves of strawberry were significantly higher than those of rice. The temperatures of petals, stigmas, anthers and leaves of strawberry decreased by 13, 12, 13 and 14 °C, respectively, when the air velocity was increased from 0.1 to 1.0 ms⁻¹. These results show that air movement is necessary to reduce the temperatures of plant reproductive organs in plant growth facilities.     

Development of autonomous control in a closed microbial bioreactor.

Space-based life support systems which include ecological components will rely on sophisticated hardware and software to monitor and control key system parameters. Autonomous closed artificial ecosystems are useful for research in numerous fields. We are developing a bioreactor designed to study both microbe-environment interactions and autonomous control systems. Currently we are investigating N-cycling and N-mass balance in closed microbial systems. The design features of the system involve real-time monitoring of physical parameters (e.g. temperature, light), growth solution composition (e.g. pH, NOx, CO2), cell density and the status of important hardware components. Control of key system parameters is achieved by incorporation of artificial intelligence software tools that permit autonomous decision-making by the instrument. These developments provide a valuable research tool for terrestrial microbial ecology, as well as a testbed for implementation of artificial intelligence concepts. Autonomous instrumentation will be necessary for robust operation of space-based life support systems, and for use on robotic spacecraft. Sample data acquired from the system, important features of software components, and potential applications for terrestrial and space research will be presented.     

Modeling gas exchange in a closed plant growth chamber.

Fluid transport models for fluxes of water vapor and CO2 have been developed for one crop of wheat and three crops of soybean grown in a closed plant growth chamber. Correspondence among these fluxes is discussed. Maximum fluxes of gases are provided for engineering design requirements of fluid recycling equipment in growth chambers. Furthermore, to investigate the feasibility of generalized crop models, dimensionless representations of water vapor fluxes are presented. The feasibility of such generalized models and the need for additional data are discussed.     

Potential vascular damage from radiation in the space environment.

Cultured endothelial cells of blood vessels have a Do of 2 Gy for X-rays. A dose of 0.5 Gy of X-rays has an acute effect on vessel diameter. The vessels may show other acute effects such as change in permeability including a change in the blood brain barrier. Changes occurring from late effects of chronic exposure in vascular architecture include telangiectasia and decrease in vascular density. Changes in the perivascular connective tissue particularly collagen may play a role in these changes. After charged particle exposure of 15 and 30 Gy, radiation changes in the blood brain barrier and vascular changes are noted in the nervous system. These long term changes are recorded by PET, MRI, and CT imaging. Chronic exposure to alpha particles causes vascular damage in compact bone resulting in bone infarcts. Using tandem scanning confocal microscopy in-situ imaging of the capillaries and collagen of the papillary dermis provides a non-invasive method of serial recording of changes in irradiated microvasculature.     

Injuries to plants from controlled environment contaminants.

The use of controlled environments is subject to problems from contaminants emitted from materials of the system and from plants. Many contaminants are difficult to identify because injurious dosages are very low, there is a lack of information on what compounds injure plants, because species and cultivars differ greatly in their sensitivity to injury and injury symptoms often are not distinctive. Plastics have been shown to emit many different volatile compounds. The compound, di-butyl phthalate, contained in certain flexible plastics, has been shown to be very toxic to plants. Other injuries have been produced by caulking compounds and bonded screening. Paints have been shown to release xylene that is toxic to plants. Steam for humidification can cause problems because of hydroxylamines and other compounds added to steam used for heating to control fungal growth in return lines. Mercury, from broken thermometers is a particular problem in growth chambers because small quantities can collect in cracks and slowly volatilize to slow growth of plants. Plants themselves release large quantities of volatile hydrocarbons, with ethylene being the commonly recognized chemical that can be damaging when allowed to accumulate. People release large quantities of carbon dioxide which can cause variations in the rate of growth of plants. Contaminant problems can be controlled through filtering of the air or ventilation with make-up air, however the potential for problems is always present and careful testing should be undertaken with the particular species and cultivars being grown insure that there are no toxic agents altering growth in each particular controlled environment being utilized.     

Controlling denitrification in closed artificial ecosystems.

Denitrification, the dissimilatory reduction of NO3- to N2O and N2, is found in a wide variety of organisms. In closed artificial systems, especially closed plant growth chambers, a significant loss of fixed-N occurs through denitrification, thereby decreasing the efficiency of the system and fouling the atmosphere with N2O. Denitrification is a form of anaerobic respiration. Whenever available, however, denitrifiers preferentially use O2 as their terminal electron acceptor. As a result, rates of denitrification and growth are a function of O2. Typically, in closed systems O2 consumption is greater than the diffusion of O2 through the medium to the cell, decreasing the O2 level near the cell and denitrification occurs. Using Pseudomonas fluorescens (ATCC # 17400) as a model organism grown in a two L bioreactor under varying levels of O2 we studied its effects on population growth and its ability to mitigate denitrification in closed systems. The results indicate that denitrification occurs in a closed system even when it is considered aerobic, that is well mixed and sparged with either air, or sufficient pure O2 to cause a complete turnover in the gaseous atmosphere in the bioreactor vessel every five minutes.     

Impaired growth of plants cultivated in a closed system: possible reasons.

Plants in experiments on "man-higher plants" closed ecosystem (CES) have been demonstrated to have inhibited growth and reduced productivity due to three basic factors: prolonged usage of a permanent nutrient solution introduction into the nutrient medium of intra-system gray water, and closure of the system. Gray water was detrimental to plants the longer the nutrient solution was used. However, higher plant growth was mostly affected by the gaseous composition of the CES atmosphere, through accumulation of volatile substances.     

The biological clock of Neurospora in a microgravity environment.

The circadian rhythm of conidiation in Neurospora crassa is thought to be an endogenously derived circadian oscillation; however, several investigators have suggested that circadian rhythms may, instead, be driven by some geophysical time cue(s). An experiment was conducted on space shuttle flight STS-9 in order to test this hypothesis; during the first 7-8 cycles in space, there were several minor alterations observed in the conidiation rhythm, including an increase in the period of the oscillation, an increase in the variability of the growth rate and a diminished rhythm amplitude, which eventually damped out in 25% of the flight tubes. On day seven of flight, the tubes were exposed to light while their growth fronts were marked. Some aspect of the marking process reinstated a robust rhythm in all the tubes which continued throughout the remainder of the flight. These results from the last 86 hours of flight demonstrated that the rhythm can persist in space. Since the aberrant rhythmicity occurred prior to the marking procedure, but not after, it was hypothesized that the damping on STS-9 may have resulted from the hypergravity pulse of launch. To test this hypothesis, we conducted investigations into the effects of altered gravitational forces on conidiation. Exposure to hypergravity (via centrifugation), simulated microgravity (via the use of a clinostat) and altered orientations (via alterations in the vector of a 1 g force) were used to examine the effects of gravity upon the circadian rhythm of conidiation.     

A combined modeling and experimental approach for achieving a simplified closed ecosystem.

In CELSS both biological and physico-chemical processes have to be used to support the main needs of the crews and to minimize the re-supply of food and air from Earth. The basic idea is to create a complete food chain (an artificial ecosystem), beginning from the crew, with its wastes, and returning to the crew to supply it with food and air. Two main other steps of this food chain are a waste treatment process and a biomass production including higher plants. We set up the connection of these key modules, which we called ECLAS (Ecosysteme Clos Artificiel Simplifie). A growth chamber containing higher plants is connected to a continuous supercritical water oxidation reactor, that converts the harvested biomass into carbon dioxide and enables the photosynthesis of the canopy. To achieve a stable coupling through optimized regulations between the modules, we programmed a modular numerical simulation of the system, in order to assess the involved fluxes and to constrain the last degrees of freedom of the experimental system already built. Simulation results and the first experimental results are here compared.     

Growth restoration in azuki bean and maize seedlings by removal of hypergravity stimuli.

Hypergravity stimuli, gravitational acceleration of more than 1 x g, decrease the growth rate of azuki bean epicotyls and maize coleoptiles and mesocotyls by decreasing the cell wall extensibility via an increase in the molecular mass of matrix polysaccharides. An increase in the pH in the apoplastic fluid is hypothesized to be involved in the processes of the increase in the molecular mass of matrix polysaccharides due to hypergravity. However, whether such physiological changes by hypergravity are induced by normal physiological responses or caused by physiological damages have not been elucidated. In the present study, we examined the effects of the removal of hypergravity stimuli on growth and the cell wall properties of azuki bean and maize seedlings to clarify whether the effects of hypergravity stimuli on growth and the cell wall properties are reversible or irreversible. When the seedlings grown under hypergravity conditions at 300 x g for several hours were transferred to 1 x g conditions, the growth rate of azuki bean epicotyls and maize coleoptiles and mesocotyls greatly increased within a few hours. The recovery of growth rate of these organs was accompanied by an immediate increase in the cell wall extensibility, a decrease in the molecular mass of matrix polysaccharides, and an increase in matrix polysaccharide-degrading activities. The apoplastic pH also decreased promptly upon the removal of hypergravity stimuli. These results suggest that plants regulate the growth rate of shoots reversibly in response to hypergravity stimuli by changing the cell wall properties, by which they adapt themselves to different gravity conditions. This study also revealed that changes in growth and the cell wall properties under hypergravity conditions could be recognized as normal physiological responses of plants. In addition, the results suggest that the effects of microgravity on plant growth and cell wall properties should be reversible and could disappear promptly when plants are transferred from microgravity to 1 x g. Therefore, plant materials should be fixed or frozen on orbit for detecting microgravity-induced changes in physiological parameters after recovering the materials to earth in space experiments.     

Energetic particle environment in near-Earth orbit.

The hazard of exposure to high doses of ionizing radiation is one of the primary concerns of extended manned space missions and a continuous threat for the numerous spacecraft in operation today. In the near-Earth environment the main sources of radiation are solar energetic particles (SEP), galactic cosmic rays (GCR), and geomagnetically trapped particles, predominantly protons and electrons. The intensity of the SEP and GCR source depends primarily on the phase of the solar cycle. Due to the shielding effect of the Earth's magnetic field, the observed intensity of SEP and GCR particles in a near-Earth orbit will also depend on the orbital parameters altitude and inclination. The magnetospheric source strength depends also on these orbital parameters because they determine the frequency and location of radiation belt passes. In this paper an overview of the various sources of radiation in the near-Earth orbit will be given and first results obtained with the Solar, Anomalous, and Magnetospheric Particle Explorer (SAMPEX) will be discussed. SAMPEX was launched on 3 July 1992 into a near polar (inclination 82 degrees) low altitude (510 x 675 km) orbit. The SAMPEX payload contains four separate instruments of high sensitivity covering the energy range 0.5 to several hundred MeV/nucleon for ions and 0.4 to 30 MeV for electrons. This low altitude polar orbit with zenith-oriented instrumentation provides a new opportunity for a systematic study of the near-Earth energetic particle environment.     

Cosmic dust analog simulation in a microgravity environment: the STARDUST program.

We have undertaken a project called STARDUST which is a collaboration with Italian and American investigators. The goals of this program are to study the condensation and coagulation of refractory materials from the vapor and to study the properties of the resulting grains as analogs to cosmic dust particles. To reduce thermal convective currents and to develop valuable experience in designing an experiment for the Gas-Grain Simulation Facility aboard Space Station Freedom we have built and flown a new chamber to study these processes under periods of microgravity available on NASA's KC-135 Research Aircraft. Preliminary results from flights with magnesium and zinc are discussed.     

Modelling of genetically engineered microorganisms introduction in closed artificial microcosms.

The possibility of introducing genetically engineered microorganisms (GEM) into simple biotic cycles of laboratory water microcosms was investigated. The survival of the recombinant strain Escherichia coli Z905 (Apr, Lux+) in microcosms depends on the type of model ecosystems. During the absence of algae blooming in the model ecosystem, the part of plasmid-containing cells E. coli decreased fast, and the structure of the plasmid was also modified. In conditions of algae blooming (Ankistrodesmus sp.) an almost total maintenance of plasmid-containing cells was observed in E. coli population. A mathematics model of GEM's behavior in water ecosystems with different level of complexity has been formulated. Mechanisms causing the difference in luminescent exhibition of different species are discussed, and attempts are made to forecast the GEM's behavior in water ecosystems.     

Bone metabolism and formation of mice bred in a 2 G environment.

The purpose of this study is to reveal the effect of chronic hypergravity exposure on the bone formation and the bone metabolism when mammals produce offspring in a 2 G environment. We measured the length and width of the thighbone, the length of the pelvis, the width of the pelvic cavity and the width of the fourth cervical vertebra on the second (F2) and the third (F3) generation mice bred in a 2 G environment every ten days from 20 days old to 60 days old in an experiment on bone formation. In an experiment on bone metabolism, we measured calcium and phosphorus in the bones of the F3 in the 2 G group. Ratios of the thighbone length, pelvis length, pelvic cavity width, and fourth cervical vertebra width versus the body length were calculated. These ratios were higher in the 2 G group than the control group during all measuring periods. Calcium and phosphorus concentrations in the thighbone and the lumbar vertebra were lower in the 2 G group than in the control group. However, the calcium and phosphorus concentrations in the cervical vertebrae of the 2G group were higher. These results suggest that the influence of gravity load may vary in the bones.     

Catalase activity as a potential indicator of the reducer component of small closed ecosystems.

Dynamics of catalase activity has been shown to reflect the growth curve of microorganisms in batch cultivation (celluloselythic bacteria Bacillus acidocaldarius and bacteria of the associated microflora Chlorella vulgaris). Gas and substrate closure of the three component ecosystems with spatially separated components "producer-consumer-reducer" (Chl. vulgaris-Paramecium caudatum-B. acidocaldarius, two bacterial strains isolated from the associated microflora Chl. vulgaris) demonstrated that the functioning of the reducer component can be estimated by the catalase activity of mciroorganisms of this component.     

High performance capillary electrophoresis in microgravity environment.

In the last two decades, capillary electrophoresis has developed to a very quick and accurate analysis technique. With micellar techniques even uncharged particles can be analysed. The field of capillary electrophoresis is broad and shows an overlap with HPLC. The equipment for capillary electrophoresis is very compact, low in weight, energy consumption and waste production.     

The "C.E.B.A.S. MINI-MODULE": a self-sustaining closed aquatic ecosystem for spaceflight experimentation.

The C.E.B.A.S. MINI-MODULE is the miniaturized space flight version of the Closed Equilibrated Biological Aquatic System (C.E.B.A.S.). It fits into a large middeck locker tray and is scheduled to be flown in the STS 85 and in the NEUROLAB missions. Its volume is about 9 liters and it consists of two animal tanks, a plant cultivator, and a bacteria filter in a monolithic design. An external sensor unit is connected to a data acquisition/control unit. The system integrates its own biological life support. The CO2 exhaled by the consumers (fishes, snails, microorganisms) is assimilated by water plants (Ceratophyllum demersum) which provide them with oxygen. The products of biomass degradation and excretion (mainly ammonia ions) are converted by bacteria into nitrite and nitrate. The latter is taken up by the plants as a nitrogen source together with other ions like phosphate. The plants convert light energy into chemical energy and their illumination is regulated via the oxygen concentration in the water by the control unit. In ground laboratory tests the system exhibited biological stability up to three month. The buffer capacity of the biological filter system is high enough to eliminate the degradation products of about one half of the dead animal biomass as shown in a "crash test". A test series using the laboratory model of the flight hardware demonstrated the biological stability and technical reliability with mission-identical loading and test duration. A comprehensive biological research program is established for the C.E.B.A.S. MINI-MODULE in which five German and three U.S.-American universities as well as the Russian Academy of Sciences are involved.