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NMR studies on Na+ transport in Synechococcus PCC 6311.

The freshwater cyanobacterium Synechococcus PCC 6311 is able to adapt to grow after sudden exposure to salt (NaCl) stress. We have investigated the mechanism of Na+ transport in these cells during adaptation to high salinity. Na+ influx under dark aerobic conditions occurred independently of delta pH or delta psi across the cytoplasmic membrane, ATPase activity, and respiratory electron transport. These findings are consistent with the existence of Na+/monovalent anion cotransport or simultaneous Na+/H(+)+anion/OH- exchange. Na+ influx was dependent on Cl-, Br-, NO3-, or NO2-. No Na+ uptake occurred after addition of NaI, NaHCO3, or Na2SO4. Na+ extrusion was absolutely dependent on delta pH and on an ATPase activity and/or on respiratory electron transport. This indicates that Na+ extrusion via Na+/H+ exchange is driven by primary H+ pumps in the cytoplasmic membrane. Cells grown for 4 days in 0.5 M NaCl medium, "salt-grown cells," differ from control cells by a lower vmax of Na+ influx and by lower steady-state ratios of [Na+]in/[Na+]out. These results indicate that cells grown in high-salt medium increase their capacity to extrude Na+. During salt adaptation Na+ extrusion driven by respiratory electron transport increased from about 15 to 50%.

Adenosine Triphosphatases↗

Biogenerative life-support system: farming on the moon.

Plants can be used to recycle food, oxygen, and water in a closed habitat (e.g., on the moon, Mars, or in a space craft. A variety of crops might be grown, probably in underground growth units to avoid harmful radiation and micrometeorites. Artificial light will be necessary although some sunlight might be brought in via fiber optics. Transpired water will be condensed in coils exposed to space and shaded from sunlight. Oxygen and CO2 levels will be maintained by controlling photosynthesis and waste oxidation. Plants will be grown hydroponically. Wheat has been produced at the rate of 60 g m-2 d-1, which could feed a human continuously from a farm only 13 m2, but nearly continuous light equivalent to sunlight is required along with ideal temperatures, enriched CO2, suitable cultivars, etc. Lower light results in more efficient photosynthesis but requires a larger farm, as do safety considerations and many crops.

Agriculture↗

Biomass recycle as a means to improve the energy efficiency of CELSS algal culture systems.

Algal cultures can be very rapid and efficient means to generate biomass and regenerate the atmosphere for closed environmental life support systems. However, as in the case of most higher plants, a significant fraction of the biomass produced by most algae cannot be directly converted to a useful food product by standard food technology procedures. This waste biomass will serve as an energy drain on the overall system unless it can be efficiently recycled without a significant loss of its energy content. We report experiments in which cultures of the algae Scenedesmus obliquus were grown in the light and at the expense of an added carbon source, which either replaced or supplemented the actinic light. As part of these experiments we tested hydrolyzed waste biomass from these same algae to determine whether the algae themselves could be made part of the biological recycling process. Results indicate that hydrolyzed algal (and plant) biomass can serve as carbon and energy sources for the growth of these algae, suggesting that the efficiency of the closed system could be significantly improved using this recycling process.

Biomass↗

Non-conventional approaches to food processing in CELSS. I--Algal proteins; characterization and process optimization.

Protein isolate obtained from green algae (Scenedesmus obliquus) cultivated under controlled conditions was characterized. Molecular weight determination of fractionated algal proteins using SDS-polyacrylamide gel electrophoresis revealed a wide spectrum of molecular weights ranging from 15,000 to 220,000. Isoelectric points of dissociated proteins were in the range of 3.95 to 6.20. Amino acid composition of protein isolate compared favorably with FAO standards. High content of essential amino acids leucine, valine, phenylalanine and lysine makes algal protein isolate a high quality component of CELSS diets. To optimize the removal of algal lipids and pigments supercritical carbon dioxide extraction (with and without ethanol as a co-solvent) was used. Addition of ethanol to supercritical CO2 resulted in more efficient removal of algal lipids and produced protein isolate with a good yield and protein recovery. The protein isolate extracted by the above mixture had an improved water solubility.

Amino Acids↗

Application of photosynthetic N2-fixing cyanobacteria to the CELSS program.

The feasibility of using photosynthetic microalgae (cyanobacteria) as a subsystem component for the CELSS program, with particular emphasis on the manipulation of the biomass (protein/carbohydrate) has been addressed. Using factors which retard growth rates, but not photosynthetic electron flux, the partitioning of photosynthetically derived reductant may be dictated towards CO2 fixation (carbohydrate formation) and away from N2 fixation (protein formation). Cold shock treatment of fairly dense cultures markedly increases the glycogen content from 1% to 35% (dry weight), and presents a useful technique to change the protein/carbohydrate ratio of these organisms to a more nutritionally acceptable form.

Biomass↗

Design of an elemental analysis system for CELSS research.

The results of experiments conducted with higher plants in tightly sealed growth chambers provide definite evidence that the physical closure of a chamber has significant effects on many aspects of a plant's biology. One of these effects is seen in the change in rates of uptake, distribution, and re-release of nutrient elements by the plant (mass balance). Experimental data indicates that these rates are different from those recorded for plants grown in open field agriculture, or in open growth chambers. Since higher plants are a crucial component of a CELSS, it is important that the consequences of these rate differences be understood with regard to the growth and yield of the plants. This paper will focus on the description of a system for elemental analysis which can be used to monitor the mass balance of nutrient elements in CELSS experiments. Additionally, data on the uptake of nutrient elements by higher plants grown in a growth chamber will be presented.

Anions↗

Utilization of potatoes in bioregenerative life support systems.

Data on the tuberization, harvest index, and morphology of 2 cvs of white potato (Solanum tuberosum L.) grown at 12, 16, 20, 24 and 28 degrees C, 250, 400 and 550 micromoles s-1 m-2 photosynthetic photon flux (PPF), 350, 1000 and 1600 microliters l-1 CO2 will be presented. A productivity of 21.9 g m-2 day-1 of edible tubers from a solid stand of potatoes grown for 15 weeks with continuous irradiation at 400 micromoles s-1 m-2, 16 degrees C and 1000 microliters l-1 CO2 has been obtained. This equates to an area of 34.3 m2 being required to provide 2800 kcal of potatoes per day for a human diet. Separated plants receiving side lighting have produced 32.8 g m-2 day-1 which equates to an area of 23.6 m2 to provide 2800 kcal. Studies with side lighting indicate that productivities in this range should be realized from potatoes. Glycoalkaloid levels in tubers of controlled-environment-grown plants are within the range of levels found in tubers of field grown plants. The use and limitation of recirculating solution cultures for potato growth is discussed.

Carbon Dioxide↗

Wheat production in controlled environments.

Our goal is to optimize conditions for maximum yield and quality of wheat to be used in a controlled-environment, life-support system (CELSS) in a Lunar or Martian base or perhaps in a space craft. With yields of 23 to 57 g m-2 d-1 of edible biomass, a minimum size for a CELSS would be between 12 and 30 m2 per person, utilizing about 600 W m-2 of electrical energy for artificial light. Temperature, irradiance, photoperiod, carbon-dioxide levels, humidity, and wind velocity are controlled in state-of-the-art growth chambers. Nutrient solutions (adjusted for wheat) are supplied to the roots via a recirculating system that controls pH by adding HNO3 and controlling the NO3/NH4 ratio in solution. A rock-wool plant support allows direct seeding and densities up to 10,000 plants per meter2. Densities up to 2000 plants m-2 appear to increase seed yield. Biomass production increases almost linearly with increasing irradiance from 400 to 1700 micromoles m-2 s-1 of photosynthetic photon flux (PPF), but the efficiency of light utilization decreases over this range. Photoperiod and temperature both have a profound influence on floral initiation, spikelet formation, stem elongation, and fertilization. High temperatures (25 to 27 degrees C) and long days shorten the life cycle and promote rapid growth, but cooler temperatures (20 degrees C) and shorter days greatly increase seed number per head and thus yield (g m-2). The life cycle is lengthened in these conditions but yield per day (g m-2 d-1) is still increased. We have evaluated about 600 cultivars from around the world and have developed several breeding lines for our controlled conditions. Some of our ultra-dwarf lines (30 to 50 cm tall) look especially promising with high yields and high harvest indices (percent edible biomass).

Biomass↗

Mass balances for a biological life support system simulation model.

Design decisions to aid the development of future space-based biological life support systems (BLSS) can be made with simulation models. Here we develop the biochemical stoichiometry for 1) protein, carbohydrate, fat, fiber, and lignin production in the edible and inedible parts of plants; 2) food consumption and production of organic solids in urine, feces, and wash water by the humans; and 3) operation of the waste processor. Flux values for all components are derived for a steady-state system with wheat as the sole food source. The large-scale dynamics of a materially-closed (BLSS) computer model is described in a companion paper. An extension of this methodology can explore multi-food systems and more complex biochemical dynamics while maintaining whole-system closure as a focus.

Biomass↗

Current and potential productivity of wheat for a Controlled Environment Life Support System.

The productivity of higher plants is determined by the incident photosynthetic photon flux (PPF) and the efficiency of the following four physiological processes: absorption of PPF by photosynthetic tissue, carbon fixation (photosynthesis), carbon use (respiration), and carbon partitioning (harvest index). These constituent processes are analyzed to determine theoretical and potentially achievable productivity. The effects of optimal environmental and cultural factors on each of these four factors is also analyzed. Results indicate that an increase in the percentage of absorbed photons is responsible for most of the improvement in wheat yields in an optimal controlled environment. Several trials confirm that there is an almost linear increase in wheat yields with increasing PPF. An integrated PPF of 150 mol m-2 d-1 (2.5 times summer sunlight) has produced 60 g m-2 d-1 of grain. Apparently, yield would continue to increase with even higher PPF's. Energy efficiency increased with PPF to about 600 micromoles m-2 s-1, then slowly decreased. We are now seeking to improve efficiency at intermediate PPF levels (1000 micromoles m-2 s-1) before further exploring potential productivity. At intermediate and equal integrated daily PPF levels, photoperiod had little effect on yield per day or energy efficiency. Decreasing temperature from 23 degrees to 17 degrees increased yield per day by 20% but increased the life cycle from 62 to 89 days. We hope to achieve both high productivity and energy efficiency.

Biomass↗

Carbon balance and productivity of Lemna gibba, a candidate plant for CELSS.

The photosynthesis and productivity of Lemna gibba were studied with a view to its use in Controlled Ecological Life Support Systems (CELSS). Photosynthesis of L. gibba floating on the nutrient solution could be driven by light coming from either above or below. Light from below was about 75% as effective as from above when the stand was sparse, but much less so with dense stands. High rates of photosynthesis (ca. 800 nanomoles CO2 g dry weight (DW)-1 s-1) were measured at 750 micromoles m-2 s-1 PPF and 1500 micromoles mol-1 CO2. This was attained at densities up to 660 g fresh weight (FW) m-2 with young cultures. After a few days growth under these conditions, and at higher densities, the rate of photosynthesis dropped to less than 25% of the initial value. This drop was only partly alleviated by thinning the stand or by introducing a short dark period at high temperature (26 degrees C). Despite the drop in the rate of photosynthesis, maximum yields were obtained in batch cultures grown under continuous light, constant temperature and high [CO2]. Plant protein content was less than reported for field grown Lemna. When the plants were harvested daily, maintaining a stand density of 600 g FW m-2, yields of 18 g DW m-2 d-1 were obtained. The total dry weight of L. gibba included 40% soluble material (sugars and amino acids), 15% protein, 5% starch, 5% ash and 35% cellulose and other polymers. We conclude that a CELSS system could be designed around stacked, alternate layers of transparent Lemna trays and lamps. This would allow for 7 tiers per meter height. Based on present data from single layers, the yield of such a system is calculated to be 135 g DW m-3 d-1 of a 100% edible, protein-rich food.

Carbon↗

Utilization of white potatoes in CELSS.

Potatoes (Solanum tuberosum) have a strong potential as a useful crop species in a functioning CELSS. The cultivar Denali has produced 37.5 g m-2 d-1 when grown for 132 days with the first 40 days under a 12-h photoperiod and a light:dark temperature cycle of 20 degrees C:16 degrees C, and then 92 days under continuous irradiance and a temperature of 16 degrees C. Irradiance was at 725 micromoles m-2 s-1 PPF and carbon dioxide at 1000 micromoles mol-1. The dried tubers had 82% carbohydrates, 9% protein and 0.6% fat. Other studies have shown that carbon dioxide supplementation (1000 micromoles mol-1) is of significant benefit under 12-h irradiance but less benefit under 24 h irradiance. Irradiance cycles of 60 minutes light and 30 minutes dark caused a reduction of more than 50% in tuber weight compared to cycles of 16 h light and 8 h dark. A diurnal temperature change of 22 degrees C for the 12-h light period to 14 degrees C during the 12-h dark period gave increased yields of 30% and 10% for two separate cultivars, compared with plants grown under a constant 18 degrees C temperature. Cultivar screening under continuous irradiance and elevated temperatures (28 degrees C) for 8 weeks of growth indicated that the cvs Haig, Denali, Atlantic, Desiree and Rutt had the best potential for tolerance to these conditions. Harvesting of tubers from plants at weekly intervals, beginning at 8 weeks after planting, did not increase yield over a single final harvest. Spacing of plants on 0.055 centers produced greater yield per m2 than spacing at 0.11 or 0.22 m2. Plants maintained 0.33 meters apart (0.111 m2 per plant) in beds produced the same yields when separated by dividers in the root matrix as when no separation was made.

Biomass↗

Transpiration during life cycle in controlled wheat growth.

We use a previously-developed model of wheat growth, which was designed for convenient incorporation into system-level models of advanced space life support systems. We apply the model to data from an experiment that grew wheat under controlled conditions and measured fresh biomass and cumulated transpiration as a function of time. We examine the adequacy of modeling the transpiration as proportional to the inedible biomass and an age factor, which varies during the life cycle. Results indicate that during the main phase of vegetative growth in the first half of the life cycle, the rate of transpiration per unit mass of inedible biomass is more than double the rate during the phase of grain development and maturation during latter half of the life cycle.

Biomass↗

Waste recycling issues in bioregenerative life support.

Research and technology development issues centering on the recycling of materials within a bioregenerative life support system are reviewed. The importance of recovering waste materials for subsequent use is emphasized. Such material reclamation will substantially decrease the energy penalty paid for bioregenerative life support systems, and can potentially decrease the size of the system and its power demands by a significant amount. Reclamation of fixed nitrogen and the sugars in cellulosic materials is discussed.

Biomass↗

Sources and processing of CELSS wastes.

The production rate and solid content of waste streams found in a life support system for a space habitat (in which plants are grown for food) are discussed. Two recycling scenarios, derived from qualitative considerations as opposed to quantitative mass and energy balances, tradeoff studies, etc., are presented; they reflect differing emphases on and responses to the waste stream formation rates and their composition, as well as indicate the required products from waste treatment that are needed in a life support system. The data presented demonstrate the magnitude of the challenge to developing a life support system for a space habitat requiring a high degree of closure.

Air Pollutants↗

Subcritical and supercritical water oxidation of CELSS model wastes.

Controlled-Ecological-Life-Support-System (CELSS) model wastes were wet-oxidized at temperatures from 250 to 500 degrees C, i.e., below and above the critical point of water (374 degrees C and 218 kg/cm2 or 21.4 MPa). A solution of ammonium hydroxide and acetic acid and a slurry of human urine, feces, and wipes were used as model wastes. Almost all of the organic matter in the model wastes was oxidized in the temperature range from 400 to 500 degrees C, i.e., above the critical conditions for water. In contrast, only a small portion of the organic matter was oxidized at subcritical conditions. Although the extent of nitrogen oxidation to nitrous oxide (N2O) and/or nitrogen gas (N2) increased with reaction temperature, most of the nitrogen was retained in solution as ammonia near 400 degrees C. This important finding suggests that most of the nitrogen in the waste feed can be retained in solution as ammonia during oxidation at low supercritical temperatures and be subsequently used as a nitrogen source for plants in a CELSS while at the same time organic matter is almost completely oxidized to carbon dioxide and water. It was also found in this study the Hastelloy C-276 alloy reactor corroded during waste oxidation. The rate of corrosion was lower above than below the critical temperature for water.

Acetates↗

Productivity and food value of Amaranthus cruentus under non-lethal salt stress.

Stress effects from the accumulation of metal salts may pose a problem for plants in closed biological systems such as spacecraft. This work examined the effects of salinity on growth, photosynthesis and carbon allocation in the crop plant, Amaranthus. Plants were germinated and grown in modified Hoagland's solution with NaCl concentrations of 0 to 1.0%. Plants received salt treatments at various times in development to assess effects on particular life history phases. For Amaranthus cruentus, germination, vegetative growth, flowering, seed development and yield were normal at salinities from 0 to 0.2%. Inhibition of these phases increased from 0.2 to 0.4% salinity and was total above 0.5%. 1.0% salinity was lethal to all developmental phases. Onset of growth phases were not affected by salinity. Plants could not be adapted by gradually increasing salinity over days or weeks. Water uptake increased, while photosynthetic CO2 uptake decreased with increasing salinity on a dry weight basis during vegetative growth. Respiration was not affected by salinity. After flowering, respiration and photosynthesis decreased markedly, such that 1.0% NaCl inhibited photosynthesis completely. Protein levels were unchanged with increasing salinity. Leaf starch levels were lower at salinities of 0.5% and above, while stem starch levels were not affected by these salinities. The evidence supports salt inhibition arising from changes in primary biochemical processes rather than from effects on water relations. While not addressing the toxic effects of specific ions, it suggests that moderate salinity per se need not be a problem in space systems.

Ecological Systems, Closed↗

Phase separated membrane bioreactor: results from model system studies.

The operation and evaluation of a bioreactor designed for high intensity oxygen transfer in a microgravity environment is described. The reactor itself consists of a zero headspace liquid phase separated from the air supply by a long length of silicone rubber tubing through which the oxygen diffuses in and the carbon dioxide diffuses out. Mass transfer studies show that the oxygen is film diffusion controlled both externally and internally to the tubing and not by diffusion across the tube walls. Methods of upgrading the design to eliminate these resistances are proposed. Cell growth was obtained in the fermenter using Saccharomyces cerevisiae showing that this concept is capable of sustaining cell growth in the terrestrial [correction of terrestial] simulation.

Carbon Dioxide↗