Plants survive rapid decompression: Implications for bioregenerative life support

Radish (Raphanus sativus), lettuce (Latuca sativa), and wheat (Triticum aestivum) plants were grown at either 98 kPa (ambient) or 33 kPa atmospheric pressure with constant 21 kPa oxygen and 0.12 kPa carbon dioxide in atmospherically closed pressure chambers. All plants were grown rockwool using recirculating hydroponics with a complete nutrient solution. At 20 days after planting, chamber pressures were pumped down as rapidly as possible, reaching 5 kPa after about 5 min and ∼1.5 kPa after about 10 min. The plants were held at 1.5 kPa for 30 min and then pressures were restored to their original settings. Temperature (22 °C) and humidity (65% RH) controls were engaged throughout the depressurization, although temperatures dropped to near 16 °C for a brief period. CO₂ and O₂ were not detectable at the low pressure, suggesting that most of the 1.5 kPa atmosphere consisted of water vapor. Following re-pressurization, plants were grown for another 7 days at the original pressures and then harvested. The lettuce, radish, and wheat plants showed no visible effects from the rapid decompression, and there were no differences in fresh or dry mass when compared to control plants maintained continuously at 33 or 98 kPa. But radish storage root fresh mass and lettuce head fresh and dry masses were less at 33 kPa compared to 98 kPa for both the controls and decompression treatment. The results suggest that plants are extremely resilient to rapid decompression, provided they do not freeze (from evaporative cooling) or desiccate. The water of the hydroponic system was below the boiling pressure during these tests and this may have protected the plants by preventing pressures from dropping below 1.5 kPa and maintaining humidity near 1.5 kPa. Further testing is needed to determine how long plants can withstand such low pressure, but the results suggest there are at least 30 min to respond to catastrophic pressure losses in a plant production chamber that might be used for life support in space.     

Biological nitrogen fixation under primordial Martian partial pressures of dinitrogen.

Early Earth and early Mars were similar enough such that past geochemical and climatic conditions on Mars may have also been favorable for the origin of life. However, one of the most striking differences between the two planets was the low partial pressure of dinitrogen (pN2) on early Mars (18 mb). On Earth, nitrogen is a key biological element and in many ecosystems the low availability of fixed nitrogen compounds is the main factor limiting growth. Biological fixation of dinitrogen on Earth is a crucial source of fixed nitrogen. Could the low availability of dinitrogen in the primordial Martian atmosphere have prevented the existence, or evolution of Martian microbiota? Azotobacter vinelandii and Azomonas agilis were grown in nitrogen free synthetic medium under various partial pressures of dinitrogen ranging from 780-0 mb (total atmosphere=1 bar). Below 400 mb the biomass, cell number, and growth rate decreased with decreasing pN2. Both microorganisms were capable of growth at a pN2 as low as 5 mb, but no growth was observed at a pN2 < or = 1 mb. The data appear to indicate that biological nitrogen fixation could have occurred on primordial Mars (pN2=18 mb) making it possible for a biotic system to have played a role in the Martian nitrogen cycle. It is possible that nitrogen may have played a key role in the early evolution of life on Mars, and that later a lack of available nitrogen on that planet (currently, pN2=0.2 mb) may have been involved in its subsequent extinction.     

Predicting the effects of gas diffusivity on photosynthesis and transpiration of plants grown under hypobaria

As part of a Bio-regenerative Life Support System (BLSS) for long-term space missions, plants will likely be grown at reduced pressure. This low pressure will minimize structural requirements for growth chambers on missions to the Moon or Mars. However, at reduced pressures the diffusivity of gases increases. This will affect the rates at which CO₂ is assimilated and water is transpired through stomata. To understand quantitatively the possible effects of reduced pressure on plant growth, CO₂ and H₂O transport were calculated for atmospheres of various total pressures (101, 66, 33, 22, 11 kPa) and CO₂ concentrations (0.04, 0.1 and 0.18 kPa). The diffusivity of a gas is inversely proportional to total pressure and shows dramatic increases at pressures below 33 kPa (1/3 atm). A mathematical relationship based on the principle of thermodynamics was applied to low pressure conditions and can be used for calculating the transpiration and photosynthesis of plants grown in hypobaria. At 33 kPa total pressure, the stomatal conductance increases by a factor of three with the boundary layer conductance increasing by a factor of ∼1.7, since the leaf conductance is a function of both stomatal and the boundary layer conductance, the overall conductance will increase resulting in significantly higher levels of transpiration as the pressure drops. The conductance of gases is also regulated by stomatal aperture in an inverse relationship. The higher CO₂ concentration inside the leaf air space during low pressure treatments may result in higher CO₂ assimilation and partial stomata closure, resulting in a decrease in transpiration rate. The results of this analysis offer guidelines for experiments in pressure and high CO₂ environments to establish ideal conditions for minimizing transpiration and maximizing the plant biomass yield in BLSS.     

Growth of wheat under one tenth of the atmospheric pressure.

Wheat plants were grown in twin closed growth chambers under normal and reduced atmospheric pressures. For the first 22 days from sowing, the reduced pressure was maintained at 200 hPa, and at 100 hPa for the remaining 27 days until harvest. These pressures were obtained by evacuation of the chamber and adding oxygen (170 and 79 hPa respectively) and carbon dioxide (0.65 and 1.0 hPa respectively; about 2 and 3 times above the control). Eighty-seven per cent of the final dry mass was produce under 100 hPa treatment. Growth and development of wheat are not negatively affected by low pressure treatment. Compared to the control, final dry mass increased by 76%, leaf number by 133%, and ear number by 35%, probably due to elevation of CO2. Shortening of shoot parts and increases in chlorophyll and proteins content are not in accordance with a predicted CO2 effect and could be attributed to the N2 removal and the subsequent alteration in gas diffusion rate.     

Growth of soybean and potato at high CO2 partial pressures.

Soybean and potato plants were grown in controlled environments at carbon dioxide (CO2) partial pressures ranging from 0.05 to 1.00 kPa. The highest yields of edible biomass occurred at 0.10 kPa for both species, with higher CO2 levels being supraoptimal, but not injurious to the plants. Stomatal conductance rates of upper canopy leaves were lowest at 0.10 kPa CO2, while conductance rates at 0.50 and 1.00 kPa were significantly greater than 0.10 kPa. Total water use by the plants was greatest at the highest CO2 pressures (i.e. 0.50 and 1.00 kPa); consequently, water use efficiencies (biomass produced/water used) were low at the highest CO2 pressures. Based on previous CO2 studies in the literature, the increased conductance and water use at the highest CO2 pressures were surprising and pose interesting challenges for managing plants in a CELSS, where CO2 pressures may exceed optimal levels.     

The effects of CO2 on growth and transpiration of radish (Raphanus sativus) in hypobaria

Plants grown on long-term space missions will likely be grown in low pressure environments (i.e., hypobaria). However, in hypobaria the transpiration rates of plants can increase and may result in wilting if the water is not readily replaced. It is possible to reduce transpiration by increasing the partial pressure of CO₂ (pCO₂), but the effects of pCO₂ at high levels (>120 Pa) on the growth and transpiration of plants in hypobaria are not known. Therefore, the effects of pCO₂ on the growth and transpiration of radish (Raphanus sativus var. Cherry Bomb II) in hypobaria were studied. The fresh weight (FW), leaf area, dry weight (DW), CO₂ assimilation rates (C_A), dark respiration rates (DR), and transpiration rates from 26 day-old radish plants that were grown for an additional seven days at different total pressures (33, 66 or 101 kPa) and pCO₂ (40 Pa, 100 Pa and 180 Pa) were measured. In general, the dry weight of plants increased with CO₂ enrichment and with lower total pressure. In limiting pCO₂ (40 Pa) conditions, the transpiration for plants grown at 33 kPa was approximately twice that of controls (101 kPa total pressure with 40 Pa pCO₂). Increasing the pCO₂ from 40 Pa to 180 Pa reduced the transpiration rates for plants grown in hypobaria and in standard atmospheric pressures. However, for plants grown in hypobaria and high pCO₂ (180 Pa) leaf damage was evident. Radish growth can be enhanced and transpiration reduced in hypobaria by enriching the gas phase with CO₂ although at high levels leaf damage may occur.     

Plant growth and gas balance in a plant and mushroom cultivation system.

In order to obtain basic data for construction of a plant cultivation system incorporating a mushroom cultivation subsystem in the CELSS, plant growth and atmospheric CO2 balance in the system were investigated. The plant growth was promoted by a high level of CO2 which resulted from the respiration of the mushroom mycelium in the system. The atmospheric CO2 concentration inside the system changed significantly due to the slight change in the net photosynthetic rate of plants and/or the respiration rate of the mushroom when the plant cultivation system combined directly with the mushroom cultivation subsystem.     

Carbon dioxide exchange of lettuce plants under hypobaric conditions.

Growth of plants in a Controlled Ecological Life Support System (CELSS) may involve the use of hypobaric pressures enabling lower mass requirements for atmospheres and possible enhancement of crop productivity. A controlled environment plant growth chamber with hypobaric capability designed and built at Ames Research Center was used to determine if reduced pressures influence the rates of photosynthesis (Ps) and dark respiration (DR) of hydroponically grown lettuce plants. The chamber, referred to as a plant volatiles chamber (PVC), has a growing area of about 0.2 m2, a total gas volume of about 0.7 m3, and a leak rate at 50 kPa of <0.1%/day. When the pressure in the chamber was reduced from ambient to 51 kPa, the rate of net Ps increased by 25% and the rate of DR decreased by 40%. The rate of Ps increased linearly with decreasing pressure. There was a greater effect of reduced pressure at 41 Pa CO2 than at 81 Pa CO2. This is consistent with reports showing greater inhibition of photorespiration (Pr) in reduced O2 at low CO2 concentrations. When the partial pressure of O2 was held constant but the total pressure was varied between 51 and 101 kPa, the rate of CO2 uptake was nearly constant, suggesting that low pressure enhancement of Ps may be mainly attributable to lowered partial pressure of O2 and the accompanying reduction in Pr. The effects of lowered partial pressure of O2 on Ps and DR could result in substantial increases in the rates of biomass production, enabling rapid throughput of crops or allowing flexibility in the use of mass and energy resources for a CELSS.     

Carbon dioxide exchange of lettuce plants under hypobaric conditions.

Growth of plants in a Controlled Ecological Life Support System (CELSS) may involve the use of hypobaric pressures enabling lower mass requirements for atmospheres and possible enhancement of crop productivity. A controlled environment plant growth chamber with hypobaric capability designed and built at Ames Research Center was used to determine if reduced pressures influence the rates of photosynthesis (Ps) and dark respiration (DR) of hydroponically grown lettuce plants. The chamber, referred to as a plant volatiles chamber (PVC), has a growing area of about 0.2 m2, a total gas volume of about 0.7 m3, and a leak rate at 50 kPa of <0.1%/day. When the pressure in the chamber was reduced from ambient to 51 kPa, the rate of net Ps increased by 25% and the rate of DR decreased by 40%. The rate of Ps increased linearly with decreasing pressure. There was a greater effect of reduced pressure at 41 Pa CO2 than at 81 Pa CO2. This is consistent with reports showing greater inhibition of photorespiration (Pr) in reduced O2 at low CO2 concentrations. When the partial pressure of O2 was held constant but the total pressure was varied between 51 and 101 kPa, the rate of CO2 uptake was nearly constant, suggesting that low pressure enhancement of Ps may be mainly attributable to lowered partial pressure of O2 and the accompanying reduction in Pr. The effects of lowered partial pressure of O2 on Ps and DR could result in substantial increases in the rates of biomass production, enabling rapid throughput of crops or allowing flexibility in the use of mass and energy resources for a CELSS.     

Effects of long-term low atmospheric pressure on gas exchange and growth of lettuce

The objectives of this research were to determine photosynthesis, evapotranspiration and growth of lettuce at long-term low atmospheric pressure. Lettuce (Lactuca sativa L. cv. Youmaicai) plants were grown at 40 kPa total pressure (8.4 kPa _pO₂$p_{{\text{O}}_2}$) or 101 kPa total pressure (20.9 kPa _pO₂$p_{{\text{O}}_2}$) from seed to harvest for 35 days. Germination rate of lettuce seeds decreased by 7.6% at low pressure, although this was not significant. There was no significant difference in crop photosynthetic rate between hypobaria and ambient pressure during the 35-day study. The crop evapotranspiration rate was significantly lower at low pressure than that at ambient pressure from 20 to 30 days after planting (DAP), but it had no significant difference before 20 DAP or after 30 DAP. The growth cycle of lettuce plants at low pressure was delayed. At low pressure, lettuce leaves were curly at the seedling stage and this disappeared gradually as the plants grew. Ambient lettuce plants were yellow and had an epinastic growth at harvest. The shoot height, leaf number, leaf length and shoot/root ratio were lower at low pressure than those at ambient pressure, while leaf area and root growth increased. Total biomass of lettuce plants grown at two pressures had no significant difference. Ethylene production at low pressure decreased significantly by 38.8% compared with ambient pressure. There was no significant difference in microelements, nutritional phytochemicals and nitrate concentrations at the two treatments. This research shows that lettuce can be grown at long-term low pressure (40 kPa) without significant adverse effects on seed germination, gas exchange and plant growth. Furthermore, ethylene release was reduced in hypobaria.     

Assessing the feasibility of involving gaseous products resulting from physicochemical oxidation of human liquid and solid wastes in the cycling of a bio-technical life support system

The study addresses the possible ways of involving gaseous products produced by “wet” incineration of human wastes mixed with H₂O₂ in an alternating electric field in the cycling of the physical model of a bio-technical life support system (BTLSS). The resulting gas mixture contains CO₂ and O₂, which are easily involved in the cycling in the closed ecosystem, and NH₃, which is unacceptable in the atmosphere of the BTLSS. NH₃ fixation has been proposed, which is followed by nitrification and involvement of the resulting products in the mass exchange of the closed system. Experiments have been performed to show that plants can be grown in the atmosphere resulting from the closing of the gas loop that includes a physicochemical installation and a growth chamber with plants representing the phototrophic compartment of the BTLSS. The results of the study suggest the conclusion that the proposed method of organic waste oxidation can be a useful tool in creating a physical model of a closed-loop integrated BTLSS.     

Light intensity and production parameters of phytocenoses cultivated on soil-like substrate under controlled [correction of controled] environment conditions.

To increase the degree of closure of biological life support systems of a new generation, we used vermicomposting to involve inedible phytomass in the intra-system mass exchange. The resulting product was a soil-like substrate, which was quite suitable for growing plants (Manukovsky et al. 1996, 1997). However, the soil like substrate can be regarded as a candidate for inclusion in a system only after a comprehensive examination of its physical, chemical, and other characteristics. An important criterion is the ability of the soil-like substrate to supply the necessary mineral elements to the photosynthesizing component under the chosen cultivation conditions. Thus, the purpose of this work was to study the feasibility of enhancing the production activity of wheat and radish crops by varying the intensity of photosynthetically active radiation, without decreasing the harvest index. The increase of light intensity from 920 to 1150 micromoles m-2 s-1 decreased the intensity of apparent photosynthesis of the wheat crops and slightly increased the apparent photosynthesis of the radish crops The maximum total and grain productivity (kg/m2) of the wheat crops was attained at the irradiance of 920 micromoles m-2 s-1. Light intensity of 1150 micromoles m-2 s-1 decreased the productivity of wheat plants and had no significant effect on the productivity of the radish crops (kg/m2) as compared to 920 micromoles m-2 s-1. The qualitative and quantitative composition of microflora of the watering solution and substrate was determined by the condition of plants, developmental phase and light intensity. By the end of wheat growth under 1150 micromoles m-2 s-1 the numbers of bacteria of the coliform family and phytopathogenic bacteria in the watering solution and substrate were an order of magnitude larger than under other illumination conditions. The obtained data suggest that the cultivation of plants in a life support system on soil-like substrate from composts has a number of advantages over the cultivation on neutral substrates, which require continual replenishment of the plant nutrient solution from the system's store to complement the macro- and micro-elements. Yet, a number of problems arise, including those related to the controlling of the production activity of the plants by the intensity of photosynthetically active radiation. It is essential to understand why the intensity of production processes is limited at higher irradiation levels and to overcome the factors responsible for this, so that the soil-like substrate could have an even better chance in the competition for the best plant cultivation technology to be used in biological life support systems.     

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.     

HCN formation under electron impact: experimental studies and application to Neptune's atmosphere.

Laboratory experiments simulating organic synthesis in Neptune's atmosphere have been performed. We have submitted to a spark discharge gaseous mixtures containing 9 mbar of molecular nitrogen and 3 mbar of methane (the p(N2)/p(CH4) ratio is compatible with upper limits in Neptune's stratosphere) with varying quantities of molecular hydrogen. The spark discharge is used to model the energetic electrons produced by the impact of cosmic rays on the high atmosphere of Neptune. HCN is synthesized in the described experimental conditions, even with a low mixing ratio of molecular nitrogen. Studying the variation of HCN production with the initial composition of the gas mixture and extrapolating to high mixing ratio of molecular hydrogen allows to estimate HCN production in Neptune's atmosphere. The computed HCN production flux is 7x10(7) m-2 s-1, which is two orders of magnitude lower than the value predicted by chemical models for an internal source of N atoms. The major uncertainty in our extrapolation is the energetic distribution of electrons, implicitly assumed comparable in the discharge and in Neptune's atmosphere. We note that this distribution is also a source of uncertainty in chemical models. The chemical mechanism responsible for the local formation of HCN in the stratosphere probably occurs in the reactor too. We propose a simple characterization of the spark discharge. We thus link the molecular nitrogen dissociation cross section by electron impact to the measured parameters of the experiments (current, voltage, initial partial pressures) and to the resulting HCN partial pressures. However, other laboratory experiments with larger hydrogen pressures, requiring a more powerful electric source, have to be performed to yield a value of the cross section.     

The physical nature of Titan's aerosols: laboratory simulations.

The atmosphere of Titan is known to contain aerosols, as evidenced by the Voyager observations of at least three haze layers. Such aerosols can have significant effects on the reflection spectrum of Titan and on the chemistry and thermal structure of its atmosphere. To investigate some of these effects, laboratory simulations of the chemistry of Titan's atmosphere have been done. The results of these studies show that photolysis of acetylene, ethylene, and hydrogen cyanide, known constituents of Titan's atmosphere, yields sub-micron sized spheres, with mean diameters ranging from 0.4 to 0.8 microns, depending on the pressures of the reactant gases. Most of the spheres are contained in near-linear aggregates. The formation of the aggregates is consistent with models of Titan's reflection spectrum and polarization, which are best fit with non-spherical particles. At room temperature, the particles are very sticky, but their properties at low temperatures on Titan are presently not known.     

Short-term and long-term effects of low total pressure on gas exchange rates of spinach.

In this study, spinach plants were grown under atmospheric and low pressure conditions with constant O2 and CO2 partial pressures, and the effects of low total pressure on gas exchange rates were investigated. CO2 assimilation and transpiration rates of spinach grown under atmospheric pressure increased after short-term exposure to low total pressure due to the enhancement of leaf conductance. However, gas exchange rates of plants grown at 25 kPa total pressure were not greater than those grown at atmospheric pressure. Stomatal pore length and width were significantly smaller in leaves grown at low total pressure. This result suggested that gas exchange rates of plants grown under low total pressure were not stimulated even with the enhancement of gas diffusion because the stomatal size and stomatal aperture decreased.     

Conceptual design of a bioregenerative life support system containing crops and silkworms

This article summarizes a conceptual design of a bioregenerative life support system for permanent lunar base or planetary exploration. The system consists of seven compartments – higher plants cultivation, animal rearing, human habitation, water recovery, waste treatment, atmosphere management, and storages. Fifteen kinds of crops, such as wheat, rice, soybean, lettuce, and mulberry, were selected as main life support contributors to provide the crew with air, water, and vegetable food. Silkworms fed by crop leaves were designated to produce partial animal nutrition for the crew. Various physical-chemical and biological methods were combined to reclaim wastewater and solid waste. Condensate collected from atmosphere was recycled into potable water through granular activated carbon adsorption, iodine sterilization, and trace element supplementation. All grey water was also purified though multifiltration and ultraviolet sterilization. Plant residue, human excrement, silkworm feces, etc. were decomposed into inorganic substances which were finally absorbed by higher plants. Some meat, ingredients, as well as nitrogen fertilizer were prestored and resupplied periodically. Meanwhile, the same amount and chemical composition of organic waste was dumped to maintain the steady state of the system. A nutritional balanced diet was developed by means of the linear programming method. It could provide 2721 kcal of energy, 375.5 g of carbohydrate, 99.47 g of protein, and 91.19 g of fat per capita per day. Silkworm powder covered 12.54% of total animal protein intakes. The balance of material flows between compartments was described by the system of stoichiometric equations. Basic life support requirements for crews including oxygen, food, potable and hygiene water summed up to 29.68 kg per capita per day. The coefficient of system material closure reached 99.40%.     

Food production and gas exchange system using blue-green alga (Spirulina) for CELSS.

In order to reduce the cultivation area required for the growth of higher plants in space adoption of algae, which have a higher photosynthetic ability, seems very suitable for obtaining oxygen and food as a useful source of high quality protein. The preliminary cultivation experiment for determining optimum cultivation conditions and for obtaining the critical design parameters of the cultivator itself has been conducted. Spirulina was cultivated in the 6-liter medium containing sodium hydrogen carbonate solution and a cultivation temperature controlled using a thermostat. Generated oxygen gas was separated using a polypropyrene porous hollow fiber membrane module. Through this experiment, oxygen gas (at a concentration of more than 46%) at a rate of 100-150 ml per minute could be obtained.     

Increase of atmospheric CO2: response patterns of a simple terrestrial man-made ecosystem.

Simple models of terrestrial ecosystems with a limited number of components are an efficient tool to study the main laws of functioning of populations, including microbial ones, and their communities, as components of natural ecosystems, under variable environmental conditions. Among other factors are the increase of carbon dioxide in the atmosphere and limitation of plants' growth by biogenic elements. The main types of ecosystems' responses to changes in environmental conditions (a change in CO2 concentration) have been demonstrated in a "plants-rhizospheric microorganisms-artificial soil" simple experimental system. The mathematical model of interactions between plants and microorganisms under normal and elevated atmospheric CO2 and limitation by nutrients (nitrogen and phosphorus) yielded a qualitative agreement between calculated and experimental values of limiting substances concentrations and release rates of exudates.