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.     

Atmospheric leakage and method for measurement of gas exchange rates of a crop stand at reduced pressure.

The variable pressure growth chamber (VPGC) was used in a 34-day functional test to grow a wheat crop using reduced pressure (70 kPa) episodes totalling 131 hours. Primary goals of the test were to verify facility and subsystem performance at 70 kPa and to determine responses of a wheat stand to reduced pressure and modified partial pressures of carbon dioxide and oxygen. Operation and maintenance of the chamber at 70 kpa involved continuous evacuation of the chamber atmosphere, leading to CO2 influx and efflux. A model for calculating CO2-exchange rates (net photosynthesis and dark respiration) was developed and tested and involved measurements of chamber leakage to determine appropriate corrections. Measurement of chamber leakage was based on the rate of pressure change over a small pressure increment (70.3 to 72.3 kPa) with the pump disabled. Leakage values were used to correct decreases and increases in chamber CO2 concentration resulting from net photosynthesis (Ps) and dark respiration (DR), respectively. Composite leakage corrections (influx and efflux) at day 7 of the test were 9% and 19% of the changes measured for Ps and DR, respectively. On day 33, composite corrections were only 3% for Ps and 4% for DR. During the test, the chamber became progressively tighter; the leak rate at 70.3 kPa decreasing from 2.36 chamber volumes/day pretest, to 1.71 volumes/day at the beginning of the test, and 1.16 volumes/day at the end of the test. Verification of the short-term leakage tests (rate of pressure rise) were made by testing CO2 leakage with the vacuum pump enabled and disabled. Results demonstrate the suitability of the VPGC or conducting gas exhange measurements of a crop stand at reduced pressure.     

Photosynthesis and respiration of a wheat stand at reduced atmospheric pressure and reduced oxygen.

A 34-day functional test was conducted in Johnson Space Center's Variable Pressure Growth Chamber (VPGC) to determine responses of a wheat stand to reduced pressure (70 kPa) and modified partial pressures of carbon dioxide and oxygen. Reduced pressure episodes were generally six to seven hours in duration, were conducted at reduced ppO2 (14.7 kPa), and were interrupted with longer durations of ambient pressure (101 kPa). Daily measurements of stand net photosynthesis (Pn) and dark respiration (DR) were made at both pressures using a ppCO2 of 121 Pa. Corrections derived from leakage tests were applied to reduced pressure measurements. Rates of Pn at reduced pressure averaged over the complete test were 14.6% higher than at ambient pressure, but rates of DR were unaffected. Further reductions in ppO2 were achieved with a molecular sieve and were used to determine if Pn was enhanced by lowered O2 or by lowered pressure. Decreased ppO2 resulted in enhanced rates of Pn, regardless of pressure, but the actual response was dependent on the ratio of ppO2/ppCO2. Over the range of ppO2/ppCO2 of 80 to 200, the rate of Pn declined linearly. Rate of DR was unaffected over the same range and by dissolved O2 levels down to 3.1 ppm, suggesting that normal rhizosphere and canopy respiration occur at atmospheric ppO2 levels as low as 11 kPa. Partial separation of effects attributable to oxygen and those related to reduced pressure (e.g. enhanced diffusion of CO2) was achieved from analysis of a CO2 drawdown experiment. Results will be used for design and implementation of studies involving complete crop growth tests at reduced pressure.     

Proximate nutritional composition of CELSS crops grown at different CO2 partial pressures.

Two CELSS candidate crops, soybean (Glycine max) and potato (Solanum tuberosum), were grown hydroponically in controlled environments maintained at carbon dioxide (CO2) partial pressures ranging from 0.05 to 1.00 kPa (500 to 10,000 ppm at 101 kPa atmospheric pressure). Plants were harvested at maturity (90 days for soybean and 105 days for potato) and all tissues analyzed for proximate nutritional composition (i.e. protein, fat, carbohydrate, crude fiber, and ash content). Soybean seed ash and crude fiber were higher and carbohydrate was lower than values reported for field-grown seed. Potato tubers showed little difference from field-grown tubers. With the exception of increased crude fiber of soybean seed with increased CO2, no trends were apparent with regard to CO2 effects on proximate composition of soybean seed and potato tubers. Crude fiber of soybean stems and leaves increased with increased CO2, as did soybean leaf protein (total nitrogen). Potato leaf and stem (combined) protein levels also increased with increased CO2, while leaf and stem carbohydrates decreased. Values for leaf and stem protein and ash were higher than values generally reported for field-grown plants for both species. Results suggest that CO2 partial pressure should have little influence on proximate composition of potato tubers or soybean seed, but that high ash and protein levels might be expected from leaves and stems of crops grown in controlled environments of a CELSS.     

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.     

Effects of modified atmosphere on crop productivity and mineral content.

Wheat, potato, pea and tomato crops were cultivated from seeding to harvest in a controlled and confined growth chamber at elevated CO2 concentration (3700 microL L-1) to examine the effects on biomass production and edible part yields. Different responses to high CO2 were recorded, ranging from a decline in productivity for wheat, to slight stimulation for potatoes, moderate increase for tomatoes, and very large enhancement for pea. Mineral content in wheat and pea seeds was not greatly modified by the elevated CO2. Short-term experiments (17 d) were conducted on potato at high (3700 microL L-1) and very high (20,000 microL L-1) CO2 concentration and/or low O2 partial pressure (approximately 20,600 microL L-1 or 2 kPa). Low O2 was more effective than high CO2 in total biomass accumulation, but development was affected: Low O2 inhibited tuberization, while high CO2 significantly increased production of tubers.     

Methods for measurement and control of leakage in CELSS and their application and performance in the Biosphere 2 facility.

Atmospheric leakage between a CELSS and its surround is driven by the differential pressure between the two. In an earth-based CELSS, both negative and positive differential pressures of atmosphere are created as the resultant of three influences: thermal expansion/contraction, transition of water between liquid and vapor phases, and external barometric pressure variations. The resultant may typically be on the order of 5000 pascals. By providing a flexible expansion chamber, the differential pressure range can be reduced two, or even three, orders of magnitude, which correspondingly reduces the leakage. The expansion chamber itself can also be used to measure the leak rate. Independent confirmation is possible by measurement of the progressive dilution of a trace gas. These methods as employed at the Biosphere 2 facility have resulted in an estimated atmospheric leak rate of less than 10 percent per year.     

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.     

Achieving and documenting closure in plant growth facilities.

As NASA proceeds with its effort to develop a Controlled Ecological Life Support System (CELSS) that will provide life support to crews during long duration space missions, it must address the question of facility and system closure. Here we discuss the concept of closure as it pertains to CELSS and describe engineering specifications, construction problems and monitoring procedures used in the development and operation of a closed plant growth facility for the CELSS program. A plant growth facility is one of several modules required for a CELSS. A prototype of this module at Kennedy Space Center is the large (7m tall x 3.5m diameter) Biomass Production Chamber (BPC), the central facility of the CELSS Breadboard Project. The BPC is atmospherically sealed to a leak rate of approximately 5% of its total volume per 24 hours. This paper will discuss the requirements for atmospheric closure in this facility, present CO2 and trace gas data from initial tests of the BPC with and without plants, and describe how the chamber was sealed atmospherically. Implications that research conducted in this type of facility will have for the CELSS program are discussed.     

A simplified closed artificial ecosystem--recycling of organic matter into wheat plants.

A simplified closed system consisting of a plant growth chamber coupled to a decomposition chamber was used to study carbon exchange dynamics. The CO2 produced via the decomposition of wheat straw was used for photosynthetic carbon uptake by wheat plants. The atmosphere of the two chambers was connected through a circuit of known flow rate. Thus, monitoring the CO2 concentrations in both compartments allowed measurement of the carbon exchange between the chambers, and estimation of the rate of respiration processes in the decomposition chamber and photosynthetic rate in the producer chamber. The objective for CELSS research was to simulate a system where a compartment producing food via photosynthesis, would be supplied by CO2 produced from respiration processes. The decomposition of biomass by the decomposer simulated both the metabolism of a crew and the result of a recycling system for inedible biomass. Concerning terrestrial ecosystems, the objective was to study organic matter decomposition in soil and other processes related to permanent grasslands.     

Evaluation of Cyanothece sp. ATCC 51142 as a candidate for inclusion in a CELSS.

Controlled ecological life support systems (CELSS) have been proposed to make long-duration manned space flights more cost-effective. Higher plants will presumably provide food and a breathable atmosphere for the crew. It has been suggested that imbalances between the CO2/O2 gas exchange ratios of the heterotrophic and autotrophic components of the system will inevitably lead to an unstable system, and the loss of O2 from the atmosphere. Ratio imbalances may be corrected by including a second autotroph with an appropriate CO2/O2 gas exchange ratio. Cyanothece sp. ATCC 51142 is a large unicellular N2-fixing cyanobacterium, exhibiting high growth rates under diverse physiological conditions. A rat-feeding study showed the biomass to be edible. Furthermore, it may have a CO2/O2 gas exchange ratio that theoretically can compensate for ratio imbalances. It is suggested that Cyanothece spp. could fulfill several roles in a CELSS: supplementing atmosphere recycling, generating fixed N from the air, providing a balanced protein supplement, and protecting a CELSS in case of catastrophic crop failure.     

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.     

Preliminary experimental results of gas recycling subsystems except carbon dioxide concentration.

Oxygen concentration and separation is an essential factor for air recycling in a CELSS. Furthermore, if the value of the plant assimilatory quotient is not coincident with that of the animal respiratory quotient, the recovery of O2 from the concentrated CO2 through chemical methods will become necessary to balance the gas contents in a CELSS. Therefore, oxygen concentration and separation equipment using Salcomine and O2 recovery equipment, such as Sabatier and Bosch reactors, were experimentally developed and tested.     

Carbon dioxide and oxygen budgets of a plant cultural system in a CELSS--a case of cultivation of lettuce and turnips.

In order to collect basic data about CO2 and O2 budgets of a plant cultural system in a CELSS, the variation of the CO2 absorption rates of lettuce and turnips were observed during the growing period, under different conditions. The O2 release rates were deduced from the CO2 absorption rates multiplied by 32/44. As a result, when the light intensity, the photoperiod and the atmospheric CO2 concentration increased, the rates also increased. The effects on the turnips were more significant than those on the lettuce. Turnips at 310 micromoles/m2/s of PPFD, 24 hours of photoperiod and 1100 ppm of CO2 concentration grew most actively in the present experimental conditions. One turnip absorbed 32.3 g CO2 and released 23.5 g O2 for 6 days between 24 days and 30 days after sowing.     

减压直流非转移弧等离子体射流速度的静电探针测量

组建了一套利用静电探针诊断技术测量减压直流非转移弧等离子体射流速度的实验系统。对以纯氩为工质的等离子体,在气流量1．25×10^-1kg/s、弧电流80A、真空室压力165Pa的条件下,测量了射流的速度及其分布。结果表明射流在发生器出口处中心最高速度约为1200m／s,在半径20mm处减小到635m／s。沿射流轴线方向的速度梯度约为10（ms^-1）／mm。射流速度随着弧电流增加而缓慢单调增加;当真空室压力从165Pa提高到2kPa时,发生器出口轴线上的射流速度从1200m／s降至570m／s。     

Controlled environments alter nutrient content of soybeans.

Information about compositional changes in plants grown in controlled environments is essential for developing a safe, nutritious diet for a Controlled Ecomological Life-Support System (CELSS). Information now is available for some CELSS candidate crops, but detailed information has been lacking for soybeans. To determine the effect of environment on macronutrient and mineral composition of soybeans, plants were grown both in the field and in a controlled environment where the hydroponic nutrient solution, photosynthetic flux (PPF), and CO2 level were manipulated to achieve rapid growth rates. Plants were harvested at seed maturity, separated into discrete parts, and oven dried prior to chemical analysis. Plant material was analyzed for proximate composition (moisture, protein, lipid, ash, and carbohydrate), total nitrogen (N), nonprotein N (NPN), nitrate, minerals, amino acid composition, and total dietary fiber. The effect of environment on composition varied by cultivar and plant part. Chamber-grown plants generally exhibited the following characteristics compared with field-grown plants: 1) increased total N and protein N for all plant parts, 2) increased nitrate in leaves and stems but not in seeds, 3) increased lipids in seeds, and 4) decreased Ca:P ratio for stems, pods, and leaves. These trends are consistent with data for other CELSS crops. Total N, protein N, and amino acid contents for 350 ppm CO2 and 1000 ppm CO2 were similar for seeds, but protein N and amino acid contents for leaves were higher at 350 ppm CO2 than at 1000 ppm CO2. Total dietary fiber content of soybean leaves was higher with 350 ppm CO2 than with 1000 ppm CO2. Such data will help in selecting of crop species, cultivars, and growing conditions to ensure safe, nutritious diets for CELSS.     

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.     

Atmospheric dynamics in the "Laboratory Biosphere" with wheat and sweet potato crops.

Laboratory Biosphere is a 40-m3 closed life system equipped with 12,000 W of high pressure sodium lamps over planting beds with 5.37 m2 of soil. Atmospheric composition changes due to photosynthetic fixation of carbon dioxide and corresponding production of oxygen or the reverse, respiration, are observed in short timeframes, e.g., hourly. To focus on inherent characteristics of the crop as distinct from its area or the volume of the chamber, we report fixation and respiration rates in mmol h-1 m-2 of planted area. An 85-day crop of USU Apogee wheat under a 16-h lighted/8-h dark regime peaked in fixation rate at about 100 mmol h-1 m-2 approximately 24 days after planting. Light intensity was about 840 micromoles m-2 s-1. Dark respiration peaked at about 31 mmol h-1 m-2 at the same time. Thereafter, both fixation and respiration declined toward zero as harvest time approached. A residual soil respiration rate of about 1.9 mmol h-1 m-2 was observed in the dark closed chamber for 100 days after the harvest. A 126-day crop of Tuskegee TU-82-155 sweet potato behaved quite differently. Under a 680 micromoles m-2 s-1, 18-h lighted/6-h dark regime, fixation during lighted hours rose to a plateau ranging from about 27 to 48 mmol h-1 m-2 after 42 days and dark respiration settled into a range of 12-23 mmol h-1 m-2. These rates continued unabated until the harvest at 126 days, suggesting that tuber biomass production might have continued at about the same rate for some time beyond the harvest time that was exercised in this experiment. In both experiments CO2 levels were allowed to range widely from a few hundred to about 3000 ppm, which permitted observation of fixation rates both at varying CO2 concentrations and at each number of days after planting. This enables plotting the fixation rate as a function of both variables. Understanding the atmospheric dynamics of individual crops will be essential for design and atmospheric management of more complex CELSS which integrate the simultaneous growth of several crops as in a sustainable remote life support system.     

Sunlight supply and gas exchange systems in the microalgal bioreactor.

The bioreactor with sunlight supply system and gas exchange systems presented here has proved feasible in ground tests and shows much promise for space use as a CELSS device. Our chief conclusions concerning the specification of total system needed for a life support system for a man in a space station are the following. (1) Sunlight supply system: compactness and low electrical consumption. (2) Bioreactor system: high density and growth rate of chlorella. (3) Gas exchange system: enough for O2 production and CO2 assimilation.     

Fundamental study on gas monitoring in CELSS.

A mass spectrometer and computer system was developed for conducting a fundamental study on gas monitoring in CELSS. Respiration and metabolism of the hamster and photosynthesis of the Spirulina were measured in a combination system consisting of a hamster chamber and a Spirulina cultivator. They are connected through a membrane gas exchanger. Some technical problems were examined. In the mass spectrometric gas monitoring, a simultaneous multi-sample measurement was developed by employing a rotating exchange valve. Long term precise measurement was obtained by employing an automatic calibration system. The membrane gas sampling probe proved to be useful for long term measurement. The cultivation rate of the Spirulina was effectively changed by controlling CO2 and light supply. The experimental results are helpful for improving the hamster-spirulina system.