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Soybean cotyledon starch metabolism is sensitive to altered gravity conditions.

We have demonstrated that etiolated soybean seedlings grown under the altered gravity conditions of clinorotation (1 rpm) and centrifugation (5xg) exhibit changes in starch metabolism. Cotyledon starch concentration was lower (-28%) in clinorotated plants and higher (+24%) in centrifuged plants than in vertical control plants. The activity of ADP-glucose pyrophosphorylase in the cotyledons was affected in a similar way, i.e. lower (-37%) in the clinorotated plants and higher (+22%) in the centrifuged plants. Other starch metabolic enzyme activities, starch synthase, starch phosphorylase and total hydrolase were not affected by the altered gravity treatments. We conclude that the observed changes in starch concentrations were primarily due to gravity-mediated differences in ADP-glucose pyrophosphorylase activity.

Centrifugation↗

Hydrotropism in pea roots in a porous-tube water delivery system.

Orientation of root growth on earth and under microgravity conditions can possibly be controlled by hydrotropism--growth toward a moisture source in the absence of or reduced gravitropism. A porous-tube water delivery system being used for plant growth studies is appropriate for testing this hypothesis since roots can be grown aeroponically in this system. When the roots of the agravitropic mutant pea ageotropum (Pisum sativum L.) were placed vertically in air of 91% relative humidity and 2 to 3 mm from the water-saturated porous tube placed horizontally, the roots responded hydrotropically and grew in a continuous arch along the circular surface of the tube. By contrast, normal gravitropic roots of Alaska' pea initially showed a slight transient curvature toward the tube and then resumed vertical downward growth due to gravitropism. Thus, in microgravity, normal gravitropic roots could respond to a moisture gradient as strongly as the agravitropic roots used in this study. Hydrotropism should be considered a significant factor responsible for orientation of root growth in microgravity.

Equipment Design↗

Light spectral quality effects on the growth of potato (Solanum tuberosum L.) nodal cuttings in vitro.

The effect of light spectral quality on the growth of in vitro nodal cuttings of potato (Solanum tuberosum L.) cultivars Norland, Superior, Kennebec, and Denali were examined. The different light spectra were provided by Vita-Lite fluorescent (VF) (a white light control), blue fluorescent (BF), red fluorescent (RF), low-pressure sodium (LPS), and a combination of low-pressure sodium plus cool-white fluorescent lamps (LPS/CWF). For all cultivars, stem lengths after 4 wk were longest under LPS, followed by RF, LPS/CWF, VF, and BF (in descending order). Microscopic studies revealed that cells were shortest when cultured in BF or VF environments, and were longest in RF or LPS lamp environments. The highest number of axillary branches occurred on plantlets grown with LPS or LPS/CWF, whereas the lowest number occurred with BF. No leaf or stem edema (callus or gall-like growths) occurred with LPS or LPS/CWF lighting, and no edema occurred on cv. Norland plantlets, regardless of lighting. Results suggest that shoot morphologic development of in vitro grown potato plants can be controlled by controlling irradiant spectral quality.

Cells, Cultured↗

Potato growth and yield using nutrient film technique (NFT).

Potato plants, cvs Denali and Norland, were grown in polyvinyl chloride (PVC) trays using a continuous flowing nutrient film technique (NFT) to study tuber yield for NASA's Controlled Ecological Life Support Systems (CELSS) program. Nutrient solution pH was controlled automatically using 0.39M (2.5% (v/v) nitric acid (HNO3), while water and nutrients were replenished manually each day and twice each week, respectively. Plants were spaced either one or two per tray, allotting 0.2 or 0.4 m2 per plant. All plants were harvested after 112 days. Denali plants yielded 2850 and 2800 g tuber fresh weight from the one- and two-plant trays, respectively, while Norland plants yielded 1800 and 2400 g tuber fresh weight from the one- and two-plant trays. Many tubers of both cultivars showed injury to the periderm tissue, possibly caused by salt accumulation from the nutrient solution on the surface. Total system water usage throughout the study for all the plants equaled 709 liters (L), or approximately 2 L m-2 d-1. Total system acid usage throughout the study (for nutrient solution pH control) equaled 6.60 L, or 18.4 ml m-2 d-1 (7.2 mmol m-2 d-1). The results demonstrate that continuous flowing nutrient film technique can be used for tuber production with acceptable yields for the CELSS program.

Culture Media↗

Effect of crop development on biogenic emissions from plant populations grown in closed plant growth chambers.

The Biomass Production Chamber at John F. Kennedy Space Center is a closed plant growth chamber facility that can be used to monitor the level of biogenic emissions from large populations of plants throughout their entire growth cycle. The head space atmosphere of a 26-day-old lettuce (Lactuca sativa cv. Waldmann's Green) stand was repeatedly sampled and emissions identified and quantified using GC-mass spectrometry. Concentrations of dimethyl sulphide, carbon disulphide, alpha-pinene, furan and 2-methylfuran were not significantly different throughout the day; whereas, isoprene showed significant differences in concentration between samples collected in light and dark periods. Volatile organic compounds from the atmosphere of wheat (Triticum aestivum cv. Yecora Rojo) were analysed and quantified from planting to maturity. Volatile plant-derived compounds included 1-butanol, 2-ethyl-1-hexanol, nonanal, benzaldehyde, tetramethylurea, tetramethylthiourea, 2-methylfuran and 3-methylfuran. Concentrations of volatiles were determined during seedling establishment, vegetative growth, anthesis, grain fill and senescence and found to vary depending on the developmental stage. Atmospheric concentrations of benzaldehyde and nonanal were highest during anthesis, 2-methylfuran and 3-methylfuran concentrations were greatest during grain fill, and the concentration of the tetramethylurea peaked during senescence.

Biomass↗

A review of recent activities in the NASA CELSS program.

A CELSS (Controlled Ecological Life Support System) is a device that utilizes photosynthetic organisms and light energy to regenerate waste materials into oxygen and food for a crew in space. The results of theoretical and practical studies conducted by investigators within the CELSS program suggest that a bioregenerative life support system can be a useful and effective method of regenerating consumable materials for crew sustenance. Experimental data suggests that the operation of a CELSS in space will be practical if plants can be made to behave predictably in the space environment. Much of the work currently conducted within the CELSS program centers on the biological components of the CELSS system. The work is particularly directed at ways of achieving high efficiency and long term stability of all components of the system. Included are explorations of the conversion of non-edible cellulose to edible materials, nitrogen fixation by biological and chemical methods, and methods of waste processing. It is the intent of the presentation to provide a description of the extent to which a bioregenerative life support system can meet the constraints of the space environment, and to assess the degree to which system efficiency and stability can be increased during the next decade.

Cosmic Radiation↗

The Breadboard Project: a functioning CELSS plant growth system.

The primary objective of the Breadboard project for the next 3-4 years is to develop, integrate and operate a Controlled Ecological Life Support System (CELSS) at a one person scale. The focus of this project over the past two years has been the development of the plant growth facility, the first module of the CELSS. The other major modules, food preparation, biomass processing, and resource recovery, have been researched at the laboratory scale during the past two years and facilities are currently under construction to scale-up these modules to an operational state. This paper will outline the design requirements for the Biomass Production Chamber (BPC), the plant growth facility for the project, and the control and monitoring subsystems which operate the chamber and will present results from both engineering and biological tests of the facility. Three production evaluations of wheat, conducted in the BPC during the past year, will be described and the data generated from these tests discussed. Future plans for the BPC will be presented along with future goals for the project as the other modules become active.

Biomass↗

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.

Atmosphere↗

Application of sunlight and lamps for plant irradiation in space bases.

The radiation sources used for plant growth on a space base must meet the biological requirements for photosynthesis and photomorphogenesis. In addition the sources must be energy and volume efficient, while maintaining the required irradiance levels, spectral, spatial and temporal distribution. These requirements are not easily met, but as the biological and mission requirements are better defined, then specific facility designs can begin to accommodate both the biological requirements and the physical limitations of a space based plant growth system.

Ecological Systems, Closed↗

Proximate composition of CELSS crops grown in NASA's Biomass Production Chamber.

Edible biomass from four crops of wheat (Triticum aestivum L.), four crops of lettuce (Lactuca sativa L.), four crops of potato (Solanum tuberosum L.), and three crops of soybean (Glycine max (L.) Merr.) grown in NASA's CELSS Biomass Production Chamber were analyzed for proximate composition. All plants were grown using recirculating nutrient (hydroponic) film culture with pH and electrical conductivity automatically controlled. Temperature and humidity were controlled to near optimal levels for each species and atmospheric carbon dioxide partial pressures were maintained near 100 Pa during the light cycles. Soybean seed contained the highest percentage of protein and fat, potato tubers and wheat seed contained the highest levels of carbohydrate, and lettuce leaves contained the highest level of ash. Analyses showed values close to data published for field-grown plants with several exceptions: In comparison with field-grown plants, wheat seed had higher protein levels; soybean seed had higher ash and crude fiber levels; and potato tubers and lettuce leaves had higher protein and ash levels. The higher ash and protein levels may have been a result of the continuous supply of nutrients (e.g., potassium and nitrogen) to the plants by the recirculating hydroponic culture.

Biomass↗

Volatile organic compounds detected in the atmosphere of NASA's Biomass Production Chamber.

Atmospheres of enclosed environments in which 20 m2 stands of wheat, potato, and lettuce were grown were characterized and quantified by gas chromatography-mass spectrometry. A large number (in excess of 90) of volatile organic compounds (VOCs) were identified in the chambers. Twenty eight VOC's were assumed to be of biogenic origin for these were not found in the chamber atmosphere when air samples were analyzed in the absence of plants. Some of the compounds found were unique to a single crop. For example, only 35% of the biogenic compounds detected in the wheat atmosphere were unique to wheat, while 36% were unique to potato and 26% were unique to lettuce. The number of compounds detected in the wheat (20 compounds) atmosphere was greater than that of potato (11) and lettuce (15) and concentration levels of biogenic and non-biogenic VOC's were similar.

Air↗

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.

Air↗

NASA's Biomass Production Chamber: a testbed for bioregenerative life support studies.

The Biomass Production Chamber (BPC) located at Kennedy Space Center, FL, USA provides a large (20 m2 area, 113 m3 vol.), closed environment for crop growth tests for NASA's Controlled Ecological Life Support System (CELSS) program. Since the summer of 1988, the chamber has operated on a near-continuous basis (over 1200 days) without any major failures (excluding temporary power losses). During this time, five crops of wheat (64-86 days each), three crops of soybean (90 to 97 days), five crops of lettuce (28-30 days), and four crops of potato (90 to 105 days were grown, producing 481 kg of dry plant biomass, 196 kg edible biomass, 540 kg of oxygen, 94,700 kg of condensed water, and fixing 739 kg of carbon dioxide. Results indicate that total biomass yields were close to expected values for the given light input, but edible biomass yields and harvest indices were slightly lower than expected. Stand photosynthesis, respiration, transpiration, and nutrient uptake rates were monitored throughout growth and development of the different crops, along with the build-up of ethylene and other volatile organic compounds in the atmosphere. Data were also gathered on system hardware maintenance and repair, as well as person-hours required for chamber operation. Future tests will include long-term crop production studies, tests in which nutrients from waste treatment systems will be used to grow new crops, and multi-species tests.

Biomass↗

Optimization of moisture content for wheat seedling germination in a cellulose acetate medium for a space flight experiment.

The Porous Tube Plant Nutrient Delivery System (PTPNDS), a hydrophilic, microporous ceramic tube hydroponic system designed for microgravity, will be tested in a middeck locker of the Space Shuttle. The flight experiment will focus on hardware operation and assess its ability to support seed germination and early seedling growth in microgravity. The water controlling system of the PTPNDS hardware has been successfully tested during the parabolic flight of the KC-135. One challenge to the development of the space flight experiment was to devise a method of holding seeds to the cylindrical porous tube. The seed-holder must provide water and air to the seed, absorb water from the porous tube, withstand sterilization, provide a clear path for shoots and roots to emerge, and be composed of flight qualified materials. In preparation for the flight experiment, a wheat seed-holder has been designed that utilizes a cellulose acetate plug to facilitate imbibition and to hold the wheat seeds in contact with the porous tube in the correct orientation during the vibration of launch and the microgravity environment of orbit. Germination and growth studies with wheat at a range of temperatures showed that optimal moisture was 78% (by weight) in the cellulose acetate seed holders. These and other design considerations are discussed.

Cellulose↗

Supraoptimal carbon dioxide effects on growth of soybean [Glycine max (L.) Merr.].

In tightly closed environments used for human life support in space, carbon dioxide (CO2) partial pressures can reach 500 to 1000 Pa, which may be supraoptimal or toxic to plants used for life support. To study this, soybeans [Glycine max (L.) Merr. cvs. McCall and Pixie] were grown for 90 days at 50, 100, 200, and 500 Pa partial pressure CO2 (500, 1000, 2000, and 5000 ppm). Plants were grown using recirculating nutrient film technique with a 12-h photoperiod, a 26 degrees C/20 degrees C thermoperiod, and approximately 300 micromoles m-2 s-1 photosynthetic photon flux (PPF). Seed yield and total biomass were greatest at 100 Pa for cv. McCall, suggesting that higher CO2 levels were supraoptimal. Seed yield and total biomass for cv. Pixie showed little difference between CO2 treatments. Average stomatal conductance of upper canopy leaves at 50 Pa CO2 approximately 500 Pa > 200 Pa > 100 Pa. Total water use over 90 d for both cultivars (combined on one recirculating system) equalled 822 kg water for 100 Pa CO2, 845 kg for 50 Pa, 879 kg for 200 Pa, and 1194 kg for 500 Pa. Water use efficiences for both cultivars combined equalled 3.03 (g biomass kg-1 water) for 100 Pa CO2, 2.54 g kg-1 for 200 Pa, 2.42 g kg-1 for 50 Pa, and 1.91 g kg-1 for 500 Pa. The increased stomatal conductance and stand water use at the highest CO2 level (500 Pa) were unexpected and pose interesting considerations for managing plants in a tightly closed system where CO2 concentrations may reach high levels.

Biomass↗

A data base of crop nutrient use, water use, and carbon dioxide exchange in a 2O square meter growth chamber: I. Wheat as a case study.

A data set is given describing the daily nutrient uptake, gas exchange, environmental conditions, and carbon (C), and nutrient partitioning at harvest for the entire canopy and root system of a wheat crop (Triticum aestivum, cv. Yecora Rojo). The data were obtained from a 20 m2 stand of wheat plants grown from planting to maturity in a closed, controlled environment, and include daily nutrient uptake [macronutrients, nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sulfur (S); and micronutrients, iron (Fe), boron (B), manganese (Mn), zinc (Zn), copper (Cu), and molybdenum (Mo)], canopy carbon dioxide (CO2) exchange rates, and transpiration. Environmental factors such as relative humidity, air temperature, nutrient solution temperature, pH and electrical conductivity, and photoperiod were controlled in the chamber to specific set points. A detailed description of biomass yield for each of the 64 plant growth trays comprising the 20 m2 of growth area is also provided, and includes dry weights of grain, straw, chaff, and roots, along with the concentration of nutrients in different plant tissues and the percent carbohydrate, fat, and protein. To our knowledge, this information represents one of the most extensive data sets available for a canopy of wheat grown from seed to maturity under controlled environmental and nutritional conditions, and thus may provide useful information for model development and validation. A methods section is included to qualify any assumptions that might be required for the use of the data in plant growth models, along with a daily event calendar indicating when adjustments in set points and occasional equipment or sensor failures occurred.

Biomass↗

Water relations, gas exchange, and nutrient response to a long term constant water deficit.

Wheat plants (Triticum aestivum) were grown for 43 days in a micro-porous tube nutrient delivery system. Roots were unable to penetrate the microporous tube, but grew on the surface and maintained capillary contact with the nutrient solution on the inside of the tube through the 5-micron pores of the porous tube. Water potential in the system was controlled at -0.4, -0.8, and -3.0 kPa by adjusting the applied pressure (hydrostatic head) to the nutrient solution flowing through the microporous tubes. A relatively small decrease in applied water potential from -0.4 to -3.0 kPa resulted in a 34% reduction of shoot growth but only a moderate reduction in the midday leaf water potential from -1.3 to -1.7 MPa. Carbon dioxide assimilation decreased and water use efficiency increased with the more negative applied water potentials, while intercellular CO2 concentration remained constant. This was associated with a decrease in stomatal conductance to water vapor from 1.90 to 0.98 mol m-2 s-1 and a decrease in total apparent hydraulic conductance from 47 to 12 micromoles s-1 MPa-1. Although the applied water potentials were in the -0.4 to -3.0 kPa range, the actual water potential perceived by the plant roots appeared to be in the range of -0.26 to -0.38 MPa as estimated by the leaf water potential of bagged plants. The amount of K, Ca, Mg, Zn, Cu, and B accumulated with each unit of transpired water increased as the applied water potential became less negative. The increase in accumulation ranged from 1.4-fold for K to 2.2-fold for B. The physiological responses observed in this study in response to small constant differences in applied water potentials were much greater than expected from either the applied water potential or the observed plant water potential. Even though the micro-porous tube may not represent natural conditions and could possibly introduce morphological and physiological artifacts, it enables a high degree of control of water potential that facilitates the investigation of many aspects of water relations not practical with other experimental systems.

Boron↗

Photomorphogenesis and photoassimilation in soybean and sorghum grown under broad spectrum or blue-deficient light sources.

The role of blue light in plant growth and development was investigated in soybean (Glycine max [L.] Merr. cv Williams) and sorghum (Sorghum bicolor [L.] Moench. cv Rio) grown under equal photosynthetic photon fluxes (approximately 500 micromoles per square meter per second) from broad spectrum daylight fluorescent or blue-deficient, narrow-band (589 nanometers) low pressure sodium (LPS) lamps. Between 14 and 18 days after sowing, it was possible to relate adaptations in photosynthesis and leaf growth to dry matter accumulation. Soybean development under LPS light was similar in several respects to that of shaded plants, consistent with an important role for blue light photoreceptors in regulation of growth response to irradiance. Thus, soybeans from LPS conditions partitioned relatively more growth to leaves and maintained higher average leaf area ratios (mean LAR) that compensated lower net assimilation rates (mean NAR). Relative growth rates were therefore comparable to plants from daylight fluorescent lamps. Reductions in mean NAR were matched by lower rates of net photosynthesis (A) on an area basis in the major photosynthetic source (first trifoliolate) leaf. Lower A in soybean resulted from reduced leaf dry matter per unit leaf area, but lower A under LPS conditions in sorghum correlated with leaf chlorosis and reduced total nitrogen (not observed in soybean). In spite of a lower A, mean NAR was larger in sorghum from LPS conditions, resulting in significantly greater relative growth rates (mean LAR was approximately equal for both light conditions). Leaf starch accumulation rate was higher for both species and starch content at the end of the dark period was elevated two- and three-fold for sorghum and soybean, respectively, under LPS conditions. Possible relations between starch accumulation, leaf export, and plant growth in response to spectral quality were considered.

Carbohydrates↗