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Plant adaptation to low atmospheric pressures: potential molecular responses.

There is an increasing realization that it may be impossible to attain Earth normal atmospheric pressures in orbital, lunar, or Martian greenhouses, simply because the construction materials do not exist to meet the extraordinary constraints imposed by balancing high engineering requirements against high lift costs. This equation essentially dictates that NASA have in place the capability to grow plants at reduced atmospheric pressure. Yet current understanding of plant growth at low pressures is limited to just a few experiments and relatively rudimentary assessments of plant vigor and growth. The tools now exist, however, to make rapid progress toward understanding the fundamental nature of plant responses and adaptations to low pressures, and to develop strategies for mitigating detrimental effects by engineering the growth conditions or by engineering the plants themselves. The genomes of rice and the model plant Arabidopsis thaliana have recently been sequenced in their entirety, and public sector and commercial DNA chips are becoming available such that thousands of genes can be assayed at once. A fundamental understanding of plant responses and adaptation to low pressures can now be approached and translated into procedures and engineering considerations to enhance plant growth at low atmospheric pressures. In anticipation of such studies, we present here the background arguments supporting these contentions, as well as informed speculation about the kinds of molecular physiological responses that might be expected of plants in low-pressure environments.

Acclimatization↗

Light-emitting diodes as an illumination source for plants: a review of research at Kennedy Space Center.

The provision of sufficient light is a fundamental requirement to support long-term plant growth in space. Several types of electric lamps have been tested to provide radiant energy for plants in this regard, including fluorescent, high-pressure sodium, and metal halide lamps. These lamps vary in terms of spectral quality, which can result in differences in plant growth and morphology. Current lighting research for space-based plant culture is focused on innovative lighting technologies that demonstrate high electrical efficiency and reduced mass and volume. Among the lighting technologies considered for space are light-emitting diodes (LEDs). The combination of red and blue LEDs has proven to be an effective lighting source for several crops, yet the appearance of plants under red and blue lighting is purplish gray, making visual assessment of plant health difficult. Additional green light would make the plant leaves appear green and normal, similar to a natural setting under white light, and may also offer psychological benefits for the crew. The addition of 24% green light (500-600 nm) to red and blue LEDs enhanced the growth of lettuce plants compared with plants grown under cool white fluorescent lamps. Coincidentally, these plants grown under additional green light would have the additional aesthetic appeal of a green appearance.

Biomass↗

Green-light supplementation for enhanced lettuce growth under red- and blue-light-emitting diodes.

Plants will be an important component of future long-term space missions. Lighting systems for growing plants will need to be lightweight, reliable, and durable, and light-emitting diodes (LEDs) have these characteristics. Previous studies demonstrated that the combination of red and blue light was an effective light source for several crops. Yet the appearance of plants under red and blue lighting is purplish gray making visual assessment of any problems difficult. The addition of green light would make the plant leave appear green and normal similar to a natural setting under white light and may also offer a psychological benefit to the crew. Green supplemental lighting could also offer benefits, since green light can better penetrate the plant canopy and potentially increase plant growth by increasing photosynthesis from the leaves in the lower canopy. In this study, four light sources were tested: 1) red and blue LEDs (RB), 2) red and blue LEDs with green fluorescent lamps (RGB), 3) green fluorescent lamps (GF), and 4) cool-white fluorescent lamps (CWF), that provided 0%, 24%, 86%, and 51% of the total PPF in the green region of the spectrum, respectively. The addition of 24% green light (500 to 600 nm) to red and blue LEDs (RGB treatment) enhanced plant growth. The RGB treatment plants produced more biomass than the plants grown under the cool-white fluorescent lamps (CWF treatment), a commonly tested light source used as a broad-spectrum control.

Biomass↗

Composition and physical properties of starch in microgravity-grown plants.

The effect of spaceflight on starch development in soybean (Glycine max L., BRIC-03) and potato (Solanum tuberosum, Astroculture-05) was compared with ground controls by biophysical and biochemical measurements. Starch grains from plants from both flights were on average 20-50% smaller in diameter than ground controls. The ratio delta X/delta rho (delta X --difference of magnetic susceptibilities, delta rho--difference of densities between starch and water) of starch grains was ca. 15% and 4% higher for space-grown soybean cotyledons and potato tubers, respectively, than in corresponding ground controls. Since the densities of particles were similar for all samples (1.36 to 1.38 g/cm3), the observed difference in delta X/delta rho was due to different magnetic susceptibilities and indicates modified composition of starch grains. In starch preparations from soybean cotyledons (BRIC-03) subjected to controlled enzymatic degradation with alpha-amylase for 24 hours, 77 +/- 6% of the starch from the flight cotyledons was degraded compared to 58 +/- 12% in ground controls. The amylose content in starch was also higher in space-grown tissues. The good correlation between the amylose content and delta X/delta rho suggests, that the magnetic susceptibility of starch grains is related to their amylose content. Since the seedlings from the BRIC-03 experiment showed elevated post-flight ethylene levels, material from another flight experiment (GENEX) which had normal levels of ethylene was examined and showed no difference to ground controls in size distribution, density, delta X/delta rho and amylose content. Therefore the role of ethylene appears to be more important for changes in starch metabolism than microgravity.

Amylopectin↗

Balancing photosynthetic light-harvesting and light-utilization capacities in potato leaf tissue during acclimation to different growth temperatures.

We investigated the effect of temperature during growth and development on the relationship between light-harvesting capacity, indicated by chlorophyll concentration, and light-utilization potential, indicated by light- and bicarbonate-saturated photosynthetic oxygen evolution, in Solanum tuberosum L. cv. Norland. Clonal plantlets were transplanted and grown at 20 degrees C for 2 weeks before transfer to 12, 16, 20, 24 and 28 degrees C for 6 weeks. After 4 weeks of the temperature treatments, leaf tissue fresh weights per area were one-third higher in plants grown at 12 degrees C vs those grown at 28 degrees C. Conversely, chlorophyll content per area in tissue grown at 12 degrees C was less than one-half of that of tissue grown at 28 degrees C at 4 weeks. Photosynthetic capacity measured at a common temperature of 20 degrees C and expressed on a chlorophyll basis was inversely proportional to growth temperature. Leaf tissue from plants grown at 12 degrees C for 4 weeks had photosynthetic rates that were 3-fold higher on a chlorophyll basis than comparable tissue from plants grown at 28 degrees C. These results suggest that the relationship between light-harvesting capacity and light-utilization potential varies 3-fold in response to the growth temperatures examined. The role of this response in avoidance of photoinhibition is discussed.

Acclimatization↗

An approach to crop modeling with the energy cascade.

Use of plants in advanced life support requires models of crop growth to analyze data, to evaluate areas for improvement, and, for design and engineering, to predict the gas exchanges of crops. We used data from experiments at Utah State University and the Kennedy Space Center for wheat (Triticum aestivum L.) and examined it for time dependence of the major three components in the energy cascade: photosynthetic photon absorption, canopy quantum yield, and carbon use efficiency. From the Utah State data, we developed a model with a total of five trends: absorption increasing until canopy closure, then constant; quantum yield as constant, then decreasing during senescence; carbon use as constant. This system probably is the lower limit of simplicity to which a model can be reduced and yet provide substantial utility. We demonstrated this utility by using the model to predict photosynthesis and respiration for experiments at Kennedy Space Center. The most uncertainty arose in predicting a start time for the senescent decrease of canopy quantum yield. The model should be generally applicable to other crops grown in controlled environments, as a generic tool for the design of life support systems.

Biomass↗

Vulnerability to dysfunction and muscle injury after unloading.

OBJECTIVE: To test whether unloading increases vulnerability to eccentric exercise-induced dysfunction and muscle injury. DESIGN: Before-after trial. SETTING: General community. PATIENTS OR OTHER PARTICIPANTS: Two women and 5 men (73 +/- 3kg [mean +/- SE]) who were active college students but were not trained in lower body resistance exercise volunteered. INTERVENTION: Five weeks of unilateral lower limb suspension (ULLS), which has been shown to decrease strength and size of the unloaded, left, but not load-bearing, right quadriceps femoris muscle group (QF) by 20% and 14%, respectively; performance of 10 sets of ten eccentric actions with each QF immediately after the ULLS strength tests with a load equivalent to 65% of the post-ULLS eccentric 1-repetition maximum. MAIN OUTCOME MEASURE(S): Concentric and eccentric 1-repetition maximum for the left, unloaded and the right, load-bearing QF measured immediately after ULLS and 1,4,7,9, and 11 days later; cross-sectional area and spin-spin relaxation time (T2) of each QF as determined by magnetic resonance imaging and measured the last day of ULLS and 3 days later. RESULTS: The mean load used for eccentric exercise was 23 +/- 2 and 30 +/- 3kg for the left, unloaded and right, load-bearing QF, respectively. The concentric and eccentric 1-repetition maximum for the unloaded and already weakened left QF was further decreased by 18% (p = .000) and 27% (p = .000), respectively, 1 day after eccentric exercise. Strength did not return to post-ULLS levels until 7 days of recovery. The right, load-bearing QF showed a 4% decrease (p = .002) in the eccentric 1-repetition maximum 1 day after eccentric exercise. The left, unloaded QF showed an increase in T2 (p = .002) in 18% of its cross-sectional area 3 days after the eccentric exercise, thus indicating muscle injury. The right, load-bearing QF showed no elevation in T2 (p = .280). CONCLUSION: Unloading increases vulnerability to eccentric exercise-induced dysfunction and muscle injury, even at relatively light loads.

Adult↗

Carbon dioxide effects on potato growth under different photoperiods and irradiance.

Carbon dioxide concentration can exert a strong influence on plant growth, but this influence can vary depending on irradiance. To study this, potato plants (Solanum tuberosum L.) cultivars Norland', Russet Burbank', and Denali' were grown in controlled-environment rooms at different levels of CO2 and irradiance. Carbon dioxide levels were maintained either at 350 or 1000 micromoles mol-1 and applied in combination with 12- or 24-h photoperiods at 400 or 800 micromoles m-2 s-1 photosynthetic photon flux. Air temperatures and relative humidity were held constant at 16 degrees C and 70%, respectively, and plants were harvested 90 d after planting. When averaged across all cultivars, CO2 enrichment increased tuber yield and total plant dry weight by 39 and 34%, respectively, under a 12-h photoperiod at 400 micromoles m-2 s-1; 27 and 19% under 12 h at 800 micromoles m-2 s-1; 9 and 9% under 24h at 400 micromoles m-2 s-1. It decreased dry weights by 9 and 9% under 24 h at 800 micromoles m-2 s-1. Tuber yield of Denali showed the greatest increase (21%) in response to increased CO2 across all irradiance treatments, while tuber yields of Russet Burbank and Norland were increased 18 and 9%, respectively. The results show a pattern of greater plant growth from CO2 enrichment under lower PPF and a short photoperiod.

Carbon Dioxide↗

Accuracy of quantum sensors measuring yield photon flux and photosynthetic photon flux.

Photosynthesis is fundamentally driven by photon flux rather than energy flux, but not all absorbed photons yield equal amounts of photosynthesis. Thus, two measures of photosynthetically active radiation have emerged: photosynthetic photon flux (PPF), which values all photons from 400 to 700 nm equally, and yield photon flux (YPF), which weights photons in the range from 360 to 760 nm according to plant photosynthetic response. We selected seven common radiation sources and measured YPF and PPF from each source with a spectroradiometer. We then compared these measurements with measurements from three quantum sensors designed to measure YPF, and from six quantum sensors designed to measure PPF. There were few differences among sensors within a group (usually <5%), but YPF values from sensors were consistently lower (3% to 20%) than YPF values calculated from spectroradiometric measurements. Quantum sensor measurements of PPF also were consistently lower than PPF values calculated from spectroradiometric measurements, but the differences were <7% for all sources, except red-light-emitting diodes. The sensors were most accurate for broad-band sources and least accurate for narrow-band sources. According to spectroradiometric measurements, YPF sensors were significantly less accurate (>9% difference) than PPF sensors under metal halide, high-pressure sodium, and low-pressure sodium lamps. Both sensor types were inaccurate (>18% error) under red-light-emitting diodes. Because both YPF and PPF sensors are imperfect integrators, and because spectroradiometers can measure photosynthetically active radiation much more accurately, researchers should consider developing calibration factors from spectroradiometric data for some specific radiation sources to improve the accuracy of integrating sensors.

Evaluation Studies as Topic↗

Effect of CO2 levels on nutrient content of lettuce and radish.

Atmospheric carbon-dioxide enrichment is known to affect the yield of lettuce and radish grown in controlled environments, but little is known about CO2 enrichment effects on the chemical composition of lettuce and radish. These crops are useful model systems for a Controlled Ecological Life-Support System (CELSS), largely because of their relatively short production cycles. Lettuce (Lactuca sativa L.) cultivar 'Waldmann's Green' and radish (Raphanus sativus L.) cultivar 'Giant White Globe' were grown both in the field and in controlled environments, where hydroponic nutrient solution, light, and temperature were regulated, and where CO2 levels were controlled at 400, 1000, 5000, or 10,000 ppm. Plants were harvested at maturity, dried, and analyzed for proximate composition (protein, fat, ash, and carbohydrate), total nitrogen (N), nitrate N, free sugars, starch, total dietary fiber, and minerals. Total N, protein N, nonprotein N (NPN), and nitrate N generally increased for radish roots and lettuce leaves when grown under growth chamber conditions compared to field conditions. The nitrate-N level of lettuce leaves, as a percentage of total NPN, decreased with increasing levels of CO2 enrichment. The ash content of radish roots and of radish and lettuce leaves decreased with increasing levels of CO2 enrichment. The levels of certain minerals differed between field- and chamber-grown materials, including changes in the calcium (Ca) and phosphorus (P) contents of radish and lettuce leaves, resulting in reduced Ca/P ratio for chamber-grown materials. The free-sugar contents were similar between the field and chamber-grown lettuce leaves, but total dietary fiber content was much higher in the field-grown plant material. The starch content of growth-chamber lettuce increased with CO2 level.

Atmosphere↗

Germination and elongation of flax in microgravity.

This experiment was conducted as part of a risk mitigation payload aboard the Space Shuttle Atlantis on STS-101. The objectives were to test a newly developed water delivery system, and to determine the optimal combination of water volume and substrate for the imbibition and germination of flax (Linum usitatissimum) seeds in space. Two different combinations of germination paper were tested for their ability to absorb, distribute, and retain water in microgravity. A single layer of thick germination paper was compared with one layer of thin germination paper under a layer of thick paper. Paper strips were cut to fit snugly into seed cassettes, and seeds were glued to them with the micropyle ends pointing outward. Water was delivered in small increments that traveled through the paper via capillary action. Three water delivery volumes were tested, with the largest (480 microliters) outperforming the 400 microliters and 320 microliters volumes for percent germination (90.6%) and root growth (mean=4.1 mm) during the 34-hour spaceflight experiment. The ground control experiment yielded similar results, but with lower rates of germination (84.4%) and shorter root lengths (mean=2.8 mm). It is not clear if the roots emerged more quickly in microgravity and/or grew faster than the ground controls. The single layer of thick germination paper generally exhibited better overall growth than the two layered option. Significant seed position effects were observed in both the flight and ground control experiments. Overall, the design of the water delivery system, seed cassettes and the germination paper strip concept was validated as an effective method for promoting seed germination and root growth under microgravity conditions.

Equipment Design↗

Skeletal muscle myosin heavy chain composition and resistance training.

We recently reported that 19 wk of heavy resistance training caused a decrease in the percentage of type IIb and an increase in the percentage of type IIa fibers as determined by qualitative histochemical analyses of myofibrillar adenosinetriphosphatase activity of biopsies of musculus vastus lateralis (Hather et al. Acta Physiol. Scand. 143: 177-185, 1991). These data were interpreted to suggest that resistance training had caused transformation among the fast-twitch fiber subtypes. To more clearly establish the influence of resistance training on muscle fiber composition, biopsies from the original study were analyzed biochemically for myosin heavy chain (MHC) composition by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and histochemically for fiber types by use of myofibrillar adenosinetriphosphatase activity. The results show that after training (n = 13), IIb MHC composition decreased (P < 0.05) from 19 +/- 4 to 7 +/- 1%. IIa MHC, in contrast, increased (P < 0.05) from 48 +/- 3 to 60 +/- 2%. These responses were essentially mirrored by alterations in fiber type distribution. The percentage of type IIb fibers decreased (P < 0.05) from 18 +/- 3 to 1 +/- 1%, whereas the percentage of type IIa fibers increased from 46 +/- 4 to 60 +/- 3% (P < 0.05). Neither I MHC composition nor type I fiber percentage changed with training. The control group (n = 4) showed no changes in MHC composition or fiber type distribution. These results suggest that heavy resistance training alters MHC composition in human skeletal muscle, presumably reflecting a change in genetic expression.

Adenosine Triphosphatases↗

Excess nutrients in hydroponic solutions alter nutrient content of rice, wheat, and potato.

Environment has significant effects on the nutrient content of field-grown crop plants. Little is known, however, about compositional changes caused by controlled environments in which plants receive only artificial radiation and soilless, hydroponic culture. This knowledge is essential for developing a safe, nutritious diet in a Controlled Ecological Life-Support System (CELSS). Three crops that are candidates for inclusion in a CELSS (rice, wheat, and white potato) were grown both in the field and in controlled environments where the hydroponic nutrient solution, photosynthetic photon flux (PPF), and CO2 level were manipulated to achieve rapid growth rates. Plants were harvested at maturity, separated into discrete parts, and dried prior to analysis. Plant materials were analyzed for proximate composition (protein, fat, ash, and carbohydrate), total nitrogen (N), nitrate, minerals, and amino-acid composition. The effect of environment on nutrient content varied by crop and plant part. Total N and nonprotein N (NPN) contents of plant biomass generally increased under controlled-environment conditions compared to field conditions, especially for leafy plant parts and roots. Nitrate levels were increased in hydroponically-grown vegetative tissues, but nitrate was excluded from grains and tubers. Mineral content changes in plant tissue included increased phosphorus and decreased levels of certain micronutrient elements under controlled-environment conditions. These findings suggest that cultivar selection, genetic manipulation, and environmental control could be important to obtain highly nutritious biomass in a CELSS.

Amino Acids↗

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↗

Computer modeling for advanced life support system analysis.

This article discusses the equivalent mass approach to advanced life support system analysis, describes a computer model developed to use this approach, and presents early results from modeling the NASA JSC BioPlex. The model is built using an object oriented approach and G2, a commercially available modeling package Cost factor equivalencies are given for the Volosin scenarios. Plant data from NASA KSC and Utah State University (USU) are used, together with configuration data from the BioPlex design effort. Initial results focus on the importance of obtaining high plant productivity with a flight-like configuration.

Atmosphere↗

Effects of bioreactor retention time on aerobic microbial decomposition of CELSS crop residues.

The focus of resource recovery research at the KSC-CELSS Breadboard Project has been the evaluation of microbiologically mediated biodegradation of crop residues by manipulation of bioreactor process and environmental variables. We will present results from over 3 years of studies that used laboratory- and breadboard-scale (8 and 120 L working volumes, respectively) aerobic, fed-batch, continuous stirred tank reactors (CSTR) for recovery of carbon and minerals from breadboard grown wheat and white potato residues. The paper will focus on the effects of a key process variable--bioreactor retention time--on response variables indicative of bioreactor performance. The goal is to determine the shortest retention time that is feasible for processing CELSS crop residues, thereby reducing bioreactor volume and weight requirements. Pushing the lower limits of bioreactor retention times will provide useful data for engineers who need to compare biological and physicochemical components. Bioreactor retention times were manipulated to range between 0.25 and 48 days. Results indicate that increases in retention time lead to a 4-fold increase in crop residue biodegradation, as measured by both dry weight losses and CO2 production. A similar overall trend was also observed for crop residue fiber (cellulose and hemicellulose), with a noticeable jump in cellulose degradation between the 5.3 day and 10.7 day retention times. Water-soluble organic compounds (measured as soluble TOC) were appreciably reduced by more than 4-fold at all retention times tested. Results from a study of even shorter retention times (down to 0.25 days), in progress, will also be presented.

Aerobiosis↗

Effects of CO2 on stomatal conductance: do stomata open at very high CO2 concentrations?

Potato and wheat plants were grown for 50 d at 400, 1000 and 10000 micromoles mol-1 carbon dioxide (CO2). and sweetpotato and soybean were grown at 1000 micromoles mol-1 CO2 in controlled environment chambers to study stomatal conductance and plant water use. Lighting was provided with fluorescent lamps as a 12 h photoperiod with 300 micromoles m-2 s-1 PAR. Mid-day stomatal conductances for potato were greatest at 400 and 10000 micromoles mol-1 and least at 1000 micromoles mol-1 CO2. Mid-day conductances for wheat were greatest at 400 micromoles mol-1 and least at 1000 and 10000 micromoles mol-1 CO2. Mid-dark period conductances for potato were significantly greater at 10000 micromoles mol-1 than at 400 or 1000 micromoles mol-1, whereas dark conductance for wheat was similar in all CO2 treatments. Temporarily changing the CO2 concentration from the native 1000 micromoles mol-1 to 400 micromoles mol-1 increased mid-day conductance for all species, while temporarily changing from 1000 to 10000 micromoles mol-1 also increased conductance for potato and sweetpotato. Temporarily changing the dark period CO2 from 1000 to 10000 micromoles mol-1 increased conductance for potato, soybean and sweetpotato. In all cases, the stomatal responses were reversible, i.e. conductances returned to original rates following temporary changes in CO2 concentration. Canopy water use for potato was greatest at 10000, intermediate at 400, and least at 1000 micromoles mol-1 CO2, whereas canopy water use for wheat was greatest at 400 and similar at 1000 and 10000 micromoles mol-1 CO2. Elevated CO2 treatments (i.e. 1000 and 10000 micromoles mol-1) resulted in increased plant biomass for both wheat and potato relative to 400 micromoles mol-1, and no injurious effects were apparent from the 10000 micromoles mol-1 treatment. Results indicate that super-elevated CO2 (i.e. 10000 micromoles mol-1) can increase stomatal conductance in some species, particularly during the dark period, resulting in increased water use and decreased water use efficiency.

Biomass↗

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↗