Effects of temperature, CO2/O2 concentrations and light intensity on cellular multiplication of microalgae, Euglena gracilis.

Microalgae culture is likely to play an important role in aquatic food production modules in bioregenerative systems for producing feeds for fish, converting CO2 to O2 and remedying water quality as well as aquatic higher plants. In the present study, the effects of culture conditions on the cellular multiplication of microalgae, Euglena gracilis, was investigated as a fundamental study to determine the optimum culture conditions for microalgae production in aquatic food production modules including both microalgae culture and fish culture systems. E. gracilis was cultured under conditions with five levels of temperatures (25-33 degrees C), three levels of CO2 concentrations (2-6%), five levels of O2 concentrations (10-30%), and six levels of photosynthetic photon flux (20-200 micromoles m-2 s-1). The number of Euglena cells in a certain volume of solution was monitored with a microscope under each environmental condition. The multiplication rate of the cells was highest at temperatures of 27-31 degrees C, CO2 concentration of 4%, O2 concentration of 20% and photosynthetic photon flux of about 100 micromoles m-2 s-1. The results demonstrate that E. gracilis could efficiently produce biomass and convert CO2 to O2 under relatively low light intensities in aquatic food production modules.     

Effects of air velocity on photosynthesis of plant canopies under elevated CO2 levels in a plant culture system.

To obtain basic data for adequate air circulation for promoting plant growth in closed plant production modules in bioregenerative life support systems in space, effects of air velocities ranging from 0.1 to 0.8 m s-1 on photosynthesis in tomato seedlings canopies were investigated under atmospheric CO2 concentrations of 0.4 and 0.8 mmol mol-1. The canopy of tomato seedlings on a plug tray (0.4 x 0.4 m2) was set in a wind-tunnel-type chamber (0.6 x 0.4 x 0.3 m3) installed in a semi-closed-type assimilation chamber (0.9 x 0.5 x 0.4 m3). The net photosynthetic rate in the plant canopy was determined with the differences in CO2 concentrations between the inlet and outlet of the assimilation chamber multiplied by the volumetric air exchange rate of the chamber. Photosynthetic photon flux (PPF) on the plant canopy was kept at 0.25 mmol m-2 s-1, air temperature at 23 degrees C and relative humidity at 55%. The leaf area indices (LAIs) of the plant canopies were 0.6-2.5 and plant heights were 0.05-0.2 m. The net photosynthetic rate of the plant canopy increased with increasing air velocities inside plant canopies and saturated at 0.2 m s-1. The net photosynthetic rate at the air velocity of 0.4 m s-1 was 1.3 times that at 0.1 m s-1 under CO2 concentrations of 0.4 and 0.8 mmol mol-1. The net photosynthetic rate under CO2 concentrations of 0.8 mmol mol-1 was 1.2 times that under 0.4 mmol mol-1 at the air velocity ranging from 0.1 to 0.8 m s-1. The results confirmed the importance of controlling air movement for enhancing the canopy photosynthesis under an elevated CO2 level as well as under a normal CO2 level in the closed plant production modules.     

The effect of gravity on surface temperature and net photosynthetic rate of plant leaves.

To clarify the effects of gravity on heat/gas exchange between plant leaves and the ambient air, the leaf temperatures and net photosynthetic rates of plant leaves were evaluated at 0.01, 1.0, 1.5 and 2.0 G of 20 seconds each during a parabolic airplane flight. Thermal images of leaves were captured using infrared thermography at an air temperature of 26 degrees C, a relative humidity of 15% and an irradiance of 260 W m-2. The net photosynthetic rates were determined by using a chamber method with an infrared gas analyzer at an air temperature of 20 degrees C, a relative humidity of 50% and a photosynthetic photon flux of 0.5 mmol m-2 s-1. The mean leaf temperature increased by 1 degree C and the net photosynthetic rate decreased by 13% with decreasing gravity levels from 1.0 to 0.01 G. The leaf temperature decreased by 0.5 degree C and the net photosynthetic rate increased by 7% with increasing gravity levels from 1.0 to 2.0 G. Heat/gas exchanges between leaves and the ambient air were more retarded at lower gravity levels. A restricted free air convection under microgravity conditions in space would limit plant growth by retarding heat and gas exchanges between leaves and the ambient air.     

Utilization of potatoes in bioregenerative life support systems.

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

Carbon dioxide interactions with irradiance and temperature in potatoes.

Separate controlled environment studies were conducted to determine the interaction of CO2 with irradiance and interaction of CO2 with temperature on growth of three potato cultivars. In the first study, an elevated CO2 concentration of 1000 micromoles mol-1 and an ambient CO2 of 350 micromoles mol-1 were maintained at the photosynthetic photon fluxes (PPF) of 17 and 34 mol m-2 d-1 with 12 h photoperiod, and at the PPF of 34 and 68 mol m-2 d-1 with 24 h photoperiod (400 and 800 micromoles m-2 s-1 PPF at each photoperiod). Tuber and total dry weights of 90-day old potatoes were significantly increased with CO2 enrichment, but the CO2 stimulation was less with higher PPF and longer photoperiod. Shoot dry weight was affected more by photoperiod than by PPF and CO2 concentrations. The elevated CO2 concentration increased leaf CO2 assimilation rates and decreased stomatal conductance with 12 h photoperiod, but had only a marginal effect with 24 h photoperiod. In the second study, four CO2 concentrations of 500, 1000, 1500 and 2000 micromoles mol-1 were combined with two air temperature regimes of 16 and 20 degrees C under a 12 h photoperiod. At harvest, 35 days after transplanting, tuber and total dry weights of potatoes reached a maximum with 1000 micromoles mol-1 CO2 at 16 degrees C, but continued to increase up to 2000 micromoles mol-1 CO2 at 20 degrees C. Plant growth was greater at 20 degrees C than at 16 degrees C under all CO2 concentrations. At 16 degrees C specific leaf weight increased substantially with increasing CO2 concentrations as compared to 500 micromoles mol-1 CO2, but increased only slightly at 20 degrees C. This suggests a carbohydrate build-up in the leaves at 16 degrees C temperature that reduces plant response to increased CO2 concentrations. The data in the two studies indicate that a PPF of 34 mol m-2 d-1, 20 degrees C temperature, and 1000-2000 micromoles mol-1 CO2 produces optimal tuber yield in potatoes.     

Effects of air current speed on gas exchange in plant leaves and plant canopies.

To obtain basic data on adequate air circulation to enhance plant growth in a closed plant culture system in a controlled ecological life support system (CELSS), an investigation was made of the effects of the air current speed ranging from 0.01 to 1.0 m s-1 on photosynthesis and transpiration in sweetpotato leaves and photosynthesis in tomato seedlings canopies. The gas exchange rates in leaves and canopies were determined by using a chamber method with an infrared gas analyzer. The net photosynthetic rate and the transpiration rate increased significantly as the air current speeds increased from 0.01 to 0.2 m s-1. The transpiration rate increased gradually at air current speeds ranging from 0.2 to 1.0 m s-1 while the net photosynthetic rate was almost constant at air current speeds ranging from 0.5 to 1.0 m s-1. The increase in the net photosynthetic and transpiration rates were strongly dependent on decreased boundary-layer resistances against gas diffusion. The net photosynthetic rate of the plant canopy was doubled by an increased air current speed from 0.1 to 1.0 m s-1 above the plant canopy. The results demonstrate the importance of air movement around plants for enhancing the gas exchange in the leaf, especially in plant canopies in the CELSS.     

CO2 crop growth enhancement and toxicity in wheat and rice.

The effects of elevated CO2 on plant growth are reviewed and the implications for crop yields in regenerative systems are discussed. There is considerable theoretical and experimental evidence indicating that the beneficial effects of CO2 are saturated at about 0.12% CO2 in air. However, CO2 can easily rise above 1% of the total gas in a closed system, and we have thus studied continuous exposure to CO2 levels as high as 2%. Elevating CO2 from 340 to 1200 micromoles mol-1 can increase the seed yield of wheat and rice by 30 to 40%; unfortunately, further CO2 elevation to 2500 micromoles mol-1 (0.25%) has consistently reduced yield by 25% compared to plants grown at 1200 micromoles mol-1; fortunately, there was only an additional 10% decrease in yield as the CO2 level was further elevated to 2% (20,000 micromoles mol-1). Yield increases in both rice and wheat were primarily the result of increased number of heads per m2, with minor effects on seed number per head and seed size. Yield increases were greatest in the highest photosynthetic photon flux. We used photosynthetic gas exchange to analyze CO2 effects on radiation interception, canopy quantum yield, and canopy carbon use efficiency. We were surprised to find that radiation interception during early growth was not improved by elevated CO2. As expected, CO2 increased quantum yield, but there was also a small increase in carbon use efficiency. Super-optimal CO2 levels did not reduce vegetative growth, but decreased seed set and thus yield. The reduced seed set is not visually apparent until final yield is measured. The physiological mechanism underlying CO2 toxicity is not yet known, but elevated CO2 levels (0.1 to 1% CO2) increase ethylene synthesis in some plants and ethylene is a potent inhibitor of seed set in wheat.     

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.     

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

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

CO2 enrichment influences yields of 'Florunner,' 'Georgia Red' and 'New Mexico' peanut cultivars.

Three peanut cultivars, 'Florunner,' 'Georgia Red,' and 'New Mexico,' were grown in reach-in chambers to determine response to CO2 enrichment. CO2 treatments were ambient (400 micromol mol-1) and 700 micromol mol-1. Growth chamber conditions included 700 micromol m-2 s-1 photosynthetic photon flux (PPF), 28/22C, 7O% RH, and 12/12 h photoperiod. Growth media consisted of a 1:1 mixture (v/v) of vermiculite and sterilized sand. Six 10 L pots of each cultivar were fertilized three times per week with 250 mL of nutrient solution containing additional Ca (10 mM) and NO3 (25 mM) and watered well. Beginning 21 days after planting (DAP) and every three weeks thereafter up to 84 days, the second leaf from the growing axis (main stem) was detached to determine CO2 effect on leaf area, specific leaf area (SLA) and dry weight. Plants were harvested 97 DAP, at which time total leaf area, leaf number, plant and root weights and pod production data were taken. Numbers of pods per plant, pod fresh and dry weights, fibrous root and plant dry weights were higher for all cultivars grown at 700 micromol mol-1 than at ambient CO2. Also, leaf area for all cultivars was larger with CO2 enrichment than at ambient. SLA tended to decline with time regardless of CO2 treatment. Percentage of total sound mature kernels (%TSMK) was similar for both treatments. Plants grown at 700 micromol mol-1 CO2 had slightly more immature pods and seeds at final harvest.     

Effect of elevated carbon dioxide on nutritional quality of tomato.

Tomato (Lycopersicon esculentum Mill.) cvs. Red Robin (RR) and Reimann Philipp (RP) were grown hydroponically for 105 d with a 12 h photoperiod, 26 degrees C/22 degrees C thermoperiod, and 500 micromol m-2 s-1 PPF at either 400, 1200, 5000, or 10,000 micromol mol-1 (0.04, 0.12, 0.50, 1.00 kPa) CO2. Harvested fruits were analyzed for proximate composition, total dietary fiber, nitrate, and elemental composition. No trends were apparent with regard to CO2 effects on proximate composition, with fruit from all treatments and both cultivars averaging 18.9% protein, 3.6% fat, 10.2% ash, and 67.2% carbohydrate. In comparison, average values for field-grown fruit are 16.6% protein, 3.8% fat, 8.1% ash, and 71.5% carbohydrate (Duke and Atchely, 1986). Total dietary fiber was highest at 10,000 micromol mol-1 (28.4% and 22.6% for RR and RP) and lowest at 1000 micromol mol-1 (18.2% and 15.9% for RR and RP), but showed no overall trend in response to CO2. Nitrate values ranged from 0.19% to 0.35% and showed no trend with regard to CO2. K, Mg, and P concentrations showed no trend in response to CO2, but Ca levels increased from 198 and 956 ppm in RR and RP at 400 micromol mol-1, to 2537 and 2825 ppm at 10,000 micromol mol-1. This increase in Ca caused an increase in fruit Ca/P ratios from 0.07 and 0.37 for RR and RP at 400 micromol mol-1 to 0.99 and 1.23 for RR and RP at 10,000 micromol mol-1, suggesting that more dietary Ca should be available from high CO2-grown fruit.     

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

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

Preparations for CELSS flight experiments with wheat.

We are planning a short-term experiment with Superdwarf wheat on the U.S. Space Shuttle and a seed-to-seed experiment on the Russian Space Station Mir. The goals of both experiments are to observe effects of microgravity on developmental steps in the life cycle and to measure photosynthesis, respiration, and transpiration by monitoring gas exchange. This requires somewhat different hardware development for the two experiments. Ground-based research aims to understand plant responses to the environments in the space growth chambers that we will use (after some modification): the Plant Growth Unit (PGU) on the shuttle and units called Svet, Svetoblock 2, or Oasis on Mir. Low irradiance levels (100 to 250 micromoles m-2 s-1 at best) pose a particular problem. Water and nutrient supply are also potentially limiting factors, especially in the long-term experiment. Our ground-based studies emphasize responses to low light levels (50 to 400 micromoles m-2 s-1); results show that all developmental steps are delayed by low light compared with plants at 400 micromoles m-2 s-1. We are also testing various rooting substrates for the shuttle experiment. A 1:1:1 mixture of peat:perlite:vermiculite appears to be the best choice.     

Crop yield and light/energy efficiency in a closed ecological system: Laboratory Biosphere experiments with wheat and sweet potato.

Two crop growth experiments in the soil-based closed ecological facility, Laboratory Biosphere, were conducted from 2003 to 2004 with candidate space life support crops. Apogee wheat (Utah State University variety) was grown, planted at two densities, 400 and 800 seeds m-2. The lighting regime for the wheat crop was 16 h of light-8 h dark at a total light intensity of around 840 micromoles m-2 s-1 and 48.4 mol m-2 d-1 over 84 days. Average biomass was 1395 g m-2, 16.0 g m-2 d-1 and average seed production was 689 g m-2 and 7.9 g m-2 d-1. The less densely planted side was more productive than the denser planting, with 1634 g m-2 and 18.8 g m-2 d-1 of biomass vs. 1156 g m-2 and 13.3 g m-2 d-1; and a seed harvest of 812.3 g m-2 and 9.3 g m-2 d-1 vs. 566.5 g m-2 and 6.5 g m-2 d-1. Harvest index was 0.49 for the wheat crop. The experiment with sweet potato used TU-82-155 a compact variety developed at Tuskegee University. Light during the sweet potato experiment, on a 18 h on/6 h dark cycle, totaled 5568 total moles of light per square meter in 126 days for the sweet potatoes, or an average of 44.2 mol m-2 d-1. Temperature regime was 28 +/- 3 degrees C day/22 +/- 4 degrees C night. Sweet potato tuber yield was 39.7 kg wet weight, or an average of 7.4 kg m-2, and 7.7 kg dry weight of tubers since dry weight was about 18.6% wet weight. Average per day production was 58.7 g m-2 d-1 wet weight and 11.3 g m-2 d-1. For the wheat, average light efficiency was 0.34 g biomass per mole, and 0.17 g seed per mole. The best area of wheat had an efficiency of light utilization of 0.51 g biomass per mole and 0.22 g seed per mole. For the sweet potato crop, light efficiency per tuber wet weight was 1.33 g mol-1 and 0.34 g dry weight of tuber per mole of light. The best area of tuber production had 1.77 g mol-1 wet weight and 0.34 g mol-1 of light dry weight. The Laboratory Biosphere experiment's light efficiency was somewhat higher than the USU field results but somewhat below greenhouse trials at comparable light levels, and the best portion of the crop at 0.22 g mol-1 was in-between those values. Sweet potato production was overall close to 50% higher than trials using hydroponic methods with TU-82-155 at NASA JSC. Compared to projected yields for the Mars on Earth life support system, these wheat yields were about 15% higher, and the sweet potato yields averaged over 80% higher.     

Volatile metabolites of higher plant crops as a photosynthesizing life support system component under temperature stress at different light intensities.

The effect of elevated temperatures of 35 and 45 degrees C (at the intensities of photosynthetically active radiation 322, 690 and 1104 micromoles m-2 s-1) on the photosynthesis, respiration, and qualitative and quantitative composition of the volatiles emitted by wheat (Triticum aestuvi L., cultivar 232) crops was investigated in growth chambers. Identification and quantification of more than 20 volatile compounds (terpenoids--alpha-pinene, delta 3 carene, limonene, benzene, alpha- and trans-caryophyllene, alpha- and gamma-terpinene, their derivatives, aromatic hydrocarbons, etc.) were conducted by gas chromatograph/mass spectrometry. Under light intensity of 1104 micromoles m-2 s-1 heat resistance of photosynthesis and respiration increased at 35 degrees C and decreased at 45 degrees C. The action of elevated temperatures brought about variations in the rate and direction of the synthesis of volatile metabolites. The emission of volatile compounds was the greatest under a reduced irradiation of 322 micromoles m-2 s-1 and the smallest under 1104 micromoles m-2 s-1 at 35 degrees C. During the repair period, the contents and proportions of volatile compounds were different from their initial values, too. The degree of disruption and the following recovery of the functional state depended on the light intensity during the exposure to elevated temperatures. The investigation of the atmosphere of the growth chamber without plants has revealed the substances that were definitely technogenic in origin: tetramethylurea, dimethylsulfide, dibutylsulfide, dibutylphthalate, and a number of components of furan and silane nature.     

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.     

Plant responses to short- and long-term exposures to high carbon dioxide levels in closed environments.

When higher plants are exposed to elevated levels of CO2 for both short- and long-term periods photosynthetic C-gain and photoassimilate export from leaves are generally increased. Water use efficiency is increased on a leaf area basis. During long-term exposures, photosynthesis rates on leaf and whole plants bases are altered in a species specific manner. The most common pattern in C3 plants is an enhanced rate of whole plant photosynthesis in a well irradiated canopy. Nevertheless, in some herbaceous species prolonged exposure to high CO2 results in remobilization of nitrogenous reserves (i.e., leaf protein degradation) and reduced rates of mature leaf photosynthesis when assayed at ambient CO2 and O2 levels. Both short- and long-term exposures to those CO2 levels (i.e., 100 to 2,000 microliter l-1) which modify photosynthesis and export, also modify both endogenous ethylene gas (C2H4) release, and substrate, 1-aminocyclopropane-1-carboxylic acid (ACC), saturated C2H4 release rates from irradiated leaves. Photosynthetically active canopy leaves contribute most of the C2H4 released from the canopy. Prolonged growth at high CO2 results in a persistent increase in the rate of endogenous C2H4 release from leaves which can, only in part, be attributed to the increase of the endogenous pools of C2H4 pathway intermediates (e.g., methionine, M-ACC, and ACC). The capacity for increasing the rate of C2H4 release in response to short-term exposures to varying CO2 levels does not decline after prolonged growth at high CO2. When leaves, whole plants, and model canopies of tomato plants are exposed to exogenous C2H4 a reduction in the rate of photosynthesis can, in each case, be attributed to the classical effects of C2H4 on plant development and morphology. The effect of C2H4 on CO2 gas exchange of plant canopies is shown to be dependent on the canopy leaf area index.     

Utilization of white potatoes in CELSS.

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

Toluene removal from air by Dieffenbachia in a closed environment.

Higher plants are likely to play a major role in bioregeneration systems for food, air and water supplies. Plants may also contribute by the removal of toxic organic substances from the air of a closed environment. Dieffenbachia amoena plants were exposed to 0 to 1.2 x 10(6) micrograms toluene m-3 at light intensities of 35 and 90 micromoles m-2 s-1 in sealed chambers. Toluene removal, photosynthesis and respiration were measured. An increased light intensity increased the rate of toluene removal five-fold over the rate at the lower intensity; the kinetics suggest active regulation by the plant. The removal rate saturated at 2700 micrograms toluene h-1 at the lower intensity and failed to saturate at the higher intensity. Toluene exposure inhibited photosynthesis and respiration only transiently and without correlation to toluene concentration. These plants can act as efficient scavengers of toluene in a contaminated environment.     

Wheat production in controlled environments.

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