Application of crop gas exchange and transpiration data obtained with CEEF to global change problem.

In order to predict carbon sequestration of vegetation with the future rise in atmospheric CO2 concentration, [CO2] and temperature, long term effects of high [CO2] and high temperature on responses of both photosynthesis and transpiration of plants as a whole community to environmental parameters need to be elucidated. Especially in the last decade, many studies on photosynthetic acclimation to elevated [CO2] at gene, cell, tissue or leaf level for only vegetative growth phase (i.e. before formation of reproductive organs) have been conducted all over the world. However, CO2 acclimation studies at population or community level for a whole growing season are thus far very rare. Data obtained from repeatable experiments at population or community level for a whole growing season are necessary for modeling carbon sequestration of a plant community. On the other hand, in order to stabilize material circulation in the artificial ecological system of Closed Ecology Experiment Facilities (CEEF), it is necessary to predict material exchange rates in the biological systems. In particular, the material exchange rate in higher plant systems is highly variable during growth periods and there is a strong dependence on environmental conditions. For this reason, dependencies of both CO2 exchange rate and transpiration rate of three rice populations grown from seed under differing conditions of [CO2] and day/night air temperature (350 microL CO2 L-1, 24/17 degrees C (population A); 700 microL CO2 L-1, 24/17 degrees C (population B) and 700 microL CO2 L-1, 26/19 degrees C (population C)) upon PPFD, leaf temperature and [CO2] were investigated every two weeks during whole growing season. Growth of leaf lamina, leaf sheath, panicle and root was also compared. From this experiment, it was elucidated that acclimation of instantaneous photosynthetic response of rice population to [CO2] occurs in vegetative phase through changes in ratio of leaf area to whole plant dry weight, LAR. But, in reproductive growth phase (i.e. after initiation of panicle formation), the difference between photosynthetic response to [CO2] of population A and that of population B decreased. Although LAR of population C was almost always less than that of population A, there was no difference between the photosynthetic response to [CO2] of population A at 24 degrees C and that of population C at 26 degrees C for its whole growth period. These results are useful to make a model to predict carbon sequestration of rice community, which is an important type of vegetation especially in Asia in future global environmental change.     

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.     

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.     

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.     

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.     

Development of a plant growth unit for growing plants over a long-term life cycle under microgravity conditions.

To study the effect of the space environment on plant growth including the reproductive growth and genetic aberration for a long-term plant life cycle, we have initiated development of a new type of facility for growing plants under microgravity conditions. The facility is constructed with subsystems for controlling environmental elements. In this paper, the concept of the facility design is outlined. Subsystems controlling air temperature, humidity, CO2 concentration, light and air circulation around plants and delivering recycled water and nutrients to roots are the major concerns. Plant experiments for developing the facility and future plant experiments with the completed facility are also overviewed. We intend to install this facility in the Japan Experiment Facility (JEM) boarded on the International Space Station.     

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.     

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.     

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

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

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.     

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.     

Carbon dioxide exchange of lettuce plants under hypobaric conditions.

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

Carbon dioxide exchange of lettuce plants under hypobaric conditions.

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

The ASTROCULTURE(TM) flight experiment series, validating technologies for growing plants in space.

A flight experiment, ASTROCULTURE(TM)-1 (ASC-1), to evaluate the operational characteristics and hardware performance of a porous tube nutrient delivery system (PTNDS) was flown on STS-50 as part of the U.S. Microgravity Laboratory-1 mission, 25 June to 9 July, 1992. This experiment is the first in a series of planned ASTROCULTURE(TM) flights to validate the performance of subsystems required to grow plants in microgravity environments. Results indicated that the PTNDS was capable of supplying water and nutrients to plants in microgravity and that its performance was similar in microgravity to that in 1g on Earth. The data demonstrated that water transfer rates through a rooting matrix are a function of pore size of the tubes, the degree of negative pressure on the 'supply' fluid, and the pressure differential between the 'supply' and 'recovery' fluid loops. A slightly greater transfer rate was seen in microgravity than in 1g, but differences were likely related to the presence of hydrostatic pressure effects at 1g. Thus, this system can be used to support plant growth in microgravity or in partial gravity as on a lunar or Mars base. Additional subsystems to be evaluated in the ASTROCULTURE(TM) flight series of experiments include lighting, humidity control and condensate recovery, temperature control, nutrient composition control, CO2 and O2 control, and gaseous contaminant control.     

Regulating plant/insect interactions using CO2 enrichment in model ecosystems.

The greenhouse environment is a challenging artificial ecosystem in which it is possible to study selected plant/insect interaction in a controlled environment. Due to a combination of "direct" and "indirect" effects of CO2 enrichment on plant photosynthesis and plant development, canopy productivity is generally increased. In this paper, we discuss the effects of daytime and nighttime CO2 enrichment protocols on gas exchange of pepper plants (Capsicum annuum L, cv Cubico) grown in controlled environments. In addition, we present the effects of thrips, a common Insect pest, on the photosynthetic and respiratory activity of these plant canopies. Carbon dioxide has diverse effects on the physiology and mortality of insects. However, our data indicate that thrips and whiteflies, at least, are not killed "directly" by CO2 levels used to enhance photosynthesis and plant growth. Together the data suggest that the insect population is affected "indirectly" by CO2 and that the primary effect of CO2 is via its effects on plant metabolism.     

Super-optimal CO2 reduces wheat yield in growth chamber and greenhouse environments.

Seven growth chamber trials (six replicate trials using 0.035, 0.12, and 0.25% CO2 in air and one trial using 0.12, 0.80, and 2.0% CO2 in air) and three replicate greenhouse trials (0.035, 0.10, 0.18, 0.26, 0.50, and 1.0% CO2 in air) compare the effects of super-optimal CO2 on the seed yield, harvest index, and vegetative growth rate of wheat (Triticum aestivum L. cvs. USU-Apogee and Veery-10). Plants in the growth chamber trials were grown hydroponically under fluorescent lamps, while the greenhouse trials were grown under sunlight and high pressure sodium lamps and in soilless media. Plants in the greenhouse trials responded similarly to those in the growth chamber trials; maximum yields occurred near 0.10 and 0.12% CO2 and decreased significantly thereafter. This research indicates that the toxic effects of elevated CO2 are not specific to only one environment and has important implications for the design of bio-regenerative life support systems in space, and for the future of terrestrial agriculture.     

The Spacelab-Mir-1 "Greenhouse-2" experiment.

The Spacelab-Mir-1 (SLM-1) mission is the first docking of the Space Shuttle Atlantis (STS-71) with the Orbital Station Mir in June 1995. The SLM-1 "Greenhouse-2" experiment will utilize the Russian-Bulgarian-developed plant growth unit (Svet). "Greenhouse-2" will include two plantings (1) designed to test the capability of Svet to grow a crop of Superdwarf wheat from seed to seed, and (2) to provide green plant material for post-flight analysis. Protocols, procedures, and equipment for the experiment have been developed by the US-Russian science team. "Greenhouse-2" will also provide the first orbital test of a new Svet Instrumentation System (SIS) developed by Utah State University to provide near real time data on plant environmental parameters and gas-exchange rates. SIS supplements the Svet control and monitoring system with additional sensors for substrate moisture, air temperature, IR leaf temperature, light, oxygen, pressure, humidity, and carbon-dioxide. SIS provides the capability to monitor canopy transpiration and net assimilation of the plants growing in each vegetation unit (root zone) by enclosing the canopy in separate, retractable, ventilated leaf chambers. Six times during the seed-to-seed experiment, plant samples will be collected, leaf area measured, and plant parts fixed and/or dried for ground analysis. A second planting initiated 30 days before the arrival of a U.S. Shuttle [originally planned to be STS-71] is designed to provide green material at the vegetative development stage for ground analysis. [As this paper is being edited, the experiment has been delayed until after the arrival of STS-71.]     

Effects of CO2 concentration and light intensity on photosynthesis of a rootless submerged plant, Ceratophyllum demersum L., used for aquatic food production in bioregenerative life support systems.

In addition to green microalgae, aquatic higher plants are likely to play an important role in aquatic food production modules in bioregenerative systems for producing feed for fish, converting CO2 to O2 and remedying water quality. In the present study, the effects of culture conditions on the net photosynthetic rate of a rootless submerged plant, Ceratophyllum demersum L., was investigated to determine the optimum culture conditions for maximal function of plants in food production modules including both aquatic plant culture and fish culture systems. The net photosynthetic rate in plants was determined by the increase in dissolved O2 concentrations in a closed vessel containing a plantlet and water. The water in the vessel was aerated sufficiently with a gas containing a known concentration of CO2 gas mixed with N2 gas before closing the vessel. The CO2 concentrations in the aerating gas ranged from 0.3 to 10 mmol mol-1. Photosynthetic photon flux density (PPFD) in the vessel ranged from 0 (dark) to 1.0 mmol m-2 s-1, which was controlled with a metal halide lamp. Temperature was kept at 28 degrees C. The net photosynthetic rate increased with increasing PPFD levels and was saturated at 0.2 and 0.5 mmol m-2 s-1 PPFD under CO2 levels of 1.0 and 3.0 mmol mol-1, respectively. The net photosynthetic rate increased with increasing CO2 levels from 0.3 to 3.0 mmol mol-1 showing the maximum value, 75 nmol O2 gDW-1 s-1, at 2-3 mmol mol-1 CO2 and gradually decreased with increasing CO2 levels from 3.0 to 10 mmol mol-1. The results demonstrate that C. demersum could be an efficient CO2 to O2 converter under a 2.0 mmol mol-1 CO2 level and relatively low PPFD levels in aquatic food production modules.